Demetallization of Enterococcus faecalis biofilm: a preliminary study

Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels.

Morphologic study of the effect of iron on pseudocyst formation in Trichomonas vaginalis and its interaction with human epithelial cells

BACKGROUND Trichomonas vaginalis is the aetiological agent of human trichomoniasis, which is one of the most prevalent sexually transmitted diseases in humans. Iron is an important element for the survival of this parasite and the colonisation of the host urogenital tract. OBJECTIVES In this study, we investigated the effects of iron on parasite proliferation in the dynamics of pseudocyst formation and morphologically characterised iron depletion-induced pseudocysts. METHODS We performed structural and ultrastructural analyses using light microscopy, scanning electron microscopy and transmission electron microscopy. FINDINGS It was observed that iron depletion (i) interrupts the proliferation of T. vaginalis, (ii) induces morphological changes in typical multiplicative trophozoites to spherical non-proliferative, non-motile pseudocysts, and (iii) induces the arrest of cell division at different stages of the cell cycle; (iv) iron is the fundamental element for the maintenance of typical trophozoite morphology; (v) pseudocysts induced by iron depletion are viable and reversible forms; and, finally, (vi) we demonstrated that pseudocysts induced by iron depletion are able to interact with human epithelial cells maintaining their spherical forms. MAIN CONCLUSIONS Together, these data suggest that pseudocysts could be induced as a response to iron nutritional stress and could have a potential role in the transmission and infection of T. vaginalis.

Effect of mitochondrial potassium channel on the renal protection mediated by sodium thiosulfate against ethylene glycol induced nephrolithiasis in rat model

Purpose: Sodium thiosulfate (STS) is clinically reported to be a promising drug in preventing nephrolithiasis. However, its mechanism of action remains unclear. In the present study, we investigated the role of mitochondrial KATP channel in the renal protection mediated by STS. Materials and Methods: Nephrolithiasis was induced in Wistar rats by administrating 0.4% ethylene glycol (EG) along with 1% ammonium chloride for one week in drinking water followed by only 0.75% EG for two weeks. Treatment groups received STS, mitochondrial KATP channel opener and closer exclusively or in combination with STS for two weeks. Results: Animals treated with STS showed normal renal tissue architecture, supported by near normal serum creatinine, urea and ALP activity. Diazoxide (mitochondria KATP channel opening) treatment to the animal also showed normal renal tissue histology and improved serum chemistry. However, an opposite result was shown by glibenclamide (mitochondria KATP channel closer) treated rats. STS administered along with diazoxide negated the renal protection rendered by diazoxide alone, while it imparted protection to the glibenclamide treated rats, formulating a mitochondria modulated STS action. Conclusion: The present study confirmed that STS render renal protection not only through chelation and antioxidant effect but also by modulating the mitochondrial KATP channel for preventing urolithiasis.

Isolated Schwannoma of the Olfactory Groove: A Case Report

Introduction Schwannoma of the olfactory groove is an extremely rare tumor that can share a differential diagnosis with meningioma or neuroblastoma. Objectives The authors present a case of giant schwannoma involving the anterior cranial fossa and ethmoid sinuses. Case Report The patient presented with a 30-month history of left nasal obstruction, anosmia, and sporadic ipsilateral bleeding. Computed tomography of the paranasal sinuses revealed expansive lesion on the left nasal cavity extending to nasopharynx up to ethmoid and sphenoid sinuses bilaterally with intraorbital and parasellar extension to the skull base. Magnetic resonance imaging scan confirmed the expansive tumor without dural penetration. Biopsy revealed no evidence of malignancy and probable neural cell. Bifrontal craniotomy was performed combined with lateral rhinotomy (Weber-Ferguson approach), and the lesion was totally removed. The tumor measured 8.0 4.3 3.7 cm and microscopically appeared as a schwannoma composed of interwoven bundles of elongated cells (Antoni A regions)mixed with less cellular regions (Antoni B). Immunohistochemical study stained intensively for vimentin and S-100. Conclusion Schwannomas of the olfactory groove are extremely rare, and the findings of origin of this tumor is still uncertain but recent studies point most probably to the meningeal branches of trigeminal nerve or anterior ethmoidal nerves. .

