EXPERIMENTAL TRIES TO ESTABLISH THE PREIMPLANTATIONAL MAMMALIAN EMBRYOS VIABILITY THROUGHOUT STAINING

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IVAN ALEXANDRA

2007-01-01

Full Text Available Presently there are more methods to assess embryo quality but, still the wieldy usedremains the morphological criteria method. In this experiment were tested twostaining methods for embryos and oocytes. The embryos were recovered from mousefemale at 72 hours after mating. The recovered embryos were first evaluated aftermorphological criteria and than by Trypan blue exclusion and Neutral red staining.Using Trypan blue exclusion were evaluated 30 embryos from which 19 (63.3 wereclassified as viable and 11 (36.7 were classified as nonviable. By Neutral redstaining were evaluated 37 embryos from which 24 (64.8 were considered viableand 13 (35.2 were considered nonviable. The oocytes recovered were alsoevaluated using the two methods: using Trypan blue exclusion were stained 10oocytes from which 9 remained uncolored and were considered viable and 1 wasstained in blue and was considered nonviable and using Neutral red 13 oocytes werestained from which 9 were evaluated as viable and 4 as nonviable.

Involvement of the Endocannabinoid System in the Development and Treatment of Breast Cancer

Science.gov (United States)

2014-04-01

chloroquine (CQ) at 5 µM in combination with the WIN2/IR combination before quantification of cell viability using trypan blue exclusion. At 96 h, CQ had no...viability was quantified using trypan blue exclusion in MCF-7 cells treated as in (A) with a co-treatment of either vehicle or 5 µM chloroquine (B...Acridine orange staining was used to image autophagic vesicles in MCF-7 cells treated with vehicle, 1 µM ADR or ADR + 5 µM chloroquine (C). In (B

Viability of mesenchymal stem cells during electrospinning

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G. Zanatta

2012-02-01

Full Text Available Tissue engineering is a technique by which a live tissue can be re-constructed and one of its main goals is to associate cells with biomaterials. Electrospinning is a technique that facilitates the production of nanofibers and is commonly used to develop fibrous scaffolds to be used in tissue engineering. In the present study, a different approach for cell incorporation into fibrous scaffolds was tested. Mesenchymal stem cells were extracted from the wall of the umbilical cord and mononuclear cells from umbilical cord blood. Cells were re-suspended in a 10% polyvinyl alcohol solution and subjected to electrospinning for 30 min under a voltage of 21 kV. Cell viability was assessed before and after the procedure by exclusion of dead cells using trypan blue staining. Fiber diameter was observed by scanning electron microscopy and the presence of cells within the scaffolds was analyzed by confocal laser scanning microscopy. After electrospinning, the viability of mesenchymal stem cells was reduced from 88 to 19.6% and the viability of mononuclear cells from 99 to 8.38%. The loss of viability was possibly due to the high viscosity of the polymer solution, which reduced the access to nutrients associated with electric and mechanical stress during electrospinning. These results suggest that the incorporation of cells during fiber formation by electrospinning is a viable process that needs more investigation in order to find ways to protect cells from damage.

Storage effect on viability and biofunctionality of human adipose tissue-derived stromal cells.

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Falah, Mizied; Rayan, Anwar; Srouji, Samer

2015-09-01

In our recent studies, the transplantation of human adipose tissue-derived stromal cells (ASCs) has shown promise for treatment of diseases related to bone and joint disorders. For the current clinical applications, ASCs were formulated and suspended in PlasmaLyte A supplemented with heparin, glucose and human serum albumin, balanced to pH 7.4 with sodium bicarbonate. This cell solution constitutes 20% of the overall transplanted mixture and is supplemented with hyaluronic acid (60%) and OraGraft particles (20%). We intended to investigate the effect of this transplantation mixture on the viability and biofunctionality of ASCs in bone formation. Freshly harvested cells were resuspended and incubated in the indicated mixture for up to 48 h at 4°C. Cell viability was assessed using trypan blue and AlamarBlue, and cell functionality was determined by quantifying their adhesion rate in vitro and bone formation in an ectopic mouse model. More than 80% of the ASCs stored in the transplantation mixture were viable for up to 24 h. Cell viability beyond 24 h in storage decreased to approximately 50%. In addition, an equal degree of bone formation was observed between the cells transplanted following incubation in transplantation mixture for up to 24 h and zero-time non-incubated cells (control). The viability and functionality of ASCs stored in the presented formulation will make such cell therapy accessible to larger and more remote populations. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Sonophotocatalytic degradation of trypan blue and vesuvine dyes in the presence of blue light active photocatalyst of Ag3PO4/Bi2S3-HKUST-1-MOF: Central composite optimization and synergistic effect study.

