Publications by authors named "Victor Gustavo Balera Brito"

11 Publications

  • Page 1 of 1

Mast cells contribute to alveolar bone loss in Spontaneously Hypertensive Rats with periodontal disease regulating cytokines production.

PLoS One 2021 4;16(3):e0247372. Epub 2021 Mar 4.

Programa Multicêntrico de Pós-graduação em Ciências Fisiológicas, SBFis, São Paulo State University (UNESP), School of Dentistry, Araçatuba, SP, Brazil.

Mast cells (MCs) play a pivotal role in inflammatory responses and had been studied in inflammatory bone disorders, however, their role in alveolar bone loss induced by periodontal disease (PD) is not yet fully understood. We, therefore, aimed to evaluate the effects of MCs depletion in the PD-induced alveolar bone loss in Wistar (W) and Spontaneously Hypertensive Rats (SHRs). PD was induced by ligating the lower first molars with silk thread one day after the MCs depletion, by the pre-treatment with compound 48/80 for 4 days. After 15 days of PD induction, the hemi-mandibles were surgically collected for qRT-PCR, histological analyses, immunostaining, and ELISA. Systolic blood pressure (SBP) was verified by tail plethysmography to confirm the hypertensive status, and SHR presented SBP >150 mmHg, and previous MC depletion alone or associated with PD did not alter this parameter. SHRs showed a more severe alveolar bone loss compared to W, and MC depletion significantly inhibited this response in both strains, with a more significant response in SHRs. MCs were less abundant in 48/80+PD groups, thus validating the previous MCs depletion in our model. PD increased the number of MC in the gingival tissue of SHR. Cytokine production (TNF-α, IL-6, IL-1β, and CXCL3) was constitutively higher in SHR and increased further after PD, which was also significantly reduced in the MCs-depleted animals. PD led to an increased expression of Opn, Rankl, Rank, Vtn, Itga5, Itgb5, Trap, and Ctsk in the mandible of W and SHRs, which was reversed in MCs-depleted animals. These results suggest that MCs significantly contributes to the PD-induced alveolar bone resorption, especially in the SHR, which is associated with a more severe PD progression compared to Wistar, partly explained by these cells contribution to the inflammatory status and mediator production, stimulating osteoclast-related response markers, which were reduced after MC depletion in our experimental model.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0247372PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7932174PMC
March 2021

Telmisartan Prevents Alveolar Bone Loss by Decreasing the Expression of Osteoclasts Markers in Hypertensive Rats With Periodontal Disease.

Front Pharmacol 2020 11;11:579926. Epub 2020 Nov 11.

Department of Basic Sciences, School of Dentistry, São Paulo State University (UNESP), Araçatuba, Brazil.

Periodontal disease (PD) is a prevalent inflammatory disease with the most severe consequence being the loss of the alveolar bone and teeth. We therefore aimed to evaluate the effects of telmisartan (TELM), an angiotensin II type 1 receptor (Agtr1) antagonist, on the PD-induced alveolar bone loss, in Wistar (W) and Spontaneous Hypertensive Rats (SHRs). PD was induced by ligating the lower first molars with silk, and 10 mg/kg TELM was concomitantly administered for 15 days. The hemimandibles were subjected to microtomography, ELISA was used for detecting tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), CXCL3, and CCL2, while qRT-PCR was used for analyzing expression of components of renin-angiotensin system (RAS) (Agt, Ace, Agt1r, Agt2r, Ace2, and Masr), and bone markers (Runx2, Osx, Catnb, Alp, Col1a1, Opn, Ocn, Bsp, Bmp2, Trap, Rank, Rankl, CtsK, Mmp-2, Mmp-9, and osteoclast-associated receptor (Oscar)). The SHR + PD group showed greater alveolar bone loss than the W + PD group, what was significantly inhibited by treatment with TELM, especially in the SHR group. Additionally, TELM reduced the production of TNF-α, IL-1β, and CXCL3 in the SHR group. The expression of Agt increased in the groups with PD, while reduced, and TELM reduced the expression of Agtr1 and increased the expression of Agtr2, in W and SHRs. PD did not induce major changes in the expression of bone formation markers, except for the expression of Alp, which decreased in the PD groups. The bone resorption markers expression, Mmp9, Ctsk, and Vtn, was higher in the SHR + PD group, compared to the respective control and W + PD group. However, TELM attenuated these changes and increased the expression of Runx2 and Alp. Our study suggested that TELM has a protective effect on the progression of PD, especially in hypertensive animals, as evaluated by the resorption of the lower alveolar bone. This can be partly explained by the modulation in the expression of Angiotensin II receptors (AT1R and AT2R), reduced production of inflammatory mediators, the reduced expression of resorption markers, and the increased expression of the bone formation markers.
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http://dx.doi.org/10.3389/fphar.2020.579926DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7751694PMC
November 2020

