Impact of an Engineered Copper-Titanium Dioxide Nanocomposite and Parent Substrates on the Bacteria Viability,Antioxidant Enzymes and Fatty Acid Profiling

Due to the systematic increase in the production of nanomaterials (NMs) and their applications in many areas of life, issues associated with their toxicity are inevitable. In particular, the performance of heterogeneous NMs, such as nanocomposites (NCs), is unpredictable as they may inherit the properties of their individual components. Therefore, the purpose of this work was to assess the biological activity of newly synthesized Cu/TiO₂-NC and the parent nanoparticle substrates Cu-NPs and TiO₂-NPs on the bacterial viability, antioxidant potential and fatty acid composition of the reference Escherichia coli and Bacillus subtilis strains. Based on the toxicological parameters, it was found that B. subtilis was more sensitive to NMs than E. coli. Furthermore, Cu/TiO₂-NC and Cu-NPs had an opposite effect on both strains, while TiO₂-NPs had a comparable mode of action. Simultaneously, the tested strains exhibited varied responses of the antioxidant enzymes after exposure to the NMs, with Cu-NPs having the strongest impact on their activity. The most considerable alternations in the fatty acid profiles were found after the bacteria were exposed to Cu/TiO₂-NC and Cu-NPs. Microscopic images indicated distinct interactions of the NMs with the bacterial outer layers, especially in regard to B. subtilis. Cu/TiO₂-NC generally proved to have less distinctive antimicrobial properties on B. subtilis than E. coli compared to its parent components. Presumably, the biocidal effects of the tested NMs can be attributed to the induction of oxidative stress, the release of metal ions and specific electrochemical interactions with the bacterial cells.     

Encapsulated yeast cells inside Paramecium primaurelia: a model system for protection capability of polyelectrolyte shells

One of the most promising applications of encapsulated living cells is their use as protected transplanted tissue into the human body. A suitable system for the protection of living cells is the use of nano‐ or microcapsules of polyelectrolytes. These shells can be deposited easily on top of the cells by means of a layer‐by‐layer technique. An interesting feature of the capsules is the possibility to control their properties on a nanometre level, tuning their wall texture via the preparation conditions. Here we introduce a model system to test the protection ability of polyelectrolyte capsules. Common bakery yeast cells were encapsulated. They were coated with a fluorescently labelled shell at conditions known to guarantee cell survival, and the cell interior was stained with DAPI. The protozoan Paramecium primaurelia was incubated with this double‐stained living yeast and visualized by means of two‐photon excitation fluorescence microscopy. Cross‐sections of the dye‐stained material as well as autofluorescence of the fixed protozoan allowed us to follow the digestion of the coated yeast with time. Our investigation reveals that capsules prepared under these deposition conditions are permeable to lysosomal enzymes, leading to degradation of the yeast inside the intact capsules. Our preliminary results indicate the suitability of the introduced model as a test system of this permeability.     

FRET measurements on fuzzy fluorescent nanostructures

In the last decade, fluorescence resonance energy transfer (FRET) has become a useful technique for studying intermolecular interactions applied to the analysis of biological systems. Although FRET measurements may be very helpful in the comprehension of different cellular processes, it can be difficult to obtain quantitative results, hence the necessity of studying FRET on controllable systems. Here, a fuzzy nanostructured system called a nanocapsule is presented as a nanometric-device allowing distance modulation, thus preserving photophysical properties of fluorescent dyes and exhibiting good potential features for improving quantitative FRET analysis. We evaluated the behavior of such a sample using four FRET methods (three of them based on steady-state fluorescence and one using lifetime measurements). Within some limitations that can be overcome, these nanodevices have the potential to serve as a benchmark system for characterizing new FRET couples and to develop quantitative approaches for FRET analysis.     

Effect of citric acid, citrate and slight a(w) decreases on the bacteriological stability of Hague liver sausage

Combinations of citric acid and citrate, with several carbohydrates and KCl, were tested in a canned Hague liver sausage model for their antimicrobial properties. pH values for the test products ranged from 5·38 to 6·48; a(w) values varied from 0·960 to 0·977. Before pasteurization, the sausage doughs were contaminated with mixtures of bacterial spores. Since the bacteriological stability sometimes appeared to be considerably improved, heated packages were challenged by incubation at 20° and 30°C; some of them for up to 2 years. Incubated packages from the standard formulation spoiled at 30°C within 3 days. Starting from this point, products with a variable, but increased, bacteriological stability were obtained. For the best stabilized batches, first symptoms of spoilage were observed after periods of incubation at 30°C, ranging from 49 to 150 days. Combinations of pH and a(w) values which can be relied upon to yield stable products consistently, cannot be derived from the accumulated data. Citric acid appeared a far more effective stabilizer than its salt. If we regard the undissociated citric acid concentration for these products (a(w) 0·960-0·972) as a significant parameter, values above approximately 13 μmkg(-1), yielded products with remarkable bacteriological stability.     

