A comparative study of different PCR-based DNA fingerprinting techniques for typing of lactic acid bacteria

A set of 92 lactic acid bacteria strains and 10 type/reference strains was analysed using three molecular techniques (randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), enterobacterial repetitive intergenic consensus (ERIC-PCR) and the polytrinucleotide (GTG)₅-PCR), in order to compare their discriminatory ability for typing of these bacteria. The results indicated that RAPD-PCR generated patterns with the largest number of bands, adequately clustering the isolates belonging to the same genus. Likewise, it showed the highest discriminatory capacity, while no discrimination between isolates of different genera was possible with ERIC-PCR and (GTG)₅-PCR. Therefore, the use of RAPD-PCR has turned out to be the most suitable procedure for typing LAB isolates belonging to different genera and species obtained from some fermented foods such as goat and Manchego cheeses, Almagro eggplant fermentations and Tempranillo wines.     

Characterization of cellulase and xylanase activities of Clostridium celerecrescens

The production of cellulases and xylanase by Clostridium celerecrescens, a new anaerobic mesophilic cellulolytic bacterium, was studied using various substrates (cellobiose, xylan and cellulose Whatman CF-11). While both cellulase (β-1,4-D-glucan glucanohydrolase) and xylanase (β-1,4-xylan xylanohydrolase) were produced on cellulose, only the latter was produced when xylan was used as the sole carbon source. A weak p-nitrophenyl-β-D cellobiohydrolase activity was detected in the extracellular filtrates when using cellulose as a substrate. Otherwise, β-glucosidase (p-nitrophenyl-β-D-glucopyranosidase) was always found to be associated with the bacteria and reached its maximum levels of growth on cellobiose. In all cases, enzyme production showed a cell growth associated profile. Activities of these enzymes had their optimal values within the ranges of temperature and pH reported for the corresponding enzymes from similar anaerobic mesophilic microorganisms, although a relatively high optimum temperature, 55°C, was found for xylanase. All enzymes showed a 90% reduction of half-life time for each 8°C increment of temperature. A 50% inhibition of xylanase and β-cellobiohydrolase activity was observed, through a competitive mechanism, by xylose (0.677 mmol dm^?3) and cellobiose (28 mmol dm^?3) respectively.     

Phenotypic and genotypic characterization of lactobacilli isolated from Spanish goat cheeses

Lactic acid bacteria from 18 Spanish goat cheeses produced by seven dairies were isolated to evaluate the genetic diversity of this bacterial community. 136 representative isolates were characterized by phenotyping and Randomly Amplified Polymorphic DNA (RAPD-PCR) analysis. Ten species were identified with predominance of Lactobacillus paracasei subsp. paracasei. The presence of L. curvatus, L. pentosus, L. cellobiosus and L. rhamnosus has not hitherto been reported in Spanish goat cheeses. A high degree of genetic diversity was found for L. paracasei subsp. paracasei, L. curvatus and L. plantarum. Some of the identified strains displayed strong acidifying and proteolytic capacities.     

Combined effect of thymol and cymene to control the growth of<Emphasis Type="Italic"> Bacillus cereus</Emphasis> vegetative cells

The antibacterial action of thymol and cymene on two Bacillus cereus strains (INRA-AVTZ415 and INRA-AVZ421) was studied. Thymol and cymene alone showed higher antibacterial effect on the growth of B. cereus in BHI at 30 °C, both in lag and in exponential growth phases, with increasing added concentrations. However, the two strains tested showed different sensitivity to these natural compounds. When thymol and cymene were applied simultaneously, it resulted in a greater antibacterial effect than when the compounds were applied separately. The synergistic behaviour of both compounds was also shown for lag phase B. cereus INRA-AVZ421 cells in food substrates (carrot juice, chicken soup and pumpkin cream) at 30 °C, though a clear food-related effect was observed. This study indicates the potential use of thymol and cymene applied simultaneously for preservation of minimally processed foods.     