Effect of pre-treatment of strength training and raloxifene in periestropause on bone healing.

Bone 2020 05 22;134:115285. Epub 2020 Feb 22.

Programa de Pós-Graduação Multicêntrico em Ciências Fisiológicas, SBFis, São Paulo State University (UNESP), Araçatuba, São Paulo, Brazil; Department of Basic Sciences, School of Dentistry, São Paulo State University (UNESP), Araçatuba, São Paulo, Brazil. Electronic address:

Background: There is evidence that strength training (ST) and raloxifene (Ral) treatment during periestropause promotes better bone quality. We wanted to determine whether the skeletal benefits of ST or Ral treatment, performed during periestropause, would persist after fracture. Therefore, the present study aimed to analyze the influence of pre-treatment with ST and administration of Ral during periestropause on bone healing after total unilateral osteotomy.

Methods: Senescent female Wistar rats between 18 and 21 months of age, performed ST on a ladder three times per week, were administered Ral by gavage (2.3 mg/kg/day), or an association of both. After 120 days, the treatments were interrupted, and a total osteotomy was performed on the left tibia in all animals. They were euthanized 1 and 8 weeks post-osteotomy.

Results: The administration of Ral during periestropause worsened the biochemical and oxidative profile, decreased gene expression of markers related to bone resorption and remodeling, which negatively affected the physicochemical properties; this lead to changes in the bone callus microarchitecture and mass, as well as a decrease in callus resistance to torsional deformation, resulting in lower tissue quality during bone healing. In contrast, ST performed prior to the osteotomy resulted in better bone healing, improvement of the biochemical and oxidative profile, alteration of the genetic profile in favor of bone formation and resorption, as well as the physic-ochemical properties of the callus. These changes led to better microarchitecture and bone mass and increased callus resistance to torsional deformation, confirming its beneficial effect on the quality of bone tissue, providing acceleration of bone consolidation. The combination of therapies at this exercise intensity and drug dosage showed a negative interaction, where the negative effect of Ral overcame the positive effect of ST, leading to decreased tissue quality in the bone healing process.

Conclusions: This study indicates that in addition to excellent non-pharmacological therapy and action in the prevention of osteoporosis, ST performed during the aging period may increase bone quality at the onset of healing and provide improved bone consolidation. Furthermore, the anti-osteoclastogenic effect of Ral shown in this model delayed the bone repair process, resulting in considerable clinical concern.
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http://dx.doi.org/10.1016/j.bone.2020.115285DOI Listing
May 2020

Soluble yerba mate (Ilex Paraguariensis) extract enhances in vitro osteoblastic differentiation of bone marrow-derived mesenchymal stromal cells.

J Ethnopharmacol 2019 Nov 1;244:112131. Epub 2019 Aug 1.

Programa Multicêntrico de Pós-Graduaçãoem Ciências Fisiológicas, SBFis/UNESP, Brazil; Laboratory of Pharmacology, Department of Basic Sciences, São Paulo State University (UNESP), School of Dentistry, Araçatuba, SP, Brazil; Department of Basic Sciences, São Paulo State University (UNESP), School of Dentistry, Araçatuba, SP, Brazil.