Antioxidant, antimicrobial, toxicity and analgesic properties of ethanol extract of Solena amplexicaulis root

BACKGROUND: This study was subjected to investigate different pharmacological properties of ethanol extract ofSolena amplexicaulis root. RESULTS: The extract contains flavonoid, alkaloid, saponin and steroid compounds. The extract exhibited excellent antioxidant activity in DPPH radical scavenging activity. The extract also showed potent activity in brine shrimp lethality bioassay. The LC50 value was found to 44.677 µg/ml. The extract showed better anti-bacterial activity against gram-negative bacteria. In antifungal assay, the maximum 79.31% of anti-mycotic activity was observed against Aspergillus ochraceus while minimum 44.2% against Rhizopus oryzae. MIC value ranged between 1500 - 3000 µg/ml. The extract was found moderately toxic with a 24-hr LD50 value of 81.47 mg/kg in Swiss albino mice. The degree of inhibition by the ethanolic extract of the root was found less than that of standard analgesic drug diclofenac sodium. The extract also showed moderate anti-inflammatory and antinociceptive activity and anti-diabetic property. Reducing power of the extract was comparable with standard ascorbic acid. Moderate in vitro thrombolytic activity, lipid peroxidation inhibition property, metal chelating ability and stress-protective activity was also observed. CONCLUSION: Ethanol extract of Solena amplexicaulis root can be valuable for treatment of different diseases.

Efeito das nanopartículas de quitosana e da variação da concentração, temperatura e pH do hipoclorito de sódio sobre biofilmes orais formados in situ / Effect of chitosan nanoparticles and the temperature and pH variations of sodium hypochlorite on biofilms formed in situ

O objetivo deste estudo foi avaliar se a variação da temperatura e pH do hipoclorito de sódio (NaOCl), incrementa sua capacidade antibacteriana e de dissolução. Foi avaliado também, se as nanopartículas de Quitosana (CNPs) inibem o crescimento bacteriano e removem a lama dentinária. Foram utilizados 260 blocos de dentina bovina infetados intra-oralmente. As soluções experimentais foram NaOCl a 1% e 2.5%, a temperatura ambiente e 37oC e acidificado a pH 5 e 7. Os tempos de exposição foram 5 e 20 min. Os espécimes foram analisados pré (controle) e pósirrigação. Após esta análise, as amostras foram incubadas em BHI por 24 horas e analisadas novamente quanto a reativação bacteriana. Estes procedimentos foram realizados em duplicado para poder determinar a porcentagem de limpeza dentinária. Para analisar o efeito quelante das CNPs sobre a dentina infetada in situ, as amostras receberam uma irrigação final com CNPs em solução e analisadas imediatamente ou infetadas intra-oralmente para avaliar o efeito antibacteriano. O NaOCl apresentou poder antibacteriano e foi capaz de dissolver significativamente o biofilme sem importar sua temperatura. O NaOCl em pHs ácidos apresentaram alto poder antibacteriano, contudo seu poder de dissolução decresceu. As CNPs foram capazes de significativamente remover a lama dentinária e interferir com o crescimento bacteriano sobre dentina. Portanto, a temperatura do NaOCl não é relevante quando é testada sobre biofilmes multi-espécies. O poder antibacteriano do NaOCl foi inversamente proporcional a seu pH, enquanto que a sua capacidade de dissolução foi diretamente proporcional. As CNPs podem ser uma alternativa para o uso de EDTA devido a suas propriedades quelantes e de interferir com a adesão inicial das bactérias sobre dentina.

