Search results for: Collagen

Authors: P. Hashim, M. S. Mohd Ridzwan, J. Bakar

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Collagen was isolated from chicken feet by using papain and pepsin enzymes in acetic acid solution at 4°C for 24h with a yield of 18.16% and 22.94% by dry weight, respectively. Chemical composition and characteristics of chicken feet collagen such as amino acid composition, SDS-PAGE patterns, FTIR spectra and thermal properties were evaluated. The chicken feet collagen is rich in the amino acids glycine, glutamic acid, proline and hydroxyproline. Electrophoresis pattern demonstrated two distinct α-chains (α1 and α2) and β chain, indicating that type I collagen is a major component of chicken feet collagen. The thermal stability of collagen isolated by papain and pepsin revealed stable denaturation temperatures of 48.40 and 53.35°C, respectively. The FTIR spectra of both collagens were similar with amide regions in A, B, I, II and III. The study demonstrated that chicken feet collagen using papain isolation method is possible as commercial alternative ingredient. 

Keywords: Chicken feet, collagen, papain, pepsin.

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Authors: Anamaria Bechir, Rodica Sirbu, Minodora Leca, Maria Maris, Dan Artenie Maris, Emilia Mihaela Cadar, Marius Maris

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This paper aims at presenting the biotechnology used to obtain collagen-based gels from shark (Squalus acanthias) and brill skin, marine fish growing in the Black Sea. Due to the structure of its micro-fibres, collagen can be considered a nanomaterial; in order to use collagen-based matrixes as biomaterial, rheological studies must be performed first, to state whether they are stable or not. For the triple-helix structure to remain stable within these gels at room or human body temperature, they must be stabilized by reticulation.

Keywords: Collagen, biotechnology, reticulation.

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Authors: Yi-Ping Fang, Hsien-Ting Cheng

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Topical photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is an alternative therapy for treating superficial cancer, especially for skin or oral cancer. ALA, a precursor of the photosensitizer protoporphyrin IX (PpIX), is present as zwitterions and hydrophilic property which make the low permeability through the cell membrane. Collagen is a traditional carrier; its molecular composed various amino acids which bear positive charge and negative charge. In order to utilize the ion-pairs with ALA and collagen, the study employed various pH values adjusting the net charge. The aim of this study was to compare a series collagen form, including solution, gel and sponge to investigate the topical delivery behavior of ALA. The in vivo confocal laser scanning microscopy (CLSM) study demonstrated that PpIX generation ability was different pattern after apply for 6 h. Gel type could generate high PpIX, and archived more deep of skin depth.

Keywords: 5-Aminolevulinic acid (ALA), Collagen, Ion-pairs, Penetration behavior

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Authors: Sirbu R., Negreanu-Pirjol T., Leca M., Bechir A., Maris M., Maris D.

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In the paper we presented the possibility of application collagen gels with active principle-s from marine algae extract and chlorhexidine salt in dental medicine. The hydro-alcoholic extracts from marine algae have been used as they have been obtained. The extracts from marine algae and chlorhexidine salt (digluconate) are incorporated in type I non-denatured fibrillar collagen matrixes. In order to obtain therapeutic effects at nanostructure level, it is important to know the rheological characteristics of the relevant mixtures of collagen gels and extracts from marine algae selected for use. In this survey we have studied mixtures made of non-denatured fibrillar collagen hydro-gels where different concentrations of marine algae have been incorporated. Based on the data obtained for the shearing tensions, we have traced the rheograms – the diagrams for shearing tensions depending on the shearing speed values – from which we have calculated the apparent viscosities as ratios between shearing tension and speed values, which have been figured in relation to the shearing speed values, with a view to levelling dependency.

Keywords: rheological properties, fibrillar collagen hydro-gel, marine algae, chlorhexidine salt, dental medicine

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Authors: B. Chouhan, A. Denesyuk, J. Heino, M. S. Johnson, K. Denessiouk

