The biocompatible membranes are widely applied in the medical field in order to stimulate tissue repair. The biological principle of this type of treatment is the repair and guided regeneration. In the literature, there are few reports of studies evaluating the effects and biological properties of norbixin in animal tissues. Thus, the present study was to evaluate the effect of polystyrene membrane with collagen and norbixin, through the micronucleus test and comet assay in rats, as part of the recommended test battery to evaluate the mutagenic potential. The research project was approved by CEP / FACID Protocol 069/2014. For this study, 15 rats were divided into 3 groups were used: A - the membrane was introduced into the peritoneum of the animals through a laparotomy; B - received cyclophosphamide at a dose of 50mg / kg intraperitoneally; C - were performed only one laparotomy. A peripheral blood sample was collected from the animals for conducting Comet assay and 72 hours after the start of the experiment were euthanized. It was collected bone marrow material of each rat to perform the micronucleus test. In conclusion, through the tests, the membrane is not genotoxic.
Protein was extracted from muscles of Channa striatus and attempts were
made to evaluate in vitro antibacterial activity against clinical bacterial isolates. The
higher concentration of protein (100μg/ml) extracts exhibited a pronounced activity
against Pseudomonas aeruginosa (21 mm), Proteus vulgaris (19 mm), Citrobacter sp
(19 mm), Klebsiella pneumoniae (18 mm), Micrococcus sp (17 mm), Bacillus subtilis (16
mm), Staphylococcus aureus (15 mm), E. coli (14 mm) and Serratia marcescens (5
mm). The minimum inhibitory concentration and minimum bactericidal concentration
were found to be 20-40 μg/ml and 80-100 μg/ml respectively for the extracts of
Channa striatus protein against test organisms. This study confirms that C. striatus fish
protein extracts possess antibacterial activity against a wide range of microbes and
justified that it could be used in the traditional medicine as a remedy for the
treatment of bacterial diseases.
Objective: To evaluate the antibacterial effects of 4 different cavity disinfectants on Streptococcus mutans, Lactobacillus acidophilus, and Enterococcus faecalis bacteria in different time periods.
Study Design: The antibacterial effects of Cavity Cleanser, Tubulicid Red Label, Chloraxid 2%, and Oxygenated Water cavity disinfectant solutions on E. faecalis (ATCC 29212), S. mutans (ATCC 25175), and L. acidophilus (RSKK 03037) bacterial strains were evaluated by disk diffusion method. In the study where vancomycin antibiogram disc constituted the positive control group, physiological saline solution was used as the negative control group. Standard, sterile, blank antibiogram discs of 5 mm in diameter, in which 15 μL of each material were added, were placed on agar plates at 2.5–3 cm intervals. The inhibition zone diameters formed around the discs that were left to incubate for 24–48 hours at 37°C were measured in millimeters. Statistical analysis of the data was performed using one-way analysis of variance, Kolmogorov-Smirnov, Levene, and Bonferroni tests.
Results: At the end of the study the solutions tested showed a statistically significant antibacterial effect on all bacterial strains used (p<0.05). Cavity Cleanser disinfectant containing 2% chlorhexidine showed the highest antibacterial effect on S. mutans and L. acidophilus, and benzalkonium-containing Tubulicid Red disinfectant on E. faecalis.
Conclusion: The antibacterial effect of all cavity disinfectants used in the study was found to be higher at the end of the 48th hour than at the end of the 24th hour, but there was no statistically significant difference (p>0.05).
Keywords: antibacterial agents; antibacterial effect; cavity disinfectants; chlorhexidine; contamination; dental caries; disinfection; disc diffusion; gram-negative bacteria; gram-positive bacteria
Objective: To identify interstitial cells of Cajal (ICC) in the common bile duct of Kunming mice.
Study Design: Common bile ducts obtained from the Kunming mice were prepared for immunohistochemical investigations using the c-kit antibody. Immunoelectron microscopy was used to detect the expression of c-kit in the ICC of the common bile duct. Transmission electron microscopy showed ultrastructure of ICC in the murine bile duct. Reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to confirm the expression of mRNA specific for the c-kit gene and production of c-kit protein in the Kunming mice common bile duct.
Results: Immunohistochemistry revealed that ICC in the murine common bile duct are c-kit positive and the ICC are located in the tela submucosa and the tunica muscularis of the murine common bile duct and do not connect with each other. Immunoelectron microscopy confirmed the expression of Kit by ICC in the murine common bile duct. Transmission electron microscopy showed that ICC in the murine common bile duct have long processes, abundant mitochondria, plenty of smooth endoplasmic reticulum (sER), a lot of lysosomes, and dense bodies. The caveolae of ICC are distinctive. At the same time, RT-PCR indicated that the Kunming mice common bile duct expressed mRNA specific for the c-kit gene, and western blot analysis showed the evidence of production of c-kit protein in the Kunming mice common bile duct.
Conclusion: ICC are found in the Kunming mice common bile duct, which is likely to lead to the development of motility study of the common bile duct.
Keywords: common bile duct; electron microscopy; immuno-electron microscopy; interstitial cells of Cajal; intestines; smooth muscle; tyrosine kinase receptor (c-kit)
Objective: To evaluate the results of the effect of nebivolol on tibial bone defect and graft application in new bone development in the rat.
Study Design: Thirty Wistar albino rats were divided into 3 groups. In the Control group, tibia bone defect was created without any treatment. In the Defect+ Graft group, allograft treatment was performed by forming a 6 mm tibial bone defect. In the Defect+Graft+ Nebivolol group, alloplastic bone graft was placed in the calvarial bone defect and then nebivolol (0.34 mg/mL solution/day) treatment was intraperitoneally applied for 28 days.
Results: Histopathological examination revealed inflammation in the defect area, congestion in the vessels, degeneration in collagen fibers, and an increase in osteoclast cells. There was an increase in inflammation and blood vessel structure in graft application, and osteoblastic activity matrix formation after reorganization nebivolol application in collagen fibers. Osteonectin expression was positive in the collagen fiber and matrix, starting in the Graft group, in osteoblasts, whereas in the Nebivolol group, osteoblasts increased in osteocytes and new bone formation.
Conclusion: Nebivolol is thought to have a positive effect on osteoinductive bone growth factors and contribute to the cell-matrix interaction, in addition to the supporting effect of the graft with its antioxidative effect.
