Disinfection of clinical materials using photo-activated tolonium chloride solution

Photo-activated disinfection is beginning to be used in dental surgery to treat deep seated bacterial infection. It works by combining a photosensitiser and light of a specific frequency to generate singlet oxygen which is toxic to many types of bacteria. It is suggested that this technique could be used as a means to help treat infection more generally. To do so, it needs to work with materials and geometries exhibiting different physical and optical characteristics to teeth. In these trials, samples of stainless steel and polymethylmethacrylate were exposed to bacterial solutions of Staphylococcus aureus and Staphylococcus epidermis. These were treated with tolonium chloride-based photo-activated disinfection regimes showing positive results with typically 4 log10 reductions in colony forming units. Tests were also carried out using slotted samples to represent geometric features which might be found on implants. These tests, showed disinfectant effect however to a much lesser degree. © 2011 Inderscience Enterprises Ltd.

Characterization of a monoclonal antibody recognizing mast cells

Immunohistochemical screening for monoclonal antibodies prepared by immunization of mice with a rat osteoblastic cell population led to identification of one antibody that reacted against a small population of cells present in the soft connective tissue compartment of 21 days fetal rat calvaria. The morphology of the cells and the immunohistochemical staining characteristics (a distinct intracellular granular pattern) suggested that the antibody might be reacting specifically against mast cells. We used combined histochemistry and immunohistochemistry to further characterize this antibody, designated RCJ102. Cryosections containing calvaria bone, soft connective tissues and skin were prepared from the top of the head of 21 days fetal rats, and from adult rats cryosections of lung, muscle, adipose tissue and small intestine were prepared. Some sections were labelled by indirect immunofluorescence with RCJ102; corresponding sections were labelled histochemically with toluidine blue. There was a direct correspondence between mast cells identified histochemically and cells labelling with RCJ102 in all tissues except intestine, in which the mast cell detectable by histochemistry were not labelled by RCJ102. These results suggest that the RCJ102 antibody will be a valuable new reagent for further elucidation of the heterogeneity described between connective tissue and intestinal mucosal mast cells.

Avaliação experimental dos efeitos do anestésico local ropivacaína em nervos ciáticos

This study was undertaken to investigate the effects of ropivacaine after intrafascicular injection into the sciatic nerves of albino rabbits. Twenty adult albine rabbits were used, following sedation with intramuscular ketamine (50 mg/kg) for nerve exposure by lateral incision. We considered three experimental groups: Group I:sciatic nerve control; Group II: intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine into the rigth nerves. The specimens were colected at 48 h after drugs administration; Group III. intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine in the rigth nerves. The specimens were colected at 7 days after drugs administration. The sciatic nerves were removed from these animals and fixed in Karnowisky solution for 24 hours. After partial dehydration up to 95% ethanol, they were embedded in historesin (Leica). The tissue was then sectioned at 1-2μm. Sections were stained with haematoxylin-eosin (HE); toluidine blue (TB) or picrosirius-haematoxylin (PSH). Comparing with control group the histological evidence of inflammatory reaction (migration of macrophagic cells and eosinophils-appeared soon after injection, with intense proliferation of perineurial cells. The results show that after 7 days of intrafascicular injection there was a severe fibrosis and an increase on perineurial vascularization. In group 2 the inflammatory reaction was noted near the local of the injection. Furthermore in this experiment we observed an increase on the number of epineurial lipoblasts and adipocytes. This study demonstrated that the toxic effects of ropivacaine are transient. In many cases there was an initial fascicular recover and axonal regeneration after 7 days of the injection.

