Plant proteases for bioactive peptides release: A review.

Proteins are a potential source of health-promoting biomolecules with medical, nutraceutical, and food applications. Nowadays, bioactive peptides production, its isolation, characterization, and strategies for its delivery to target sites are a matter of intensive research. In vitro and in vivo studies regarding the bioactivity of peptides has generated strong evidence of their health benefits. Dairy proteins are considered the richest source of bioactive peptides, however proteins from animal and vegetable origin also have been shown to be important sources. Enzymatic hydrolysis has been the process most commonly used for bioactive peptide production. Most commercial enzymatic preparations frequently used are from animal (e.g., trypsin and pepsin) and microbial (e.g., Alcalase® and Neutrase®) sources. Although the use of plant proteases is still relatively limited to papain and bromelain from papaya and pineapple, respectively, the application of new plant proteases is increasing. This review presents the latest knowledge in the use and diversity of plant proteases for bioactive peptides release from food proteins including both available commercial plant proteases as well as new potential plant sources. Furthermore, the properties of peptides released by plant proteases and health benefits associated in the control of disorders such as hypertension, diabetes, obesity, and cancer are reviewed.

Environmental risk assessment of effluents as a whole emerging contaminant: Efficiency of alternative tertiary treatments for wastewater depuration.

Emerging contaminants (ECs) and regulated compounds (RCs) from three different WWTP effluents were measured in the current study. The efficiency of two tertiary treatments, Photobiotreatment (PhtBio) and Multi-Barrier Treatment (MBT), for removing contaminants was determined. Results indicated different percentages of removal depending on the treatment and the origin of the effluent. Risk Quotients (RQs) were determined for different species of algae, Daphnia, and fish. RQ results revealed diverse risk values depending on the bioindicator species. Tonalide, galaxolide (fragrances), and ofloxacin (antibiotic) were the most persistent and harmful substances in tested effluents. "Negligible risk" category was reached since a wide diversity of ECs were removed by MBT with high removal percentages. Contrarily, PhtBio was effective only in the depuration of certain chemical compounds, and its efficiency depended on the composition of the raw effluent.

Post-treatment of refinery wastewater effluent using a combination of AOPs (H2O2 photolysis and catalytic wet peroxide oxidation) for possible water reuse. Comparison of low and medium pressure lamp performance.

The main aim of this work was to study the feasibility of multi-barrier treatment (MBT) consisting of filtration, hydrogen peroxide photolysis (H2O2/UVC) and catalytic wet peroxide oxidation (CWPO) for post-treatment of petroleum refinery effluent. Also the possibility of water reuse or safe discharge was considered. The performance of MBT using medium (MP) and low (LP) pressure lamps was compared as well as operation and maintenance (O&M) cost. Decomposition of organic compounds was followed by means of gas chromatography-mass spectrometry (GC-MS), total organic carbon (TOC) and chemical oxygen demand (COD) analysis. After filtration step (25 µm) turbidity and concentration of suspended solids decreased by 92% and 80%, respectively. During H2O2/UVC process with LP lamp at optimal conditions (H2O2:TOC ratio 8 and UVC dose received by water 5.28 WUVC s cm(-2)) removal of phenolic compounds, TOC and COD was 100%, 52.3% and 84.3%, respectively. Complete elimination of phenolic compounds, 47.6% of TOC and 91% of COD was achieved during H2O2/UVC process with MP lamp at optimal conditions (H2O2:TOC ratio 5, UVC dose received by water 6.57 WUVC s cm(-2)). In order to compare performance of H2O2/UVC treatment with different experimental set up, the UVC dose required for removal of mg L(-1) of COD was suggested as a parameter and successfully applied. The hydrophilicity of H2O2/UVC effluent significantly increased which in turn enhanced the oxidation of organic compounds during CWPO step. After H2O2/UVC treatment with LP and MP lamps residual H2O2 concentration was 160 mg L(-1) and 96.5 mg L(-1), respectively. Remaining H2O2 was fully consumed during subsequent CWPO step (6 and 3.5 min of contact time for LP and MP, respectively). Total TOC and COD removal after MBT was 94.7% and 92.2% (using LP lamp) and 89.6% and 95%, (using MP lamp), respectively. The O&M cost for MBT with LP lamp was estimated to be 0.44 € m(-3) while with MP lamp it was nearly five times higher. Toxicity assessment was performed using two marine species (Vibrio fischeri and Paracentrotus lividus sea-urchin) after each treatment step. The highest toxicity was attributed to H2O2/UVC effluent for both tested species. After MBT a drastic decrease of toxicity was achieved.

