Biocidal and synergistic effect of three types of biologically synthesised silver/silver chloride nanoparticles.

Three types of silver/silver chloride nanoparticles were obtained by green synthesis from three types of microbial biomass. Their biocidal capacity was tested against six microorganisms. Two filamentous fungi were used that had previously demonstrated the ability to synthesise nanoparticles, Penicillium sp. 8L2 and Botryosphaeria rhodina MAMB-05. Also, the synthesis capacity of a yeast, Rhodotorula mucilaginosa 1S1, was evaluated. The original protocols underwent slight modifications. At the same time, the fractional inhibitory concentration was obtained. The interaction between specific antibiotics and the nanoparticles that showed the greatest biocidal capacity came from Penicillium sp.8L2, and it was studied further. All nanoparticles were characterised by UV-vis spectrophotometry and transmission electron microscopy (TEM). Also, their size distribution was analysed, which was in the range of 4 to 34 nm. The biocidal capacity of the nanoparticles for a group of bacteria and fungi was studied, presenting very low values in the range of 2.5-10 µg/mL for bacteria and 4-256 µg/mL for fungi. The interactions between the nanoparticles synthesised by Penicillium sp. 8L2 and a group of specific antibiotics for the tested microorganisms were also studied, proving that there was a synergistic interaction with vancomycin and ciprofloxacin and Staphylococcus epidermidis CECT 4183 and Escherichia coli CECT 101 bacteria, respectively.

Biotechnological use of the ubiquitous fungus Penicillium sp. 8L2: Biosorption of Ag(I) and synthesis of silver nanoparticles.

In this work, the efficiency of the ubiquitous fungus Penicillium sp. 8L2 to remove Ag(I) ions from synthetic solutions and its potential to synthesize silver nanoparticles (AgNPs) was evaluated. Using a Rotatable Central Composite Design pH and biomass concentration were optimized. Maximum biosorption capacity 51.53 mg/g, by Langmuir model, comparing favourably with other reports. The fungal biomass was characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and analyzed before and after the biosorption process by different techniques: X-ray diffraction (XRD), Field Emission Scanning Electron Microscopy (FE-SEM), Ultra-High Resolution Transmission Electron Microscopy and Energy Dispersive X-ray (HR-TEM-EDX) and Ultraviolet-Visible Spectrophotometry (UV-vis). The results showed that the fungus applied several mechanisms to remove Ag(I) ions from the solution and that some of them induced the synthesis of AgNPs. This fact could be verified in the synthesis tests from the cell extract in which aqueous suspensions with high concentrations of AgNPs were obtained. These nanoparticles had diameters between 2 and 9 nm and therefore a high potential for their use as biocidal agents. The results indicated that the synthesis of nanoparticles could be an added value to the heavy metal biosorption process.

Optimization with Response Surface Methodology of Microwave-Assisted Conversion of Xylose to Furfural.

The production of furfural from renewable sources, such as lignocellulosic biomass, has gained great interest within the concept of biorefineries. In lignocellulosic materials, xylose is the most abundant pentose, which forms the hemicellulosic part. One of the key steps in the production of furfural from biomass is the dehydration reaction of the pentoses. The objective of this work was to assess the conditions under which the concentration of furfural is maximized from a synthetic, monophasic, and homogeneous xylose medium. The experiments were carried out in a microwave reactor. FeCl3 in different proportions and sulfuric acid were used as catalysts. A two-level, three-factor experimental design was developed for this purpose. The results were further analyzed through a second experimental design and optimization was performed by response surface methodology. The best operational conditions for the highest furfural yield (57%) turned out to be 210 °C, 0.5 min, and 0.05 M FeCl3.

Assessment of By-Product from Botryosphaeria rhodina MAMB-05 as an Effective Biosorbent of Pb(II).