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Mosleh, S; Rahimi, M R; Ghaedi, M; Dashtian, K

2016-09-01

An efficient simultaneous sonophotocatalytic degradation of trypan blue (TB) and vesuvine (VS) using Ag3PO4/Bi2S3-HKUST-1-MOF as a novel visible light active photocatalyst was carried out successfully in a continuous flow-loop reactor equipped to blue LED light. Ag3PO4/Bi2S3-HKUST-1-MOF with activation ability under blue light illumination was synthesized and characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX), photoluminescence (PL) and diffuse reflectance spectra (DRS). The effect of operational parameters such as the initial TB and VS concentration (5-45mg/L), flow rate (30-110mL/min), irradiation and sonication time (10-30min), pH (3-11) and photocatalyst dosage (0.15-0.35g/L) has been investigated and optimized using central composite design (CCD) combined with desirability function (DF). Maximum sonophotodegradation percentage (98.44% and 99.36% for TB and VS, respectively) was found at optimum condition set as: 25mg/L of each dye, 70mL/min of solution flow rate, 25min of irradiation and sonication time, pH 6 and 0.25g/L of photocatalyst dosage. At optimum conditions, synergistic index value was obtained 2.53 that indicated the hybrid systems including ultrasound irradiation and photocatalysis have higher efficiency compared with sum of the individual processes. Copyright © 2016. Published by Elsevier B.V.

Observations Regardin Oocyte in Vitro Maturation after Recovery from Slaughter House Females

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Valeriu Carabă

2011-05-01

Full Text Available The oocytes viability must be taken as an important selection parameter for successful in vitro cultivation. The ovaries were collected from the slaughterhouse and maintained at 4°C for 7 days. Fallowing cumulus -oocytes complexes recovery the viability was tested using two staining methods. For the first experiment we used 27 cumulus - oocytes complexes, stained with Neutral red and for the second experiment we used 11 cumulus - oocytes complexes stained with Trypan blue. Fallowing staining with Neutral red 23 cumulus - oocytes complexes were assessed as viable (were stained in red – enzymatic activity within the cells and for the Trypan blue staining 11 cumulus - oocytes complexes were assessed as viable (remained unstained – integers cellular membranes.

Cytotoxicity study of plant Aloe vera (Linn

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Atul N Chandu

2012-01-01

Full Text Available Background: The objective of this study has been to evaluate the in-vitro antitumor activity of Aloe vera extract of in cultured B16F10 melanoma cell line by measuring cell viability using "Trypan blue exclusion assay" method. Aim: To find out such kind of anticancer drug which is a cheap, safe, less toxic, and more potent drug compared to chemotherapy drug. Materials and Methods: In-vitro antitumor activity cell culture1, drug treatment (standard and test extract and Trypan blue exclusion assay growth and viability test 1 were used. Treatment of Aloe vera extract against B16F10 melanoma cell line, in all concentration range, showed decrease in percent cell viability, as compared to that of negative when examined by "Trypan blue exclusion assay". Results: In overall variation of test samples, Aloe vera extract showed its best activity in the concentration of 300 μg/ml, which was approximately equal to the activity of standard drug doxorubicin. Evaluation of in-vitro antitumor activity revealed that Aloe vera extract exhibits good cytotoxic activity. The best cytotoxic activity by Aloe vera was shown at 200 μg/ml concentration. Conclusion: The study of cytoprotection against normal cells by micronucleus assay has shown that the herbal extracts have less toxic effects to the normal blood lymphocytes, as compared to that of standard anticancer drug.

