Lactobacilli isolated from kefir grains: evidence of the presence of S-layer proteins

In the present study we report for the first time the presence of S-layer proteins in Lactobacillus kefir and Lactobacillus parakefir isolated from kefir grains. Soluble whole-cell protein profile obtained either by mechanical disruption (X-press) or by a combined treatment with lysozyme and SDS on whole cells, showed a significant band of apparent molecular mass of 66-71 kDa as measured by SDS-PAGE. The intensity of this band was considerably reduced when cells were treated with 5 M-LiCl. The above mentioned proteins were recovered in the LiCl extracts. After dialysis and concentration, the proteins extracted were able to reassemble in a regular array. Negative staining of these protein preparations were analysed by transmission electron microscopy and a paracrystalline arrangement was seen. Thin sections of bacteria analysed by transmission electron micrographs showed an outermost layer over the bacterial cell wall, that was lost after the LiCl treatment. The production of this surface structure under different culture conditions was also evaluated. Finally, the relationship between the presence of S-layer proteins and surface properties (e.g. adhesion to Caco-2 cells, autoaggregation, and hemagglutination) was investigated.     

Novel isolates of lactobacilli from fermented Portuguese olive as potential probiotics

The purpose of this work was to screen for and characterize the potential probiotic features of strains of lactic acid bacteria isolated from Galega cultivar fermented olives, to eventually develop an improved probiotic food from plant origin. From 156 isolated strains, 10 were acid – and bile salt tolerant, and exhibited survival rates up to 48%, following simulated digestion. All strains exhibited auto- (4–12%) and co-aggregation features (≥30%), as well as hydrophobicity (5–20%) and exopolysaccharide-producing abilities, while no strain possessed haemolytic capacity or ability to hydrolyse mucin. Antibiotic resistance, oleuropein degradation, proteolytic activity and antimicrobial activity were strain-dependent features. Overall, 10 strains – belonging to Lactobacillus plantarum and Lactobacillus paraplantarum, appear to possess a probiotic value.     

Poly(3-methylthiophene)/MnO2 composite electrodes as electrochemical capacitors

Composite electrodes prepared by electrodeposition of manganese oxide on titanium substrates modified with poly(3-methylthiophene) (PMeT) were investigated and compared with Ti/MnO₂ electrodes. The polymer films were prepared by galvanostatic deposition at 2 mA cm⁻² with different deposition charges (250 and 1500 mC cm⁻²). The electrodes were characterized by cyclic voltammetry in 1 mol L⁻¹ Na₂SO₄ and by scanning electron microscopy. The results show a very significant improvement in the specific capacitance of the oxide due the presence of the polymer coating. For Ti/MnO₂ the specific capacitance was of 122 F g⁻¹, while Ti/PMeT₂₅₀/MnO₂ and Ti/PMeT₁₅₀₀/MnO₂ displayed values of 218 and 66 F g⁻¹, respectively. If only oxide mass is considered, the capacitances of the composite electrode increases to 381 and 153 F g⁻¹, respectively. The micrographs of samples show that the polymer coating leads to very significant changes in the morphology of the oxide deposit, which in consequence, generate the improvement observed in the charge storage property.     

Amination of Polymeric Braid Structures to Improve Tendon Healing: An Experimental Comparison

Several polymers are researched for tendon repair as polyethylene terephthalate (PET) and polylactic acid (PLA). These are biocompatible and useful in scaffolding repair though with minimal success due to long-term failure. There is a need to improve such scaffolds' design and physical–chemical nature. This work concerns surface functionalization of polymeric braids (PET and PLA) that fulfill the high mechanical demands of tissues such as tendons. The functionalization aims to incorporate amine groups in the braids' surface, improve cell adhesion, and consequently, the poor healing rate of these tissues and the biointegration of the braids. Two approaches are compared: the direct application of NH₃ plasma and the surface grafting of EDA after O₂ plasma activation. X-ray photoelectron spectroscopy (XPS) shows that amine groups are effectively introduced onto the samples' surfaces. Besides, the plasma parameters chosen do not compromise the topography and tensile behavior of the braids. Resazurin assay and scanning electron microscopy show that the NH₃ treatment improves cell–biomaterial interaction as improved cell adhesion and proliferation are observed. Both approaches are safe for biomedical applications. The NH₃ plasma approach is more environmentally friendly, faster, and easier to scale-up, showing potential for application in the final hybrid medical device.     

Bioavailability of Quercetin in Humans with a Focus on Interindividual Variation

After consumption of plant‐derived foods or beverages, dietary polyphenols such as quercetin are absorbed in the small intestine and metabolized by the body, or they are subject to catabolism by the gut microbiota followed by absorption of the resulting products by the colon. The resulting compounds are bioavailable, circulate in the blood as conjugates with glucuronide, methyl, or sulfate groups attached, and they are eventually excreted in the urine. In this review, the various conjugates from different intervention studies are summarized and discussed. In addition, the substantial variation between different individuals in the measured quercetin bioavailability parameters is assessed in detail by examining published human intervention studies where sources of quercetin have been consumed in the form of food, beverages, or supplements. It is apparent that most reported studies have examined quercetin and/or metabolites in urine and plasma from a relatively small number of volunteers. Despite this limitation, it is evident that there is less interindividual variation in metabolites which are derived from absorption in the small intestine compared to catabolites derived from the action of microbiota in the colon. There is also some evidence that a high absorber of intact quercetin conjugates could be a low absorber of microbiota‐catalyzed phenolics, and vice versa. From the studies reported so far, the reasons or causes of the interindividual differences are not clear, but, based on the known metabolic pathways, it is predicted that dietary history, genetic polymorphisms, and variations in gut microbiota metabolism would play significant roles. In conclusion, quercetin bioavailability is subject to substantial variation between individuals, and further work is required to establish if this contributes to interindividual differences in biological responses.     