In this work, two types of biomass preparations (VMSM and M3) from the filamentous fungus Botryosphaeria rhodina MAMB-05, which were previously used in a process of production of ß-glucan, were assessed as biosorbents of lead. The operating conditions, optimized through response surface methodology and experimental design, were shown to be pH 5.29 and a biosorbent dose of 0.23 g/L for the VMSM biomass type; and pH 5.06 and a dose of biosorbent of 0.60 g/L for the M3 biomass type, at a constant temperature of 27 °C. Fourier transform-infrared spectroscopy analyzed the presence of functional groups on the biomass surface. In addition to give an extra value to the by-product biomass, the VMSM-type from B. rhodina MAMB-05 showed an excellent lead biosorption capacity (qm) with a value of 403.4 mg/g for the Langmuir model, comparing favorably with literature results, while the M3 subtype biomass showed a value of 96.05 mg/g.

Ag(I) Biosorption and Green Synthesis of Silver/Silver Chloride Nanoparticles by Rhodotorula mucilaginosa 1S1.

The efficiency of Rhodotorula mucilaginosa 1S1 as an Ag(I) biosorbent and at the same time its ability to biosynthesize recoverable silver nanoparticles is evaluated. Kinetic, equilibrium and thermodynamic tests are carried out for 19 °C, 27 °C and 37 °C, from which the process is adjusted to a pseudo second-order kinetics and to the Freundlich model, while optimal operational conditions are determined at 27 °C. The thermodynamic study shows positive values for enthalpy (ΔH: 133.23 kJ/mol) and entropy (ΔS: 0.4976 kJ/(mol K)), while the Gibbs free energy (ΔG) value is 12.136 kJ/mol. For a metal concentration of 459 mg/L, a maximum biosorption capacity (qm) of 137.2 mg/g at 19 °C is obtained, while for 100 mg/L concentration a qm value of 60.44 mg/g is obtained at the same temperature. The mechanisms involved in the biosorption process are studied by infrared spectroscopy, X-ray diffraction and scanning and transmission electron microscopy, while the nanoparticle synthesis is evaluated by ultraviolet-visible spectrophotometry (UV-vis) and transmission electron microscopy. The results indicate that the biomass is a good biosorbent and also has the ability to synthesize silver nanoparticles (Ag/AgCl) with sizes between 12 nm and 20 nm.

Clinical Outcomes of a Monofocal, Optimized, Aspheric, Hydrophobic Acrylic Intraocular Lens Implant.

Purpose: To evaluate the refractive and visual outcomes of eyes implanted with monofocal, optimized, aspheric, hydrophobic acrylic intraocular lenses (IOL) following cataract surgery. Methods: The study assessed 55 eyes implanted with CT LUCIA® 621P IOLs (Carl Zeiss Meditec AG, Jena, Germany) at 3-months postsurgery. The main outcome measures were refractive error, monocular corrected distance visual acuity (CDVA), monocular uncorrected distance visual acuity (UDVA), photopic and mesopic contrast sensitivity, and wavefront aberrations. Patients were asked to complete the Catquest-9SF questionnaire. Results: At 3 months, the cumulative Snellen visual acuity was 20/20 in 94.55% of eyes and 20/25 or better in 100%. The difference between the UDVA and CDVA was either the same (70.91%) or better (29.09%) in all eyes. The mean Snellen decimal UDVA and DCVA were 1.07 ± 0.15 and 1.13 ± 0.11, respectively. The safety and efficacy indexes were 1.48 and 1.40, respectively. 47.27% of eyes showed a spherical equivalent ±0.13 D, with 92.73% of the eyes were within ±0.50 D and all eyes were within ±1.00 D. The mean spherical equivalent was -0.03 ± 0.30 D and the mean refractive cylinder -0.36 ± 0.34 D. The log photopic and mesopic contrast sensitivity functions were good and similar for all spatial frequencies. All patients reported being either fairly satisfied (score 3) or very satisfied (score 4) with their vision (mean = 3.64 ± 0.49). No intra- and postoperative complications were reported during the 3 months of follow-up. Conclusion: The current study demonstrates that excellent visual and refractive outcomes, as well as patient satisfaction, can be achieved after cataract surgery with optimized, aspheric CT LUCIA 621P monofocal IOLs.

Hematic Antegrade Repriming Reduces Emboli on Cardiopulmonary Bypass: A Randomized Controlled Trial.