Are combined AOPs effective for toxicity reduction in receiving marine environment? Suitability of battery of bioassays for wastewater treatment plant (WWTP) effluent as an ecotoxicological assessment.

Ecotoxicological assessment of three different wastewater treatment plant (WWTP) effluents D1, D2 and D3 was performed before and after tertiary treatment using combination of advanced oxidation processes (AOPs). A multibarrier treatment (MBT) consisting of microfiltration (MF), hydrogen peroxide photolysis (H2O2/UVC) and catalytic wet peroxide oxidation (CWPO) was applied for all effluents. Sparus aurata, Paracentrotus lividus, Isochrysis galbana and Vibrio fischeri, representing different trophic levels, constituted the battery of bioassays. Different acute toxicity effects were observed in each WWTP effluents tested. The percentage of sea urchin larval development and mortality fish larvae were the most sensitive endpoints. Significant reduction (p < 0.05) of effluent's toxicity was observed using a classification pT-method after MBT process. Base on obtained results, tested battery of bioassays in pT-method framework can be recommended for acute toxicity preliminary evaluation of WWTP effluents for the marine environment.

Post-treatment of biologically treated wastewater containing organic contaminants using a sequence of H2O2 based advanced oxidation processes: photolysis and catalytic wet oxidation.

In this paper the feasibility of a multi-barrier treatment (MBT) for the regeneration of synthetic industrial wastewater (SIWW) was evaluated. Industrial pollutants (orange II, phenol, 4-chlorophenol and phenanthrene) were added to the effluent of municipal wastewater treatment plant. The proposed MBT begins with a microfiltration membrane pretreatment (MF), followed by hydrogen peroxide photolysis (H2O2/UVC) and finishing, as a polishing step, with catalytic wet peroxide oxidation (CWPO) using granular activated carbon (GAC) at ambient conditions. During the microfiltration step (0.7 µm) the decrease of suspended solids concentration, turbidity and Escherichia coli in treated water were 88, 94 and 99%, respectively. Also, the effluent's transmittance (254 nm) was increased by 14.7%. Removal of more than 99.9% of all added pollutants, mineralization of 63% of organic compounds and complete disinfection of total coliforms were reached during the H2O2/UVC treatment step (H2O2:TOC w/w ratio = 5 and an UVC average dose accumulated by wastewater 8.80 WUVC s cm(-2)). The power and efficiency of the lamp, the water transmittance and photoreactor geometry are taken into account and a new equation to estimate the accumulated dose in water is suggested. Remaining organic pollutants with a higher oxidation state of carbon atoms (+0.47) and toxic concentration of residual H2O2 were present in the effluent of the H2O2/UVC process. After 2.3 min of contact time with GAC at CWPO step, 90 and 100% of total organic carbon and residual H2O2 were removed, respectively. Also, the wastewater toxicity was studied using Vibrio fischeri and Sparus aurata larvae. The MBT operational and maintenance costs (O&M) was estimated to be 0.59 € m(-3).

Unsupervised cardiac image segmentation via multiswarm active contours with a shape prior.

This paper presents a new unsupervised image segmentation method based on particle swarm optimization and scaled active contours with shape prior. The proposed method uses particle swarm optimization over a polar coordinate system to perform the segmentation task, increasing the searching capability on medical images with respect to different interactive segmentation techniques. This method is used to segment the human heart and ventricular areas from datasets of computed tomography and magnetic resonance images, where the shape prior is acquired by cardiologists, and it is utilized as the initial active contour. Moreover, to assess the performance of the cardiac medical image segmentations obtained by the proposed method and by the interactive techniques regarding the regions delineated by experts, a set of validation metrics has been adopted. The experimental results are promising and suggest that the proposed method is capable of segmenting human heart and ventricular areas accurately, which can significantly help cardiologists in clinical decision support.

Chemical degradation of PCB in the contaminated soils slurry: direct Fenton oxidation and desorption combined with the photo-Fenton process.

