Effect of iodonium salt and chitosan on the physical and antibacterial properties of experimental infiltrants

Abstract Resinous infiltrants are indicated in the treatment of incipient carious lesions, and further development of these materials may contribute to greater control of these lesions. The aim of this study was to analyze the physical and antibacterial properties of experimental infiltrants containing iodonium salt and chitosan. Nine experimental infiltrants were formulated by varying the concentration of the diphenyliodonium salt (DPI) at 0, 0.5 and 1 mol%; and chitosan at 0, 0.12 and 0.25 g%. The infiltrants contained the monomeric base of triethylene glycol dimethacrylate and bisphenol-A dimethacrylate ethoxylate in a 75 and 25% proportion by weight, respectively; 0.5 mol% camphorquinone and 1 mol% ethyl 4-dimethylaminobenzoate. The degree of conversion was evaluated using Fourier transformer infrared spectroscopy, and the flexural strength and elastic modulus using the three-point bending test. Sorption and solubility in water, and antibacterial analysis (minimum inhibitory concentration and minimum bactericidal concentration) were also analyzed. Data was analyzed statistically by two-way ANOVA and Tukey's test (p<0.05), with the exception of the antibacterial test, which was evaluated by visual inspection. In general, the infiltrant group containing 0.5% DPI and 0.12% chitosan showed high values of degree of conversion, higher values of elastic modulus and flexural strength, and lower sorption values in relation to the other groups. Antibacterial activity was observed in all the groups with DPI, regardless of the concentration of chitosan. The addition of DPI and chitosan to experimental infiltrants represents a valid option for producing infiltrants with desirable physical and antibacterial characteristics.

Comportamiento antibacteriano de partículas de cobre frente a microorganismos obtenidos de úlceras crónicas infectadas y su relación con la resistencia a antimicrobianos de uso común / Antibacterial activity of copper salts against microorganisms isolated from chronic infected wounds

Background: The antimicrobial activity of copper (Cu+2) is recognized and used as an antimicrobial agent. Aim: To evaluate the antimicrobial activity of copper against microorganisms obtained from chronic cutaneous wound infections. Material and Methods: Five chemical products that contained copper particles in their composition were tested (zeolite, silica, acetate, nitrate and nanoparticle of copper). The antimicrobial activity against antibiotic resistant strains usually isolated from chronic cutaneous wound infections was determined for two of the products with better performance in copper release. Results: The minimal inhibitory and minimal bactericidal concentrations of copper acetate and nitrate were similar, fluctuating between 400-2,000 µg/ml. Conclusions: The studied copper salts show great potential to be used to control both gram positive and gram negative, antibiotic resistant bacteria isolated from wound infections.

Inactivation of Listeria monocytogenes ATCC 7644 on fresh-cut tomato using nisin in combinations with organic salts

ABSTRACT The inhibition of Listeria monocytogenes ATCC 7644 on fresh-cut tomato was investigated using nisin alone, and in combinations with organic salts. Nisin at a concentration of 5000 UI/mL was introduced alone or in combination with an organic salt (sodium citrate or sodium acetate each at 3 and 5 g/100 mL each) on fresh-cut tomato previously inoculated with 108 CFU/mL of L. monocytogenes ATCC 7644. Chlorine at 200 ppm was used as a control. The inoculated samples were incubated at different temperatures (4, 10 and 25 °C) and examined at 0, 24, 48 and 72 h. The effects of the antimicrobial treatments on quality parameters of tomato (pH, soluble solids, titratable acidity and vitamin C) were also evaluated, and colour parameters were observed at the lowest storage temperature for 10 days. Both nisin and the organic salts inhibited growth of L. monocytogenes, but the combinations of two compounds were more effective. The nisin-sodium citrate (5%) combination was significantly (p ≤ 0.05) effective, while chlorine was least effective against L. monocytogenes. The quality parameters were substantially retained, especially at 4 °C, suggesting good shelf stability at a low temperature. These results substantiate the use of the cheap and eco-friendly approach to reducing this pathogen of health concern in common fresh produce.