Optical, magnetic and electronic properties of Ln2O2Te (Ln = La, Sm and Gd)

The synthesis, the optical and magnetic properties and the electronic structure of the rare-earth oxytellurides of formula Ln₂O₂Te (Ln = La, Sm and Gd) are reported. The magnetic measurements show that La₂O₂Te exhibit a diamagnetic behavior. The value of the magnetization M of Gd₂TeO₂ at our highest available field (50 kOe) is about 5.5μ_B, well below the expected value for two independent Gd ions (2gJ = 14μ_B) confirming the antiferromagnetic character of this compound. The magnetic properties of Sm₂TeO₂ reveal that Sm is in its 3+ oxidation state with a large Van Vleck contribution. The electronic structure calculations, studied by means of a first principles DFT approach, are consistent with the optical measurements and suggest an indirect band gap semiconductor behavior.     

Virtual work bench for electronic instrumentation teaching

This article describes the design and implementation of a virtual laboratory, computer-generated for the theoretical and practical teaching of electronic instrumentation. The virtual laboratory is implemented in a windows framework for which the teacher or student requires no prior knowledge to manage the program. The Virtual Instrumentation Workbench for Instrumentation allows visualization of the time and frequency effects produced in analog to digital conversion and in digital signal processing as well as visualizing the effects of 16 different teaching topics. It incorporates ten different waveforms, eight windows algorithms, five digital filters, and five discrete/fast Fourier transforms (DFT/FFT). The use of the virtual laboratory has allowed more than 500 students to understand the functioning of the new devices that are flooding the instrumentation market. Although it has been developed as an aid for electronic instrumentation teaching, it can also be used for the teaching of digital signal treatment     

IKKα Induces Epithelial–Mesenchymal Changes in Mouse Skin Carcinoma Cells That Can Be Partially Reversed by Apigenin

NMSC (non-melanoma skin cancer) is a common tumor in the Caucasian population, accounting for 90% of skin cancers. Among them, squamous cell carcinomas (SCCs) can metastasize and, due to its high incidence, constitute a severe health problem. It has been suggested that cutaneous SCCs with more risk to metastasize express high levels of nuclear IKKα. However, the molecular mechanisms that lead to this enhanced aggressiveness are largely unknown. To understand in depth the influence of nuclear IKKα in skin SCC progression, we have generated murine PDVC57 skin carcinoma cells expressing exogenous IKKα either in the nucleus or in the cytoplasm to further distinguish the tumor properties of IKKα in both localizations. Our results show that IKKα promotes changes in both subcellular compartments, resembling EMT (epithelial–mesenchymal transition), which are more pronounced when IKKα is in the nucleus of these tumor cells. These EMT-related changes include a shift toward a migratory phenotype and induction of the expression of proteins involved in cell matrix degradation, cell survival and resistance to apoptosis. Additionally, we have found that apigenin, a flavonoid with anti-cancer properties, inhibits the expression of IKKα and attenuates most of the pro-tumoral EMT changes induced by IKKα in mouse tumor keratinocytes. Nevertheless, we have found that apigenin only inhibits the expression of the IKKα protein when it is localized in the cytoplasm.     

Characterization of retanned chrome bovine leather by thermomechanical analysis

The thermomechanical analysis is applied to characterize the shrinking process of bovine chrome leather. The interpretation of the curves obtained is performed and a value of glass transition temperature is estimated. The influence of the sampling zone on the thermomechanical parameters is discussed. The influence of different “retanning agents,” which act as crosslinking compounds on the fibrous structure of collagen, is also evaluated. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 82: 314–322, 2001     

NLRP3 Inflammasome: Potential Role in Obesity Related Low-Grade Inflammation and Insulin Resistance in Skeletal Muscle

Among multiple mechanisms, low-grade inflammation is critical for the development of insulin resistance as a feature of type 2 diabetes. The nucleotide-binding oligomerization domain-like receptor family (NOD-like) pyrin domain containing 3 (NLRP3) inflammasome has been linked to the development of insulin resistance in various tissues; however, its role in the development of insulin resistance in the skeletal muscle has not been explored in depth. Currently, there is limited evidence that supports the pathological role of NLRP3 inflammasome activation in glucose handling in the skeletal muscle of obese individuals. Here, we have centered our focus on insulin signaling in skeletal muscle, which is the main site of postprandial glucose disposal in humans. We discuss the current evidence showing that the NLRP3 inflammasome disturbs glucose homeostasis. We also review how NLRP3-associated interleukin and its gasdermin D-mediated efflux could affect insulin-dependent intracellular pathways. Finally, we address pharmacological NLRP3 inhibitors that may have a therapeutical use in obesity-related metabolic alterations.