Lethal photosensitization in microbiological treatment of ligature-induced peri-implantitis: a preliminary study in dogs

This pilot study evaluated, by culture testing, the effectiveness of lethal photosensitization for the microbiological treatment of peri-implantitis in dogs. Experimental peri-implantitis was induced by ligature placement for 2 months. Following ligature removal, plaque control was instituted by scrubbing with 0.12% chlorhexidine daily for 12 months. Subsequently, mucoperiosteal flaps were elevated for scaling the implant surface. Microbial samples were obtained with paper points before and after treatment of implant surfaces by means of 100 microg/ml toluidine blue O (TBO,) and were exposed, for 80 s, to light with a wavelength of 685 nm from a 50 mW GaAlAs diode laser. The mean initial and final bacterial counts were 7.22 +/- 0.20 and 6.84 +/- 0.44 CFU/ml, respectively for TVC (P < 0.0001); 6.19 +/- 0.45 and 3.14 +/- 3.29 CFU/ml for P. intermedia/nigrescens (P = 0.001); 5.98 +/- 0.38 and 1.69 +/- 2.90 CFU/ml for Fusobacterium spp. (P = 0.001); and 6.07 +/- 0.22 to 1.69 +/- 2.94 CFU/ml for beta-hemolytic Streptococcus (P = 0.0039). It may be concluded that lethal photosensitization resulted in a reduction of the bacterial count. Complete elimination of bacteria was achieved in some samples.

Organização celular dos testículos em Hylidae e leptodactylidae, no Pantanal (Estado do Mato Grosso do Sul, Brasil)

The objective of this work was to make a comparative analysis of germ cell organization at different stages of cellular differentiation in adult males of Dendropsophus nanus (Boulenger, 1889), Pseudis limellum (Cope, 1862), P. paradoxa (Linnaeus, 1758), and Scinax acuminatus (Cope, 1862), belonging to the family Hylidae; and Leptodactylus chaquensis (Cei, 1950) and L. podicipinus (Cope, 1862), belonging to the family Leptodactylidae, collected in the Pantanal and in Serra da Bodoquena, Mato Grosso do Sul State, Brazil. The testes were removed and fixed, dehydrated in a graded series of ethanol and embedded in methacrylate glycol resin (Historesin Leica®). The sections were stained by 1% toluidine blue and observed under light microscope. It was detected that all individual of the Hylidae family show, throughout the year, the presence of all germ cell types of spermatogenesis. However, all Leptodactylidae family individuals only show the presence of all germ cell types during the rainy season. The variations of characteristics in seminiferous epithelium organization, as well as the evident difference in the amount of spermatozoa inside the tubules, are evidence that the anurans in this work show different forms of spermatogenesis development throughout the year: the cycle is continuous for the Hylidae family, and discontinuous with explosive release of spermatozoa for the Leptodactylidae family.

Treatment of experimental periodontitis in rats using repeated adjunctive antimicrobial photodynamic therapy

The aim of this study was to histologically and histometrically evaluate the influence of repeated adjunctive antimicrobial photodynamic therapy (aPDT) on bone loss (BL) in furcation areas in rats. Periodontitis was induced by placing a ligature around the mandibular molar in 75 rats. The animals were divided into five groups: the SS group was treated with saline solution (SS); the SRP group received scaling and root planing (SRP); the aPDT1 group received SRP as well as toluidine blue (TBO) and low-level laser therapy (LLLT; InGaAlP, 660 nm; 4.94 J/cm2/point) postoperatively at 0 h; the aPDT2 group received SRP as well as TBO and LLLT postoperatively at 0, 24, 28, and 72 h; and the aPDT3 group received SRP, TBO, and LLLT postoperatively at 0, 48, 96, and 144 h. The area of BL in the furcation region of the molar was histometrically analyzed. Data were analyzed statistically (P < 0.05). Animals treated with a single episode of aPDT showed less BL at days 7 and 30 than those who received only SRP treatment. No significant differences were found among the aPDT groups (P > 0.05). Repeated aPDT did not improve BL reduction when compared to a single episode of aPDT. © 2012 Springer-Verlag London Ltd.

Biogenic Silver Nanoparticles Associated With Silver Chloride Nanoparticles (ag@agcl) Produced By Laccase From Trametes Versicolor.

In the present study, semi-purified laccase from Trametes versicolor was applied for the synthesis of silver nanoparticles, and the properties of the produced nanoparticles were characterized. All of the analyses of the spectra indicated silver nanoparticle formation. A complete characterization of the silver nanoparticles showed that a complex of silver nanoparticles and silver ions was produced, with the majority of the particles having a Ag(2+) chemical structure. A hypothetical mechanistic scheme was proposed, suggesting that the main pathway that was used was the interaction of silver ions with the T1 site of laccase, producing silver nanoparticles with the concomitant inactivation of laccase activity and posterior complexing with silver ions.