Micropropagation of Pluchea sagittalis (Lam. ) Cabrera / Micropropagação do quitoco (Pluchea sagittalis (Lam. ) Cabrera)

ABSTRACT: The objective of this study was to develop an in vitro protocol for the micropropagation of Pluchea sagittalis (Lam.) Cabrera. Plants were regenerated in vitro from stem segments. The procedure employed includes: 1) surface sterilization of shoots by immersion in 70% ethanol for 10 s followed by 1.0% NaOCl for 10 min, and subsequent immersion in 0.05% HgCl2 for 3 min and two washes with sterile distilled water; 2) induction of root and shoot by culture on hormone-free Murashige and Skoog medium (MS); 3) acclimatization of 60 day-old-plantlets in soil under ex vitro conditions. Minimum contamination was observed for apical shoot explants (10%). However, independently of the explant position in the stem, all explants regenerated new shoots. Various successive cultivations from stem explants every 60 days during more than 1 year have been shown to be a suitable method to propagate P. sagittalis in vitro. Low salt concentration (25% of the normal concentration) in the medium promoted greater growth of plantlets because the plants had a higher number of roots and longer roots in such an environment. Our protocol for the micropropagation of P. sagittalis can be accomplished as a two-step procedure within a short period of time (two months) before transplanting.

RESUMO: O Objetivo deste estudo foi desenvolver um protocolo para a micropropagação in vitro da Pluchea sagittalis (Lam.) Cabrera. Plantas foram regeneradas in vitro a partir de segmentos de ramo. O procedimento empregado incluiu: 1) esterilização da superfície de ramos pela imersão em etanol 70% por 10 s seguida pela de NaOCl 1.0% por 10 min e, subsequentemente, em HgCl2 0.05% por 3 min e duas lavagens em água destilada e esterilizada; 2) indução de raízes e parte aérea pelo cultivo em meio Murashige & Skoog (MS) isento de hormônio; 3) aclimatização de plantas com 60 dias de idade em solo sob condições ex vitro. Contaminação mínima foi observada em explantes caulinares do ápice (10%). Entretanto, independentemente da posição do segmento no caule, todos explantes regeneraram novos ramos. Vários cultivos sucessivos a cada 60 dias durante mais de um ano tem mostrado ser um método adequado para a propagação in vitro de P. sagittalis. A baixa concentração de sais no meio (25% da concentração normal) promoveu maior crescimento das plântulas devido às mesmas apresentarem maior número e comprimento de raízes. O protocolo para a micropropagação da P. sagittalis pode ser executado em procedimento de duas etapas dentro de um período de tempo curto (dois meses) antes do transplantio.

Incorporation of Na+/H+ antiporter gene from Aeluropus littoralis confers salt tolerance in soybean (Glycine max L.).

To develop a salt-tolerant soybean (Glycine max L.) cultivar, a minimal linear Na+/H+ antiporter gene cassette (35S CaMV promoter, open-reading-frame of AlNHX1 from Aeluropus littoralis and NOS terminator) was successfully expressed in soybean cultivar TF-29. Southern and Northern blot analysis showed that AlNHX1 was successfully incorporated into the genome and expressed in the transgenic plants. The AlNHX1 transgenic plant lines exhibited improved growth in severe saline condition (150 mM NaCl). The transgenic lines accumulated a lower level of Na+ and a higher level of K+ in the leaves than wild-type plants under saline condition (150 mM NaCl). Observations on the chlorophyll content, photosynthetic rates, malondialdehyde and relative electrical conductivity indicated that transgenic plants exhibited tolerance to salt stress, growing normally at salt concentrations up to 150 mM. These results demonstrated that AlNHX1 was successfully transferred into soybean and the salt-tolerance was improved by the overexpression of AlNHX1.

