Environmental Friendly Deproteinization and Saccharification of Industrial Fungal Biomass by Enzymatic Processing

( Cecilia Perez-cruz ),( Carlos N. Cano-gonzalez ),( Jose Fuentes ),( Nagamani Balagurusamy ),( Carolina E. Vita ),( Roque A. Hours ),( Cristobal N. Aguilar ),( Sebastian F. Cavalitto ),( Juan C. Cont 한국키틴키토산학회 2018 한국키틴키토산학회지 Vol.23 No.3

Aspergillus niger biomass, an industrial by-product of citric acid fermentation is an emergent source of glycoderivatives with applications in biofuel, cosmetics, feed, energy, food, medicine, and nanotechnology. In this study, the effect of purified neutral protease for deprotenization of fungal biomass studied at various levels (0, 5, 10, 20 and 40 U/100 mg of biomass) and the saccharification of fungal biomass was evaluated with amylolytic enzymes and chitosanases. The efficiency of deproteinization of fungal biomass was based on the enzyme concentration and contact time. Protease at a concentration of 20 U/100 mg of dry biomass and with a contact time of 8 h achieved 30% final deproteinization. No effect on saccharification of A. niger biomass was observed by treatment with purified amylolytic enzymes. Meanwhile, the endo- and exo-chitosanases treatment yielded 54 g of g reducing sugars (equivalent to amino sugars)/ kg of fungal biomass, which can be employed for tailor-made carbohydrate production.

Bioelectronics-on-a-chip for cardio myoblast proliferation enhancement using electric field stimulation

Ángel Aragón,María Cebro-Márquez,Eliseo Perez,Antonio Pazos,Ricardo Lage,José Ramón González-Juanatey,Isabel Moscoso,Carmen Bao-Varela,Daniel Nieto 한국생체재료학회 2020 생체재료학회지 Vol.24 No.3

Background: Cardio myoblast generation from conventional approaches is laborious and time-consuming. We present a bioelectronics on-a-chip for stimulating cells cardio myoblast proliferation during culture. Method: The bioelectronics chip fabrication methodology involves two different process. In the first step, an aluminum layer of 200 nm is deposited over a soda-lime glass substrate using physical vapor deposition and selectively removed using a Q-switched Nd:YVO4 laser to create the electric tracks. To perform the experiments, we developed a biochip composed of a cell culture chamber fabricated with polydimethylsiloxane (PDMS) with a glass coverslip or a cell culture dish placed over the electric circuit tracks. By using such a glass cover slip or cell culture dish we avoid any toxic reactions caused by electrodes in the culture or may be degraded by electrochemical reactions with the cell medium, which is crucial to determine the effective cell-device coupling. Results: The chip was used to study the effect of electric field stimulation of Rat ventricular cardiomyoblasts cells (H9c2). Results shows a remarkable increase in the number of H9c2 cells for the stimulated samples, where after 72 h the cell density double the cell density of control samples. Conclusions: Cell proliferation of Rat ventricular cardiomyoblasts cells (H9c2) using the bioelectronics-on-a-chip was enhanced upon the electrical stimulation. The dependence on the geometrical characteristics of the electric circuit on the peak value and homogeneity of the electric field generated are analyzed and proper parameters to ensure a homogeneous electric field at the cell culture chamber are obtained. It can also be observed a high dependence of the electric field on the geometry of the electrostimulator circuit tracks and envisage the potential applications on electrophysiology studies, monitoring and modulate cellular behavior through the application of electric fields.