The aim of this study was to evaluate if the variation of the sodium hypochlorite (NaOCl) temperature and pH increase its antibacterial and dissolution abilities. It was also evaluated if the chitosan nanoparticles (CNPs) inhibit the bacterial growth and remove smear layer. Two hundred-sixty bovine dentin blocks were infected intraorally. The experimental solutions were 1% and 2.5% NaOCl at room temperature and 37oC. the solution was acidified at pH 5 and 7. The exposure times were 5 and 20 min. The specimens were analyzed pre- (control) and postirrigation. After that, the samples were incubated in BHI and analyzed again to evaluate the bacterial recolonization. These procedures were performed in duplicate to access the percentage of dentinal cleaning. The samples were rinsed with a final irrigation of CNPs and analyzed immediately to determine the chelating effect, or infected intraorally to evaluate its antibacterial ability. NaOCl showed antibacterial power and was able to significantly dissolve the biofilm regardless its temperature. NaOCl at acid pHs showed great antibacterial properties, however its dissolution ability decreased. The CNPs were able to remove significantly the smear layer and interfere with the bacterial growth on dentin. Therefore, the NaOCl temperature is not relevant when the solution was tested on multi-specie biofilms. The antibacterial power of NaOCl was inversely proportional to its pH, while its dissolution capacity was directly proportional. CNPs could be an alternative instead of EDTA due to its chelating properties and ability to interfere with the earlier bacterial adhesion on dentin.

Efeito das nanopartículas de quitosana e da variação da concentração, temperatura e pH do hipoclorito de sódio sobre biofilmes orais formados in situ / Effect of chitosan nanoparticles and the temperature and pH variations of sodium hypochlorite on biofilms formed in situ

O objetivo deste estudo foi avaliar se a variação da temperatura e pH do hipoclorito de sódio (NaOCl), incrementa sua capacidade antibacteriana e de dissolução. Foi avaliado também, se as nanopartículas de Quitosana (CNPs) inibem o crescimento bacteriano e removem a lama dentinária. Foram utilizados 260 blocos de dentina bovina infetados intra-oralmente. As soluções experimentais foram NaOCl a 1% e 2.5%, a temperatura ambiente e 37oC e acidificado a pH 5 e 7. Os tempos de exposição foram 5 e 20 min. Os espécimes foram analisados pré (controle) e pósirrigação. Após esta análise, as amostras foram incubadas em BHI por 24 horas e analisadas novamente quanto a reativação bacteriana. Estes procedimentos foram realizados em duplicado para poder determinar a porcentagem de limpeza dentinária. Para analisar o efeito quelante das CNPs sobre a dentina infetada in situ, as amostras receberam uma irrigação final com CNPs em solução e analisadas imediatamente ou infetadas intra-oralmente para avaliar o efeito antibacteriano. O NaOCl apresentou poder antibacteriano e foi capaz de dissolver significativamente o biofilme sem importar sua temperatura. O NaOCl em pHs ácidos apresentaram alto poder antibacteriano, contudo seu poder de dissolução decresceu. As CNPs foram capazes de significativamente remover a lama dentinária e interferir com o crescimento bacteriano sobre dentina. Portanto, a temperatura do NaOCl não é relevante quando é testada sobre biofilmes multi-espécies. O poder antibacteriano do NaOCl foi inversamente proporcional a seu pH, enquanto que a sua capacidade de dissolução foi diretamente proporcional. As CNPs podem ser uma alternativa para o uso de EDTA devido a suas propriedades quelantes e de interferir com a adesão inicial das bactérias sobre dentina...