Novel organic solvent-tolerant esterase isolated by metagenomics: insights into the lipase/esterase classification / Nueva esterasa tolerante a los solventes orgánicos aislada por metagenómica: ideas sobre la clasificación de las esterasas/lipasas

In order to isolate novel organic solvent-tolerant (OST) lipases, a metagenomic library was built using DNA derived from a temperate forest soil sample. A two-step activity-based screening allowed the isolation of a lipolytic clone active in the presence of organic solvents. Sequencing of the plasmid pRBest recovered from the positive clone revealed the presence of a putative lipase/esterase encoding gene. The deduced amino acid sequence (RBest1) contains the conserved lipolytic enzyme signature and is related to the previously described OST lipase from Lysinibacillus sphaericus 205y, which is the sole studied prokaryotic enzyme belonging to the 4.4 a/ß hydrolase subgroup (abH04.04). Both in vivo and in vitro studies of the substrate specificity of RBest1, using triacylglycerols or nitrophenyl-esters, respectively, revealed that the enzyme is highly specific for butyrate (C4) compounds, behaving as an esterase rather than a lipase. The RBest1 esterase was purified and biochemically characterized. The optimal esterase activity was observed at pH 6.5 and at temperatures ranging from 38 to 45 °C. Enzymatic activity, determined by hydrolysis of p-nitrophenyl esters, was found to be affected by the presence of different miscible and non-miscible organic solvents, and salts. Noteworthy, RBest1 remains significantly active at high ionic strength. These findings suggest that RBest1 possesses the ability of OST enzymes to molecular adaptation in the presence of organic compounds and resistance of halophilic proteins.

Con el fin de aislar nuevas variantes de lipasas tolerantes a solventes organicos (OST), se construyo una libreria metagenomica a partir de ADN obtenido de una muestra de suelo de bosque templado. A traves de un monitoreo en dos etapas, basado en la deteccion de actividades, se aislo un clon con actividad lipolitica en presencia de solventes organicos. La secuenciacion del plasmido pRBest recuperado del clon positivo revelo la presencia de un gen codificante de una hipotetica lipasa/esterasa. La secuencia deducida de amino acidos (RBest1) contiene los motivos conservados de enzimas lipoliticas y esta relacionada con la lipasa OST previamente descrita de Lysinibacillus sphaericus 205y, que es la unica enzima procariota estudiada perteneciente al subgrupo 4.4 de a/ß hidrolasas (abH4.04). Estudios in vivo e in vitro sobre la especificidad de sustratos de RBest1, utilizando triacil-gliceroles o p-nitrofenil-esteres, respectivamente, revelaron que la enzima es altamente especifica para compuestos butiricos (C4), comportandose como una esterasa y no como una lipasa. La esterasa RBest1 fue purificada y caracterizada bioquimicamente. La actividad optima de esterasa fue observada a pH 6,5 y las temperaturas optimas fueron entre 38 y 45 °C. Se establecio que la actividad enzimatica, determinada por hidrolisis de p-nitrofenil esteres, es afectada en presencia de diferentes solventes organicos miscibles y no miscibles, y tambien sales. Notoriamente, RBest1 permanece significativamente activa a elevadas fuerzas ionicas. Estos hallazgos sugieren que RBest1 posee la capacidad de las enzimas OST de la adaptacion molecular en presencia de compuestos organicos, asi como la resistencia de las proteinas halofilas.

Taurine prevents collagen abnormalities in high fructose-fed rats.

BACKGROUND & OBJECTIVE: Accumulation of collagen and changes in its physiochemical properties contribute to the development of secondary complications of diabetes. We undertook this study to see the effects of taurine on the content and characteristics of collagen from tail tendon of rats fed with high fructose diet. METHODS: The rats were divided into four groups of six each: control group (CON), taurine-supplemented control group (CON+TAU), taurine supplemented (FRU+TAU) and not supplemented fructose-fed group (FRU). The physico-chemical properties of collagen isolated from the tail tendon were studied. RESULTS: Fructose administration caused accumulation of collagen in tail tendon. Enhanced glycation and advanced glycation end products (AGE)-linked fluorescence together with alterations in aldehyde content, solubility pattern, susceptibility to denaturing agents and shrinkage temperature were observed in fructose-fed rats. Elevated b component of type I collagen was evidenced from the SDS gel pattern of collagen from the fructose-fed rats. Simultaneous administration of taurine alleviated these changes. INTERPRETATION & CONCLUSION: Taurine administration to fructose-rats had a positive influence on both quantitative and qualitative properties of collagen. The results of the present study suggested a role for the action of taurine in delaying diabetic complications and the possible use of taurine as an adjuvant therapeutic measure in the management of diabetes and its complications.

