The U.S. catfish industry is a major industry that has been declining over the years due to imports competition and growing operational costs. Catfish processing wastewater management and high energy requirement put a large financial burden on catfish processing facilities. Recovered protein-based solids have provided some value-added co-products, however, more co-products are needed to offset processing costs. Anaerobic digestion is a proven waste treatment method that produces methane, which is an energetic co-product that can be used within the processing facilities. This study was conducted to evaluate the potential of anaerobic digestion as an alternative to the currently used aerobic biotreatment of catfish processing wastewater. Initial assessments indicated the recalcitrance of the full-strength wastewater to anaerobic digestion, yielding only ~4 m3 per ton (U.S.) of input chemical oxygen demand (CODinput). Thus, several strategies were evaluated to improve the methane yield fr... [more]

The aim of this study was to characterize in detail 25 sour cherry cultivars and provide data on their industrial processing into high-quality sour cherry cloudy juices (ScCJ). Anthocyanin composition was identified and quantified by LC-PDA-ESI-MS QTof, UPLC-PDA. Kinetic degradation (k × 103, t1/2, D value) and color (CIE La*b*) were measured before and after 190 days of storage at 4 °C and 30 °C. A total of five anthocyanins, four cyanidins (-3-O-sophoroside, -3-O-glucosyl-rutinoside, -3-O-glucoside, and -3-O-rutinoside) and one peonidin-3-O-rutinoside were detected across all investigated juices. Total anthocyanin content ranged from ~590 to ~1160 mg/L of juice, with the highest levels in Skierka, Nagy Erdigymulscu, Wilena, Wiblek, and Safir cvs., and the lowest in Dradem and Nanaones. During 190 days of storage a significant change was observed in the content of anthocyanins. Their degradation depended rather on the storage conditions (time and temperature) than on the type of antho... [more]

In the present work, essential oils (EOs) extracted from different parts of sour orange Citrus aurantium (green leaves/twigs, small branches, wooden branches, and branch bark) were studied through gas chromatography coupled with mass spectrometry (GC/MS). Furthermore, the EOs in the amounts of 5, 10, 15, 20, and 25 µL were studied for their antibacterial activity against three pathogenic bacteria, Agrobacterium tumefaciens, Dickeya solani, and Erwinia amylovora. The main EO compounds in the leaves/twigs were 4-terpineol (22.59%), D-limonene (16.67%), 4-carvomenthenol (12.84%), and linalool (7.82%). In small green branches, they were D-limonene (71.57%), dodecane (4.80%), oleic acid (2.72%), and trans-palmitoleic acid (2.62%), while in branch bark were D-limonene (54.61%), γ-terpinene (6.68%), dodecane (5.73%), and dimethyl anthranilate (3.13%), and in branch wood were D-limonene (38.13%), dimethyl anthranilate (8.13%), (-)-β-fenchol (6.83%), and dodecane (5.31%). At 25 µL, the EO from... [more]

The objective of this study was to evaluate effects of dark brown sugar on acrylamide and 5-hydroxymethylfurfural (HMF) levels in brown sugar cookies. Dark brown sugar was used as a raw material instead of sucrose, and chitosan, chitooligosaccharides, or calcium carbonate were added to investigate their effect on acrylamide and HMF mitigation. The results demonstrated that the higher the content of acrylamide in the dark brown sugar, the higher the amount of acrylamide produced in baked brown sugar cookies. The addition of dark brown sugar significantly increased the diameter and decreased the thickness of cookies, which induced more acrylamide formation. Therefore, the sucrose control cookies were harder and thicker than dark brown sugar cookies. The addition of 1% chitosan, chitooligosaccharide, or calcium carbonate did not reduce the acrylamide formation of the brown sugar cookies. The content of acrylamide and HMF in the sucrose control group was lower than that in dark brown sugar... [more]

In this study, six macroporous resins were screened on their adsorption and de-adsorption characteristics for the amygdalin in the debitterizing wastewater concentrate (DWC). The results indicate that the XDA-1 resin exerts good adsorption and de-adsorption capacities on the amygdalin. In order to further confirm its feasibility, the factors affecting the capacity of adsorption and de-adsorption, and its adsorption mechanisms were also investigated. The results suggest that the optimum purification conditions were as follows: loading concentration of samples with 78.05 mg/mL, flow rate of 2 mL/min, and de-adsorption with 80% ethanol solution. The recovery rate was 88.75% and the relative content achieved 61.58% after purification by XDA-1 resin. The Freundlich model can be used to describe the entirety of the exothermic and physical adsorption processes. In summary, the conclusion which can be made from this research is that the wastewater generated from the debitterizing of apricot ke... [more]

