Distribution and elimination of levamisole in eggs and tissues after oral administration to laying hens, determined by LC-MS/MS.

Levamisole was administered to laying hens, and concentrations in eggs and tissues (thigh muscle, breast muscle, liver and kidney) were determined by a newly developed liquid chromatography tandem mass spectrometry method, which allowed trace level quantification of levamisole. The adopted analytical method showed good sensitivity, repeatability and percentage of recovery from spiked matrices. Maximum concentrations of levamisole were found on the first day after the administration (531.1 µg/kg in liver, 164.3 µg/kg in egg yolk, 130.7 µg/kg in kidney, 78.0 µg/kg in breast muscle, 70.7 µg/kg in thigh muscle and 64.0 µg/kg in egg white), after which there is a decline. The compound was rapidly eliminated from eggs, with a half-life of 1.3 days. Elimination appeared to be slower in thigh muscle (3.5 days), breast muscle (3.4 days) and liver (3.3 days). According to this experiment, the levamisole withdrawal periods calculated for eggs, liver, kidney, breast muscle and thigh muscle in laying hens were 14.1, 6.1, >4.0, 14.5 and 13.0 days, respectively. The longest time for levamisole residues to be completely released from tissues was seen in liver samples (37.4 days), followed by thigh muscle, breast muscle and kidney. Elimination from eggs was fastest (16.4 days for levamisole residues to drop below the method quantification limit).

Dynamic finite-element model for efficient modelling of electric currents in electroporated tissue.

In silico experiments (numerical simulations) are a valuable tool for non-invasive research of the influences of tissue properties, electrode placement and electric pulse delivery scenarios in the process of electroporation. The work described in this article was aimed at introducing time dependent effects into a finite element model developed specifically for electroporation. Reference measurements were made ex vivo on beef liver samples and experimental data were used both as an initial condition for simulation (applied pulse voltage) and as a reference value for numerical model calibration (measured pulse current). The developed numerical model is able to predict the time evolution of an electric pulse current within a 5% error over a broad range of applied pulse voltages, pulse durations and pulse repetition frequencies. Given the good agreement of the current flowing between the electrodes, we are confident that the results of our numerical model can be used both for detailed in silico research of electroporation mechanisms (giving researchers insight into time domain effects) and better treatment planning algorithms, which predict the outcome of treatment based on both spatial and temporal distributions of applied electric pulses.

Acid resistance and response to pH-induced stress in two Lactobacillus plantarum strains with probiotic potential.

Two new Lactobacillus plantarum strains, KR6-DSM 28780 and M5 isolated from sour turnip and traditional dried fresh cheese, respectively, were evaluated for species identity, antibiotic susceptibility, resistance to gastrointestinal conditions and adaptive response to low pH. Resistance mechanisms involved in the adaptation to acid-induced stress in these two strains were investigated by quantitative PCR of the atpA, cfa1, mleS and hisD genes. In addition to absence of antibiotic resistance, the two L. plantarum strains showed excellent survival rates at pH values as low as 2.4. Adaptive response to low pH was clearly observed in both strains; strain KR6 was superior to M5, as demonstrated by its ability to survive during 3 h incubation at pH 2.0 upon adaptation to moderately acidic conditions. In contrast, acid adaptation did not significantly affect the survival rate during simulated passage through the gastrointestinal tract. In both strains, induction of histidine biosynthesis (hisD) was upregulated during the acid adaptation response. In addition, significant upregulation of the cfa1 gene, involved in modulation of membrane fatty acid composition, was observed during the adaptation phase in strain KR6 but not in strain M5. Cells adapted to moderately acidic conditions also showed a significantly increased viability after the lyophilisation procedure, a cross-protection phenomenon providing additional advantage in probiotic application.

Electrochemotherapy: a new technological approach in treatment of metastases in the liver.