The aim of this study was to evaluate if the variation of the sodium hypochlorite (NaOCl) temperature and pH increase its antibacterial and dissolution abilities. It was also evaluated if the chitosan nanoparticles (CNPs) inhibit the bacterial growth and remove smear layer. Two hundred-sixty bovine dentin blocks were infected intraorally. The experimental solutions were 1% and 2.5% NaOCl at room temperature and 37oC. the solution was acidified at pH 5 and 7. The exposure times were 5 and 20 min. The specimens were analyzed pre- (control) and postirrigation. After that, the samples were incubated in BHI and analyzed again to evaluate the bacterial recolonization. These procedures were performed in duplicate to access the percentage of dentinal cleaning. The samples were rinsed with a final irrigation of CNPs and analyzed immediately to determine the chelating effect, or infected intraorally to evaluate its antibacterial ability. NaOCl showed antibacterial power and was able to significantly dissolve the biofilm regardless its temperature. NaOCl at acid pHs showed great antibacterial properties, however its dissolution ability decreased. The CNPs were able to remove significantly the smear layer and interfere with the bacterial growth on dentin. Therefore, the NaOCl temperature is not relevant when the solution was tested on multi-specie biofilms. The antibacterial power of NaOCl was inversely proportional to its pH, while its dissolution capacity was directly proportional. CNPs could be an alternative instead of EDTA due to its chelating properties and ability to interfere with the earlier bacterial adhesion on dentin...

Etidronate from Medicine to Endodontics: effects of different irrigation regimes on root dentin roughness

An increase in dentin roughness, associated with surface composition, contributes to bacterial adherence in recontaminations. Surface roughness is also important for micromechanical interlocking of dental materials to dentin, and understanding the characteristics of the surface is essential to obtain the adhesion of root canal sealers that have different physico-chemical characteristics. OBJECTIVES: To evaluate the effects of sodium hypochlorite (NaOCl), ethylenediaminetetraacetic (EDTA), etidronic (HEBP), and citric acid (CA) associated with different irrigation regimens on root dentin roughness. MATERIAL AND METHODS: Forty-five root halves of anterior teeth were used. The root parts were sectioned in thirds, embedded in acrylic resin and polished to a standard surface roughness. Initially, the samples of each third were randomly assigned into 3 groups and treated as follows: G1 - saline solution (control); G2 - 5% NaOCl+18% HEBP mixed in equal parts; and G3 - 2.5% NaOCl. After initial measuments, the G3 samples were distributed into subgroups G4, G5 and G6, which were subjected to 17% EDTA, 10% CA and 9% HEBP, respectively. Following the new measuments, these groups received a final flush with 2.5% NaOCl, producing G7, G8 and G9. The dentin surface roughness (Ra) was determined before and after treatments using a profilometer. The Wilcoxon test (α<0.05) was used to compare the values before and after treatments, and the Friedman test (α<0.05) to detect any differences among root thirds. RESULTS: (i) NaOCl did not affect the surface roughness; (ii) there was a significant increase in roughness after the use of chelating agents (P<0.01); and (iii) only the G3 group showed a difference in surface roughness between apical third and other thirds of the teeth (P<0.0043). CONCLUSION: Only the irrigation regimens that used chelating agents altered the roughness of root dentin. .

Co-administration of trientine and flaxseed oil on oxidative stress, serum lipids and heart structure in diabetic rats.

The administration of flaxseed oil or flaxseed oil plus trientine in diabetic rats reduced triglyceride, very low density lipoprotein, and total cholesterol. Furthermore, the combined treatment significantly increased superoxide dismutase activity and attenuated serum Cu2+. The results suggest that the administration of flaxseed oil plus trientine is useful in controlling serum lipid abnormalities, oxidative stress, restoring heart structure, and reducing serum Cu2+ in diabetic rats.