Triphenyltin (TPT) has severely polluted the environment, and it often coexists with metal ions, such as Cu2+. This paper describes the cell’s molecular recognition of TPT, the interaction between TPT recognition and Cu2+ biosorption, and their effect on cell permeability. We studied the recognition of TPT by Bacillus thuringiensis cells and the effect of TPT recognition on Cu2+ biosorption by using atomic force microscopy to observe changes in cell surface mechanical properties and cellular morphology and by using flow cytometry to determine the cell growth status and cell permeability. The results show that B. thuringiensis can quickly recognize different media. The adhesion force of cells in contact with Tween 80 was significantly reduced to levels that were much lower than that of cells in contact with PBS. Conversely, the cell surface adhesion force increased as TPT became more degraded. B. thuringiensis cells maintained their original morphology after 48 h of TPT treatment. The a... [more]

Angiogenesis, the formation of new blood vessels from pre-existing ones, is essential for both normal development and numerous pathologies. Systems biology has offered a unique approach to study angiogenesis by profiling tyrosine kinase receptors (RTKs) that regulate angiogenic processes and computationally modeling RTK signaling pathways. Historically, this systems biology approach has been applied on ex vivo angiogenesis assays, however, these assays are difficult to quantify and limited in their potential of temporal analysis. In this study, we adopted a simple two-dimensional angiogenesis assay comprised of human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs) and examined temporal dynamics of a panel of six RTKs and cell heterogeneity up to 17 days. We observed ~2700 VEGFR1 (vascular endothelial growth factor receptor 1) per cell on 24-h-old cocultured HDF plasma membranes, which do not express VEGFR when cultured alone. We observed 4000−8100 VEGFR2 p... [more]

Active pharmaceutical ingredient (API) particle size distribution is important for both downstream processing operations and in vivo performance. Crystallization process parameters and reactor configuration are important in controlling API particle size distribution (PSD). Given the large number of parameters and the scale-dependence of many parameters, it can be difficult to design a scalable crystallization process that delivers a target PSD. Population balance modeling is a useful tool for understanding crystallization kinetics, which are primarily scale-independent, predicting PSD, and studying the impact of process parameters on PSD. Although population balance modeling (PBM) does have certain limitations, such as scale dependency of secondary nucleation, and is currently limited in commercial software packages to one particle dimension, which has difficulty in predicting PSD for high aspect ratio morphologies, there is still much to be gained from applying PBM in API crystallizat... [more]

The addition of Fe2+ is considered an effective method for increasing methane production, but the added Fe2+ may not be absorbed by anaerobic microorganisms due to complex chemical reactions. In this study, ethylenediaminetetraacetic acid (EDTA) was used as a ligand of Fe2+ (EDTA-Fe) to promote the dissolution of Fe, and the anaerobic performances of kitchen wastes (KWs) in a semi-continuous reactor were studied. The results indicated that the biogas yields and methane contents were enhanced to 594−613 mL·g−1VSadd·d−1 and 63.6−64.4% at an organic loading rate (OLR) of 2.5 gVSadd·L−1·d−1 due to EDTA-Fe addition. Simultaneously, the EDTA-Fe was more effective than Fe2+ in preventing the acidification of KWs with a high OLR (5.0 gVSadd·L−1·d−1). In addition, the sequential extraction results showed that the water-soluble fraction of Fe in the R3 (EDTA-Fe addition) was 1.49-fold of that in the R2 with Fe2+ addition. The contents of coenzymes F420 and F430 were also improved 1.09 and 1.11 t... [more]