Amelioration of arsenic toxicity by phosphate salts in mungbean seedlings.

Sodium arsenate (Na2HAsO4.7H2O) is a potent inhibitor of mungbean seed germination and seedling growth. Germination is totally stopped at or above 50 microM Na2HAsO4.7H2O. Inhibition of seedling elongation started at a lower concentration of 5 microM As(V) and was drastically reduced at 20 microM As(V). Nutrients like salts of macroelements viz., NaH2PO4.2H2O, KH2PO4, K2SO4, MgSO4.7H2O, CaCl2.2H2O, (NH4)2SO4 NH4NO3 solutions at a concentration of 10mM and microelements viz., ZnSO4, CuSO4.5H2O, Na2MoO4.2H2O, MnCl2.4H2O, CoCl2.6H2O, FeSO4.7H2O solutions at a concentration of 1mM could help to ameliorate the toxic effects of As(V) to different degrees. Amelioration of As(V) toxicity was possible only when the mungbean seeds were pretreated with the above mentioned nutrients for 24 hr and then transferred to sodium arsenate. Simultaneous treatment of nutrients with As(V) or using nutrient solutions following As(V) treatment were of no help to reverse the toxic effects of sodium arsenate.

Protective effect of various chemicals on the impairment of sperm motility by Staphylococcus aureus.

In female genital tract spermatozoa remain active for several days, however, their activity is of much shorter duration if they remain in the seminal fluid outside the body. Majority of spermatozoa cease to move within first 24 hrs. This may be due to presence of bacteria or excretion of bacterial toxic products; To check whether the addition of certain chemicals can enhance the motility of spermatozoa in presence of bacteria, in vitro studies were conducted. 72h old culture supernatant of Staphylococcus aureus isolated from infertile male causing 100% immobilization of spermatozoa was incubated with semen samples in presence of various chemicals. The results showed that sodium chloride, magnesium sulfate, sodium citrate, folic acid, nicotinic acid, H2O2 and glutathione had protective effect on the impairment of motility. This study implicates that even if bacteria causing 100% immobilization of spermatozoa are present, supplementation of certain chemicals is beneficial.

Estudio microbiológico de ajo(Allium sativum L.) y cebolla (Allium cepa L.) deshidratados / Microbiological study of garlic (Allium sativum L.) and onion (Allium cepaL.) dehydrated

En este trabajo se efectuó un estudio microbiológico durante la deshidratación y el almacenamiento de ajo (Allium sativum L.) y de cebolla (Allium cepa L.). Al ajo se le efectuó un proceso de escaldado y a la cebolla un salmuereado previo a la deshidratación. En las materias primas los recuentos promedio expresados en UFC/g fueron: bacterias aerobias mesófilas entre 1,2 x 102y 1,6 x 103, mohos y levaduras entre 60 y 1,6 x 103, Lactobacillus spp. y Leuconostocmesenteroidessubesp. mesenteroides entre 10 y 50.Se identificaron: Penicillium spp., Monilia spp. Lactobacillus brevis,Leuconostoc mesenteroides subesp. mesenteroides y levaduras en ajo; Mucor spp., Penicillium spp., Monilia spp., Lactobacillus brevis y levaduras en dos tipos de cebolla. Leuconostoc mesenteroides subesp. mesenteroides sólo se detectó en un tipo de cebolla. En ajos deshidratados y durante el almacenamiento se hallaron Penicillium spp., Monilia spp., Lactobacillus brevis y levaduras mientras que los mismos con escaldado no presentaron desarrollo. Mucor spp., Penicillium spp., Monilia spp. y Lactobacillus brevis se identificaron en los dos tipos de cebolla deshidratadas. Con la incorporación del salmuereado la microflora se redujo significativamente hallándose solamente Penicillium spp. La utilización de barreras adicionales de control microbiano tales como escaldado y salmuereado produce un aporte importante a la estabilidad microbiológica de los productos.