Keywords: allograft; bone; bone regeneration; disease models, animal; nebivolol; orthopedic procedures; osteonectin; rats; tibia; tibial defect
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
Protein was extracted from muscles of Channa striatus and attempts were
made to evaluate in vitro antibacterial activity against clinical bacterial isolates. The
higher concentration of protein (100μg/ml) extracts exhibited a pronounced activity
against Pseudomonas aeruginosa (21 mm), Proteus vulgaris (19 mm), Citrobacter sp
(19 mm), Klebsiella pneumoniae (18 mm), Micrococcus sp (17 mm), Bacillus subtilis (16
mm), Staphylococcus aureus (15 mm), E. coli (14 mm) and Serratia marcescens (5
mm). The minimum inhibitory concentration and minimum bactericidal concentration
were found to be 20-40 μg/ml and 80-100 μg/ml respectively for the extracts of
Channa striatus protein against test organisms. This study confirms that C. striatus fish
protein extracts possess antibacterial activity against a wide range of microbes and
justified that it could be used in the traditional medicine as a remedy for the
treatment of bacterial diseases.
Objective: To evaluate the antibacterial effects of 4 different cavity disinfectants on Streptococcus mutans, Lactobacillus acidophilus, and Enterococcus faecalis bacteria in different time periods.
Study Design: The antibacterial effects of Cavity Cleanser, Tubulicid Red Label, Chloraxid 2%, and Oxygenated Water cavity disinfectant solutions on E. faecalis (ATCC 29212), S. mutans (ATCC 25175), and L. acidophilus (RSKK 03037) bacterial strains were evaluated by disk diffusion method. In the study where vancomycin antibiogram disc constituted the positive control group, physiological saline solution was used as the negative control group. Standard, sterile, blank antibiogram discs of 5 mm in diameter, in which 15 μL of each material were added, were placed on agar plates at 2.5–3 cm intervals. The inhibition zone diameters formed around the discs that were left to incubate for 24–48 hours at 37°C were measured in millimeters. Statistical analysis of the data was performed using one-way analysis of variance, Kolmogorov-Smirnov, Levene, and Bonferroni tests.
Results: At the end of the study the solutions tested showed a statistically significant antibacterial effect on all bacterial strains used (p<0.05). Cavity Cleanser disinfectant containing 2% chlorhexidine showed the highest antibacterial effect on S. mutans and L. acidophilus, and benzalkonium-containing Tubulicid Red disinfectant on E. faecalis.
Conclusion: The antibacterial effect of all cavity disinfectants used in the study was found to be higher at the end of the 48th hour than at the end of the 24th hour, but there was no statistically significant difference (p>0.05).
Keywords: antibacterial agents; antibacterial effect; cavity disinfectants; chlorhexidine; contamination; dental caries; disinfection; disc diffusion; gram-negative bacteria; gram-positive bacteria
Objective: To identify interstitial cells of Cajal (ICC) in the common bile duct of Kunming mice.
Study Design: Common bile ducts obtained from the Kunming mice were prepared for immunohistochemical investigations using the c-kit antibody. Immunoelectron microscopy was used to detect the expression of c-kit in the ICC of the common bile duct. Transmission electron microscopy showed ultrastructure of ICC in the murine bile duct. Reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to confirm the expression of mRNA specific for the c-kit gene and production of c-kit protein in the Kunming mice common bile duct.
Results: Immunohistochemistry revealed that ICC in the murine common bile duct are c-kit positive and the ICC are located in the tela submucosa and the tunica muscularis of the murine common bile duct and do not connect with each other. Immunoelectron microscopy confirmed the expression of Kit by ICC in the murine common bile duct. Transmission electron microscopy showed that ICC in the murine common bile duct have long processes, abundant mitochondria, plenty of smooth endoplasmic reticulum (sER), a lot of lysosomes, and dense bodies. The caveolae of ICC are distinctive. At the same time, RT-PCR indicated that the Kunming mice common bile duct expressed mRNA specific for the c-kit gene, and western blot analysis showed the evidence of production of c-kit protein in the Kunming mice common bile duct.
Conclusion: ICC are found in the Kunming mice common bile duct, which is likely to lead to the development of motility study of the common bile duct.
Keywords: common bile duct; electron microscopy; immuno-electron microscopy; interstitial cells of Cajal; intestines; smooth muscle; tyrosine kinase receptor (c-kit)
Objective: To evaluate the results of the effect of nebivolol on tibial bone defect and graft application in new bone development in the rat.
Study Design: Thirty Wistar albino rats were divided into 3 groups. In the Control group, tibia bone defect was created without any treatment. In the Defect+ Graft group, allograft treatment was performed by forming a 6 mm tibial bone defect. In the Defect+Graft+ Nebivolol group, alloplastic bone graft was placed in the calvarial bone defect and then nebivolol (0.34 mg/mL solution/day) treatment was intraperitoneally applied for 28 days.
Results: Histopathological examination revealed inflammation in the defect area, congestion in the vessels, degeneration in collagen fibers, and an increase in osteoclast cells. There was an increase in inflammation and blood vessel structure in graft application, and osteoblastic activity matrix formation after reorganization nebivolol application in collagen fibers. Osteonectin expression was positive in the collagen fiber and matrix, starting in the Graft group, in osteoblasts, whereas in the Nebivolol group, osteoblasts increased in osteocytes and new bone formation.
Conclusion: Nebivolol is thought to have a positive effect on osteoinductive bone growth factors and contribute to the cell-matrix interaction, in addition to the supporting effect of the graft with its antioxidative effect.
Keywords: allograft; bone; bone regeneration; disease models, animal; nebivolol; orthopedic procedures; osteonectin; rats; tibia; tibial defect
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Biomaterials for tissue engineering slideshareBukar Abdullahi
An overview of Tissue Engineering with some basics in Biomaterials and Synthetic Polymers. Further references should be considered as I presented this a specific target audience.
Objective: To investigate the protective effect of lo- sartan, an angiotensin II type 1 receptor blocker with antioxidative effect on intestinal ischemia-reperfusion (I/R) injury in rats, against inflammation and apoptotic development.
Study Design: Forty male Wistar albino rats with a mean weight of 200–250 g each were divided into 4 groups: (1) Sham operation (laparotomy only, sham surgical preparation including isolation of the superior mesenteric artery [SMA] without occlusion), (2) Ischemia model with SMA closure for 2 hours, (3) I/R group (2 hours of ischemia followed by 3-hour reperfusion (SMA occlusion for 120 minutes followed by 240 minutes reperfusion), and (4) Losartan group (2 hours of ischemia, 40 mg/kg losartan was administered to the animals; losartan was dissolved in 1 mL distilled water and administered intraperitoneally after 2 hours of ischemia). Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels were examined in jejunum tissue.