Particulate and gaseous microemboli (GME) are side effects of cardiac surgery that interfere with postoperative recovery by causing endothelial dysfunction and vascular blockages. GME sources during surgery are multiple, and cardiopulmonary bypass (CPB) is contributory to this embolic load. Hematic antegrade repriming (HAR) is a novel procedure that combines the benefits of repriming techniques with additional measures, by following a standardized procedure to provide a reproducible hemodilution of 300 ml. To clarify the safety of HAR in terms of embolic load delivery, a prospective and controlled study was conducted, by applying Doppler probes to the extracorporeal circuit, to determine the number and volume of GME released during CPB. A sample of 115 patients (n = 115) was considered for assessment. Both groups were managed under strict normothermia, and similar clinical conditions and protocols, receiving the same open and minimized circuit. Significant differences in GME volume delivery (control group [CG] = 0.28 ml vs. HAR = 0.08 ml; p = 0.004) and high embolic volume exposure (>1 ml) were found between the groups (CG = 30.36% vs. HAR = 4.26%; p = 0.001). The application of HAR did not represent an additional embolic risk and provided a four-fold reduction in the embolic volume delivered to the patient (coefficient, 0.24; 95% CI, 0.08-0.72; p = 0.01), which appears to enhance GME clearance of the oxygenator before CPB initiation.

Effect of first ball possession on partial and final scores in 2003, 2005, and 2007 Water Polo World Championships.

The purpose was to analyze the influence of winning the first ball possession on the partial and final score in male and female water polo. The 288 matches disputed by the teams participating in the 2003, 2005, and 2007 Water Polo World Championship were recorded. The results reflect statistically significant differences for the influence of gaining the first ball possession on the partial and final scoreboard of each period and for the influence between the total number of first possessions obtained and the final result.

Biosorption mechanisms of Ag(I) and the synthesis of nanoparticles by the biomass from Botryosphaeria rhodina MAMB-05.

Two biomass types of Botryosphaeria rhodina MAMB-05 (VMSM and M3) were evaluated to determine their effectiveness in removing Ag(I) ions from synthetic solutions. Both biomass types obtained good results in the biosorption process with maximum biosorption capacities (qm) for the Langmuir model of 34.67 and 39.23 mg Ag(I)/g dry biomass for M3 and VMSM, respectively. The biomass was characterized by X-ray microfluorescence and Fourier-transform-infrared spectroscopy (FT-IR). After the biosorption process, the mechanisms involved in biosorption were studied by FT-IR, X-ray diffraction (XRD), Field Emission Scanning Microscopy/Energy Dispersive X-ray Analysis (FESEM/EDX) and Ultraviolet-Visible Spectrophotometry. The results demonstrated the participation of various mechanisms in the retention of silver on biomass (bioadsorption, complexation, ion exchange, covalent bonding) that resulted in the formation of silver chloride nanoparticles (AgCl-NPs) and silver nanoparticles (AgNPs). The sizes of AgCl-NPs (chlorargyrite) according to the Debye-Scherrer equation were 19.29 nm (VMSM biomass) and 24.9 nm for the M3 type. For AgNPs the crystal size was between 1.5 and 0.8 nm for VMSM and M3 respectively. Furthermore, it was found that an undetermined fraction of the silver nanoparticles after biosorption remained in solution, which could be advantageous for their recovery.

Optimization of ultrasound-assisted extraction of biomass from olive trees using response surface methodology.