Determination of the maximum inhibitory dilution of cetylpyridinium chloride-based mouthwashes against staphylococcus aureus: an in vitro study

The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)-based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Juá+Romã+Propolis (Natural Honey®) and CPC (Cepacol®), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37ºC for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Juá+Romã+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol® showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol® was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect.

Physiological implications of the regulation of vacuolar H+-ATPase by chloride ions

Vacuolar H+-ATPase is a large multi-subunit protein that mediates ATP-driven vectorial H+ transport across the membranes. It is widely distributed and present in virtually all eukaryotic cells in intracellular membranes or in the plasma membrane of specialized cells. In subcellular organelles, ATPase is responsible for the acidification of the vesicular interior, which requires an intraorganellar acidic pH to maintain optimal enzyme activity. Control of vacuolar H+-ATPase depends on the potential difference across the membrane in which the proton ATPase is inserted. Since the transport performed by H+-ATPase is electrogenic, translocation of H+-ions across the membranes by the pump creates a lumen-positive voltage in the absence of a neutralizing current, generating an electrochemical potential gradient that limits the activity of H+-ATPase. In many intracellular organelles and cell plasma membranes, this potential difference established by the ATPase gradient is normally dissipated by a parallel and passive Cl- movement, which provides an electric shunt compensating for the positive charge transferred by the pump. The underlying mechanisms for the differences in the requirement for chloride by different tissues have not yet been adequately identified, and there is still some controversy as to the molecular identity of the associated Cl--conducting proteins. Several candidates have been identified: the ClC family members, which may or may not mediate nCl-/H+ exchange, and the cystic fibrosis transmembrane conductance regulator. In this review, we discuss some tissues where the association between H+-ATPase and chloride channels has been demonstrated and plays a relevant physiologic role.

Photolysis of ferric ions in the presence of sulfate or chloride ions: implications for the photo-Fenton process

The photo-Fenton process (Fe(2+)/Fe(3+), H(2)O(2), UV light) is one of the most efficient and advanced oxidation processes for the mineralization of the organic pollutants of industrial effluents and wastewater. The overall rate of the photo-Fenton process is controlled by the rate of the photolytic step that converts Fe(3+) back to Fe(2+). In this paper, the effect of sulfate or chloride ions on the net yield of Fe(2+) during the photolysis of Fe(3+) has been investigated in aqueous solution at pH 3.0 and 1.0 in the absence of hydrogen peroxide. A kinetic model based on the principal reactions that occur in the system fits the data for formation of Fe(2+) satisfactorily. Both experimental data and model prediction show that the availability of Fe(2+) produced by photolysis of Fe(3+) is inhibited much more in the presence of sulfate ion than in the presence of chloride ion as a function of the irradiation time at pH 3.0.

Sisal, sugarcane bagasse and microcrystalline celluloses: Influence of the composition of the solvent system N,N-dimethylacetamide/lithium chloride on the solubility and acetylation of these polysaccharides

The present work describes an investigation concerning the acetylation of celluloses extracted from short-life-cycle plant sources (i.e. sugarcane bagasse and sisal fiber) as well as microcrystalline cellulose. The acetylation was carried out under homogeneous conditions using the solvent system N,N-dimethylacetamide/lithium chloride. The celluloses were characterized, and the characterizations included an evaluation of the amount of hemicellulose present in the materials obtained from lignocellulosics sources (sugarcane and sisal). The amount of LiCl was varied and its influence on the degree of acetate substitution was analyzed. It was found that the solvent system composition and the nature of the cellulose influenced both the state of chain dissolution and the product characteristics. The obtained results demonstrated the importance of developing specific studies on the dissolution process as well as on the derivatization of celluloses from various sources.