Results: Losartan treatment reduced the I/R-induced increase in MDA levels in the gut. Statistically, while SOD, CAT, and GSH activities decreased significantly in the I/R group, they increased in the I/R+Losartan group. Villus loss and increase in inflammation after ischemia persisted after reperfusion. Losartan treatment played a role in the reduction of inflammation and apoptosis and in the regulation of TNF-α and caspase-9 activity.
Conclusion: It has been thought that losartan in I/R may reduce mucosal damage and cell apoptosis in the direction of inflammation and may stabilize caspase-9 activity by inhibiting TNF-α stimulus.
Keywords: caspase-9, ischemia, ischemia/reperfusion, rat, reperfusion injury, TNF-α, tumor necrosis factor-alpha
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
TISSUE DEVELOPMENT WITH TISSUE ENGINEERING APPROACHFelix Obi
Tissue Engineering is the development and practice of combining scaffolds, cells, and suitable biochemical factors (regulatory factors or Signals) into functional tissues. The goal of tissue engineering is to assemble functional constructs that restore, maintain, or improve damaged tissues or whole organs.
Cells are the building blocks of tissue, and tissues are the basic unit of function in the body. Generally, groups of cells make and secrete their own support structures, called extracellular matrix. This matrix, or scaffold, does more than just support the cells; it also acts as a relay station for various signaling molecules. Thus, cells receive messages from many sources that become available from the local environment. Each signal can start a chain of responses that determine what happens to the cell. By understanding how individual cells respond to signals, interact with their environment, and organize into tissues and organisms, Tissue Engineers are now able to manipulate these processes to amend damaged tissues or even create new ones.
A presentation on Tissue Engineering made by Deepak Rajput. It was presented as a seminar requirement at the University of Tennessee Space Institute in Spring 2009.
Before hiring any baby care servant, ensure they have clean background, VVScore provide services like baby sitter background check, Address check, Criminal Check etc.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Biomaterials for tissue engineering slideshareBukar Abdullahi
An overview of Tissue Engineering with some basics in Biomaterials and Synthetic Polymers. Further references should be considered as I presented this a specific target audience.
Objective: To investigate the protective effect of lo- sartan, an angiotensin II type 1 receptor blocker with antioxidative effect on intestinal ischemia-reperfusion (I/R) injury in rats, against inflammation and apoptotic development.
Study Design: Forty male Wistar albino rats with a mean weight of 200–250 g each were divided into 4 groups: (1) Sham operation (laparotomy only, sham surgical preparation including isolation of the superior mesenteric artery [SMA] without occlusion), (2) Ischemia model with SMA closure for 2 hours, (3) I/R group (2 hours of ischemia followed by 3-hour reperfusion (SMA occlusion for 120 minutes followed by 240 minutes reperfusion), and (4) Losartan group (2 hours of ischemia, 40 mg/kg losartan was administered to the animals; losartan was dissolved in 1 mL distilled water and administered intraperitoneally after 2 hours of ischemia). Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels were examined in jejunum tissue.
Results: Losartan treatment reduced the I/R-induced increase in MDA levels in the gut. Statistically, while SOD, CAT, and GSH activities decreased significantly in the I/R group, they increased in the I/R+Losartan group. Villus loss and increase in inflammation after ischemia persisted after reperfusion. Losartan treatment played a role in the reduction of inflammation and apoptosis and in the regulation of TNF-α and caspase-9 activity.
Conclusion: It has been thought that losartan in I/R may reduce mucosal damage and cell apoptosis in the direction of inflammation and may stabilize caspase-9 activity by inhibiting TNF-α stimulus.
Keywords: caspase-9, ischemia, ischemia/reperfusion, rat, reperfusion injury, TNF-α, tumor necrosis factor-alpha
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
TISSUE DEVELOPMENT WITH TISSUE ENGINEERING APPROACHFelix Obi
Tissue Engineering is the development and practice of combining scaffolds, cells, and suitable biochemical factors (regulatory factors or Signals) into functional tissues. The goal of tissue engineering is to assemble functional constructs that restore, maintain, or improve damaged tissues or whole organs.
Cells are the building blocks of tissue, and tissues are the basic unit of function in the body. Generally, groups of cells make and secrete their own support structures, called extracellular matrix. This matrix, or scaffold, does more than just support the cells; it also acts as a relay station for various signaling molecules. Thus, cells receive messages from many sources that become available from the local environment. Each signal can start a chain of responses that determine what happens to the cell. By understanding how individual cells respond to signals, interact with their environment, and organize into tissues and organisms, Tissue Engineers are now able to manipulate these processes to amend damaged tissues or even create new ones.
A presentation on Tissue Engineering made by Deepak Rajput. It was presented as a seminar requirement at the University of Tennessee Space Institute in Spring 2009.
Before hiring any baby care servant, ensure they have clean background, VVScore provide services like baby sitter background check, Address check, Criminal Check etc.
Existance Theory for First Order Nonlinear Random Dfferential Equartioninventionjournals
In this paper, the existence of a solution of nonlinear random differential equation of first order is proved under Caratheodory condition by using suitable fixed point theorem. 2000 Mathematics Subject Classification: 34F05, 47H10, 47H4
Tuition Reduction Determination Using Fuzzy Tsukamotoinventionjournals
The process of determining cuts tuition for students are usually given with the same nominal. And in this paper is the determination of the pieces tuition for students who are less able to be different, depending on how much income parents and the number of children covered. For income parents who get discounted tuition fee of IDR Rp.1,500,000 and for the number of children in these families also determine the number of pieces obtained. Tsukamoto Fuzzy system is the model used in this paper. Each input variable is divided into three membership functions. In this paper, Nine Tsukamoto Fuzzy model rules have been applied. The system also provides a consequent change of parameters if the current parameter values to be changed. The smaller the parent's income, the greater the pieces obtained. The more children insured the greater the college acquired pieces.