Olive tree pruning biomass (OTP) and olive mill leaves (OML) are the main residual lignocellulosic biomasses that are generated from olive trees. They have been proposed as a source of value-added compounds and biofuels within the biorefinery concept. In this work, the optimization of an ultrasound-assisted extraction (UAE) process was performed to extract antioxidant compounds present in OTP and OML. The effect of the three parameters, ethanol/water ratio (20, 50, 80% of ethanol concentration), amplitude percentage (30, 50, 70%) and ultrasonication time (5, 10, 15 min), on the responses of total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities (DPPH, ABTS and FRAP) were evaluated following a Box-Behnken experimental design. The optimal conditions obtained from the model, taking into account simultaneously the five responses, were quite similar for OTP and OML, with 70% amplitude and 15 min for both biomasses and a slight difference in the optimum concentration of ethanol. (54.5% versus 51.3% for OTP and OML, respectively). When comparing the antioxidant activities obtained with OTP and OML, higher values were obtained for OML (around 40% more than for OTP). The antioxidant activities reached experimentally under the optimized conditions were 31.6 mg of TE/g of OTP and 42.5 mg of TE/g of OML with the DPPH method, 66.5 mg of TE/g of OTP and 95.9 mg of TE/g of OML with the ABTS method, and 36.4 mg of TE/g of OTP and 49.7 mg of TE/g of OML with the FRAP method. Both OTP and OML could be a potential source of natural antioxidants.

Genetic Susceptibility in Head and Neck Squamous Cell Carcinoma in a Spanish Population.

Despite classical environmental risk factors like tobacco, alcohol or viral infection, not all individuals develop head and neck cancer. Therefore, identification of the genetic susceptibility produced by single nucleotide polymorphisms (SNPs) is an important task. A total of 296 human papillomavirus negative head and neck cancer (HNC) patients (126 laryngeal, 100 pharyngeal and 70 oral cavity) were included in the study, involving 29 candidate SNPs in genes within important carcinogenic pathways (oncogenesis and tumour suppression, DNA repair, inflammation, oxidation and apoptosis). Genotyping was performed using TaqMan probes or restriction fragment length assays in peripheral blood DNA. In addition, 259 paired controls were also evaluated with the same risk factors for each specific location. Nine SNPs in DNA repair (ERCC1 rs11615, ERCC2 rs13181), inflammatory (IL2 rs2069762, IL6 rs1800795), oxidative (NFE2L2 rs13035806 and rs2706110) and apoptotic genes (TP53 rs1042522, MDM2 rs2279744, BCL2 rs2279115) were differently associated with HNSCC susceptibility by location. Some of these SNPs were not described before in this tumour type. In conclusion, we describe several SNPs associated with HNC in a Spanish population.

Evaluation of steam explosion pre-treatment for enzymatic hydrolysis of sunflower stalks.

Sunflower stalks, a largely available and cheap agricultural residue lacking of economic alternatives, were subjected to steam explosion pre-treatment, the objective being to optimize pre-treatment temperature in the range 180-230°C. Enzymatic hydrolysis performed on the pre-treated solids by a cellulolytic complex (Celluclast 1.5L) and analysis of filtrates were used to select the best pre-treatment temperature. Temperature selection was based on the susceptibility to enzymatic hydrolysis of the cellulose residue and both the cellulose recovery in the solid and the hemicellulose-derived sugars recoveries in the filtrate. After 96h of enzymatic action, a maximum hydrolysis yield of 72% was attained in the water-insoluble fiber obtained after pre-treatment at 220°C, corresponding to a glucose concentration of 43.7g/L in hydrolysis media. Taking into account both cellulose recovery and hydrolysis yield, the maximum value of glucose yield referred to unpretreated raw material was also found when using steam pre-treated sunflower stalks at 220°C, obtaining 16.7g of glucose from 100g of raw material. With regard to the filtrate analysis, most of the hemicellulosic-derived sugars released during the steam pre-treatment were in oligomeric form, the highest recovery being obtained at 210°C pre-treatment temperature. Moreover, the utilisation of hemicellulosic-derived sugars as a fermentation substrate would improve the overall bioconversion of sunflower stalks into fuel ethanol.

Conversion of olive tree biomass into fermentable sugars by dilute acid pretreatment and enzymatic saccharification.