Ethnopharmacological Relevance: Yerba mate (Ilex paraguariensis) consumption has been associated with beneficial effects on bone health.

Aim Of The Study: The purpose of this study was to evaluate the mechanism by which soluble yerba mate (SYM) stimulates osteoblast differentiation of bone marrow-derived mesenchymal stromal cells (BM-MSCs).

Materials And Methods: BM-MSCs from male Wistar rats were induced towards osteoblastic differentiation with different concentrations of SYM (10, 20, and 50 μg/mL). Osteoblastic differentiation was evaluated by measuring proliferation rates, alkaline phosphatase activity, MMP-2 activity, mineralization, and gene expression of Runx2, Osterix, β-catenin (Catnb), collagen type I (Col1a1), osteopontin (Opn), osteocalcin (Ocn), bone sialoprotein (Bsp), bone morphogenetic protein-2 (Bmp2), osteoprotegerin (Opg), and Rankl. We also analyzed cytokine production and MAP kinase pathways.

Results: SYM (10 μg/mL) did not show a cytotoxic effect and induced a slight increase in ALP activity; however, a great increase in mineralization was observed. SYM was also able to reduce TNF-α and IL-10 production; increase the expression of transcription factors Runx2, Osterix, and Catnb; and increase matrix proteins Opn, Bsp, Ocn, and Bmp2. We also observed a decrease in intracellular signaling of ERK, JNK, and p38 MAPK, which seemed to be related to the SYM response.

Conclusions: Together, these results help to explain the promoting effect on osteoblast differentiation produced by a low SYM concentration. However, a higher SYM concentration presented deleterious effects, including cytotoxicity, decreased ALP activity, increased cytokine production, decreased bone marker gene expression, increased MAPK signaling, and significant mineralization reduction. In conclusion, our results suggest a concentration-specific direct stimulatory effect of SYM on osteoblastic differentiation in vitro.
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http://dx.doi.org/10.1016/j.jep.2019.112131DOI Listing
November 2019

Aliskiren Attenuates the Inflammatory Response and Wound Healing Process in Diabetic Mice With Periodontal Disease.

Front Pharmacol 2019 4;10:708. Epub 2019 Jul 4.

Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo (USP), Bauru, Brazil.

The aim of this study was to characterize the role of local RAS (renin-angiotensin system) in the inflammatory response of normal (N) and diabetic (D) mice with periodontal disease (PD). Diabetes Mellitus (DM) was induced by peritoneal injection of streptozotocin in Balb/c mice. PD was induced by ligature around the first molar in both N and D, irrespective of whether they were treated with aliskiren (50 mg/kg, Alisk). Mandibles were harvested for histomorphometric analyses, and gingival tissue (GT) was collected to evaluate gene expression and extracellular matrix components (ECM). Immunohistochemical (IHC) analyses were used to localize RAS in GT. The production of C-reactive protein (CRP), IL-1β, CXCL2, and CCL8 was evaluated by enzyme-linked immunosorbent assay (ELISA). Renin was found to exacerbate the inflammation and periodontal bone loss at 14 days after PD, and Alisk inhibited this process in GT of N and D. PD increased CRP, CXCL2, CCL8, and IL-1β production in both animals. Alisk could inhibit CRP, CXCL2, and CCL8 primarily in D animals. However, only CCL8 was decreased in N animals after Alisk pretreatment. PD enhanced expression and production of AGT, ACE, AT1R, and AT2R in both N and D. AT1R expression was higher in D with PD, and AT2R expression was higher in N with PD. ACE2 and receptor Mas (MasR) expression and production was elevated in the control group of both animals. PD inhibited ACE2 in N but not in D. MasR expression was unaffected in both N and D with PD. Alisk reduced expression and production of all RAS components in GT of both animals, except for ACE2 in N. RAS staining was observed in all layers of epithelium, basal cell layer, and lamina propria and was higher in N with PD. Col1a1, Col1a2, Col3a1, and fibronectin (Fn1) were increased in both animals with PD. Alisk inhibited Col1a1 and Fn in both animals, Col1a2 was decreased only in D, while levels of Col3a1 remained unchanged in all animal groups. In conclusion, these data demonstrated the presence and functional role of local RAS in GT, exacerbating the inflammatory response, periodontal bone loss, and wound healing processes in both N and D animal groups. In addition, Alisk was able to significantly reduce gingival inflammation, excessive wound healing processes, and periodontal bone loss.
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http://dx.doi.org/10.3389/fphar.2019.00708DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6620569PMC
July 2019

Effect of maternal periodontitis on GLUT4 and inflammatory pathway in adult offspring.