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Integrins are a large family of multidomain α/β cell signaling receptors. Some integrins contain an additional inserted I domain, whose earliest expression appears to be with the chordates, since they are observed in the urochordates Ciona intestinalis (vase tunicate) and Halocynthia roretzi (sea pineapple), but not in integrins of earlier diverging species. The domain-s presence is viewed as a hallmark of integrins of higher metazoans, however in vertebrates, there are clearly three structurally-different classes: integrins without I domains, and two groups of integrins with I domains but separable by the presence or absence of an additional αC helix. For example, the αI domains in collagen-binding integrins from Osteichthyes (bony fish) and all higher vertebrates contain the specific αC helix, whereas the αI domains in non-collagen binding integrins from vertebrates and the αI domains from earlier diverging urochordate integrins, i.e. tunicates, do not. Unfortunately, within the early chordates, there is an evolutionary gap due to extinctions between the tunicates and cartilaginous fish. This, coupled with a knowledge gap due to the lack of complete genomic data from surviving species, means that the origin of collagen-binding αC-containing αI domains remains unknown. Here, we analyzed two available genomes from Callorhinchus milii (ghost shark/elephant shark; Chondrichthyes – cartilaginous fish) and Petromyzon marinus (sea lamprey; Agnathostomata), and several available Expression Sequence Tags from two Chondrichthyes species: Raja erinacea (little skate) and Squalus acanthias (dogfish shark); and Eptatretus burgeri (inshore hagfish; Agnathostomata), which evolutionary reside between the urochordates and osteichthyes. In P. marinus, we observed several fragments coding for the αC-containing αI domain, allowing us to shed more light on the evolution of the collagen-binding integrins.

Keywords: Integrin αI domain, integrin evolution, collagen binding, structure, αC helix

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Authors: Ratchada Sopakayang, Gerhard A. Holzapfel

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In this study, a new constitutive model is developed to describe the hyperelastic behavior of collagenous tissues with a parallel arrangement of collagen fibers such as ligaments and tendons. The model is formulated using a continuum approach incorporating the structural changes of the main tissue components: collagen fibers, proteoglycan-rich matrix and fiber-matrix interaction. The mechanical contribution of the interaction between the fibers and the matrix is simply expressed by a coupling term. The structural change of the collagen fibers is incorporated in the constitutive model to describe the activation of the fibers under tissue straining. Finally, the constitutive model can easily describe the stress-stretch nonlinearity which occurs when a ligament/tendon is axially stretched. This study shows that the interaction between the fibers and the matrix contributes to the mechanical tissue response. Therefore, the model may lead to a better understanding of the physiological mechanisms of ligaments and tendons under axial loading.

Keywords: Hyperelasticity, constitutive model, fiber-matrix interaction, ligament, tendon.

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Authors: Yue Du, Sahan C. B. Herath, Qing-Guo Wang, Harry Asada, Peter C. Y. Chen

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Mechanical interaction between endothelial cells (ECs) and the extracellular matrix (or collagen gel) is known to influence the sprouting response of endothelial cells during angiogenesis. This influence is believed to impact on the capability of endothelial cells to sense soluble chemical cues. Quantitative analysis of endothelial-cell-mediated displacement of the collagen gel provides a means to explore this mechanical interaction. Existing analysis in this context is generally limited to 2D settings. In this paper, we investigate the mechanical interaction between endothelial cells and the extracellular matrix in terms of the endothelial-cellmediated displacement of the collagen gel in both 2D and 3D. Digital image correlation and Digital volume correlation are applied on confocal reflectance image stacks to analyze cell-mediated displacement of the gel. The skeleton of the sprout is extracted from phase contrast images and superimposed on the displacement field to further investigate the link between the development of the sprout and the displacement of the gel.

Keywords: Angiogenesis, digital image correlation, digital volume correlation, interaction between ECs and ECM.

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Authors: P. Vosynek, T. Návrat, M. Peč, J. Pořízka, P. Diviš

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Bone properties and response behavior after static or dynamic activation (loading) are still interesting topics in many fields of the science especially in the biomechanical problems such as bone loss of astronauts in space, osteoporosis, bone remodeling after fracture or remodeling after surgery (endoprosthesis and implants) and in osteointegration. This contribution deals with the relation between physiological, demineralized and deproteinized state of the turkey long bone – tibia. Three methods for comparison were used: 1) densitometry, 2) three point bending and 3) frequency analysis. The main goal of this work was to describe the decrease of the protein (collagen) or mineral of the bone with relation to the fracture in three point bending. The comparison is linked to the problem of different bone mechanical behavior in physiological and osteoporotic state.

Keywords: Bone properties, long bone, osteoporosis, response behavior.