The production of fermentable sugars from olive tree biomass was studied by dilute acid pretreatment and further saccharification of the pretreated solid residues. Pretreatment was performed at 0.2%, 0.6%, 1.0% and 1.4% (w/w) sulphuric acid concentrations while temperature was in the range 170-210 degrees C. Attention is paid to sugar recovery both in the liquid fraction issued from pretreatment (prehydrolysate) and that in the water-insoluble solid (WIS). As a maximum, 83% of hemicellulosic sugars in the raw material were recovered in the prehydrolysate obtained at 170 degrees C, 1% sulphuric acid concentration, but the enzyme accessibility of the corresponding pretreated solid was not very high. In turn, the maximum enzymatic hydrolysis yield (76.5%) was attained from a pretreated solid (at 210 degrees C, 1.4% acid concentration) in which cellulose solubilization was detected; moreover, sugar recovery in the prehydrolysate was the poorest one among all the experiments performed. To take account of fermentable sugars generated by pretreatment and the glucose released by enzymatic hydrolysis, an overall sugar yield was calculated. The maximum value (36.3 g sugar/100 g raw material) was obtained when pretreating olive tree biomass at 180 degrees C and 1% sulphuric acid concentration, representing 75% of all sugars in the raw material. Dilute acid pretreatment improves results compared to water pretreatment.

Valorisation of olive agro-industrial by-products as a source of bioactive compounds.

A large amount of olive-derived biomass is generated yearly in Spain, which could be used as a potential source of bioactive compounds. The present work evaluates the recovery of natural antioxidants from olive tree pruning (OTP) and olive mill leaves (OML). For this purpose, the effect of different solvents on the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity was evaluated. The solvent was found to have a significant effect (p < 0.05) on the TPC, TFC, and the DPPH, ABTS, and FRAP activity, affording similar results for the extracts from the two by-products. The extracts obtained using 50% ethanol showed high TPC (23.85 and 27.54 mg GAE/gdw for OTP and OML, respectively) and TFC (52.82 and 52.39 mg RE/gdw for OTP and OML, respectively). Also, the OTP and OML extracts exhibited notable antioxidant activity as measured by the ABTS method (45.96 and 42.71 mg TE/gdw, respectively). Using pyrolysis-gas chromatography/mass spectrometry, 30 bioactive compounds were detected in both extracts. Additionally, UPLC-DAD-ESI-MS allowed the identification of 15 compounds in the samples. Furthermore, the antioxidant extracts were found to inhibit the growth of several food pathogenic bacteria. This research demonstrates that these by-products from olive grove farming are a good source of antioxidant compounds with antibacterial properties, which have potential applications in the food and pharmaceutical industries.

Biosorption of Ag(I) from aqueous solutions by Klebsiella sp. 3S1.

This study investigated the potential ability of Klebsiella sp. 3S1 to remove silver cations from aqueous solutions. The selected strain is a ubiquitous bacterium selected from among several microorganisms that had been isolated from wastewaters. To optimise the operating conditions in the biosorption process, a Rotatable Central Composite Experimental Design was developed establishing pH, temperature and biomass concentration as independent variables. Interaction mechanisms involved were analysed through kinetic and equilibrium studies. The experimental results suit pseudo-second order kinetics with two biosorption stages, being the first almost instantly. The Langmuir equilibrium model predicted a maximum capacity of biosorption (q) of 114.1mg Ag/g biomass. The study of the mechanisms involved in the biosorption was completed by employing advanced techniques which revealed that both bacterium-surface interactions and intracellular bioaccumulation participate in silver removal from aqueous solutions. The ability of Klebsiella sp. 3S1 to form silver chloride nanoparticles with interesting potential applications was also discussed.

Bifidobacterial growth stimulation by oligosaccharides generated from olive tree pruning biomass.

This work aims to evaluate the prebiotic potential of oligosaccharides (OS) obtained from autohydrolysis of olive tree pruning biomass (OTPB). Two selected fractions (F1 and F2) were characterized and used in in vitro fermentations by two Bifidobacterium spp. (B. adolescentis and B. longum) and one fecal inoculum. The fraction F1 presented a lower average degree of polymerization (DP) mainly with OS ranging from 3 to 6 DP, whereas the fraction F2 corresponded to a pool of unsubstituted and acetylated oligomers with DP between 4 and 19. In the fermentation by Bifidobacterium, F1 supported a higher biomass formation, OS consumption and organic acids production than F2. With the fecal inoculum, the accumulation of organic acids, as the sum of acetate, propionate and butyrate, was similar for F1 and F2 (107 and 101mM, respectively). The bifidobacteria counts also increased during the incubation time for both OS fractions.