Assessment of the anterior loop of the mandibular canal: A study using cone-beam computed tomography

Eduarda Helena Leandro do Nascimento,Maria Luiza dos Anjos Pontual,Andréa dos Anjos Pontual,Danyel Elias da Cruz Perez,José Natal Figueiroa,Marco Antônio Gomes Frazão,Flávia Maria de Moraes Ramos-Pere 대한영상치의학회 2016 Imaging Science in Dentistry Vol.46 No.2

Purpose: Sufficient area in the interforaminal region is required for dental implant placement, and the anterior loop of the mandibular canal is located within the limits of this area. The aim of this study was to evaluate the prevalence and extent of the anterior loop in a Brazilian sample population using cone-beam computed tomography (CBCT). Materials and Methods: CBCT images from 250 patients (500 hemimandibles) obtained for various clinical indications were randomly selected and evaluated to determine the presence and length of the anterior loop. The length of the anterior loop was then compared based on gender, age, and the side of the mandible. The data were analyzed using the Pearson chi-square test and linear regression analysis. Results: An anterior loop was identified in 41.6% of the cases, and its length ranged from 0.25 mm to 4.00 mm (mean, 1.1±0.8 mm). The loop had a greater mean length and was significantly more prevalent in males (p=0.014). No significant differences were found between the right and left sides regarding length (p=0.696) or prevalence (p=0.650). Conclusion: In this study, a high prevalence of the anterior loop of the mandibular canal was found, and although its length varied greatly, in most cases it was less than 1 mm long. Although this is a prevalent anatomical variation, safety limits for the placement of implants in this region cannot be established before an accurate evaluation using imaging techniques in order to identify and preserve the neurovascular bundles. Materials and Methods CBCT images from 250 patients (500 hemimandibles) obtained for various clinical indications were randomly selected and evaluated to determine the presence and length of the anterior loop. The length of the anterior loop was then compared based on gender, age, and the side of the mandible. The data were analyzed using the Pearson chi-square test and linear regression analysis. Anatomic Variation; Cone-Beam Computed Tomography; Mandible Results An anterior loop was identified in 41.6% of the cases, and its length ranged from 0.25 mm to 4.00 mm (mean, 1.1±0.8 mm). The loop had a greater mean length and was significantly more prevalent in males (p=0.014). No significant differences were found between the right and left sides regarding length (p=0.696) or prevalence (p=0.650). Conclusion In this study, a high prevalence of the anterior loop of the mandibular canal was found, and although its length varied greatly, in most cases ... Purpose: Sufficient area in the interforaminal region is required for dental implant placement, and the anterior loop of the mandibular canal is located within the limits of this area. The aim of this study was to evaluate the prevalence and extent of the anterior loop in a Brazilian sample population using cone-beam computed tomography (CBCT). Materials and Methods: CBCT images from 250 patients (500 hemimandibles) obtained for various clinical indications were randomly selected and evaluated to determine the presence and length of the anterior loop. The length of the anterior loop was then compared based on gender, age, and the side of the mandible. The data were analyzed using the Pearson chi-square test and linear regression analysis. Results: An anterior loop was identified in 41.6% of the cases, and its length ranged from 0.25 mm to 4.00 mm (mean, 1.1±0.8 mm). The loop had a greater mean length and was significantly more prevalent in males (p=0.014). No significant differences were found between the right and left sides regarding length (p=0.696) or prevalence (p=0.650). Conclusion: In this study, a high prevalence of the anterior loop of the mandibular canal was found, and although its length varied greatly, in most cases it was less than 1 mm long. Although this is a prevalent anatomical variation, safety limits for the placement of implants in this region cannot be established before an accurate evaluation using imaging techniques in order to identify and preserve the neurovascular bundles. Materials and Methods CBCT images from 250 patients (500 hemimandibles) obtained for various clinical indications were randomly selected and evaluated to determine the presence and length of the anterior loop. The length of the anterior loop was then compared based on gender, age, and the side of the mandible. The data were analyzed using the Pearson chi-square test and linear regression analysis. Results An anterior loop was identified in 41.6% of the cases, and its length ranged from 0.25 mm to 4.00 mm (mean, 1.1±0.8 mm). The loop had a greater mean length and was significantly more prevalent in males (p=0.014). No significant differences were found between the right and left sides regarding length (p=0.696) or prevalence (p=0.650). Conclusion In this study, a high prevalence of the anterior loop of the mandibular canal was found, and although its length varied greatly, in most cases it was less than 1 mm long. Although this is a prevalent anatomical variation, safety limits for the placement of implants in this region cannot be established before an accurate evaluation using imaging techniques in order to identify and preserve the neurovascular bundles.