The effect of gestational diabetes on proliferation capacity and viability of human umbilical cord-derived stromal cells.

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Wajid, Nadia; Naseem, Rashida; Anwar, Sanam Saiqa; Awan, Sana Javaid; Ali, Muhammad; Javed, Sara; Ali, Fatima

2015-09-01

Stomal cells derived from Wharton's jelly of human umbilical cord (WJMSCs) are considered as the potential therapeutic agents for regeneration and are getting famous for stem cell banking. Our study aims to evaluate the effects of gestational diabetes on proliferation capacity and viability of WJMSCs. Mesenchymal stromal cells were isolated from Wharton's jelly of human umbilical cords from normal and gestational diabetic (DWJMSCs) mothers. Growth patterns of both types of cells were analyzed through MTT assay and population doubling time. Cell survival, cell death and glucose utilization were estimated through trypan blue exclusion assay, LDH assay and glucose detection assay respectively. Angiogenic ability was evaluated by immunocytochemistry and ELISA for VEGF A. Anti-cancerous potential was analyzed on HeLa cells. DWJMSCs exhibited low proliferative rate, increased population doubling time, reduced cell viability and increased cell death. Interestingly, DWJMSCs were found to have a reduced glucose utilization and anti-cancerous ability while enhanced angiogenic ability. Gestational diabetes induces adverse effects on growth, angiogenic and anti-cancerous potential of WJMSCs.

Evaluation of Periodontal Ligament Cell Viability in Three Different Storage Media: An in Vitro Study

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Meenakshi Sharma

2016-01-01

Full Text Available Objectives: This study was undertaken to evaluate the viability of periodontal ligament (PDL cells of avulsed teeth in three different storage media.Materials and Methods: Forty-five premolars extracted for orthodontic therapeutic purposes were randomly and equally divided into three groups based on storage media used [Group I: milk (control; Group II: aloe vera (experimental; Group III: egg white (experimental]. Following extractions, the teeth were placed in one of the three different storage media for 30 minutes, following which the scrapings of the PDL from these teeth were collected in Falcon tubes containing collagenase enzyme in 2.5 mL of phosphate buffered saline. The tubes were subsequently incubated for 30 minutes and centrifuged for five minutes at 800 rpm. The obtained PDL cells were stained with Trypan Blue and were observed under optical microscope. The percentage of viable cells was calculated.Results: Aloe vera showed the highest percentage of viable cells (114.3±8.0, followed by egg white (100.9±6.3 and milk (101.1±7.3.Conclusion: Within the limitations of this study, it appears that aloe vera maintains PDL cell viability better than egg white or milk.

In vitro cytotoxicity and induction of apoptosis by multiwalled carbon ...

African Journals Online (AJOL)

obtained from Frascati Laboratory, Sigma Co., Sun Nanotech Co. and Shenzhen Co.) were used in this study. Cell viability and apoptotic activity were evaluated by trypan blue exclusion test and Annexin-V/PI staining in peripheral blood ...

7-Piperazinethylchrysin inhibits melanoma cell proliferation by ...

African Journals Online (AJOL)

PEC) on melanoma cell lines. Methods: Cell viability was analyzed by trypan blue exclusion assays and the cell cycle by flow cytometry using ModFit LT software. Specifically, cells were stained with propidium iodide (0.5 mg/mL) supplemented ...

Group B streptococcal beta-hemolysin/cytolysin directly impairs cardiomyocyte viability and function.