Electrochemotherapy is now in development for treatment of deep-seated tumors, like in bones and internal organs, such as liver. The technology is available with a newly developed electric pulse generator and long needle electrodes; however the procedures for the treatment are not standardized yet. In order to describe the treatment procedure, including treatment planning, within the ongoing clinical study, a case of successful treatment of a solitary metastasis in the liver of colorectal cancer is presented. The procedure was performed intraoperatively by inserting long needle electrodes, two in the center of the tumor and four around the tumor into the normal tissue. The insertion of electrodes proved to be feasible and was done according to the treatment plan, prepared by numerical modeling. After intravenous bolus injection of bleomycin the tumor was exposed to electric pulses. The delivery of the electric pulses did not interfere with functioning of the heart, since the pulses were synchronized with electrocardiogram in order to be delivered outside the vulnerable period of the ventricles. Also the post treatment period was uneventful without side effects. Re-operation of the treated metastasis demonstrated feasibility of the reoperation, without secondary effects of electrochemotherapy on normal tissue. Good antitumor effectiveness with complete tumor destruction was confirmed with histological analysis. The patient is disease-free 16 months after the procedure. In conclusion, treatment procedure for electrochemotherapy proved to be a feasible technological approach for treatment of liver metastasis. Due to the absence of the side effects and the first complete destruction of the treated tumor, treatment procedure for electrochemotherapy seems to be a safe method for treatment of liver metastases with good treatment effectiveness even in difficult-to-reach locations.

Heap leaching of lead contaminated soil using biodegradable chelator [S,S]-ethylenediamine disuccinate.

The feasibility of heap leaching Pb contaminated soil using biodegradable chelator [S,S]-ethylenediamine disuccinate ([S,S]-EDDS) in concentrations ranging from 10 to 40 mmol kg(-1) was assessed in a laboratory study. Treatment with 40 mmol kg(-1) [S,S]-EDDS was the most efficient, removing 31.3% of soil Pb and producing a waste washing solution with a peak Pb concentration slightly exceeding 10 mM. The removal of Pb from simulated washing solution containing 10 mM Pb by biodegradation of the Pb-[S,S]-EDDS complex and sorption of the released Pb on a vermiculite/soya meal/Slovakite based permeable bed was completed in 7.5 days. Lead recovery from washing solutions with higher molarity was significantly slower. Lead fractionation and bioavailability were assessed 6-months after soil treatment. Heap leaching removed Pb proportionally from carbonates (approx. 25% Pb) and organic matter (> 50% Pb), the two main Pb-bearing soil fractions,and increased the small pool of Pb soluble in the soil solution. The latter had no effect on Pb phytoavailability. Accumulation of Pb in leaves of the reported hyperaccumulator species Thlaspi goesingense was < 20 mg kg(-1) in all treatments, comparable to the Pb concentration in lettuce (Latuca sativa). Soil treatment with 40 mmol kg(-1) [S,S]-EDDS reduced concentrations of Pb orally-accessible in the stomach and intestine phase from initial 234.0 and 52.5 mg kg(-1) to 157.8 and 29.7 mg kg(-1), respectively, according to Ruby's test. Our results indicate that heap leaching of Pb with [S,S]-EDDS is not a viable remediation option for the tested soil and conditions used.

Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92.

AIMS: To investigate the functional role of surface layer proteins (S-layer) in probiotic strain Lactobacillus acidophilus M92, especially its influence on adhesiveness to mouse ileal epithelial cells. METHODS AND RESULTS: Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, ca at 45 kDa in L. acidophilus M92. Southern blot with pBK1 plasmid, containing slpA gene, gave a positive signal, suggesting that L. acidophilus M92 has a slpA gene coding for the S-layer proteins. S-layer proteins of this strain are present during all phases of growth. The S-layer proteins appeared when cells treated with 5 mol l(-1) LiCl were allowed to grow again. Removal of the S-layer proteins reduced adhesion of L. acidophilus M92 to mouse ileal epithelial cells. Furthermore, the viability of cells without S-layer were reduced in simulated gastric juice at low pH range (2, 2.5, 3) and simulated pancreatic juice with bile salts (1.5 and 3 g l(-1)). S-layer proteins of L. acidophilus M92 were resistant to pepsin and pancreatin, in contrast, the treatment with proteinase K led to a significant proteolysis of the S-layer proteins. CONCLUSIONS: These results demonstrated functional role of S-layer; it is responsible for adhesiveness of Lactobacillus acidophilus M92 to mouse ileal epithelial cells and has a protective role for this strain. SIGNIFICANCE AND IMPACT OF THE STUDY: S-layer proteins have an important role in the establishment of probiotic strain Lactobacillus acidophilus M92 in the gastrointestinal tract.

Washing of Pb contaminated soil using [S,S] ethylenediamine disuccinate and horizontal permeable barriers.