The Role of Religious Sensibilities on The Relationship Between Religious Rul...inventionjournals
No matter what the religion is the impact of the religious beliefs cannot be neglected on consumer behavior. In Islamic countries or wherever Islamic population has a large market, the Muslim consumer is ultra-sensitive on their beliefs. With a global point of view “Halal food” looks like the main concern of Muslim consumers but there are several other rules that especially global companies should strictly follow in Muslim target markets. Islamic rules which can be applied in marketing vary from bribery to hoarding. Countries governing under Islamic Shari'ah apply these rules as law articles which mandate people to follow. In other countries with major Islamic populations like Turkey which adopts modern law systems do not directly follow Islamic rules in business and marketing. But in these countries also Islamic rules has an impact on consumer behavior. Ethical systems and religious lifestyle in these countries reveals Islamic sensitivities other than law. In this study it is examined the impact of Muslim consumers’ religious rules on hedonic consumption and the mediating role of religioussensibilities on this relationship. A qualitative approach has been followed in the research and focus group interview method has been conducted.
Examining the Usability of Message Reading Features on Smartwatchesinventionjournals
This study examined smart watch message reading features by considering the following assessment points: simplicity, comprehensibility, perceived usefulness, and overall satisfaction. Three smart watches (Pebble Watch, Samsung Galaxy Gear2, and Sony Smartwatch2) were subjected to user tests, and indepth user interviews were conducted. The results showed that 83.3% of participants experienced difficulty in checking and reading messages. Screen size, text size, and accessing messages were the top three factors causing inconvenience. The participants believed that the most useful feature of the smart watch was message notification, and message reading was also considered useful. The message notification function could be further developed for smartwatch design.
An Analogical Study of the Narrative Techniques Used In the Film Paradesi (20...inventionjournals
Narrations in films have always taken the primary place. Understanding the narrative culture of a medium becomes the prime motive of the narrator while working on adaptations. There is a strong connection between the verbal and visual representation is sequence, since literary and filmic signs are apprehended consecutively through time. The success and acceptability of a film is based on the narration style and techniques. This is very much true in Tamil cinema narrative patterns. While trying to adapt the story from a medium the director of the film Bala, has focused on trying to maintain the narrative syntax of the original, but has also closely followed the semantics of visual design. The Tamil film Paradesi is an adaptation of the Tamil translation Yerium Panikkadu of the novel ‘Red Tea’, which was has been an inspiration for the director.
In Causality Principle as the Framework to Contextualize Time in Modern Physicsinventionjournals
Since the moment Boethius meditated on the nature of time in his fifth book on The Consolation of Philosophy, we have more tools to reflect on the subject. The onset of relativity and quantum physics provides us with the best insight, to date, that guides our reflections on the philosophical debates that attempt to theorize a definition of time. To clearly address the problems related to the theoretical models that account for the nature of time, adjustments to our interpretation of the contextual issues involved in special relativity are in order if we are going to preserve our notion of causal reality. As the construction of string theory emerges as the reigning theory for quantum gravity, a precise picture of causal reality can be accounted for through theories such as Dyson’s Chronological Protection Agency, Hořava’s theory of gravity, and new insight to how simultaneity is interpreted in relativity theory. With this model, the question about time in the philosophical debates can now be clearly defined through the Presentists’ view of the universe. Thus, if we are going to accept the premise of quantum mechanics (QM) and the theory of relativity, we can safely say that string theory (ST) is a reasonable theory of quantum gravity and that its conclusions about time must be taken seriously.
“Desquamative Gingivitis Treated By An Antioxidant Therapy- A Case Report”inventionjournals
Desquamative gingivitis is described as an erythematous, desquamated or eroded gingival lesion. Various etiologic factors are present for the appearance of such lesions. Despite of considering etiology, treatment is oftenly provided by systemic or topical corticosteroids. Apart from steroid application, another optionable treatment is antioxidant therapy which provides rapid healing of the tissue. As antioxidants posses various advantageous properties, it can be considered as a first treatment option for desquamative gingivitis. The presented case report of desquamative gingivitis is successfully treated using systemic antioxidants in the form of commercially available „oxitard capsule‟.
Examination of Total Precipitable Water using MODIS measurements and Comparis...inventionjournals
In this research, precipitable water vapor, as the most effective character in the production of biomass is estimated using remote sensing techniques. Total Precipitable Water (TPW) was estimated using measurements in the Near Infrared bands of the MODIS. To examine the level of confidence in TPW deriving, a simultaneous in situ measurement by Radiosonde and ground-based Global Positioning System (GPS) was carried out. The TPW as results in Radiosonde and GPS was accomplished using the relevant physical equations and base on wet delay troposphere, respectively. Results showed a high correlation among the values of TPW derived from MODIS banding ratio, Radiosonde and GPS data at the Mehrabad station. Also, Using the ratio of the apparent reflectance in the water vapor absorption band to reflectance in non-absorbing band, the atmospheric water vapor transparency was mapped, that the maps showed a high correlation between apparent reflectance and TPW MODIS as their statistical results showed an inverse negative relationship(R²= -0.97).
Synthesis And Evaluation of Different Numbers of Phosphonate Group Containing...inventionjournals
Oil fields were seriously affected by the mineral scale formation, mainly calcium deposition. This paper focused on the synthesis and scale prevention of oilfield scaling, especially carbonates scale and sulfate scale. Scale can cause loss of production due to blockages in the near-well formation, in the tubing or in the surface facilities. Phosphonates are widely accepted as the most effective inhibitors for calcium carbonate and calcium sulfate, also are widely used in many water treatment applications. Most good scale inhibitors should work at dosage levels between 5 and 25 ppm on most production systems depending on calcium concentration. The treatment rates can be affected by contaminants which take the scale inhibitor out of solution. Six different compounds of phosphonate containing groups have been synthesized and evaluated for scale inhibition property. Evaluation of synthetic phosphonate compounds as scale inhibitors has been done by using NACE Standard TM-0374-2001. From the results clear that one and two phosphonate group containing compounds do not show any characteristic of prevention of scales of either calcium carbonate or calcium sulphate. The compounds which have phosphonate group three or more show characteristics of prevention of both types of scales.
Entrepreneurship is considered as a source of wealth creation, economic growth, social progress, and technological development. The current paper seeks to shed light on obstacles that are impeding business creators to start their businesses. To identify the observed constraints, we developed a questionnaire that we addressed to 120 new entrepreneurs drawing on the theoretical and empirical literature. The application of factor analysis has revealed that business creators’ decision to launch their ventures is hindered mainly by the following factors: lack of managerial and business skills, poor training programs, and risk aversion. The study also suggests some recommendations to alleviate obstacles facing new entrepreneurs when deciding to launch their projects
Taller ErgoCV en el XII Congreso Internacional de Prevención de Riesgos Labor...ErgoCV
Desde ErgoCV se han realizado a lo largo del ejercicio 2013 talleres de trabajo, entre otras materias sobre ergonomía en inclusión de personas con discapacidad, procedimientos de violencia en el trabajo, y retos de la ergonomia en la internacionalización de las Empresas.