J Periodontol 2019 08 21;90(8):884-893. Epub 2019 Feb 21.

Multicenter Post-Graduate Program in Physiological Sciences (SBFis), Department of Basic Sciences, School of Dentistry, São Paulo State University (UNESP), Araçatuba, Brazil.

Background: Maternal periodontal disease leads to low birth weight (LBW), insulin resistance (IR), increased TNF-α levels, and alterations in insulin signaling in adult offspring. TNF-α has been associated with the stimulation of IKKβ/NF-κB, resulting in the decreased expression of GLUT4. Another mechanism that may be involved in decreasing GLUT4 expression is DNA methylation. This study aimed to evaluate in the adult offspring of rats with periodontal disease: IR, inflammatory pathways, DNA methylation, and expression of GLUT4.

Methods: Female Wistar rats were distributed into control and experimental periodontal disease groups. Seven days after induction of periodontal disease, both groups were mated with healthy male rats. After weaning, male offspring were distributed into control offspring (CN-o) and periodontal disease offspring (PED-o) groups. Body weights were measured from 0-75 days of age. At day 75, the following were measured in the offspring: IR (HOMA-IR index); TNF-α and NF-κBp65 content in the gastrocnemius muscle (GM) by western blotting; IKKα/β, JNK, ERK 1/2, NF-κBp65, and NF-κBp50 phosphorylation status in the GM by western blotting; DNA methylation by restriction digest and real-time PCR(qAMP); and expression of GLUT4 mRNA in the GM by real-time PCR.

Results: LBW, IR, increases in TNF-α, IKKα/β, ERK 1/2, NF-κBp65, and NF-κBp50 decreased expression of GLUT4 mRNA were observed in the PED-o rats. No differences were identified in JNK phosphorylation status and DNA methylation in the evaluated regions of the GLUT4-encoding gene Slc2a4.

Conclusion: Maternal periodontal disease causes LBW, IR, activation of inflammatory pathways, and decreased GLUT4 expression in the GM of adult offspring.
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http://dx.doi.org/10.1002/JPER.18-0568DOI Listing
August 2019

Influence of Preparation and Exposure Periods of Eluates from Ocular Prosthesis Acrylic Resin in Human Conjunctival Cell Line

Iran Biomed J 2019 01 17;23(1):78-86. Epub 2018 Jul 17.

Department of Dental Materials and Prosthodontics, Aracatuba Dental School, Sao Paulo State University (UNESP), Aracatuba, Sao Paulo, Brazil.

Background: This study was undertaken to analyze if different preparation and exposure periods of eluates from ocular prosthesis acrylic resin influence the cytotoxicity for conjunctival cells.

Methods: Twenty-four acrylic resin specimens were divided, according to the period of eluate exposure to Chang conjunctival cells (24 and 72 hours). Eluates were prepared in four different ways: 24, 48, and 72 hours of resin specimen immersion in medium and 24 hours of immersion in water, followed by 24 hours of immersion in medium. MTT assay was used to evaluate the cytotoxic effect. The production of IL-1β, IL-6, TNF-α, and chemokine macrophage inflammatory protein 1α was evaluated by ELISA, while the mRNA expression of type IV collagen (COL IV), transforming growth factor β (TGF-β), and matrix metalloproteinase 9 (MMP9) were evaluated by real-time RT-PCR technique. The statistical analysis was carried out using ANOVA with Bonferroni post-hoc test and the student’s t-test (p < 0.05).