Combined acid/alkaline-peroxide pretreatment of olive tree biomass for bioethanol production.

Olive tree biomass (OTB) can be used for producing second generation bioethanol. In this work, extracted OTB was subjected to fractionation using a sequential acid/alkaline oxidative pretreatment. In the first acid stage, the effects of sulfuric acid concentration and reaction times at 130°C were investigated. Up to 71% solubilization of hemicellulosic sugars was achieved under optimized conditions (2.4% H2SO4, 84min). In the second stage, the influence of hydrogen peroxide concentration and process time were evaluated at 80°C. Approximately 80% delignification was achieved under the best operational conditions (7% H2O2, 90min) within the experimental range studied. This pretreatment produced a substrate with 72% cellulose that was highly accessible to enzymatic attack, yielding 82g glucose/100g glucose in delignified OTB. Ethanol production from both hemicellulosic sugars solubilized in the acid pretreatment and glucose from enzymatic hydrolysis of delignified OTB yielded 15g ethanol/100g OTB.

Analysis of autophagy gene polymorphisms in Spanish patients with head and neck squamous cell carcinoma.

Head and neck squamous cell carcinoma (HNSCC) is the sixth cancer on incidence worldwide. Tobacco and alcohol consumption are the most classical risk factors associated with its development. Autophagy process has a dual effect both in tumourigenesis and tumour suppressing activity. To investigate the importance of this pathway in HNSCC susceptibility, a risk factor matched case-control association study was performed with four candidate polymorphisms in autophagy genes (ATG2B, ATG5, ATG10, ATG16L1). We found an association between the variant in ATG10 rs1864183 and a higher susceptibility to develop laryngeal cancer, ATG2B rs3759601 and pharyngeal cancer and ATG16L1 rs2241880 and oral carcinoma. ATG5 rs2245214 SNP was not associated with any location. Overall, our results indicate the importance of the autophagy pathway in the susceptibility of head and neck squamous cell carcinoma and demonstrate the heterogeneity between its locations encompassed under a single terminology.

Ethanol production from pretreated olive tree wood and sunflower stalks by an SSF process.

Olive tree wood and sunflower stalks are agricultural residues largely available at low cost in Mediterranean countries. As renewable lignocellulosic materials, their bioconversion may allow both obtaining a value-added product, for fuel ethanol, and facilitating their elimination. In this work, the ethanol production from olive tree wood and sunflower stalks by a simultaneous saccharification and fermentation (SSF) process is studied. As a pretreatment, steam explosion at different temperatures was applied. The water insoluble fractions of steam-pretreated sunflower stalks and steamed, delignified olive tree wood were used as substrates at 10% w/v concentration for an SSF process by a cellulolytic commercial complex and Saccharomyces cerevisiae. After 72-h fermentation, ethanol concentrations up to 30 g/L were obtained in delignified steam-pretreated olive tree wood at 230 degrees C and 5 min. Sunflower stalks pretretated at 220 degrees C and 5 min gave maximum ethanol concentrations of 21 g/L in SSF experiments.

Obtaining sugars and natural antioxidants from olive leaves by steam-explosion.

In this work, steam-explosion treatment was evaluated as a procedure to recover sugars and natural antioxidants from olive tree leaves. The treatment was carried out following a Box-Behnken experimental design, with three factors, temperature (180-220°C), process time (2-10min) and milling time (0-15s). Response surface methodology showed that temperature was the most influential factor, followed by process time, while the best results were achieved with whole leaves. The operational conditions for simultaneously maximizing the sugars and natural antioxidants recoveries resulted to be 180°C, 8.3min and whole leaf; under these conditions 18.39g and 1950mg were obtained from 100g dry olive leaves, respectively. This is equivalent to 70% recovery of the initial sugars present in olive leaves, with a very low formation of inhibitory compounds and an important amount of natural products with antioxidant capacity such as oleuropein, hydroxytyrosol and flavonoids.