Confocal laser scanning microscopy to study formation and properties of polyelectrolyte nanocapsules derived from CdCO3 templates

Three-dimensional confocal laser scanning microscopy (CLSM) was used as an essential investigation method to obtain information about the formation and morphological characteristics of nanocapsules. Nanocapsules are built by layer-by-layer deposition of alternatively charged polyelectrolytes on templates forming nanostructured hollow shells. CLSM is unique in allowing for monitoring of the core dissolution process in real time and for studying nanocapsule functioning in hydrated conditions within a three-dimensional and temporal framework. Since we are also interested in the identification of other possible templates, we briefly report on the use of yeast cells as biocolloidal cores monitored by means of two-photon microscopy. Here we focus our attention on the use of CdCO(3) crystals as template candidates for the preparation of stable capsules. Both cubic and spherical CdCO(3) cores have been produced. Cubic cores exhibit higher monodispersity and smaller size compared to spherical ones. Capsules templated on these cores have a higher surface-to-volume ratio that is valuable for applications related to drug delivery, functional properties of the shells and adsorption of proteins, and other biologically relevant molecules. Microsc. Res. Tech. 59:536-541, 2002.     

The investigations of gas‐powder two phase flow in packed bed

Gas‐powder two‐phase flow in packed beds was investigated and the results are presented in this paper. The experimental system, in which the glass powder and air gas were injected into the lower part of the packed bed allowed the estimation of the influence of gas velocity, powder feed rate, powder and packed particles diameter on pressure loss and total hold up of powder in the packed bed. On the base of the experimental results, a one‐dimensional mathematical model for gas‐powder two‐phase flow in the packed bed was developed. This model allows satisfactory prediction of the pressure loss and the hold ups of powders. The maximum deviation between calculated and measured values was less than ±15%. Futhermore, the conditions when the blockade of the flow occurs were defined. The additional pressure loss (expressed by F_k) due to the gravitional force of powders and the collision and the friction between powders and packed particles, was correlated with Froude number as: for the void fraction in the packed bed ?₀ of between 0.36 to 0.41 or .     

Efficacy and safety of acarbose in insulin-treated patients with type 2 diabetes

OBJECTIVE: To demonstrate the efficacy, tolerability, and safety of acarbose compared with placebo in patients with type 2 diabetes inadequately controlled with diet and insulin. RESEARCH DESIGN AND METHODS: A multicenter randomized double-blind placebo-controlled parallel-group comparison study was conducted. The trial was 26 weeks with a 2-week screening period and a 24-week period of treatment with acarbose or placebo, with forced titration from 25 mg t.i.d. to 50 mg t.i.d. after 4 weeks, and titration of 50 mg t.i.d. to 100 mg t.i.d. after 12 weeks based on glucose control. The dosage of insulin was to remain stable. The primary efficacy variable was mean change from baseline in HbA1c, and secondary efficacy variables included mean changes in fasting and postprandial plasma glucose and triglyceride levels. RESULTS: The addition of acarbose to the treatment of patients receiving background insulin and diet therapy resulted in a statistically significant reduction in mean HbA1c of 0.69% compared with placebo. There were statistically significant reductions in postprandial plasma glucose and glucose area under the curve, and in postprandial serum triglyceride levels in the acarbose-treated patients. Gastrointestinal side effects were more frequently reported in the acarbose-treated patients. There were no significant differences in hypoglycemic events or liver transaminase elevations between groups. CONCLUSIONS: This study demonstrated that the addition of acarbose to patients with type 2 diabetes who are inadequately controlled with insulin and diet is safe and generally well tolerated and that it significantly lowers HbA1c and postprandial glucose levels.     

Functionalized gold nanoparticles: a detailed in vivo multimodal microscopic brain distribution study

In the present study, the in vivo distribution of polyelectrolyte multilayer coated gold nanoparticles is shown, starting from the living animal down to cellular level. The coating was designed with functional moieties to serve as a potential nano drug for prion disease. With near infrared time-domain imaging we followed the biodistribution in mice up to 7 days after intravenous injection of the nanoparticles. The peak concentration in the head of mice was detected between 19 and 24 h. The precise particle distribution in the brain was studied ex vivo by X-ray microtomography, confocal laser and fluorescence microscopy. We found that the particles mainly accumulate in the hippocampus, thalamus, hypothalamus, and the cerebral cortex.