This paper reports the results of the direct treatment of PCBs sorbed to solid particles (glass beads and sand) employing the Fenton process. The results obtained with contaminated sandy soils show 98% removal of the original PCB structure and 82% dechlorination, all within a reaction time of 72 hours. The degree of removal was observed to be dependent on the level of congener chlorination. The optimized conditions were: 5% H(2)O(2); 100 ppm Fe(3+); sandy soil mass/volume of oxidizing solution ratio (m/V) of 1/3 g/mL, vigorous agitation and dispensed with the need for heat. Results obtained by applying an integrated desorption treatment followed by photo-Fenton oxidation to sandy soils contaminated with PCBs are also present in this paper. Desorption of PCBs with surfactant solutions took place to an extent of around 90% (92.2% with K-perfluoroalkyl sulfonate, FT800, and 87% with Lineal Alkyl benzene Sulfonate, LAS), while photo-Fenton oxidation at 254 nm achieved degradation percentages close to 100% in 30 minutes for PCBs in solution, both with FT800 (98%) and LAS (97%) surfactants.

Optimization of the solution parameters during the degradation of Orange II in a photo-reactor mediated by Fe-Nafion membranes.

The Fenton and photo-Fenton mediated degradation process of Orange II was investigated in a flow photo-reactor. The degradation was monitored as a function of the wavelength of the applied light, recirculation flow rate, amount of H(2)O(2) and the initial concentration of Orange II. Optimization of the photo-Fenton degradation mediated by Fe-Nafion membranes indicated that an Orange II (0.25 M) solution discolored above 95% within 2.5 hours at an H(2)O(2)/Orange II ratio of 20. A concomitant mineralization of 40% of Orange II was observed after 5 h reaction. Homogeneous photo-Fenton processes were able to fully discolore Orange II within 1 hour and concomitantly fully mineralize the dye in the presence of Fe(III) (2 ppm) and an H(2)O(2)/Orange II ratio of 20. Surfactants such as linear alkylbenzene sulphonates (LAS) and K-perfluoroalkyl sulphonate (FT 800) slowed down the Orange II abatement in photo-Fenton processes.

Using solar and ultraviolet light to degrade PCBs in sand and transformer oils.

The present study describes the effect of the operating variables in the sensitized solar photolysis of sandy sediments contaminated by polychlorinated biphenyls (PCBs) in alkaline isopropanol. PCBs were almost completely removed in two weeks with stoichiometric release of chloride and biphenyl (BP) by a mechanism of reductive dechlorination. The concentration of the sensitizer (acetone) was the variable found to have most influence on the process. Under the same conditions, only partial dechlorination of the PCBs (34%) was observed in dielectric fluids without generating biphenyl to any appreciable extent. Direct photolysis (ultraviolet light at 254nm) of dielectric oils dissolved in alkaline isopropanol lead to 92% removal of PCBs in 60min. The observed mechanism was again that of reductive dechlorination, with stoichiometric release of chloride, and the accumulation of biphenyl, quaterphenyl and condensation products of biphenyl with isopropanol.

Determination of short-chain free fatty acids in lipolyzed milk fat by capillary electrophoresis.

The objective of this study was to monitor the release of short-chain free fatty acids (FFA) from milk fat during hydrolysis with lipase using capillary electrophoresis. Sample and run buffer allowed FFA to be maintained in solution by using cyclodextrin and methanol. Indirect UV detection at 270 nm was used, employing p-anisate as a chromophore. Calibration curves constructed for each individual FFA followed linear relationships with highly significant (p < 0.01) correlation coefficients. Electrophoretic FFA profiles of fresh milk fat and lipolyzed milk fat showed marked qualitative and quantitative differences. Butanoic acid (C4) was found in a concentration of 64 ppm, while hexanoic (C6) and octanoic (C8) acids were found in concentrations of 3.8 ppm in fresh milk fat. After a 60-min hydrolysis with commercial lipase, FFA released from milk fat consisted mainly of high concentrations (ppm) of butanoic (C4) (900), followed by hexanoic (C6) (427), octanoic (C8) (282), decanoic (C10) (92), pentanoic (C5) (47), and dodecanoic (C12) (37.5) acids. Ratios of FFA that were associated with flavor balance were calculated. The application of CE for lipolysis monitoring in milk fat offers a simple and fast method for the determination of FFA. Quantitative data can be obtained in 20 min, including sample preparation. The lengthy and laborious steps required in traditional chromatographic techniques, such as lipid extraction, FFA isolation, and derivatization, were not required in this CE method. The implementation of CE for milk fat lipolysis monitoring may be a useful quality control tool for dairy flavor development and production.

Retrograde amnesia in rats with dorsomedial thalamic damage.