Results: Significant quantities of IL-6 (4.594 pg/mL) and mRNA expression of COL IV (1.58) were verified at 72 hours of eluate exposure to cells, as compared to 24 hours. After the 72-hour exposure of eluates to cells, lower cell proliferation (88.4%) and higher IL-6 quantities (12.374 pg/mL), as well as mRNA expression of COL IV (2.21), TGF-β (2.02), and MMP9 (5.75) were observed, which corresponded to 72 hours of a specimen immersed in medium.

Conclusion: Longer periods of eluate preparation and exposure from the acrylic resin to cells are related to higher production of proinflammatory cytokines and extracellular matrix proteins.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6305826PMC
January 2019

Biocompatibility of primers and an adhesive used for implant-retained maxillofacial prostheses: An in vitro analysis.

J Prosthet Dent 2017 Jun 9;117(6):799-805. Epub 2016 Nov 9.

Associate Professor, Department of Dental Materials and Prosthodontics, Aracatuba Dental School, São Paulo State University, São Paulo, Brazil. Electronic address:

Statement Of Problem: Implant-retained maxillofacial prostheses should be biocompatible, regardless of the primers and adhesives used to bond the acrylic resin and facial silicone. The authors are unaware of any study evaluating the influence of these primers and adhesives on the biocompatibility of maxillofacial prostheses.

Purpose: The purpose of this in vitro study was to evaluate the cytotoxic effect of primers and an adhesive used to bond acrylic resin and facial silicone during the fabrication of implant-retained maxillofacial prostheses.

Material And Methods: Twenty-eight circular specimens made of resin and silicone were fabricated, either bonded or nonbonded with primer and adhesive. The specimens were divided into 7 groups: resin; silicone; resin+silastic medical adhesive type A+silicone; resin+DC 1205 primer silicone; resin+Sofreliner primer+silicone; resin+DC 1205 primer+silastic medical adhesive type A+silicone; and resin+Sofreliner primer+silastic medical adhesive type A+silicone. Eluates of the materials tested were prepared by setting 4 specimens of each experimental group in Falcon tubes with medium and incubating at 37°C for 24 hours. The eluate cytotoxicity was evaluated by an assay of survival/proliferation ((3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide [MTT] test) in cultures of human keratinocytes. The levels of IL1, IL6, TNFα, and the chemokine MIP-1α were evaluated by enzyme-linked immunosorbent assay. The mRNA expressions for MMP-9, TGF-β, and collagen type IV were analyzed by the real time polymerase chain reaction. Data were submitted to analysis of variance with Bonferroni post hoc tests (α=.05).

Results: An increased cell proliferation was observed for the RAS group, with statistically significant differences (P<.001) compared with the unstimulated group. The RDCpS group showed the highest IL6 concentration values (P<.001). No significant statistical difference was found in the relative quantification of mRNA for collagen type IV, MMP9, or TGFβ between the groups (P>.05).

Conclusions: The RAS group showed the highest cell proliferation percentage, while the RDCpS group exhibited the highest IL6 concentration values. No detectable levels of IL1β, TNF α, or CCL3/MIP1α were observed. The tested materials showed no toxic effects on the HaCaT cell line.
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http://dx.doi.org/10.1016/j.prosdent.2016.09.002DOI Listing
June 2017

Effect of different methods of polymerizing ocular prosthesis acrylic resin on a human conjunctival cell line.

J Prosthet Dent 2016 Nov 14;116(5):818-823. Epub 2016 Jul 14.

Professor, Department of Basic Sciences, Aracatuba Dental School, São Paulo State University, São Paulo, Brazil.

Statement Of Problem: Ocular prosthesis acrylic resins should be biocompatible regardless of the polymerization method. The authors are unaware of a study that evaluated the biocompatibility of ocular prostheses.

Purpose: The purpose of this in vitro study was to evaluate the cytotoxicity of different methods of polymerizing ocular prosthesis acrylic resin. This was accomplished by analyzing the cell proliferation, production of proinflammatory cytokines, and expression of extracellular matrix proteins related to tissue remodeling and repair of a human conjunctival cell line.