Structural and Optical Properties of Zn(1-x)CdxO Solid Solutions Grown on ZnO Substrates by Using MO-CVD

A. Lusson,N. Haneche,V. Sallet,P. Galtier,V. Munoz-Sanjose,J. Zuniga-Perez,S. Agouram,J. A. Bastos Segura,E. Leroy 한국물리학회 2008 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.53 No.1

Zn(1-x)CdxO solid solutions with a composition ranging from pure ZnO up to x = 0.07 have been grown on ZnO substrates by using metalorganic chemical vapor deposition (MO-CVD). The alloy concentration was carefully measured using a Castaing electron microprobe. The measured lateral homogeneity of the alloy was around 0.1 %. Two dierent photoluminescence lines were observed on Zn(1-x)CdxO alloys. At low temperature, a localized exciton line was identied and a second line appeared at higher energy for intermediate temperatures. This eect is interpreted as a delocalization eect of excitons trapped in potential uctuations related to a local alloy disorder. Zn(1-x)CdxO solid solutions with a composition ranging from pure ZnO up to x = 0.07 have been grown on ZnO substrates by using metalorganic chemical vapor deposition (MO-CVD). The alloy concentration was carefully measured using a Castaing electron microprobe. The measured lateral homogeneity of the alloy was around 0.1 %. Two dierent photoluminescence lines were observed on Zn(1-x)CdxO alloys. At low temperature, a localized exciton line was identied and a second line appeared at higher energy for intermediate temperatures. This eect is interpreted as a delocalization eect of excitons trapped in potential uctuations related to a local alloy disorder.

Novel therapeutic core-shell hydrogel scaffolds with sequential delivery of cobalt and bone morphogenetic protein-2 for synergistic bone regeneration

Perez, R.A.,Kim, J.H.,Buitrago, J.O.,Wall, I.B.,Kim, H.W. Elsevier BV 2015 ACTA BIOMATERIALIA Vol.23 No.-

Enabling early angiogenesis is a crucial issue in the success of bone tissue engineering. Designing scaffolds with therapeutic potential to stimulate angiogenesis as well as osteogenesis is thus considered a promising strategy. Here, we propose a novel scaffold designed to deliver angiogenic and osteogenic factors in a sequential manner to synergize the bone regeneration event. Hydrogel fibrous scaffolds comprised of a collagen-based core and an alginate-based shell were constructed. Bone morphogenetic protein 2 (BMP2) was loaded in the core, while the shell incorporated Co ions, enabled by the alginate crosslinking in CoCl<SUB>2</SUB>/CaCl<SUB>2</SUB> solution. The incorporation of Co ions was tunable by altering the concentration of Co ions in the crosslinking solution. The incorporated Co ions, that are known to play a role in angiogenesis, were released rapidly within a week, while the BMP2, acting as an osteogenic factor, was released in a highly sustainable manner over several weeks to months. The release of Co ions significantly up-regulated the in vitro angiogenic properties of cells, including the expression of angiogenic genes (CD31, VEGF, and HIF-1α), secretion of VEGF, and the formation of tubule-like networks. However, BMP2 did not activate the angiogenic processes. Osteogenesis was also significantly enhanced by the release of Co ions as well as BMP2, characterized by higher expression of osteogenic genes (OPN, ALP, BSP, and OCN), and OCN protein secretion. An in vivo study on the designed scaffolds implanted in rat calvarium defect demonstrated significantly enhanced bone formation, evidenced by new bone volume and bone density, due to the release of BMP2 and Co ions. This is the first study using Co ions as an angiogenic element together with the osteogenic factor BMP2 within scaffolds, and the results demonstrated the possible synergistic role of Co ions with BMP2 in the bone regeneration process, suggesting a novel potential therapeutic scaffold system. Statement of Significance: This is the first report that utilizes Co ion as a pro-angiogenic factor in concert with osteogenic factor BMP-2 in the fine-tuned core-shell hydrogel fiber scaffolds, and ultimately achieves osteo/angiogenesis of MSCs and bone regeneration through the sequential delivery of both biofactors. This novel approach facilitates a new class of therapeutic scaffolds, aiming at successful bone regeneration with the help of angiogenesis.