En dichos talleres han participado empresas, técnicos de prevención y departamentos de recursos humanos.
En el ORP Zaragoza se presentaron las conclusiones de esos talleres, completándose con las aportaciones de otros participantes durante el taller de PERU, ECUADOR, VENEZUELA, CHILE y ARGENTINA.
Practical Guidelines to Improve Defect Prediction Model – A Reviewinventionjournals
Defect prediction models are used to pinpoint risky software modules and understand past pitfalls that lead to defective modules. The predictions and insights that are derived from defect prediction models may not be accurate and reliable if researchers do not consider the impact of experimental components (e.g., datasets, metrics, and classifiers) of defect prediction modeling. Therefore, a lack of awareness and practical guidelines from previous research can lead to invalid predictions and unreliable insights. Through case studies of systems that span both proprietary and open-source domains, find that (1) noise in defect datasets; (2) parameter settings of classification techniques; and (3) model validation techniques have a large impact on the predictions and insights of defect prediction models, suggesting that researchers should carefully select experimental components in order to produce more accurate and reliable defect prediction models.
EWMA 2013 - Ep563 - A new anti-biofilm dressing: in vivo investigationEWMAConference
David Parsons1 PhD, MRSC CChem
Akhil Seth2 MD, BS
Thomas Mustoe2 MD, FACS
1.ConvaTec Research & Development, Flintshire, Wales, UK
2.Division of Plastic Surgery, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.
This presentation is for qualified registered small animal veterinary surgeons who are interested in using Lipogems Canine. The presentation outlines the science behind the concept, an outline of the procedure, some case study data and links for further reading. Please be aware this presentation contains videos and therefore the file is very large and may take some time to download. For a quick download please request the link for the quick PDF download.
Cosmetic shop management system project report.pdfKamal Acharya
Buying new cosmetic products is difficult. It can even be scary for those who have sensitive skin and are prone to skin trouble. The information needed to alleviate this problem is on the back of each product, but it's thought to interpret those ingredient lists unless you have a background in chemistry.
Instead of buying and hoping for the best, we can use data science to help us predict which products may be good fits for us. It includes various function programs to do the above mentioned tasks.
Data file handling has been effectively used in the program.
The automated cosmetic shop management system should deal with the automation of general workflow and administration process of the shop. The main processes of the system focus on customer's request where the system is able to search the most appropriate products and deliver it to the customers. It should help the employees to quickly identify the list of cosmetic product that have reached the minimum quantity and also keep a track of expired date for each cosmetic product. It should help the employees to find the rack number in which the product is placed.It is also Faster and more efficient way.
Sachpazis:Terzaghi Bearing Capacity Estimation in simple terms with Calculati...Dr.Costas Sachpazis
Terzaghi's soil bearing capacity theory, developed by Karl Terzaghi, is a fundamental principle in geotechnical engineering used to determine the bearing capacity of shallow foundations. This theory provides a method to calculate the ultimate bearing capacity of soil, which is the maximum load per unit area that the soil can support without undergoing shear failure. The Calculation HTML Code included.
Courier management system project report.pdfKamal Acharya
It is now-a-days very important for the people to send or receive articles like imported furniture, electronic items, gifts, business goods and the like. People depend vastly on different transport systems which mostly use the manual way of receiving and delivering the articles. There is no way to track the articles till they are received and there is no way to let the customer know what happened in transit, once he booked some articles. In such a situation, we need a system which completely computerizes the cargo activities including time to time tracking of the articles sent. This need is fulfilled by Courier Management System software which is online software for the cargo management people that enables them to receive the goods from a source and send them to a required destination and track their status from time to time.
CFD Simulation of By-pass Flow in a HRSG module by R&R Consult.pptxR&R Consult
CFD analysis is incredibly effective at solving mysteries and improving the performance of complex systems!
Here's a great example: At a large natural gas-fired power plant, where they use waste heat to generate steam and energy, they were puzzled that their boiler wasn't producing as much steam as expected.
R&R and Tetra Engineering Group Inc. were asked to solve the issue with reduced steam production.
An inspection had shown that a significant amount of hot flue gas was bypassing the boiler tubes, where the heat was supposed to be transferred.
R&R Consult conducted a CFD analysis, which revealed that 6.3% of the flue gas was bypassing the boiler tubes without transferring heat. The analysis also showed that the flue gas was instead being directed along the sides of the boiler and between the modules that were supposed to capture the heat. This was the cause of the reduced performance.
Based on our results, Tetra Engineering installed covering plates to reduce the bypass flow. This improved the boiler's performance and increased electricity production.
It is always satisfying when we can help solve complex challenges like this. Do your systems also need a check-up or optimization? Give us a call!
Work done in cooperation with James Malloy and David Moelling from Tetra Engineering.
More examples of our work https://www.r-r-consult.dk/en/cases-en/
Welcome to WIPAC Monthly the magazine brought to you by the LinkedIn Group Water Industry Process Automation & Control.
In this month's edition, along with this month's industry news to celebrate the 13 years since the group was created we have articles including
A case study of the used of Advanced Process Control at the Wastewater Treatment works at Lleida in Spain
A look back on an article on smart wastewater networks in order to see how the industry has measured up in the interim around the adoption of Digital Transformation in the Water Industry.
About
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
• Remote control: Parallel or serial interface.
• Compatible with MAFI CCR system.
• Compatible with IDM8000 CCR.
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
• Easy in configuration using DIP switches.
Technical Specifications
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
Key Features
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
• Remote control: Parallel or serial interface
• Compatible with MAFI CCR system
• Copatiable with IDM8000 CCR
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
Application
• Remote control: Parallel or serial interface.
• Compatible with MAFI CCR system.
• Compatible with IDM8000 CCR.
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
• Easy in configuration using DIP switches.