The feasibility of in situ washing of soil contaminated with Pb (6.83 mmol kg(-1)) using biodegradable chelator, [S,S] stereoisomere of ethylenediamine disuccinate ([S,S]-EDDS) and horizontal permeable barriers was examined in soil columns. After 4-cycles of 10 mmol kg(-1) soil [S,S]-EDDS applications, followed by irrigation, 24.7% of total initial Pb was washed from the contaminated soil and accumulated into the barrier. Sequential extractions indicated that washing removed most of the Pb from the organic soil fraction. Barriers were positioned 20 cm deep in the soil and consisted of a 2 cm layer of nutrient enriched vermiculite. Barriers reduced leaching of Pb in the first cycle of [S,S]-EDDS addition by more than 500-times compared to columns with no barrier. After four cycles of chelator addition, a total of 0.24% of the initial Pb was leached from the columns with barriers. Four cycles of in situ soil washing in soil columns were less effective than simulated ex situ soil washing with 40 mmol kg(-1) [S,S]-EDDS, where 51.0% of the Pb was removed after 48-h extraction. Ex situ soil washing with 10 mmol kg(-1) [S,S]-EDDS was equally effective as the first cycle of in situ soil washing (15.5% and 14.5% of removed Pb, respectively).

Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92.

AIMS: To investigate aggregation and adhesiveness of Lactobacillus acidophilus M92 to porcine ileal epithelial cells in vitro, and the influence of cell surface proteins on autoaggregation and adhesiveness of this strain. METHODS AND RESULTS: Lactobacillus acidophilus M92 exhibits a strong autoaggregating phenotype and manifests a high degree of hydrophobicity determined by microbial adhesion to xylene. Aggregation and hydrophobicity were abolished upon exposure of the cells to pronase and pepsin. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, approximated at 45 kDa, in L. acidophilus M92. The relationship between autoaggregation and adhesiveness to intestinal tissue was investigated by observing the adhesiveness of L. acidophilus M92 to porcine ileal epithelial cells. Removal of the S-layer proteins by extraction with 5 mol l-1 LiCl reduced autoaggregation and in vitro adhesion of this strain. CONCLUSIONS: These results demonstrate that there is relationship between autoaggregation and adhesiveness ability of L. acidophilus M92, mediated by proteinaceous components on the cell surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation has shown that L. acidophilus M92 has the ability to establish in the human gastrointestinal tract, which is an important determinant in the choice of probiotic strains.

[Respiratory function in street cleaners and garbage collectors]. / Respiracijska funkcija radnika na ciscenju ulica i odvazanju otpada.

The prevalence of chronic impairment of the respiratory system and ventilatory capacity was studied in 81 sanitation workers. In addition, the prevalence of chronic respiratory symptoms was studied in 65 non-exposed control workers. Among the sanitation workers the prevalence of most of the chronic respiratory symptoms was statistically higher than among the controls. The sanitation workers--smokers had a significantly higher prevalence of chronic cough, chronic phlegm and chest tightness than the non-smoking sanitation workers (P < 0.05). The sanitation smokers exposed for more than 10 years had significantly higher prevalences of nasal catarrh (P < 0.01) and sinusitis (P < 0.05) than those with a shorter exposure. FVC and FEV1 were significantly reduced in sanitation workers exposed for more than 10 years in relation to predicted normal values. These differences were not significant for FEF50 and FEF25. Our results suggest that sanitation workers may develop chronic respiratory symptoms and lung function changes, in the first place those of FVC and FEV1.

Ocorrência da infiltraçäo marginal de oito materiais seladores provisórios: estudo "in vitro" / Ocurrence of marginal leakage of eight temporary sealing materials in vitro study

Os autores realizaram um pesquisa "in vitro" avaliando dentre oito diferentes materiais seladores provisórios disponíveis no mercado aqueles que apresentam melhor capacidade seladora e fornecem maior proteçäo para o curativo de demora nas terapias pulpares

Avulsäo dentária - reimplante e apicificaçäo: relato de um caso / Dental avulsion, replantation and apical formation: report of a case

A avulsäo dos dentes anteriores é uma séria situaçäo de emergência e o reimplante um processo de enorme valor funcional, estético e psicológico. Os autores relatam um caso de avulsäo, reimplante e apicificaçäo, após realizar uma breve revisäo da literatura, enfatizando a importância de conduta adequada para obtençäo de sucesso