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Authors: Sirilak Namwong , Wonnop Visessanguan, Soottawat Benjakul, Somboon Tanasupawat

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The halophilic proteinase showed a maximal activity at 50°C and pH 9~10, in 20% NaCl and was highly stabilized by NaCl. It was able to hydrolyse natural actomyosin (NAM), collagen and anchovy protein. For NAM hydrolysis, the myosin heavy chain was completely digested by halophilic proteinase as evidenced by the lowest band intensity remaining, but partially hydrolysed actin. The SR5-3 proteinase was also capable hydrolyzing two major components of collagen, β- and α-compounds, effectively. The degree of hydrolysis (DH) of the halophilic proteinase and commercial proteinases (Novozyme, Neutrase, chymotrypsin and Flavourzyme) on the anchovy protein, were compared, and it was found that the proteinase showed a greater degree of hydrolysis towards anchovy protein than that from commercial proteinases. DH of halophilic proteinase was sharply enhanced according to the increase in the concentration of enzyme from 0.035 U to 0.105 U. The results warranting that the acceleration of the production of fish sauce with higher quality, may be achieved by adding of the halophilic proteinase from this bacterium.

Keywords: Halophilic proteinase, Halobacillus sp. SR5-3, anchovy (Spolephorus spp.) proteins, fish sauce

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Authors: Adel Dalilottojari, Bahman Delalat, Frances J. Harding, Michaelia P. Cockshell, Claudine S. Bonder, Nicolas H. Voelcker

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Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.

Keywords: Cardiovascular disease, cell microarray platform, cell therapy, endothelial progenitor cells, high throughput screening.

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Authors: A. A. Vasin, N. V. Klassen, A. M. Likhter

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Water suspensions of in-organic (metals and oxides) and organic nano-objects (chitozan and collagen) were subjected to the treatment of direct and alternative electrical fields. In addition to quasi-periodical spatial patterning resonance-like performance of spatial distributions of these suspensions has been found at low frequencies of alternating electrical field. These resonances are explained as the result of creation of equilibrium states of groups of charged nano-objects with opposite signs of charges at the interparticle distances where the forces of Coulomb attraction are compensated by the repulsion forces induced by relatively negative polarization of hydrated regions surrounding the nanoparticles with respect to pure water. The low frequencies of these resonances are explained by comparatively big distances between the particles and their big masses with t\respect to masses of atoms constituting molecules with high resonance frequencies. These new resonances open a new approach to detailed modeling and understanding of mechanisms of the influence of electrical fields on the functioning of internal organs of living organisms at the level of cells and neurons.

Keywords: Bio-polymers, chitosan, collagen, nanoparticles, coulomb attraction, polarization repulsion, periodical patterning, electrical low frequency resonances, transformations.

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Authors: S. Jaramillo, Y. Montoya, W. Agudelo, J. Bustamante

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Cardiovascular diseases (CVD) are related to affectations of the heart and blood vessels, within these are pathologies such as coronary or peripheral heart disease, caused by the narrowing of the vessel wall (atherosclerosis), which is related to the accumulation of Low-Density Lipoproteins (LDL) in the arterial walls that leads to a progressive reduction of the lumen of the vessel and alterations in blood perfusion. Currently, the main therapeutic strategy for this type of alteration is drug treatment with statins, which inhibit the enzyme 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase), responsible for modulating the rate of cholesterol production and other isoprenoids in the mevalonate pathway. This enzyme induces the expression of LDL receptors in the liver, increasing their number on the surface of liver cells, reducing the plasma concentration of cholesterol. On the other hand, when the blood vessel presents stenosis, a surgical procedure with vascular implants is indicated, which are used to restore circulation in the arterial or venous bed. Among the materials used for the development of vascular implants are Dacron® and Teflon®, which perform the function of re-waterproofing the circulatory circuit, but due to their low biocompatibility, they do not have the ability to promote remodeling and tissue regeneration processes. Based on this, the present research proposes the development of a hydrolyzed collagen and polyethylene oxide electrospun membrane reinforced with medium and high-intensity statins, so that in future research it can favor tissue remodeling processes from its microarchitecture.

Keywords: atherosclerosis, medium and high-intensity statins, microarchitecture, electrospun membrane

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Authors: N. Namviriyachote, N. Bang, P. Aramwit

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Silk sericin (SS) is a glue-like protein from silkworm cocoon. With its outstanding moisturization and activation collagen synthesis properties, silk protein is applied for wound healing. Since wound dressing in film preparation can facilitate patients- convenience and reduce risk of wound contraction, SS and polyvinyl alcohol (PVA) films were prepared with various concentrations of SS. Their physical properties such as surface density, light transmission, protein dissolution and tensile modulus were investigated. The results presented that 3% SS with 2% PVA is the best ingredient for SS film forming.