Chitosan: effect of a new chelating agent on the microhardness of root dentin

The effect of solutions of 0.2% chitosan, 15% EDTA and 10% citric acid on the microhardness of root dentin was evaluated comparatively in this study. Thirteen sound human maxillary central incisors were selected and decoronated at the cementoenamel junction. Ten roots were set into rapid polymerization acrylic resin and the root/resin block was fitted to the cutting machine to obtain slices from the cervical third. The first slice was discarded and the second slice was divided into four quadrants. Each quadrant was used to construct a sample, so that 4 specimens were obtained from each root slice, being one for each chelating solution to be tested: 15% EDTA, 10% citric acid, 0.2% chitosan and distilled water (control). The specimens were exposed to 50 μL of the solution for 5 min, and then washed in distilled water. A microhardness tester (Knoop hardness) with a 10 g load was used for 15 s. Data were analyzed statistically by one-way ANOVA and Tukey-Kramer test (α=0.05). The other 3 roots had the canals instrumented and irrigated at the end of the biomechanical preparation with the test solutions, and then examined by scanning electron microscopy (SEM) for qualitative analysis. All solutions reduced the microhardness of root dentin in a way that was statistically similar to each other (p>0.05) but significantly different from the control (p>0.05). The SEM micrographs showed that the three solutions removed smear layer from the middle third of the root canal. In conclusion, 0.2% chitosan, 15% EDTA and 10% citric acid showed similar effects in reducing dentin microhardness.

Avaliou-se o efeito das soluções de quitosana 0,2%, EDTA 15% e ácido cítrico 10% sobre a microdureza da dentina radicular. Foram utilizados 13 incisivos centrais superiores humanos, os quais tiveram suas coroas seccionadas transversalmente e desprezadas. Dez raízes foram incluídas em resina acrílica de rápida polimerização e o bloco formado raiz/resina adaptado à maquina de corte. Desprezou-se o primeiro corte transversal da porção cervical e dividiu-se o segundo, em 4 quadrantes. Cada quarto foi destinado à confecção do corpo-de-prova obtendo-se 4 espécimes para cada raiz, um para cada solução (n=10): EDTA a 15%, ácido cítrico a 10%, quitosana a 0,2% e água destilada (controle). Os espécimes receberam 50 μ L da solução por 5 min, sendo em seguida, lavados com água destilada. Utilizou-se um microdurômetro (dureza Knoop) com carga de 10 g durante 15 s. Os dados foram avaliados por meio do teste ANOVA e Tukey-Kramer (α=0,05). Três incisivos centrais superiores foram instrumentados e irrigados, ao final da biomecânica, com uma das soluções estudadas. Os espécimes foram levados para MEV e posterior análise qualitativa. Todas as soluções avaliadas reduziram a microdureza da dentina radicular de forma semelhante entre si (p>0,05) e estatisticamente diferente do controle (p<0,01). As fotomicrografias mostraram que as 3 soluções removeram a smear layer do terço médio do canal radicular. Concluiu-se que as soluções de quitosana 0,2%, EDTA 15% e ácido cítrico 10% apresentam efeito semelhante na redução da microdureza dentinária.

A comparative scanning electron microscopy evaluation of smear layer removal with apple vinegar and sodium hypochlorite associated with EDTA

OBJECTIVE: The purpose of this study was to evaluate by scanning electron microscopy (SEM) the removal of smear layer from the middle and apical root thirds after use of different irrigating solutions. MATERIAL AND METHODS: Forty roots of permanent human teeth had their canals instrumented and were randomly assigned to 4 groups (n=10), according to the irrigating solution: apple vinegar (group A), apple vinegar finished with 17 percent ethylenediaminetetraacetic acid (EDTA) (group B), 1 percent sodium hypochlorite (NaOCl) finished with 17 percent EDTA (group C) and saline (group D - control). After chemomechanical preparation, the roots were cleaved longitudinally and their middle and apical thirds were examined by SEM at ×1,000 magnification. Two calibrated examiners (kappa=0.92) analyzed the SEM micrographs qualitatively attributing scores that indicated the efficacy of the solutions in removing the smear layer from the surface of the dentin tubules (1 - poor, 2 - good and 3 - excellent). Data from the control and experimental groups were analyzed by the Kruskal-Wallis and Dunn's test, while the Wilcoxon test was used to compare the middle and apical thirds of the canals within the same group (a=0.05). RESULTS: The middle third presented less amount of smear layer than the apical third, regardless of the irrigant. There was statistically significant difference (p=0.0402) among the groups in the middle third. In the apical third, the apple vinegar/EDTA group showed the greatest removal of smear layer (p=0.0373). CONCLUSION: Apple vinegar associated or not with EDTA was effective in removing smear layer when used as an endodontic irrigant.