COLLEGE BUS MANAGEMENT SYSTEM PROJECT REPORT.pdfKamal Acharya
The College Bus Management system is completely developed by Visual Basic .NET Version. The application is connect with most secured database language MS SQL Server. The application is develop by using best combination of front-end and back-end languages. The application is totally design like flat user interface. This flat user interface is more attractive user interface in 2017. The application is gives more important to the system functionality. The application is to manage the student’s details, driver’s details, bus details, bus route details, bus fees details and more. The application has only one unit for admin. The admin can manage the entire application. The admin can login into the application by using username and password of the admin. The application is develop for big and small colleges. It is more user friendly for non-computer person. Even they can easily learn how to manage the application within hours. The application is more secure by the admin. The system will give an effective output for the VB.Net and SQL Server given as input to the system. The compiled java program given as input to the system, after scanning the program will generate different reports. The application generates the report for users. The admin can view and download the report of the data. The application deliver the excel format reports. Because, excel formatted reports is very easy to understand the income and expense of the college bus. This application is mainly develop for windows operating system users. In 2017, 73% of people enterprises are using windows operating system. So the application will easily install for all the windows operating system users. The application-developed size is very low. The application consumes very low space in disk. Therefore, the user can allocate very minimum local disk space for this application.
NO1 Uk best vashikaran specialist in delhi vashikaran baba near me online vas...Amil Baba Dawood bangali
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Student information management system project report ii.pdfKamal Acharya
Our project explains about the student management. This project mainly explains the various actions related to student details. This project shows some ease in adding, editing and deleting the student details. It also provides a less time consuming process for viewing, adding, editing and deleting the marks of the students.
TECHNICAL TRAINING MANUAL GENERAL FAMILIARIZATION COURSEDuvanRamosGarzon1
AIRCRAFT GENERAL
The Single Aisle is the most advanced family aircraft in service today, with fly-by-wire flight controls.
The A318, A319, A320 and A321 are twin-engine subsonic medium range aircraft.
The family offers a choice of engines
Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by Micronucleus Test and Comet Assay
1. International Journal of Pharmaceutical Science Invention
ISSN (Online): 2319 – 6718, ISSN (Print): 2319 – 670X
www.ijpsi.org Volume 5 Issue 4 ‖July 2016 ‖PP.07-11
www.ijpsi.org 7 | P a g e
Genotoxicity Evaluation of Polystyrene Membrane with Collagen
and Norbixin by Micronucleus Test and Comet Assay
Samylla Miranda Monte1
, Antônio Luiz Martins Maia Filho1
, Fabricio Pires de
Moura do Amaral1
, Danniel Cabral Leão Ferreira1
, Wcleubianne Matias
Nascimento2
, Vicente Galber Freitas Viana3
, Zulmira Lúcia Oliveira Monte4
,
Charllyton Luis Sena Costa5
, Vivaldo Moura Neto6
.
1
Center for Research in Biotechnology and Biodiversity (NPBio) of the State University of Piauí
(UFPI) Teresina, PI, Brazil.
2
Pharmacy course at the College Integral Differential (FACID / DeVry) Teresina, PI, Brazil.
3
Federal Institute of Piauí (IFPI), Teresina, PI, Brazil.
4
Federal University of Piauí (FSA), Teresina, PI, Brazil.
5
College St. Augustine (FSA), Caxias, MA, Brazil.
6
Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brazil.
ABSTRACT: The biocompatible membranes are widely applied in the medical field in order to stimulate tissue repair.
The biological principle of this type of treatment is the repair and guided regeneration. In the literature, there are few
reports of studies evaluating the effects and biological properties of norbixin in animal tissues. Thus, the present study was
to evaluate the effect of polystyrene membrane with collagen and norbixin, through the micronucleus test and comet assay in
rats, as part of the recommended test battery to evaluate the mutagenic potential. The research project was approved by
CEP / FACID Protocol 069/2014. For this study, 15 rats were divided into 3 groups were used: A - the membrane was
introduced into the peritoneum of the animals through a laparotomy; B - received cyclophosphamide at a dose of 50mg / kg
intraperitoneally; C - were performed only one laparotomy. A peripheral blood sample was collected from the animals for
conducting Comet assay and 72 hours after the start of the experiment were euthanized. It was collected bone marrow
material of each rat to perform the micronucleus test. In conclusion, through the tests, the membrane is not genotoxic.
Keywords: Genotoxicity, Mutagenicity, Biomaterials, Norbixin.
I. Introduction
Articular cartilage is a tissue with no vascularization and no nerve endings and low capacity for self-repair, a
damaged time has no restorability, thus resulting in osteoarthritis [1]. Osteoarthritis (OA) is a chronic
degenerative disease which involves the articular cartilage, synovial fluid, ligaments, bone, meniscus, tendon
and periarticular muscle, however, the destruction of cartilage is a common feature of OA progression and
results in inefficient operation the affected joint.
The normal joint cartilage is composed of large amounts of extracellular matrix (mainly type II collagen),
produced and maintained by chondrocytes. Currently, there is no effective therapy in the treatment of OA,
except those that relieve the symptoms of the disease until the time that the joints need replacement surgery [2].
The lack of repair mechanism is attributed to the lack of cell recruitment to the injured area [1]. To overcome
this problem, a lot of research on cartilage lesions articular treatments are discussed, but until now no study
reports the regeneration of hyaline cartilage altogether. Several alternatives are practiced, including,
arthroscopic interventions, techniques, cell-based, autograft/allograft cartilage and gene therapy technique,
however, compensation is always performed by fibrocartilage that does not have the same biomechanical
characteristics that the hyaline cartilage, necessary for the normal functioning of joints [3, 4].
There is therefore a clear need for alternative strategies for the production of curative treatments. Tissue
engineering is an interdisciplinary field of rapid development that involves both materials science and
engineering to medical research to address clinical problems. Most tissue engineering strategies is based on the
use of biocompatible materials to facilitate tissue regeneration [5].
In recent years, it has increased the search for substances that can be used in tissue engineering in order to
develop biomaterials that can be applied to renovate or restore the function of diseased or traumatized tissues in
the body. Generally, naturally occurring biomaterials are characterized with biocompatibility, ie, have a lower
rate of rejection by the living tissue allowing the adhesion and migration of cells within their structures in
relation to synthetic polymers and also a lower cost manufacturing [6 ].
Methods based on the principles of tissue engineering have emerged as the most promising approaches for the
repair of joint damage. These methods utilize a combination of cell signaling molecules and scaffolds for
increasing the regenerative capacity of the natural joint with the aim of restoring healthy structure and function
2. Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by Micronucleus..
www.ijpsi.org 8 | P a g e
of the damaged tissue [4]. Tissue engineering has successfully produced a number of applications in soft tissue,
but regeneration of the articular surface is a formidable challenge, in view of the articular cartilage has a limited
capacity to regenerate and mechanical stimuli play a major role in structure and function of cartilage and
subchondral bone [4, 6].