Influence of Ionophore Supplementation on Growth Performance, Dietary Energetics and Carcass Characteristics in Finishing Cattle during Period of Heat Stress

Barreras, A.,Castro-Perez, B.I.,Lopez-Soto, M.A.,Torrentera, N.G.,Montano, M.F.,Estrada-Angulo, A.,Rios, F.G.,Davila-Ramos, H.,Plascencia, A.,Zinn, R.A. Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.11

Forty-eight crossbred heifers ($378.1{\pm}18$ kg) were used in a 56-d feeding trial (four pens per treatment in a randomised complete block design) to evaluate the influence of ionophore supplementation on growth performance, dietary energetics and carcass characteristics in finishing cattle during a period of heat stress. Heifers were fed a diet based on steam-flaked corn (2.22 Mcal $NE_m/kg$) with and without an ionophore. Treatments were: i) control, no ionophore; ii) 30 mg/kg monensin sodium (RUM30); iii) 20 mg/kg lasalocid sodium (BOV20), and iv) 30 mg/kg lasalocid sodium (BOV30). Both dry matter intake (DMI) and climatic variables were measured daily and the temperature humidity index (THI) was estimated. The maximum THI during the study averaged 93, while the minimum was 70 (THI average = $79.2{\pm}2.3$). Compared to controls, monensin supplementation did not influence average daily gain, the estimated NE value of the diet, or observed-to-expected DMI, but tended (p = 0.07) to increase (4.8%) gain to feed. Compared to controls, the group fed BOV30 increased ($p{\leq}0.03$) daily gain (11.8%), gain to feed (8.3%), net energy of the diet (5%), and observed-to-expected DMI (5.2%). Daily weight gain was greater (7.6%, p = 0.05) for heifers fed BOV30 than for heifers fed MON30. Otherwise, differences between the two treatments in DMI, gain to feed, and dietary NE were not statistically significant (p>0.11). Plotting weekly intakes versus THI, observed intake of controls was greater (p<0.05) at THI values ${\leq}77$ than ionophore groups. When THI values were greater than 79, DMI of control and MON30 were not different (p = 0.42), although less than that of groups fed lasalocid (p = 0.04). Variation in energy intake was lower (p>0.05) in the ionophores group (CV = 1.7%) than in the control group (CV = 4.5%). Inclusion of ionophores in the diet resulted in relatively minor changes in carcass characteristics. It is concluded that ionophore supplementation did not exacerbate the decline of DM intake in heat-stressed cattle fed a high-energy finishing diet; on the contrary, it stabilised feed intake and favoured feed efficiency. Ionophore supplementation reduced estimated maintenance coefficients around 10% in finishing cattle during a period of heat stress. This effect was greatest for heifers supplemented with 30 mg lasalocid/kg of diet.

Influence of Quaternary Benzophenantridine and Protopine Alkaloids on Growth Performance, Dietary Energy, Carcass Traits, Visceral Mass, and Rumen Health in Finishing Ewes under Conditions of Severe Temperature-humidity Index

Estrada-Angulo, A.,Aguilar-Hernandez, A.,Osuna-Perez, M.,Nunez-Benitez, V.H.,Castro-Perez, B.I.,Silva-Hidalgo, G.,Contreras-Perez, G.,Barreras, A.,Plascencia, A.,Zinn, R.A. Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.5