Effect of aquatine endodontic cleanser on smear layer removal in the root canals of ex vivo human teeth

OBJECTIVES: The purpose of this study was to measure and compare the root canal cleanliness and smear layer removal effectiveness of Aquatine Endodontic Cleanser (Aquatine EC) when used as an endodontic irrigating solution in comparison with 6 percent sodium hypochlorite (NaOCl). MATERIAL AND METHODS: Forty-five human teeth were randomly allocated to five treatment groups; the pulp chamber was accessed, cleaned, and shaped by using ProTaper and ProFile rotary instrumentation to an ISO size #40. The teeth were then processed for scanning electron microscopy, and the root canal cleanliness and removal of smear layer were examined. RESULTS: The most effective removal of smear layer occurred with Aquatine EC and NaOCl, both with a rinse of EDTA. CONCLUSIONS: Aquatine EC appears to be the first hypochlorous acid approved by the FDA to be a possible alternative to the use of NaOCl as an intracanal irrigant. Further research is needed to identify safer and more effective alternatives to the use of NaOCl irrigation in endodontics.

Antioxidant properties of a dry product from Vitis vinifera seeds (Leucoselect) in rat liver endoplasmic reticulum / Propiedades antioxidantes de un producto seco de semillas de Vitis vinifera (Leucoselect) en retículo endoplásmico de hígado de rata

Leucoselect is a commercial dry product obtained from grape seeds and enriched in procyanidins, which display antioxidant activity in virtue to their ability to scavenge oxygen free radicals and to chelate transition metal ions. The hypoxanthine/xanthine oxidase and Cu2+/ascorbate systems are capable of generating reactive oxygen species; the latter system can also promote non-specific binding of copper ions to proteins. Therefore, we assessed the ability of Leucoselect to inhibit oxidative phenomena elicited by both oxidative systems on rat liver microsomes: lipid peroxidation, oxidation of protein thiols, and inhibition of the cytochrome P450 system. The antioxidant activity of Leucoselect was a reflection of its ability to scavenge oxygen free radicals, chelate copper ions, and protect microsomal membranes through direct interaction. These mechanisms were displayed in a dependent manner with the type of biomolecule studied and also with the oxidative system employed, which is an interesting phenomenon to consider when evaluating the antioxidant activity of herbal products.

Leucoselect es un producto comercial seco obtenido de semillas de uva y enriquecido en procianidinas, las cuales presentan actividad antioxidante debido a su capacidad para atrapar radicales libres y quelar metales de transición. Los sistemas hipoxantina/xantina oxidasa y Cu2+/ascorbato generan especies reactivas del oxígeno; este último sistema también promueve la unión inespecífica de iones cobre a proteínas. Por lo tanto, evaluamos la capacidad de Leucoselect para inhibir los fenómenos oxidativos producidos por ambos sistemas oxidantes en microsomas hepáticos de rata: lipoperoxidación, oxidación de tioles proteicos e inhibición de la actividad del sistema citocromo P450. La actividad antioxidante de Leucoselect fue un reflejo de su capacidad de atrapar radicales libres del oxígeno, quelar iones cobre y proteger membranas microsómicas por interacción directa. Dichos mecanismos se manifestaron en forma dependiente del tipo de biomolécula estudiada y del sistema oxidante empleado, fenómeno interesante de considerar al evaluar la actividad antioxidante de preparados herbales.

DNA damage by ribose: Inhibition at high ribose concentrations.