Thus, it was developed a polystyrene membrane with collagen and norbixin following the biological principle of
such treatment, ie, tissue engineering, which is guided in repair and regeneration. Norbixin is a compound
extracted from the surface of the annatto seeds, fruit Urucuzeiro, plant species Bixa Orellana L., native to
Central and South America, which has great potential in obtaining biodegradable and biocompatible polymers.
The norbixin (C24H2804) is a hydrophilic dicarboxylic carotenoid, antioxidant. In the literature, there are few
studies of reports that evaluate the effects and biological properties of norbixin in animal tissues [8].
Little is known, however, about the biocompatibility and cytotoxicity in vivo of such biomaterial. Therefore, in
this study, our aim was to evaluate the biocompatibility and cytotoxicity of this membrane using invivo methods
such as the micronucleus rat bone marrow cells test and the comet assay using peripheral blood of animals [9
10].
II. Material And Methods
2.1 Polystyrene membrane and norbixin with collagen
The membrane components were dissolved in chloroform in a ratio of 0.20 grams of polystyrene for 0.05 g of
norbixin and 0.05 g of collagen. After 2 hours of magnetic stirring the mixture was placed in a petri dish at
room temperature to evaporate the solvent forming the membrane.
2.2 ForMicronucleus Test
The experimental procedure was carried out in Experimentation and Mutagenicity Laboratory (LABEMUT) and
Laboratory of Molecular Biology and Biological Studies Injuries (LABMINBIO) of the State University of
Piauí (UESPI) as approved (Protocol - 069/2014) in the Research Ethics Committee (CEP/FACID). The
exposed group (E) consisting of 05 rats received the membrane introduced into the peritoneum through a
laparotomy and were fully anesthetized. They were euthanized 72 hours after the beginning of the experiment. It
was collected bone marrow material from each rat after euthanasia to perform the micronucleus test in
polychromatic erythrocyte, to evaluate the degree of genotoxicity.
2.2.1 Negative Control (NC)
Consisting of 5 rats in which were performed only one laparotomy. The duration and application are processed
in parallel with the comet assay.
2.2.2 Positive Control (PC)
Five rats received injections via the I. P. with a single dose of cyclophosphamide at a concentration of 50 mg/kg
per animal. The duration and application are processed in parallel with the comet assay.
2.2.3 Processing, preparation of the slide and staining
Bone marrow cells were collected immediately after the sacrifice of animals. For this, we use a 01 ml syringe
for collection. This syringe was filled with Fetal Bovine Serum (FBS). We introduce the needle into the opening
of a femur and FBS injected through the channel, pushing the medullary component toward the other end
positioned in a Falcon tube, previously marked with the animal code. Resuspend the FBS in bone marrow
material, to achieve homogeneity. The suspension was centrifuged for 5 minutes at 1,000 rpm and the
supernatant discarded at the end of the procedure with Pasteur pipette. Completed the sample with 0.5 ml of
FBS and resuspend by homogenization. Dripping the prepared suspension 02 drops in smears dull edge of a
slide (previously marked with the animal's code) and with the aid of other slide bent at a 45 degree angle, we
smear. After making the smear, let air dry. Were prepared two slides per animal. Staining was performed 24
hours after preparation of the slides for 3 minutes in Giemsa.
2.2.4 Analysis of the slides
The analysis was performed in blind field in an increase of 100x (immersion objective) in a short time by the
same observer. We determined the frequency of micronuclei in 2000 PCEs cells per animal. The entire protocol
is based in several publications [11-15].
2.3 For Comet Assay
The experimental procedure was performed in the LABEMUT and LABMINBIO UESPI laboratories. The
exposed group (E) consisted of 5 animals that received the membrane introduced into the peritoneum through a
laparotomy.
3. Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by Micronucleus..
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2.3.1 Negative Control (NC)
Consisting of 5 rats in which were performed only one laparotomy. The duration and application are processed
in parallel with the micronucleus test.
2.3.2 Positive Control (PC)
Five rats received injections via the I. P. with a single dose of cyclophosphamide at a concentration of 50 mg /
kg per animal. The duration and application are processed in parallel with the micronucleus test. Samples were
processed 4h and 24h after exposure.
2.3.3 Processing, preparation of the slide and staining
The comet assay was performed on peripheral blood taken from the tail of each animal after 4 hours of exposure
(check the damage) and 24 hours of exposure (check the repair).At the end of each period it is collected40 uL,
transferring them to microtubes containing120 uL of low melting-point agarose (1.5%) at 37ºC.The mixture was
homogenized and transferred topre-covered with agarose slides,getting coverslips (to spread the content
homogeneously) and stored at 4ºC for 30 minutes [16]. Then the coverslips were removed and immersed
invertical glass cuvettes containing lysing solution [NaCl (2.5M); EDTA (100 mM) and 1.2 g of TRIS
(10mM)].Upon use, was added 1% Triton X-100 and 10% of DMSO.The slides were placed in a vessel
containing electrophoretic buffer pH> 13.0 (300 mM NaOH and 1 mM EDTA, prepared from a stock solution
of 10N NaOH and 200 mM EDTA, pH 10.0), remaining at rest for 20 minutes.The "race" electrophoretic was
performed with 25 V and 300 mA at a temperature of 4ºC lasting 15 minutes in the darkconditions.After
electrophoresis, the slides were removed from the chamber and immersed in neutralizing solution (0.4 M Tris,
pH 7.5, for 5 min).This process was repeated 3 times.Finally, they were rinsed with distilled water and air dried,
flushing them with Gel Red (2.0 Dilution: 10,000 uL per 10 minutes).All slides were analyzed in
immunofluorescence microscopy (increase of 40X) equipped with excitation filter (420-490 nm) and barrier
filter (520 nm).The images were obtained by a Opton System (digital camera CCD 5.0 mega pixel for
immunofluorescence).It evaluated the damage to DNA by DNA percentage of measurement in the tail (% DNA-
measurement of the proportion of the total DNA present in the tail) and the time of the tail (TM- tail length
times the percentage of DNA in the tail) [ 17].These parameters were calculated on 100 nucleoids/sample (two
slides per individual). For this, we used the software OpenComet [18].