Material And Methods: Nine acrylic resin specimens were divided into 3 groups: polymerization in a water bath, by microwave, or by autopolymerization. Eluates (prepared for 72 hours) were exposed to cells for 72 hours. A medium without specimens served as negative control (nonstimulated group). The tetrazolium dye MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was performed to evaluate the cytotoxic effect, and an enzyme-linked immunosorbent assay was executed for analysis of interleukin 1 β (IL1β), IL6, tumor necrosis factor α (TNFα), and CCL3/MIP1α production. Also, real-time reverse transcriptase (RT)-PCR was performed for analysis of mRNA expression of type IV collagen (COL IV), TGFβ, and MMP9, and data were tested using ANOVA with Bonferroni post hoc test (α=.05).

Results: Microwave-processed resin showed slight cytotoxicity due to a significant reduction in cell proliferation and an increase in IL6 quantity. Higher levels of mRNA expression of COL IV, MMP9, and TGFβ were verified in water bath-processed resin, which were similar to those in the nonstimulated group.

Conclusions: Microwave-processed resin showed a significant reduction in cell proliferation and an increase in IL6 quantity. Heat-polymerized resin exhibited a higher mRNA expression of COL IV, MMP9, and TGFβ; this result was similar to that in the nonstimulated group.
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http://dx.doi.org/10.1016/j.prosdent.2016.06.001DOI Listing
November 2016

Osteogenic markers are reduced in bone-marrow mesenchymal cells and femoral bone of young spontaneously hypertensive rats.

Life Sci 2016 Feb 13;146:174-83. Epub 2016 Jan 13.

School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Campus Araçatuba, Department of Basic Sciences, São Paulo, Brazil; Programa de Pós-graduação Multicêntrico em Ciências Fisiológicas - SBFIS/FOA - Araçatuba, Department of Basic Sciences School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, Department of Basic Sciences, São Paulo, Brazil. Electronic address:

Aims: Spontaneously hypertensive rats (SHR) and normotensive rats (W) has significant changes in bone metabolism. The purpose of this study was to investigate whether, the genetic predisposition, is sufficient to induce changes in the osteoblast differentiation and osteogenic markers in the BMSCs or in the femoral bone. For this we use young SHR rats without hypertension, but, with genetic predisposition in compared with young W.

Main Methods: BMSCs were cultured in a proliferation medium (MEM) or osteogenic medium. Osteogenic differentiation was analyzed by proliferation, total protein, alkaline phosphatase, mineralization, and the mRNA expression of RUNX-2, β-cathenin, osterix, bone morphogenetic protein-2(BMP-2), osteocalcin (OCN), bone sialoprotein (BSP), collagen type I (Col I), and osteopontin (OPN).

Key Findings: Osteoblast differentiation in SHR BMSCs (SHRC) had an increased proliferation compared with W BMSCs (WC). After osteogenic induction, there was greater reduction in proliferation in SHR (SHROM) than in W, in the same condition (WOM). On day 7, although no significant difference in the ALP activity was observed between SHROM and WOM, poor mineralization and osteoblast differentiation was noted in SHROM. The Osterix and β-catenin are involved in the reduced osteoblast differentiation in SHROM. The decreased expression of osteoblast-associated proteins such as OCN, BSP, COL I and OPN revealed poor quality of extracellular matrix (ECM) in SHROM. In the femoral bone, the immunostaining of COL1, BALP, OPN and OCN in SHR was decreased compared with the W. TRAP-positive immunoreactions were observed in major extension in the SHR femur.

Significance: This study is the first to compare osteoblast differentiation in vitro and femoral bone from SHR and W rats. Our results demonstrated that young SHR (4weeks old), without hypertension, but with genetic predisposition, had alterations in osteoblast differentiation of BMSCs and in the femoral bone when compared with their progenitor strain, W.
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http://dx.doi.org/10.1016/j.lfs.2016.01.015DOI Listing
February 2016