Nucleobases and DNA react non-enzymatically with sugars, and generate DNA-advanced glycation end products (DNA-AGEs). Incubation of plasmid pBr322 with ribose for 3-7 days caused the transformation of the supercoiled plasmid to the open circular and linear forms. Removal of sugar after an initial 24 h incubation resulted in marked enhancement in the transformation rate. Enhancement in transformation was also observed when bovine serum albumin (BSA) exposed to ribose for short durations was incubated with plasmid in absence of the sugar. The effect on DNA was attenuated when excess ribose remained in the incubation mixture of ribose and protein. The data suggested that an increase in ribose concentration in the vicinity of DNA could be damaging and the damage exacerbated, if sugar levels fell subsequently. Incubation of herring sperm DNA with ribose resulted in a concentration and time-dependent increase of N2-carboxyethyl-2’-deoxyguanosine (CEdGA,B). The concentration of CEdGA,B, however, did not increase further when ribose was removed from the reaction mixture.

Hydrogen peroxide induces a specific DNA base change profile in the presence of the iron chelator 2, 2’ dipyridyl in Escherichia coli

Pretreatment of Escherichia coli cultures with the iron chelator 2,2’-dipyridyl (1 mM) protects against the lethal effects of low concentrations of hydrogen peroxide (<15 mM). However, at H2O2 concentrations equal to or greater than 15 mM, dipyridyl pretreatment increases lethality and mutagenesis, which is attributed to the formation of different types of DNA lesions. We show here that pretreatment with dipyridyl (1 mM) prior to challenge with high H2O2 concentrations (≥15 mM) induced mainly G:C→A:T transitions (more than 100X with 15 mM and more than 250X with 20 mM over the spontaneous mutagenesis rate) in E. coli. In contrast, high H2O2 concentrations in the absence of dipyridyl preferentially induced A:T→T:A transversions (more than 1800X and more than 300X over spontaneous mutagenesis for 15 and 20 mM, respectively). We also show that in the fpg nth double mutant, the rpoB gene mutation (RifS-RifR) induced by 20 mM H2O2 alone (20X higher) was increased in 20 mM H2O2 and dipyridyl-treated cultures (110X higher), suggesting additional and/or different lesions in cells treated with H2O2 under iron deprivation. It is suggested that, upon iron deprivation, cytosine may be the main damaged base and the origin of the pre-mutagenic lesions induced by H2O2.

Antioxidant activity of mojabanchromanol, a novel chromene, isolated from brown alga Sargassum siliquastrum.

Silica gel chromatography HPLC, El-Mass, and NMR analyses were performed in order to determine the structure obtained from the separation and refinement of the main components of an ethanol extraction of Sargassum siliquastrum, which indicated its antioxidant activity An ethanol extraction of Sargassum siliquastrum demonstrated the strongest antioxidant activities of ethyl acetate when isolated by silica gel chromatography This amounts to 84.9 +/- 1.2% of its 0.5 mg ml(-1) concentration. The thiobarbituric acid reactive substances (TBARS) measurements from 3 isolations of an HPLC separation of ethyl acetate showed that the antioxidant activity of peak 2 was the most dominant at 84.08% in a 0.5 mg ml(-1) concentration. Peak 2 was verified as a type of chromene through El-Mass and NMR analysis. Its metal sealing characteristic was low while its characteristic of TBARS and DPPH erasure indicated similar or higher levels of metal sealing characteristic. The NMR spectroscopic data was used to elucidate the structure of this new compound, which showed strong antioxidant activity in the assay.

High susceptibility of activated lymphocytes to oxidative stress-induced cell death

The present study provides evidence that activated spleen lymphocytes from Walker 256 tumor bearing rats are more susceptible than controls to tert-butyl hydroperoxide (t-BOOH)-induced necrotic cell death in vitro. The iron chelator and antioxidant deferoxamine, the intracellular Ca2+ chelator BAPTA, the L-type Ca2+ channel antagonist nifedipine or the mitochondrial permeability transition inhibitor cyclosporin A, but not the calcineurin inhibitor FK-506, render control and activated lymphocytes equally resistant to the toxic effects of t-BOOH. Incubation of activated lymphocytes in the presence of t-BOOH resulted in a cyclosporin A-sensitive decrease in mitochondrial membrane potential. These results indicate that the higher cytosolic Ca2+ level in activated lymphocytes increases their susceptibility to oxidative stress-induced cell death in a mechanism involving the participation of mitochondrial permeability transition.