Twenty $Pelibuey{\times}Katahdin$ ewes ($35{\pm}2.3kg$) were used to determine the effects of the consumption of standardized plant extract containing a mixture of quaternary benzophenanthridine alkaloids and protopine alkaloids (QBA+PA) on growth performance, dietary energetics, visceral mass, and ruminal epithelial health in heat-stressed ewes fed with a high-energy corn-based diet. The basal diet (13.9% crude protein and 2.09 Mcal of net energy [NE] of maintenance/kg of dry matter) contained 49.7% starch and 15.3% neutral detergent fiber. Source of QBA+PA was Sangrovit RS (SANG) which contains 3 g of quaternary benzophenathridine and protopine alkaloids per kg of product. Treatments consisted of a daily consumption of 0 or 0.5 g SANG/ewe. Ewes were grouped by weight and assigned to 10 pens (5 pens/treatment), with two ewes per pen. The experimental period lasted 70 days. The mean temperature humidity index during the course of this experiment was $81.7{\pm}1.0$ (severe heat stress). There were no treatment effects on water intake. Dry matter intake was not affected (p = 0.70) by treatments, but the group fed SANG had a numerically (11.2%) higher gain in comparison to the control group, SANG improved gain efficiency (8.3%, p = 0.04), dietary NE (5.2%, p<0.01) and the observed-to-expected NE (5.9%, p<0.01). Supplemental SANG did not affect ($p{\geq}0.12$) carcass characteristics, chemical composition of shoulder, and organ weights (g/kg empty body weight) of stomach complex, intestines, and heart/lung. Supplemental SANG decreased liver weight (10.3%, p = 0.02) and increased visceral fat (16.9%, p = 0.02). Rumen epithelium of ewes fed SANG had lower scores for cellular dropsical degeneration (2.08 vs 2.34, p = 0.02), parakeratosis (1.30 vs 1.82, p = 0.03) and neutrophil infiltration (2.08 vs 2.86, p = 0.05) than controls. It is concluded that SANG supplementation helped ameliorate the negative effects of severe heat on growth performance of feedlot ewes fed high-energy corn-based diets. Improvement in energetic efficiency may have been mediated, in part, by anti-inflammatory effects of supplemental SANG and corresponding enhancement of nutrient uptake.

Simple Fractional Order Controller Combined with a Smith Predictor for Temperature Control in a Steel Slab Reheating Furnace

Vicente Feliu-Batlle,Raul Rivas-Perez,Fernando J. Castillo-García 제어·로봇·시스템학회 2013 International Journal of Control, Automation, and Vol.11 No.3

This paper proposes a simple fractional order controller combined with a Smith predictor scheme for controlling the temperature of a steel slab reheating furnace. The dynamic model of the preheating zone of this process is obtained from an identification procedure applied in an industrial furnace. This identification procedure yields a second order plus time delay transfer function which undergoes large time delay changes. A fractional order integral controller combined with a Smith predictor is therefore designed. Simulated results compare the performances of the proposed fractional order controller with a standard PI controller, also combined with a Smith predictor, an LQR controller, and an H∞ robust controller, in the case of the nominal process, and when the time delay varies. Four performance indexes have been used in this comparison: three related to the output performance (settling time, overshooting, and integral absolute error (IAE)), and a fourth one related to the control effort (TV). The analysis of these indexes shows that the simple fractional order controller provides lower values of the compared indexes when time delay becomes much higher than the nominal value.

Surface guidance of stem cell behavior: Chemically tailored co-presentation of integrin-binding peptides stimulates osteogenic differentiation in vitro and bone formation in vivo

Fraioli, R.,Dashnyam, K.,Kim, J.H.,Perez, R.A.,Kim, H.W.,Gil, J.,Ginebra, M.P.,Manero, J.M.,Mas-Moruno, C. Elsevier BV 2016 Acta Biomaterialia: structure-property-function re Vol.43 No.-