Keywords: Sericin, silk protein, film, wound healing.

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Authors: Aroonsri Priprem, Sucharat Limsitthichaikoon, Suttasinee Thappasarapong

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Anthocyanins are natural pigments with effective UV protection but their topical use could be limited due to their physicochemical characteristics. An attempt to overcome such limitations by complexation of 2 major anthocyanin-rich sources, C. ternatea and Z. mays, has potentiated its use as topical antiinflammatory. Cell studies indicate no cytotoxicity of the anthocyanin complex (AC) up to 1 mg/ml tested in HaCaT and human fore head fibroblasts by MTT. Croton oil-induced ear edema in Wistar rats suggests an effective dose of 5 mg/cm2 of AC as a topical anti-inflammatory in comparison to 0.5 mg/cm2 of fluocinolone acetonide. Niosomal encapsulation of the AC significantly prolonged the anti-inflammatory activity particularly at 8 h after topical application (p = 0.0001). The AC was not cytotoxic and its anti-inflammatory and activity was dose-dependent and prolonged by niosomal encapsulation. It has also shown to promote collagen type 1 production in cell culture. Thus, AC could be a potential candidate for topical anti-inflammatory agent from natural resources.

Keywords: Anthocyanin complex, ear edema, inflammation, niosomes, skin.

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Authors: J. Franková, V. Pivodová, F. Růžička, J. Ulrichová

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An implant elicits a biological response in the surrounding tissue which determines the acceptance and long-term function of the implant. Dental implants have become one of the main therapy methods in clinic after teeth lose. A successful implant is in contact with bone and soft tissue represent by fibroblasts. In our study we focused on the interaction between six different chemically and physically modified titanium implants (Tis-MALP, Tis-O, Tis- OA, Tis-OPAAE, Tis-OZ, Tis-OPAE) with alveolar fibroblasts as well as with five type of microorganisms (S. epidermis, S.mutans, S. gordonii, S. intermedius, C.albicans). The analysis of microorganism adhesion was determined by CFU (colony forming unite) and biofilm formation. The presence of α3β1 and vinculin expression on alveolar fibroblasts was demonstrated using phospho specific cell based ELISA (PACE). Alveolar fibroblasts have the highest expression of these proteins on Tis-OPAAE and Tis-OPAE. It corresponds with results from bacterial adhesion and biofilm formation and it was related to the lowest production of collagen I by alveolar fibroblasts on Tis-OPAAE titanium disc.

Keywords: titanium disc, alveolar fibroblasts, microorganisms, adhesion

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Authors: M R Asadi, G Torkaman, M Hedayati

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We evaluated the effect of sensory (direct current (DC), 600μA) and motor (monophasic current, pulse duration 300μs, 100 Hz, 2.5-3mA) intensities of cathodal electrical stimulation (ES) current to release VEGF and biomechanical properties of wound. 54 male Sprague-dawley rats were randomly assigned into one control and two experimental groups. A full thickness skin incision was made on animals- dorsal region. The experimental groups received ES for 1h/day and every other day. VEGF expression was measured in skin on the 7th day after surgical incision and tensile strength was measured on 21st day. On the 7th day, the values of skin VEGF in the sensory group were significantly greater than those of the other groups (p < 0.05). Sensory and Motor intensity stimulation, can not improve the biomechanical properties of the repaired wounds. It seems the mechanical environment induced by sensory and motor intensity of electrical stimulation, could not simulate the role of normal daily stress and strain to maturation of collagen fibers and their cross links. Further work is needed to determine the relationship between VEGF expression after ES and its effect on tensile strength of healed wound.