The present research addresses the effects of dorsomedial thalamic lesions on remembering a delayed alternation problem learned before damage. Male rats were trained on a delayed alternation task. Successful performance of the test first required learning of the general principle that Go and No Go trials alternate with each other and, second, in every trial, recalling the specific consequences of the responses in the previous trial after a relatively short or long interval. The animals which had reached the learning criteria were paired by the learning level shown during the training and assigned semi-randomly to one unoperated control and one dorsomedial thalamic lesioned group. Lesions of the dorsomedial nucleus of thalamus produced a moderate and transient impairment of remembering of the delayed alternation learned before the damage for delays ranging from 0 to 40 s, and a severe impairment of the postoperative performance when delays were increased to 80 s. Detailed analysis of the animals' performance throughout the postoperative retraining confirms the role of the dorsomedial nucleus of the thalamus in memory and suggests that this thalamic region plays a role in recalling specific response-related events. Further research concerning the extension of retrograde amnesia associated with thalamic damage and the nature of the memory loss is proposed as necessary.

Amygdala, hippocampus and associative memory in rats.

Male wistar rats received either electrolytic or sham lesions into the amygdala, hippocampus or amygdala plus hippocampus, or were assigned to an unoperated control group. In Experiment 1, all lesioned and control animals were tested for the ability to master an associative memory test in which recall was assessed over delays ranging between 10 and 180 s. The goal of Experiment 2 was to study the susceptibility to proactive interference following the above mentioned types of damage. The role of the amygdala and hippocampus in remembering stimulus-magnitude of reward associations was evaluated in Experiment 3. Lesions of the dorsal and ventral hippocampus had no effect on acquisition of the associative memory test, but disrupted the animals' performance in the task after 120 and 180 s delays. The same lesions increased the sensitivity to interference but did not impair the performance of several stimulus/magnitude of reward discriminations. By contrast, amygdala lesions impaired the acquisition of the associative memory paradigm and the animals' performance over the successive delays. Moreover, the animals with these lesions were not able to learn the stimulus/magnitude of reward discriminations, although they did not show an increased susceptibility to interference. Combined damage to the amygdala plus hippocampus severely disrupted the acquisition of the associative memory paradigm and the animals' performance over successive delays. The same damage increased the susceptibility of the animals to interference and impaired the performance in the stimulus-magnitude of reward discriminations.

The role of different nuclei of the thalamus in processing episodic information.

Male rats received either electrolytic or sham lesions bilaterally into the dorsomedial nucleus or the anterior nuclei of the thalamus or were assigned to an unoperated control group. After the postoperative recovery period all lesioned and control animals were tested for the ability to master a delayed alternation task. It was characterized by the alternation of Go and No Go trials, which were separated by intervals ranging between 0 and 80 s. Its successful performance required, firstly, the learning of the general principle that Go and No Go trials alternate with each other and secondly, in every trial, recalling the specific consequences of the responses in the previous trial after a short or long delay. Bilateral damage of the thalamus including the dorsomedial nucleus or anterior nuclei severely disrupted the performance of a delayed alternation task. A detailed analysis of the performance throughout the training suggests that the impairment associated with both thalamic lesions could be interpreted as a deficit in recalling specific information. This analysis also reveals a failure in the early stages of learning. Further research concerning the role of the dorsomedial nucleus and the anterior nuclei of the thalamus in learning and memory is proposed as necessary.

The role of the amygdala and the hippocampus in working memory for spatial and non-spatial information.

Male rats received either electrolytic or sham lesions bilaterally into the amygdala, hippocampus or amygdala plus hippocampus, or were assigned to an unoperated control group. After the postoperative recovery period all lesioned and control animals were tested for the ability to master a spatial delayed non-matching-to-sample (DNMS), a visual DNMS and a visuo-tactile DNMS. Retention of these paradigms was evaluated 24 h after the last respective training session. Bilateral lesions of the amygdala severely disrupted the acquisition and retention of a DNMS paradigm with visual and visuo-tactile cues as discriminative stimuli and had no effect on the acquisition and retention of a spatial DNMS. On the contrary, bilateral lesions of the hippocampus impaired the acquisition and retention of spatial DNMS, but the animals with these lesions showed an acquisition and retention of the visual and visuo-tactile DNMS paradigms significantly better than those of animals with amygdala lesions. Combined lesions of the amygdala and hippocampus severely disrupted the acquisition and retention of the 3 paradigms. The contribution of the amygdala and the hippocampus in the working memory for spatial and non-spatial information is discussed.