An expert flow system involving in-line prior assay for turbidimetric determination of chloride and sulphate in natural waters

A multi-pumping flow system exploiting prior assay is proposed for sequential turbidimetric determination of sulphate and chloride in natural waters. Both methods are implemented in the same manifold that provides facilities for: in-line sample clean-up with a Bio-Rex 70 mini-column with fluidized beads: addition of low amounts of sulphate or chloride ions to the reaction medium for improving supersaturation; analyte precipitation with Ba(2+) or Ag(+); real-time decision on the need for next assay. The sample is initially run for chloride determination, and the analytical signal is compared with a preset value. If higher, the sample is run again, now for sulphate determination. The strategy may lead to all increased sample throughput. The proposed system is computer-controlled and presents enhanced figures of merit. About 10 samples are run per hour (about 60 measurements) and results are reproducible and Unaffected by the presence of potential interfering ions at concentration levels usually found in natural waters. Accuracy was assessed against ion chromatography. (C) 2008 Elsevier B.V. All rights reserved.

Low nanomolar concentration of mercury chloride increases vascular reactivity to phenylephrine and local angiotensin production in rats

Exposure to mercury at nanomolar level affects cardiac function but its effects on vascular reactivity have yet to be investigated. Pressor responses to phenylephrine (PHE) were investigated in perfused rat tail arteries before and after treatment with 6 nM HgCl2 during 1 h,,in the presence (E+) and absence (E-) of endothelium, after L-NAME (10(-4) M), indomethacin (10(-5) M), enalaprilate (1 mu M), tempol (1 mu M) and deferoxamine (300 mu M) treatments. HgCl2 increased sensitivity (pD(2)) without modifying the maximum response (Em) to PHE, but the pD(2) increase was abolished after endothelial damage. L-NAME treatment increased pD(2) and Emax. However, in the presence of HgCl2, this increase was smaller, and it did not modify Emax. After indomethacin treatment, the increase of pD(2) induced by HgCl2 was maintained. Enalaprilate, tempol and deferoxamine reversed the increase of pD(2) evoked by HgCl2. HgCl2 increased the angiotensin converting enzyme (ACE) activity explaining the result obtained with enalaprilate. Results suggest that at nanomolar concentrations HgCl2 increase the vascular reactivity to PHE. This response is endothelium mediated and involves the reduction of NO bioavailability and the action of reactive oxygen species. The local ACE participates in mercury actions and depends on the angiotensin 11 generation. (c) 2007 Elsevier Inc. All rights reserved.

Influence of chloride ion concentration and temperature on the electrochemical properties of passive films formed on a superduplex stainless steel

Polarization measurements were conducted to monitor the corrosion behavior of superduplex stainless steel ASTM A995M-Gr.SA/EN 10283-Mat#1.4469(GX2CrNiMo26-7-4) when exposed to a) an electrolyte containing 22,700 parts per million (ppm) of chloride ions at seven different temperatures and b) an electrolyte at 25 GC and different chloride ion concentrations (5800, 22,700, 58,000 and 80,000 ppm of Cl(-)). The polarization curves indicate that the passive films formed are only slightly affected by NaCl concentration, but the pitting potential decreases drastically increasing the temperature, in particular >60 degrees C. The image analysis of the microstructure after potentiodynamic polarization showed that the pitting number and size vary in function of the temperature of the tested medium. Nyquist diagrams were determined by electrochemical impedance spectroscopy to characterize the resistance of the passive layer. According to Nyquist plots, the arc polarization resistance decreases increasing the temperature due to a catalytic degradation of the oxide passive films. (C) 2009 Elsevier Inc. All rights reserved.

Influence of the slice position on chloride migration tests for concrete in marine conditions

Chloride migration tests are used to measure the concrete capacity to inhibit chloride attack. Many researchers carry out this test in a slice of concrete extracted from the central part of cylindrical specimens, discarding about 75% of the concrete used to mold the specimens. This fact generated the question: would it be possible to extract more slices from a same specimen without losing the confidence in the results? The main purpose of this work is to answer this question. Moreover, another aim of this study was to show the difference of chloride penetration between finished faces and the formwork surfaces of concrete beams and slabs. The results indicated that it is possible to use more slices of a single specimen for a chloride migration test. Moreover, it was demonstrated that there is a significant difference of chloride penetration between the finished surface and the formwork surface of the specimens. (C) 2008 Elsevier Ltd. All rights reserved.