Keywords: Biomechanical properties Direct current, Monophasic current, Skin, VEGF

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Authors: Addion Nizori, Lan T.T. Bui, Darryl M. Small

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Ascorbic acid (AA), commonly known as vitamin C, is essential for normal functioning of the body and maintenance of metabolic integrity. Among its various roles are as an antioxidant, a cofactor in collagen formation and other reactions, as well as reducing physical stress and maintenance of the immune system. Recent collaborative research between the Australian Defence Science and Technology Organisation (DSTO) in Scottsdale, Tasmania and RMIT University has sought to overcome the problems arising from the inherent instability of ascorbic acid during processing and storage of foods. The recent work has demonstrated the potential of microencapsulation by spray drying as a means to enhance retention. The purpose of this current study has been focused upon the influence of spray drying conditions on the properties of encapsulated ascorbic acid. The process was carried out according to a central composite design. Independent variables were: inlet temperature (80-120° C) and feed flow rate (7-14 mL/minute). Process yield, ascorbic acid loss, moisture content, water activity and particle size distribution were analysed as responses. The results have demonstrated the potential of microencapsulation by spray drying as a means to enhance retention. Vitamin retention, moisture content, water activity and process yield were influenced positively by inlet air temperature and negatively by feed flow rate.

Keywords: Microencapsulation, spray drying, ascorbic acid.

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Authors: Jelena S. Kiurski, Vesna S. Kecić, Snežana M. Aksentijević

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The identification and evaluation of organic and inorganic pollutants were performed in a flexographic facility in Novi Sad, Serbia. Air samples were collected and analyzed in situ, during 4-hours working time at five sampling points by the mobile gas chromatograph and ozonometer at the printing of collagen casing. Experimental results showed that the concentrations of isopropyl alcohol, acetone, total volatile organic compounds and ozone varied during the sampling times. The highest average concentrations of 94.80 ppm and 102.57 ppm were achieved at 200 minutes from starting the production for isopropyl alcohol and total volatile organic compounds, respectively. The mutual dependences between target hazardous and microclimate parameters were confirmed using a multiple linear regression model with software package STATISTICA 10. Obtained multiple coefficients of determination in the case of ozone and acetone (0.507 and 0.589) with microclimate parameters indicated a moderate correlation between the observed variables. However, a strong positive correlation was obtained for isopropyl alcohol and total volatile organic compounds (0.760 and 0.852) with microclimate parameters. Higher values of parameter F than F_critical for all examined dependences indicated the existence of statistically significant difference between the concentration levels of target pollutants and microclimates parameters. Given that, the microclimate parameters significantly affect the emission of investigated gases and the application of eco-friendly materials in production process present a necessity.

Keywords: Flexographic printing, indoor air, multiple regression analysis, pollution emission.

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Authors: P. C. Boisteanu, M. G. Usturoi, Roxana Lazar, B. V. Avarvarei

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The paper is included within the framework of a complex research program, which was initiated from the hypothesis arguing on the existence of a correlation between pineal indolic and peptide hormones and the somatic development rhythm, including thus the epithalamium-epiphysis complex involvement. At birds, pineal gland contains a circadian oscillator, playing a main role in the temporal organization of the cerebral functions. The secretion of pineal indolic hormones is characterized by a high endogenous rhythmic alternation, modulated by the light/darkness (L/D) succession and by temperature as well. The research has been carried out using 100 chicken broilers - “Ross" commercial hybrid, randomly allocated in two experimental batches: Lc batch, reared under a 12L/12D lighting schedule and Lexp batch, which was photic pinealectomised through continuous exposition to light (150 lux, 24 hours, 56 days). Chemical and physical features of the meat issued from breast fillet and thighs muscles have been studied, determining the dry matter, proteins, fat, collagen, salt content and pH value, as well. Besides the variations of meat chemical composition in relation with lighting schedule, other parameters have been studied: live weight dynamics, feed intake and somatic development degree. The achieved results became significant since chickens have 7 days of age, some variations of the studied parameters being registered, revealing that the pineal gland physiologic activity, in relation with the lighting schedule, could be interpreted through the monitoring of the somatic development technological parameters, usually studied within the chicken broilers rearing aviculture practice.

Keywords: lighting schedule, physic-chemical characteristics ofmeat, pineal gland at birds.

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Authors: M. Jarrar, S. Behl, N. Shaheen, A. Fatima, R. Nasab

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Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is central on aging of skin especially by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photodamage is exceedingly valued. Retinoids and alpha hydroxy acids have been endorsed by some researchers as possible candidates for protecting and or repairing the effect of UV damaged skin. For consolidating a better system of anti- and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblast’s elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30 -35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in maintaining elastin concentration in UV exposed cells. We assume this preservation could be the result of increased tropo-elastin gene expression stimulated by retinol whereas lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: Alpha Hydroxy Acid, Elastin, Retinol, Ultraviolet radiations.