Non-Functionalized Gold Nanoparticles Inhibit Human Papillomavirus (HPV) Infection

The spontaneous interaction between human papillomavirus type 16 (HPV16) L1 virus-like particles (VLPs) and non-functionalized gold nanoparticles (nfGNPs) interferes with the nfGNPs’ salt-induced aggregation, inhibiting the red–blue color shift in the presence of NaCl. Electron microscopy and competition studies showed that color-shift inhibition is a consequence of direct nfGNP–VLP interaction and, thus, may produce a negative impact on the virus entry cell process. Here, an in vitro infection system based on the HPV16 pseudovirus (PsV) was used to stimulate the natural infection process in vitro. PsVs carry a pseudogenome with a reporter gene, resulting in a fluorescent signal when PsVs infect a cell, allowing quantification of the viral infection process. Aggregation assays showed that nfGNP-treated PsVs also inhibit color shift in the presence of NaCl. High-resolution microscopy confirmed nfGNP–PsV complex formation. In addition, PsVs can interact with silver nanoparticles, suggesting a generalized interaction of metallic nanoparticles with HPV16 capsids. The treatment of PsVs with nfGNPs produced viral infection inhibition at a higher level than heparin, the canonical inhibitor of HPV infection. Thus, nfGNPs can efficiently interfere with the HPV16 cell entry process and may represent a potential active component in prophylactic formulations to reduce the risk of HPV infection.     

Combining Radiation- with Immunotherapy in Prostate Cancer: Influence of Radiation on T Cells

Radiation of tumor cells can lead to the selection and outgrowth of tumor escape variants. As radioresistant tumor cells are still sensitive to retargeting of T cells, it appears promising to combine radio- with immunotherapy keeping in mind that the radiation of tumors favors the local conditions for immunotherapy. However, radiation of solid tumors will not only hit the tumor cells but also the infiltrated immune cells. Therefore, we wanted to learn how radiation influences the functionality of T cells with respect to retargeting to tumor cells via a conventional bispecific T cell engager (BiTE) and our previously described modular BiTE format UNImAb. T cells were irradiated between 2 and 50 Gy. Low dose radiation of T cells up to about 20 Gy caused an increased release of the cytokines IL-2, TNF and interferon-γ and an improved capability to kill target cells. Although radiation with 50 Gy strongly reduced the function of the T cells, it did not completely abrogate the functionality of the T cells.     

ASALBP: the alternative subgraphs assembly line balancing problem

Assembly line balancing problems basically consist of assigning a set of tasks to a group of workstations while maintaining the tasks’ precedence relations, which are represented by a predetermined precedence graph. However, one or more parts of a product's assembly process may admit alternative precedence subgraphs, which represent possible assembly variants. In general, because of the great difficulty of the problem and the impossibility of representing alternative subgraphs in a precedence graph, the system designer will decide to select, a priori, one of such alternative subgraphs. This paper presents, characterizes and formulates a new general assembly line balancing problem with practical relevance: the alternative subgraphs assembly line balancing problem (ASALBP). Its novel characteristic is that it considers the possibility of having alternative assembly subgraphs, with the processing times and/or the precedence relations of certain tasks dependent on the assembly subgraph selected. Therefore, solving this problem implies simultaneously selecting an assembly subgraph for each part of the assembly that allows alternatives and balancing the line. The potentially positive effects of this on the solution of the problem are shown in a numerical example. Finally, a simple mathematical programming model is described and the results of a brief computational experiment are presented.     

Systemic Dietary Hesperidin Modulation of Osteoclastogenesis,Bone Homeostasis and Periodontal Disease in Mice

This study aimed to evaluate the effects of hesperidin (HE) on in vitro osteoclastogenesis and dietary supplementation on mouse periodontal disease and femoral bone phenotype. RAW 264.7 cells were stimulated with RANKL in the presence or absence of HE (1, 100 or 500 µM) for 5 days, and evaluated by TRAP, TUNEL and Western Blot (WB) analyses. In vivo, C57BL/6 mice were given HE via oral gavage (125, 250 and 500 mg/kg) for 4 weeks. A sterile silk ligature was placed between the first and second right maxillary molars for 10 days and microcomputed tomography (μCT), histopathological and immunohistochemical evaluation were performed. Femoral bones subjected or not to dietary HE (500 mg/kg) for 6 and 12 weeks were evaluated using μCT. In vitro, HE 500 µM reduced formation of RANKL-stimulated TRAP-positive(+) multinucleated cells (500 µM) as well as c-Fos and NFATc1 protein expression (p < 0.05), markers of osteoclasts. In vivo, dietary HE 500 mg/kg increased the alveolar bone resorption in ligated teeth (p < 0.05) and resulted in a significant increase in TRAP+ cells (p < 0.05). Gingival inflammatory infiltrate was greater in the HE 500 mg/kg group even in the absence of ligature. In femurs, HE 500 mg/kg protected trabecular and cortical bone mass at 6 weeks of treatment. In conclusion, HE impaired in vitro osteoclastogenesis, but on the contrary, oral administration of a high concentration of dietary HE increased osteoclast numbers and promoted inflammation-induced alveolar bone loss. However, HE at 500 mg/kg can promote a bone-sparing effect on skeletal bone under physiological conditions.