A microbiological study during the process and the storage of garlic (Allium sativum L.) and onion (Allium cepa L.) dehydrated, with the additional barriers of blanching or brine immersion, was made. In all raw materials the average counts of aerobic mesophilic bacteria expressed in CFU/g ranged from 1.2 x 102to 1.6 x 103, molds and yeasts from 60 to 1.6 x 103, Lactobacillus spp. and Leuconostocmesenteroides subsp. mesenteroides between 10 and 50. Microorganisms identified were Penicillium spp., Monilia spp., Lactobacillus brevis, Leuconostoc mesenteroides subsp. mesenteroides and yeasts in garlic; Mucor spp., Penicillium spp., Monilia spp., Lactobacillus brevis and yeasts in both types of onions. Leuconostoc mesenteroides subsp. mesenteroides was detected in only kind of onion. In dehydrated garlic storage, Penicillium spp., Monilia spp., Lactobacillus brevis and yeasts were detected. In garlic, when a blanching step was carried out no microflora was detected. Mucor spp., Penicillium spp., Monilia spp. and Lactobacillus brevis were identified in both types of dehydrated onions. When brine immersion was included the microflora detected was significantly lower and only Penicillium spp. were found. The use of additional barriers such as blanching or brine immersion produces an important effect on the microbiological stability in these products.

Adrenocortical involvement during diverse stress in soft-shelled turtle Lissemys p. punctata Bonnoterre.

Adrenocortical responses to diverse stressful situations (dehydration, formaldehyde treatment and salt loading) were studied in the adult female soft-shelled turtle, Lissenmys p. punctata. Dehydration, formaldehyde treatment (formalin, 1%: 0.1 ml/100 g body weight daily) or salt loading (NaCl, 1%: 0.1 ml/100 g body weight daily) treatments consecutively for 7 days caused hypertrophy of the adrenocortical cells with their nuclear diameter increased, and depletions of adrenal cholesterol and ascorbic acid concentrations followed by decreased acid phosphatase and alkaline phosphatase activities in turtles. Corticosterone levels were elevated in both the adrenal gland and serum of turtles after dehydration and formalin stress, but the hormone level remained unaltered after salt loading in turtles. The results suggest active involvement of adrenal cortex in stress for homeostasis in Lissemys turtles.

Methylene blue binding to DNA with alternating GC base sequence: continuum treatment of salt effects.

Methylene blue (MB), an efficient singlet oxygen generating photoactive dye, binds to DNA and allows photosensitized reactions to be used for sequence-specific cleavage of the DNA backbone. Intercalation and groove binding are possible binding modes of the dye, depending on base sequences and environmental conditions. In a recent modeling study of methylene blue binding to a double stranded DNA decamer with an alternating GC sequence, six structural models for intercalation structures and for minor and major groove binding have been obtained. By estimating the binding energies (including electrostatic reaction field contributions of a salt-free aqueous solvent), symmetric intercalation at the 5'-CpG-3' and 5'-GpC-3' steps was found as the predominant binding mode, followed by a slightly weaker binding of the dye in the minor groove. In this study, the stability of the modeled structures has been analysed as a function of salt concentration. The results of finite difference numerical solutions of the non-linear Poisson-Boltzmann equation show that the stabilizing effect of salt is larger for free DNA than for the modeled MB-DNA complexes. Accordingly, the estimated binding energies decrease with increasing ionic strength. A slightly higher stabilization of the groove binding complexes results in comparable binding energies for symmetric intercalation and minor groove binding at high salt concentration. Both results are in qualitative agreement with experimental data.

Effect of inorganic salts, soaps and detergents on dissolution and larvicidal activity of alginate formulation of Bacillus sphaericus.

Various inorganic salts and commonly used soaps and detergents were tested in the laboratory for their effect on the dissolution and larvicidal residual activity of a slow-release alginate encapsulated granular formation of Bacillus sphaericus. Fluoride, chloride and sulphate salts and a detergent powder affected the residual activity of this formulation drastically by rupturing it but did not effect its larvicidal activity. Nitrates and phosphates of sodium and potassium also had the same effect but to a moderate level. The safest concentration of these water impurities for effective functioning of the alginate encapsulated B. sphaericus formulation have been determined.