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Authors: M. Qwamizadeh, K. Zhou, Z. Zhang, YW. Zhang

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Typical load-bearing biological materials like bone, mineralized tendon and shell, are biocomposites made from both organic (collagen) and inorganic (biomineral) materials. This amazing class of materials with intrinsic internally designed hierarchical structures show superior mechanical properties with regard to their weak components from which they are formed. Extensive investigations concentrating on static loading conditions have been done to study the biological materials failure. However, most of the damage and failure mechanisms in load-bearing biological materials will occur whenever their structures are exposed to dynamic loading conditions. The main question needed to be answered here is: What is the relation between the layout and architecture of the load-bearing biological materials and their dynamic behavior? In this work, a staggered model has been developed based on the structure of natural materials at nanoscale and Finite Element Analysis (FEA) has been used to study the dynamic behavior of the structure of load-bearing biological materials to answer why the staggered arrangement has been selected by nature to make the nanocomposite structure of most of the biological materials. The results showed that the staggered structures will efficiently attenuate the stress wave rather than the layered structure. Furthermore, such staggered architecture is effectively in charge of utilizing the capacity of the biostructure to resist both normal and shear loads. In this work, the geometrical parameters of the model like the thickness and aspect ratio of the mineral inclusions selected from the typical range of the experimentally observed feature sizes and layout dimensions of the biological materials such as bone and mineralized tendon. Furthermore, the numerical results validated with existing theoretical solutions. Findings of the present work emphasize on the significant effects of dynamic behavior on the natural evolution of load-bearing biological materials and can help scientists to design bioinspired materials in the laboratories.

Keywords: Load-bearing biological materials, nanostructure, staggered structure, stress wave decay.

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Authors: Manish Kumar Gautam, Raj Kumar Goel

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Delayed wound healing in diabetes is primarily associated with hyperglycemia, over-expression of inflammatory marker, oxidative stress and delayed collagen synthesis. This unmanaged wound is producing high economic burden on the society. Thus research is required to develop new and effective treatment strategies to deal with this emerging issue. Our present study incorporates the evaluation of wound healing effects of 50% ethanol extract of Ocimum sanctum (OSE) in streptozotocin (45mg/kg)-induced diabetic rats with concurrent wound ulcer. The animals showing diabetes (Blood glucose level >140 and <250 mg/dL) will be selected for wound healing study using standard dead space wound model. Wounds were created by implanting two polypropylene tubes (0.5 x 2.5 cm2 each), one on either side in the lumbar region on the dorsal surface of each rat. On the 10th postwounding day, the animals were sacrificed and granulation tissue formed on the implanted tubes was carefully dissected out and study the status of antioxidants (Superoxide dismutase, SOD and Glutathione, GSH) free radicals (Lipid peroxidation, LPO and nitric oxide, NO) acute inflammatory marker (myeloperoxidase, MPO) connective tissue determinants, hydroxyproline, hexosamine and hexuronic acid, which play a major role in wound healing and diabetes. Besides the anti-diabetic parameters (estimation of serum blood glucose, triglycerides and total cholesterol), the above parameters for wound healing were studied both in normal, untreated and OSE treated diabetic rats. The effects of extract on above parameters will be compared with known standard antioxidant (Vitamin E) and anti-diabetic (Glybenclamide) drugs. OSE 400 mg/kg substantiated by significantly decreased serum blood glucose, triglycerides and total cholesterol. OSE also decrease granulation tissue free radicals (LPO, 58.1% and NO, 52.7%) and myeloperoxidase (MPO, 63.3%), and enhanced antioxidants (GSH, 116.4% and SOD, 201.1%)

Keywords: Wound healing, diabetes, Ocimum sanctum, Antioxidant, Free radical, Myeloperoxidase

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Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: Mesenchymal stem cells, Cryopreservation, Stemness, Senescence.