Surface modification stands out as a versatile technique to create instructive biomaterials that are able to actively direct stem cell fate. Chemical functionalization of titanium has been used in this work to stimulate the differentiation of human mesenchymal stem cells (hMSCs) into the osteoblastic lineage, by covalently anchoring a synthetic double-branched molecule (PTF) to the metal that allows a finely controlled presentation of peptidic motifs. In detail, the effect of the RGD adhesive peptide and its synergy motif PHSRN is studied, comparing a random distribution of the two peptides with the chemically-tailored disposition within the custom made synthetic platform, which mimics the interspacing between the motifs observed in fibronectin. Contact angle measurement and XPS analysis are used to prove the efficiency of functionalization. We demonstrate that, by rationally designing ligands, stem cell response can be efficiently guided towards the osteogenic phenotype: In vitro, PTF-functionalized surfaces support hMSCs adhesion, with higher cell area and formation of focal contacts, expression of the integrin receptor α5β1 and the osteogenic marker Runx2, and deposition a highly mineralized matrix, reaching values of mineralization comparable to fibronectin. Our strategy is also demonstrated to be efficient in promoting new bone growth in vivo in a rat calvarial defect. These results highlight the efficacy of chemical control over the presentation of bioactive peptides; such systems may be used to engineer bioactive surfaces with improved osseointegrative properties, or can be easily tuned to generate multi-functional coatings requiring a tailored disposition of the peptidic motifs. Statement of significance: Organic coatings have been proposed as a solution to foster osseointegration of orthopedic implants. Among them, extracellular matrix-derived peptide motifs are an interesting biomimetic strategy to harness cell-surface interactions. Nonetheless, the combination of multiple peptide motifs in a controlled manner is essential to achieve receptor specificity and fully exploit the potentiality of synthetic peptides. Herein, we covalently graft to titanium a double branched molecule to guide stem cell fate in vitro and generate an osseoinductive titanium surface in vivo. Such synthetic ligand allows for the simultaneous presentation of two bioactive motifs, thus is ideal to test the effect of synergic sequences, such as RGD and PHSRN, and is a clear example of the versatility and feasibility of rationally designed biomolecules.

Comparative evaluation of supplemental zilpaterol hydrochloride sources on growth performance, dietary energetics and carcass characteristics of finishing lambs

A. Rivera-Villegas,A. Estrada-Angulo,B.I. Castro-Perez,J.D. Urias-Estrada,F.G. Rios-Rincon,D. Rodriguez-Cordero,A. Barreras,A. Plascencia,V.M.Gonzalez-Vizcarra,J.F. Sosa-Gordillo,R.A. Zinn 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.2

Objective: We compare the effects of three different approved sources of supplemental zilpaterol on growth-performance responses and carcass characteristics of finishing lambs. Methods: Twenty four Pelibuey×Katahdin lambs (46.75±2.43 kg) were used in a 33-day feeding trial. Lambs were fed a dry rolled corn-based finishing diet. Treatments consisted of the non-supplemental basal diet (Control) versus the basal diet supplemented with 125 mg zilpaterol/kg of diet (as fed basis) from three commercial sources marketed in Mexico: Zilmax (ZIL), Grofactor, and Zipamix. Results: Compared to controls, zilpaterol (ZH) supplementation did not affect dry matter intake (DMI), but increased carcass adjusted daily weight gain (ADG, 36.7%), gain efficiency (34.2%), and dietary net energy (26.0%), and decreased (23.4%) the ratio of observed:expected DMI. Compared to controls, supplemental ZH increased hot carcass weight (6.4%), dressing percentage (3.2%), m. longissimus thoracis (LM) area (15.6%), and shoulder muscle:fat ratio (28.7%), but decreased kidney-pelvic-heart fat, and fat thickness. Supplemental ZH increased 10.9% and 14.3% whole cut weight of loin and leg, respectively, and the proportion (as percentage of cold carcass weight) of leg (4.3%). These increases were reflected in greater forequarter and hindquarter weights. Lambs fed ZH increased (4.6%) empty body weight (EBW) and reduced (14.7%) liver/spleen weight (as g/kg EBW). Likewise, ZH supplementation tended (p = 0.08) to lower (8.9%) visceral fat. Growth performance, energetic efficiency, hot carcass weight, dressing percentage, LM area and whole cuts were not different across supplemental ZH sources. However, compared with non-supplemented controls, only ZIL appreciably decreased carcass fat distribution, including fat thickness, percentage kidney pelvic and heart fat, shoulder fat, and visceral fat. Conclusion: Supplemental ZH increases ADG, gain efficiency, carcass dressing percentage, and LM area. The magnitude of these responses was similar among ZH sources. Nevertheless, compared with non-supplemented controls, only ZIL appreciably decreases carcass fat. The basis for this is uncertain, but indicative that some practical differences in zilpaterol bio-equivalency may exist across commercial sources tested.