Purification and characterization of ribulose-1,5-bisphosphate carboxylase from triticale.

Ribulose-1,5-bisphosphate carboxylase has been isolated from a synthetic cereal triticale and purified using a newly developed rapid procedure involving precipitation with ammonium sulphate (35-55% saturation), DEAE-cellulose (DE-52) chromatography and filtration through Sepharose CL-68. Molecular weights of the enzyme subunits are 15.5 and 52 kDa which corresponds to 540 kDa for the hexadecameric holoenzyme. Isoelectric focussing showed that the enzyme has a pI of 4.2. Various kinetic constants determined under aerobic conditions are: Km (CO2), 118 microM; Km (RuBP), 220 microM (at 20 mM NaHCO3) and Vmax, 690 nmole CO2 fixed/mg enzyme/min.

Açäo de diferentes sais sobre algumas propriedades texturais do gel (kamaboko) de surimi de sardinha / Action of differents salts on some properties of sardine surimi characteristics

O efeito de diferentes concentraçoes de alguns sais de cloro e de pirofosfato de sódio em algumas propriedades do gel de surimi de sardinha foi estudado. O emprego de MgCl2 incrementou a força do gel do surimi comparativamente ao NaCl (na mesma força iônica), enquanto o emprego de KCl resultou em gel de pior qualidade. Este comportamento foi observado para ambos os tratamentos térmicos empregados: cozimento direto (90 graus Celsius/30 min.) ou "incubaçao + cozimento (40 graus Celsius/30 min + 90 graus Celsius/30 min.) A força do gel mostrou ser fortemente influenciada pelo tratamento térmico empregado, enquanto a capacidade de retençao de água do gel mostrou-se dependente do tipo de sal empregado, porém, nao foi afetada pelo tipo de tratamento térmico. A substituiçao parcial do NaCl por MgCl2 resultou em gel de boas características texturais, e a adiçao de polifosfato de sódio, nos níveis empregados neste trabalho, nao resultou em melhora das propriedades estudadas, sendo mesmo prejudicial ao nível de capacidade de retençao de água.

Isolation of a salt-inducible gene from rice and its molecular cloning.

The genomic sequences of a salt inducible gene of rice have been isolated from both Indica and Japonica varieties of rice. The cDNA of this gene, SalT, characterized earlier shows sequence identity with both the genomic sequences. Approximately 750 base pairs of intervening sequence is present in the 1.2kb genomic sequence, obtained by PCR. There is a high degree of homology between the Indica and Japonica SalT genes except for the presence of an extra HindIII site in the former. A genomic library of Indica rice was screened and a phage clone of the SalT gene was partially characterized. Different species of monocot and dicot plant DNA showed sequence homology with SalT gene.

GABA/BZ-and NMDA-receptor interaction in digoxin-induced convulsions in rats.

Digoxin (7.5 micrograms icv) induced 'pop-corn' type of convulsions and 100% mortality. The GABA-ergic agents produced varying degree of protection against digoxin-induced neurotoxicity. Diazepam (4 mg/kg) offered significant protection whereas pentobarbital (5 mg/kg) and baclofen (5 mg/kg) markedly reduced per cent mortality, but ethanol (2 g/kg), progabide (50 mg/kg) and muscimol (0.5 mg/kg) as well as GABA (50 mg/kg) could not offer significant protection in doses used. GABA-ergic agonists; GABA, baclofen, diazepam and pentobarbital when administered along with MK-801 (0.5 mg/kg) a non-competitive NMDA antagonist, a potentiation of anticonvulsant action of MK-801 was observed. MK-801 showed potent anticonvulsant profile in dose range (0.25-1 mg/kg) studied. A synergistic influence of Mg2+ and K+ ions on NMDA receptor antagonism was also observed. A role of GABA-ergic facilitation and NMDA antagonism as a potential anticonvulsant approach in digoxin-induced convulsions in rats has been suggested.