Hyaluronan (HA), a glycosaminoglycan with important medical applications, is commercially produced from pathogenic microbial sources. The metabolism of HA-producing recombinant generally regarded as safe (GRAS) systems needs to be more strategically engineered to achieve yields higher than native producers. Here, we use a genome-scale model (GEM) to account for the entire metabolic network of the cell while predicting strategies to improve HA production. We analyze the metabolic network of Lactococcus lactis adapted to produce HA and identify non-conventional strategies to enhance HA flux. We also show experimental verification of one of the predicted strategies. We thus identified an alternate route for enhancement of HA synthesis, originating from the nucleoside inosine, that can function in parallel with the traditionally known route from glucose. Adopting this strategy resulted in a 2.8-fold increase in HA yield. The strategies identified and the experimental results show that the... [more]

Development of biomaterials for the diagnosis and treatment of neurotraumatic ailments has been significantly advanced with our deepened knowledge of the pathophysiology of neurotrauma. Canadian research in the fields of biomaterial-based contrast agents, non-invasive axonal tracing, non-invasive scaffold imaging, scaffold patterning, 3D printed scaffolds, and drug delivery are conquering barriers to patient diagnosis and treatment for traumatic injuries to the nervous system. This review highlights some of the highly interdisciplinary Canadian research in biomaterials with a focus on neurotrauma applications.

The improvement of the loading content of hydrophilic drugs by polymer nanoparticles (NPs) recently has received increased attention from the field of controlled release. We developed a novel, simply modified, drop-wise nanoprecipitation method which separated hydrophilic drugs and polymers into aqueous phase (continuous phase) and organic phase (dispersed phase), both individually and involving a mixing process. Using this method, we produced ciprofloxacin-loaded NPs by Poly (d,l-lactic acid)-Dextran (PLA-DEX) and Poly lactic acid-co-glycolic acid-Polyethylene glycol (PLGA-PEG) successfully, with a considerable drug-loading ability up to 27.2 wt% and an in vitro sustained release for up to six days. Drug content with NPs can be precisely tuned by changing the initial drug feed concentration of ciprofloxacin. These studies suggest that this modified nanoprecipitation method is a rapid, facile, and reproducible technique for making nano-scale drug delivery carriers with high drug-loadin... [more]

CHO cells are often used to produce monoclonal antibodies in mammalian cell expression systems. In the process of large-scale cell culture, apoptosis is related to cell survival and product quality. Over-expressing an anti-apoptotic gene to delay apoptosis and improve cell growth is one of the strategies for improving productivity of monoclonal antibodies. Autophagy inducer rapamycin can extend the culture duration of CHO cells and affect the yield of antibodies. A method was developed for transient co-transfection of anti-apoptotic genes and genes of interest combined with rapamycin to increase the transient expression of the anti-PD1 antibody. Under the optimal transfection conditions, the combination of Bcl-xL and rapamycin can significantly delay cell apoptosis, inhibit cell proliferation, and prolong cell life-time. As a result, anti-PD1 monoclonal antibody expression levels are increased by more than 2 times.

Enzymatic hydration of oleic acid into 10-hydroxystearic acid (10-HSA) represents a theme of substantial scientific and practical interest. In this study, a fatty acid hydratase (OHase) from Lactococcus garvieae was cloned and expressed in Escherichia coli. The recombinantly expressed enzyme was identified as oleate hydratase (EC 4.2.1.53) confirming its highest hydration activity for oleic acid. The optimally yielded enzyme fraction was purified and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A solitary band on SDS-PAGE confirmed the molecular weight of 65 kDa. Gas chromatography-mass spectrometry (GC-MS) analysis scrutinized the silylated hydroxy fatty acid products acquired from the hydration of oleic acid by the oleate hydratase from L. garvieae. Optimal reaction conditions for the enzymatic production of 10-HSA from oleic acid using the purified oleate hydratase were pH 7.5, 30 °C, 105.49 U/mL enzyme solution and 30 g/L oleic acid. In the pre... [more]

Determining the optimum levels of macroalgal density and salinity for removing aquaculture effluent has gained increasing research interest in recent years because of the growing concerns over environmental sustainability. Here, we determined the effects of macroalgal density and salinity on the uptake of NO2−, NO3−, NH3, and PO43− by Caulerpa lentillifera from the effluent of Poecilia latipinna using spectrophotometry. Laboratory experiments were conducted to measure nutrient uptake at five different macroalgal density levels (10, 20, 30, 40, and 50 g/L) and three salinity levels (20, 30, and 40 ppt) with and without aeration. Quadratic regression analysis revealed significant nonlinear and linear effects of macroalgal density on the uptake of NO2−, NO3−, NH3, and PO43−, where the maximum uptake was predicted to occur at the macroalgal densities of 31.6, 33.3, 50.0, and 20.0 g/L, respectively. Likewise, the effects of salinity on the uptake of NO2−, NO3−, NH3, and PO43− were significa... [more]