O presente estudo demonstra que linfócitos ativados de baço de ratos portadores do tumor de Walker 256 são mais susceptíveis à morte celular necrótica induzida por tert-butil hidroperóxido (t-BOOH) in vitro quando comparados aos controles. O quelante de ferro e antioxidante deferoxamina, o quelante intracelular de Ca2+ BAPTA, o antagonista de canal de Ca2+ nifedipina ou o inibidor da transição de permeabilidade mitocondrial ciclosporina-A, mas não o inibidor de calcineurina FK-506, inibiram de maneira similar a morte celular induzida por t-BOOH em linfócitos ativados e controles. Os linfócitos ativados apresentaram redução do potencial de membrana mitocondrial induzida por t-BOOH num mecanismo sensível a ciclosporina-A. Nossos resultados indicam que o aumento da concentração de Ca2+ citosólico em linfócitos ativados aumenta a susceptibilidade dos mesmos à morte celular induzida por estresse oxidativo, num mecanismo envolvendo a participação do poro de transição de permeabilidade mitocondrial.

Cross talk between P2 purinergic receptors modulates extracellular ATP-mediated interleukin-10 production in rat microglial cells

Previously we demonstrated that ATP released from LPS-activated microglia induced IL-10 expression in a process involving P2 receptors, in an autocrine fashion. Therefore, in the present study we sought to determine which subtype of P2 receptor was responsible for the modulation of IL-10 expression in ATP-stimulated microglia. We found that the patterns of IL-10 production were dose-dependent (1, 10, 100, 1,000 micrometer) and bell-shaped. The concentrations of ATP, ATP-gammaS, ADP, and ADP-beta S that showed maximal IL-10 release were 100, 10, 100, and 100 micrometer respectively. The rank order of agonist potency for IL-10 production was 2'-3'-O-(4-benzoyl)-benzoyl ATP (BzATP) = dATP > 2-methylthio-ADP (2-meSADP). On the other hand, 2-methylthio-ATP (2-meSATP), alpha,beta-methylene ATP (alpha,beta-meATP), UTP, and UDP did not induce the release of IL-10 from microglia. Further, we obtained evidence of crosstalk between P2 receptors, in a situation where intracellular Ca2+ release and/or cAMP-activated PKA were the main contributors to extracellular ATP-(or ADP)-mediated IL-10 expression, and IL-10 production was down- regulated by either MRS2179 (a P2Y1 antagonist) or 5'-AMPS (a P2Y11 antagonist), indicating that both the P2Y1 and P2Y11 receptors are major receptors involved in IL-10 expression. In addition, we found that inhibition of IL-10 production by high concentrations of ATP-gammaS (100 micrometer) was restored by TNP-ATP (an antagonist of the P2X1, P2X3, and P2X4 receptors), and that IL-10 production by 2-meSADP was restored by 2meSAMP (a P2Y12 receptor antagonist) or pertusis toxin (PTX; a Gi protein inhibitor), indicating that the P2X1, P2X3, P2X4 receptor group, or the P2Y12 receptor, negatively modulate the P2Y11 receptor or the P2Y1 receptor, respectively.

Role of chelating agents in thalassaemia major patients

To evaluate current chelation in thalassaemia major patients. It is a retrospective study. This study was conducted at Sundus Foundation, Shadman, Lahore Pakistan from March 2006 to August 2006. One hundred and seventy patients with transfusion dependent thalassaemia major were evaluated for chelation practice and iron overload. 98.2% were found to be either non-chelators or inadequate chelators whereas 82.3% patients had serum ferritin levels above 2500 ng/ml. Growth failure and hepatosplenomegaly were also common. Survival appears to be limited as only 6 patients were older than 20 years. Patients face risks and complications during treatment of thalassaemia major. Urgent and effective measures should be taken to remove the obstacles for improvement in quality of life in these patients. Involvement of physicians about current concepts in treatment of thalassaemia major can produce promising results