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Authors: Amira E. Derbalah, Doaa M. Zaghloul

Abstract:

10 clinically healthy hemal nodes were collected from male bulls aged 2-3 years. Light microscopy revealed a capsule of connective tissue consisted mainly of collagen fiber surrounding hemal node, numerous erythrocytes were found in wide subcapsular sinus under the capsule. The parenchyma of the hemal node was divided into cortex and medulla. Diffused lymphocytes, and lymphoid follicles, having germinal centers were the main components of the cortex, while in the medulla there was wide medullary sinus, diffused lymphocytes and few lymphoid nodules. The area occupied with lymph nodules was larger than that occupied with non-nodular structure of lymphoid cords and blood sinusoids. Electron microscopy revealed the cellular components of hemal node including elements of circulating erythrocytes intermingled with lymphocytes, plasma cells, mast cells, reticular cells, macrophages, megakaryocytes and endothelial cells lining the blood sinuses. The lymphocytes were somewhat triangular in shape with cytoplasmic processes extending between adjacent erythrocytes. Nuclei were triangular to oval in shape, lightly stained with clear nuclear membrane indentation and clear nucleoli. The reticular cells were elongated in shape with cytoplasmic processes extending between adjacent lymphocytes, rough endoplasmic reticulum, ribosomes and few lysosomes were seen in their cytoplasm. Nucleus was elongated in shape with less condensed chromatin. Plasma cells were oval to irregular in shape with numerous dilated rough endoplasmic reticulum containing electron lucent material occupying the whole cytoplasm and few mitochondria were found. Nuclei were centrally located and oval in shape with heterochromatin emarginated and often clumped near the nuclear membrane. Occasionally megakaryocytes and mast cells were seen among lymphocytes. Megakaryocytes had multilobulated nucleus and free ribosomes often appearing as small aggregates in their cytoplasm, while mast cell had their characteristic electron dense granule in the cytoplasm, few electron lucent granules were found also, we conclude that, the main function of the hemal node of cattle is proliferation of lymphocytes. No role for plasma cell in erythrophagocytosis could be suggested.

Keywords: Cattle, Electron microscopy, Hemal node, Histology, Immune system.

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Authors: Carmiña L. Vargas-Zapata, María A. Conde-Sarmiento, Maria Consuelo Maestre-Vargas

Abstract:

Calcium is an essential element for good growth and development of the organism, and its requirement is increased at school age. Low socio-economic populations of developing countries such as Colombia may have food deficiency of this mineral in schoolchildren that could be reflected in calcium biochemical indicators, bone alterations and anthropometric indicators. The objective of this investigation was to evaluate some calcium biochemical indicators in a group of schoolchildren of low socioeconomic level from Barranquilla city and to correlate with body mass index. 60 schoolchildren aged 7 to 15 years were selected from Jesus’s Heart Educational Institution in Barranquilla-Atlántico, apparently healthy, without suffering from infectious or gastrointestinal diseases, without habits of drinking alcohol or smoking another hallucinogenic substance and without taking supplementation with calcium in the last six months or another substance that compromises bone metabolism. The research was approved by the ethics committee at Universidad del Atlántico. The selected children were invited to donate a blood and urine sample in a fasting time of 12 hours, the serum was separated by centrifugation and frozen at ˗20 ℃ until analyzed and the same was done with the urine sample. On the day of the biological collections, the weight and height of the students were measured to determine the nutritional status by BMI using the WHO tables. Calcium concentrations in serum and urine (SCa, UCa), alkaline phosphatase activity total and of bone origin (SAPT, SBAP) and urinary creatinine (UCr) were determined by spectrophotometric methods using commercial kits. Osteocalcin and Cross-linked N-telopeptides of type I collagen (NTx-1) in serum were measured with an enzyme-linked inmunosorbent assay. For statistical analysis the Statgraphics software Centurium XVII was used. 63% (n = 38) and 37% (n = 22) of the participants were male and female, respectively. 78% (n = 47), 5% (n = 3) and 17% (n = 10) had a normal, malnutrition and high nutritional status, respectively. The averages of evaluated indicators levels were (mean ± SD): 9.50 ± 1.06 mg/dL for SCa; 181.3 ± 64.3 U/L for SAPT, 143.8 ± 73.9 U/L for SBAP; 9.0 ± 3.48 ng/mL for osteocalcin and 101.3 ± 12.8 ng/mL for NTx-1. UCa level was 12.8 ± 7.7 mg/dL that adjusted with creatinine ranged from 0.005 to 0.395 mg/mg. Considering serum calcium values, approximately 7% of school children were hypocalcemic, 16% hypercalcemic and 77% normocalcemic. The indicators evaluated did not correlate with the BMI. Low values ​​were observed in calcium urinary excretion and high in NTx-1, suggesting that mechanisms such as increase in renal retention of calcium and in bone remodeling may be contributing to calcium homeostasis.

Keywords: Calcium, calcium biochemical, indicators, school children, low socioeconomic status.

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