Motivation: Immune cell dynamics is a critical factor of disease-associated pathology (immunopathology) that also impacts the levels of mRNAs in diseased tissue. Deconvolution algorithms attempt to infer cell quantities in a tissue/organ sample based on gene expression profiles and are often evaluated using artificial, non-complex samples. Their accuracy on estimating cell counts given temporal tissue gene expression data remains not well characterized and has never been characterized when using diseased lung. Further, how to remove the effects of cell migration on transcript counts to improve discovery of disease factors is an open question. Results: Four cell count inference (i.e., deconvolution) tools are evaluated using microarray data from influenza-infected lung sampled at several time points post-infection. The analysis finds that inferred cell quantities are accurate only for select cell types and there is a tendency for algorithms to have a good relative fit (R 2 ) but... [more]

The presented work describes the method of preparation of thermosensitive chitosan hydrogels using calcium β-glycerophosphate salt as the only pH neutralizing agent and supporting the crosslinking process. The presence of calcium ions instead of sodium ions is particularly important in the case of scaffolds in bone tissue engineering. Rheological and physicochemical properties of low concentrated chitosan solutions with the addition of calcium β-glycerophosphate were investigated using rotational rheometry techniques, Zeta potential (by electrophoresis), XPS, and SEM analysis together with an EDS detector. It was found to be possible to prepare colloidal solutions of chitosan containing only calcium β-glycerophosphate (without sodium ions) undergoing a sol-gel phase transition at the physiological temperature of the human body. It has also been shown that it is possible to further enrich the obtained cellular scaffolds with calcium ions. Using the addition of calcium carbonate, hydroge... [more]

Recombinant proteins produced by the baculovirus expression vector system (BVES) have been widely applied in the agricultural and medical fields. However, the procedure for protein expression is inefficient and needs to be improved. Herein, we propose a simple construct that incorporates a selectable marker (enhanced green fluorescent protein, EGFP) and a picorna viral-derived “self-cleaving” 2A-like peptide to separate the EGFP and target proteins in a monocistronic baculovirus vector to facilitate isolation of the recombinant baculovirus in the BVES. In this study, porcine adiponectin (ADN), a secreted, multimeric protein with insulin-sensitizing properties, was used to demonstrate its utility in our EGFP-2A-based expression system. EGFP and ADN were simultaneously expressed by a recombinant alphabaculovirus. Co-expression of EGFP facilitates the manipulation of the following processes, such as determining expression kinetics and harvesting ADN. The results showed that the 2A “self-c... [more]

The increasing rate of diabetes is found across the planet. Therefore, the diagnosis of pre-diabetes and diabetes is important in populations with extreme diabetes risk. In this study, a machine learning technique was implemented over a data mining platform by employing Rule classifiers (PART and Decision table) to measure the accuracy and logistic regression on the classification results for forecasting the prevalence in diabetes mellitus patients suffering simultaneously from other chronic disease symptoms. The real-life data was collected in Nigeria between December 2017 and February 2019 by applying ten non-intrusive and easily available clinical variables. The results disclosed that the Rule classifiers achieved a mean accuracy of 98.75%. The error rate, precision, recall, F-measure, and Matthew’s correlation coefficient MCC were 0.02%, 0.98%, 0.98%, 0.98%, and 0.97%, respectively. The forecast decision, achieved by employing a set of 23 decision rules (DR), indicates that age, ge... [more]

Tyrosine kinase receptor (RTK) ligation and dimerization is a key mechanism for translating external cell stimuli into internal signaling events. This process is critical to several key cell and physiological processes, such as in angiogenesis and embryogenesis, among others. While modulating RTK activation is a promising therapeutic target, RTK signaling axes have been shown to involve complicated interactions between ligands and receptors both within and across different protein families. In angiogenesis, for example, several signaling protein families, including vascular endothelial growth factors and platelet-derived growth factors, exhibit significant cross-family interactions that can influence pathway activation. Computational approaches can provide key insight to detangle these signaling pathways but have been limited by the sparse knowledge of these cross-family interactions. Here, we present a framework for studying known and potential non-canonical interactions. We construct... [more]