Figure 01. (A) Photomicrograph of comets. (B 'and B' ') processing the Open Comet software. Analysis of 100
comets per exposed individual.
2.4Statistical analysis
Data were analyzed using One-Way ANOVA and Tukey test. For all tests was considered a 5% significance
level. The results were expressed as mean, or standard deviation of three independent experiments. The result
was considered positive if there was a statistically significant increase (p <0.05).
III. Results
3.1 For micronucleus test
In this paper we evaluate the genotoxic potential ofpolystyrene membrane with collagen and norbixina on bone
marrow cells from Rattus norvegicus. Table 01 shows the mean micronuclei group E (Laparotomy + membrane)
was 10.80 ± 7.56, PC group (cyclophosphamide) was 42.40 ± 11.93 NC group (laparotomy) was 3.6 ± 0.89.
Process of exposure to the membrane did not increase significantly the mutagenicity when compared to the
negative control (p> 0.05). When compared to the positive control, it detected a statistically significant
difference (p> 0.001) (Fig. 02).
4. Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by Micronucleus..
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Table 01. Frequency MN in polychromatic erythrocytes in bone marrow Rattus norvegicus exposed to
polystyrene membrane with collagen and norbixina
Group Number of PCEs Analyzed PCEMNs Mean± standard deviation
N0
%
CN 2.000 18 0,9 3,6 ± 0,89 a
CP 2.000 212 10,6 42,40 ± 11,93
E 2.000 178 8,9 10,80 ± 7,56 a
Legend: NC = negative control; PC = positive control (cyclophosphamide 50 mg/kg); E = Exposed; PCE =
polychromatic erythrocytes and PCEMNs = Micronuclei in Erythrocytes polychromatic. "A" indicates a statistically
significant difference (P <0.001) when compared to CP.
Figure 02.In (A) and (B) polychromatic erythrocyte with micronucleus in bone marrow of Rattus
norvegicus(arrow). (C) Results found after the MN test: meanpolychromatic erythrocyte with a micronucleus.
3.2 For Comet Assay
The results presented in Table 02 show that the membrane did not induce DNA damage and genotoxicity of
animal cells. The toxic potential of the membrane was investigated and reviewed comet indicated an increase of
damage to the 1st 4h exposure evaluated by the% DNA in the tail. However, after 24 h, there was a statistically
significant decrease compared to the first few hours, indicating a repair of the initial genotoxic effect. This
finding is consistent with other parameters analyzed, as the% of DNA in the head, TailMoment and the length of
the tail.
Table 02. DNA damage found per group after exposure to polystyrene membrane with collagen and norbixina.
Parameters Positive control Negative Control Membrane 4h Membrane 24h
Head DNA % 3,2 ± 0,4 12,3 ± 0,89 a
5,0 ± 0,42 a
13,2 ± 1,2 a
Tail DNA % 4,5 ± 0,8 0,8 ± 0,02 a
7,3 ± 1,2 a
0,3 ± 0,07 a
Tail length (μM) 16,0 ± 1,7 14,0 ± 2,0 a
18,0 ± 2,0 a
3,0 ± 1,3 a
TailMoment 1.4 ± 0,56 0,8 ± 0,63 a
1,7 ± 0,21 a
0,93 ± 0,25 a
Results expressed as mean and standard deviation. "A" indicates a statistically significant difference (P <0.001) when
compared to CP.
IV. Discussion
A variety of biomaterials formed by natural polymers, synthetic polymers and derivatives of acellular matrix is
huge [19], however, they are only classified as biomaterials are free of causing local or systemic adverse
biological response may not be toxic, carcinogenic, antigenic and mutagen [20]. Under the biotechnological
perspective obtaining water-soluble derivatives of carotenoids is interesting in commercial perspective. The use
of synthetic materials for replacement or repair of biological tissues, has always been a major concern in the
medical field, which justifies the investigation of new materials in tissue engineering.
The selection of the biomaterial to be used should take into account the physical, chemical and mechanical
properties of the material [21, 22]. The membranes are designed to provide a favorable microenvironment of
repair of articular cartilage, formed to the diffusion of nutrients, growth factors and other biologically active
agents in the field of tissue regeneration [23].
In this study, toxicity was assessed inpolystyrene membrane with collagen and norbixin through in vivo
experiments in rats. To determine the genotoxic potential of the membrane, two experiments were conducted,
5. Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by Micronucleus..
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micronucleus test and comet assay. The micronucleus test showed that extracts from the membrane does not
alter the incidence of polychromatic erythrocyte suggesting that the membrane has no genetic toxicity in
rats.The Comet Assay showed an increase of damage in the first 4 hours, and for 24 hours of exposure time,
showed a statistically significant decrease from the early hours suggesting repair genotoxic effect. Taken
together, our results suggest that the membrane does not have biological toxicity. It is known that DNA repair is
essential to providing the maintenance of homeostasis and, therefore, cell and tissue survival. In most cases, this
is correctly handles, restoring the original molecule [24].
In summary, the findings indicated no damage after exposure generated thepolystyrene membrane with collagen
and norbixin,suggesting that there was adequate or sufficient time to repair the same, showing that the
membrane is not cytotoxic, and therefore, is safe for in vivo applications. Thus, future studies will be conducted
to investigate the effects of the membrane in the repair of articular cartilage.
Referências
[1] J. Y. Chung, M. Song, C.-W. Ha, J.-A. Kim, C.-H. Lee, and Y.-B, Park, Comparison of articular cartilage repair with different
hydrogel-human umbilical cord blood-derived mesenchymal stem cell composites in a rat model,Stem Cell Res Ther, 5(2), 2014,
39.
[2] T. P. Lozito, P. G. Alexander, H. Lin, R. Gottardi, AW-M Cheng, and R. S. Tuan, Three-dimensional osteochondral microtissue
to model pathogenesis of osteoarthritis. Stem Cell Research & Therapy,4 (1), 2013, 1-6.
[3] M. Falah, G. Nierenberg, M. Soudry, M. Hayden, and G. Volpin, Treatment of articular cartilage lesions of the knee.
International Orthopaedics (SICOT), 34, 2010, 621–30.
[4] A. K.Lynn, R. A. Brooks, W. Bonfield, and N. Rushton, Repair of defects in articular joints: prospects for material-based
solutions intissue engineering. J Bone Joint Surg [Br],86-B (8), 2004, 1093-9.
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