Genome-scale models have become indispensable tools for the study of cellular growth. These models have been progressively improving over the past two decades, enabling accurate predictions of metabolic fluxes and key phenotypes under a variety of growth conditions. In this work, an efficient computational method is proposed to incorporate genome-scale models into superstructure optimization settings, introducing them as viable growth models to simulate the cultivation section of biorefinaries. We perform techno-economic and life-cycle analyses of an algal biorefinery with five processing sections to determine optimal processing pathways and technologies. Formulation of this problem results in a mixed-integer nonlinear program, in which the net present value is maximized with respect to mass flowrates and design parameters. We use a genome-scale metabolic model of Chlamydomonas reinhardtii to predict growth rates in the cultivation section. We study algae cultivation in open ponds, in... [more]

Many drugs have a propensity for agglomeration, resulting in poor flowability. Spherical crystallization can be used to improve product properties including flowability and particle size. In this work, two methods were developed and utilized to successfully make two kinds of azithromycin spherical particles, namely solid and hollow spheres. The resultant product exhibited regular spherical shape, large particle size, narrow particle size distribution and excellent flowability. The formation mechanism of these different spherical crystals was investigated with the help of a particle vision microscope (PVM). The immersion mechanism and the counter diffusion mechanism were proposed as the formation mechanisms for solid and hollow spheres, respectively. The effects of crystallization parameters on the spherical crystallization processes were investigated systematically. Furthermore, the tablet properties were evaluated to verify that the spherical particles obtained in this work can be dir... [more]

Ultrasonic-assisted extraction (UAE) was optimized using response surface methodology (RSM) to maintain the cyto-protective activity of M.toringoides against oxidative stress. The optimal conditions for UAE were a 58 mL/g liquid-solid ratio, a 38 °C extraction temperature, an 85% solvent concentration, and a 19-min extraction time, which resulted in a protection rate of 54.57% against hydrogen peroxide-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). These results were comparable to the predicted value of 53.75%. The extracts showed excellent antioxidant activity, and phlorizin was detected in the dried leaves of Malus.toringoides. The highest yield of phlorizin (101.239 mg/g) was also obtained using these conditions. Taken together, these results showed that the method successfully integrated RSM and partial least squares regression methods to optimize M.toringoides extraction to yield the highest cyto-protective activity and effectively increase the yield... [more]

A simple method for obtaining pure and well-defined oligosaccharides was established by hydrolyzing agar with β-agarase from Vibrio natriegens. The conditions for enzymolysis were optimized as follows: a temperature of 45 °C, a pH of 8.5, a substrate concentration of 0.3%, an enzyme amount of 100 U/g and an enzymolysis time of 20 h. Neoagaro-oligosaccharides with different degrees of polymerization were obtained by hydrolyzing agar with β-agarase for different lengths of time. After removing pigments using activated carbon and salts by dialyzing, the enzyme hydrolysis solution was separated with Bio-Gel P2 column chromatography. Neoagaro-oligosaccharides with different degrees of polymerization were acquired. By comparing with authentic standard substances, along with further confirmation by FTIR, MS and NMR, structures of the purified neoagaro-oligosaccharides were identified as neoagarobiose (NA2), neoagaroteraose (NA4), neoagarohexaose (NA6), neoagarooctaose (NA8), neoagaro-decaose... [more]

The constraint-based rMeCBM-KU50 model of cassava storage root growth was analyzed to evaluate its sensitivity, with respect to reaction flux distribution and storage root growth rate, to changes in model inputted data and constraints, including sucrose uptake rate-related data—photosynthetic rate, total leaf area, total photosynthetic rate, storage root dry weight, and biomass function-related data. These mainly varied within ±90% of the model default values, although exceptions were made for the carbohydrate (−90% to 8%) and starch (−90% to 9%) contents. The results indicated that the predicted storage root growth rate was highly affected by specific sucrose uptake rates through the total photosynthetic rate and storage root dry weight variations; whereas the carbon flux distribution, direction and partitioning inclusive, was more sensitive to the variation in biomass content, particularly the carbohydrate content. This study showed that the specific sucrose uptake rate based on the... [more]