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Mary E Hensler

Full Text Available BACKGROUND: Group B Streptococcus (GBS is a leading cause of neonatal sepsis where myocardial dysfunction is an important contributor to poor outcome. Here we study the effects of the GBS pore-forming beta-hemolysin/cytolysin (Bh/c exotoxin on cardiomyocyte viability, contractility, and calcium transients. METHODOLOGY/PRINCIPAL FINDINGS: HL-1 cardiomyocytes exposed to intact wild-type (WT or isogenic Deltabeta h/c mutant GBS, or to cell-free extracts from either strain, were assessed for viability by trypan blue exclusion and for apoptosis by TUNEL staining. Functionality of exposed cardiomyocytes was analyzed by visual quantitation of the rate and extent of contractility. Mitochondrial membrane polarization was measured in TMRE-loaded cells exposed to GBS beta h/c. Effects of GBS beta h/c on calcium transients were studied in fura-2AM-loaded primary rat ventricular cardiomyocytes. Exposure of HL-1 cardiomyocytes to either WT GBS or beta h/c extracts significantly reduced both rate and extent of contractility and later induced necrotic and apoptotic cell death. No effects on cardiomyocyte viability or function were observed after treatment with Deltabeta h/c mutant bacteria or extracts. The beta h/c toxin was associated with complete and rapid loss of detectable calcium transients in primary neonatal rat ventricular cardiomyocytes and induced a loss of mitochondrial membrane polarization. These effects on viability and function were abrogated by the beta h/c inhibitor, dipalmitoyl phosphatidylcholine (DPPC. CONCLUSIONS/SIGNIFICANCE: Our data show a rapid loss of cardiomyocyte viability and function induced by GBS beta h/c, and these deleterious effects are inhibited by DPPC, a normal constituent of human pulmonary surfactant.. These findings have clinical implications for the cardiac dysfunction observed in neonatal GBS infections.

Cellular effects of halogen blue light from dental curing unit

International Nuclear Information System (INIS)

Trosic, I.; Pavicic, I.; Jukic, S.

2008-01-01

Full text: Halogen curing lights are the most frequently used polymerization source in dental offices. Light-cured bonding systems have become increasingly popular among clinicians because they offer a number of advantages over self-cured adhesives. The effort to increase polymerization quality releases the commercially available high power light density dental curing units. Emitted visible blue light belongs to the range of nonionizing radiation. Common concern in both, patients and dentist grows with regard to the unfavorable effects on the pulp tissue. The aim of study was to evaluate the time and dose dependence effect of halogen light curing unit (Elipar TriLight, ESPE Dental AG, Germany) at the disposed condition modes in vitro. A quartz-tungsten-halogen light source emits radiation of the wavelengths between 400 and 515 nm. This halogen blue light source operates in the three illumination modes, medium (M), exponential (E) and standard (S), and five illumination times. The total irradiance or the light intensity was measured by the light intensity control area on the control panel of device and mean light intensity given by manufacturer was 800 m W/cm 2 . Continuous culture of V79 cells was illuminated in triplicate. The influence of medium mode (M), exponential (E) and standard (S) illumination during 20, 40 and 80 sec on the cell viability, colony forming ability and proliferation of V79 cell culture was investigated. Trypan blue exclusion test was used to determine cell viability, both, in the treated and control cell samples. Colony forming ability was assessed for each exposure time and mode by colony count on post-exposure day 7. Cell proliferation was determined by cell counts for each time and mode of exposure during five post-exposure days. Statistical difference were determined at p<0.05 (Statistica 7.0, StatSoft Inc., USA). Viability of cells was not affected by blue light in view of exposure time and modes. Regardless to exposure or illumination

Coconut milk and probiotic milk as storage media to maintain periodontal ligament cell viability: an in vitro study.

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Saini, Divya; Gadicherla, Prahlad; Chandra, Prakash; Anandakrishna, Latha

2017-06-01

The viability of periodontal ligament (PDL) cells is a significant determinant of the long-term prognosis of replanted avulsed teeth. A storage medium is often required to maintain the viability of these cells during the extra-alveolar period. Many studies have been carried out to search for the most suitable storage medium for avulsed teeth, but an ideal solution has not yet been found. The purpose of the study was to compare and analyze the ability of coconut milk and probiotic milk to maintain PDL cell viability. In an in vitro setting, 69 caries free human premolars with normal periodontium that had been extracted for orthodontic purposes were randomly divided into two experimental groups on the basis of storage media used (i.e., coconut milk or probiotic milk) and a Hanks' balanced salt solution (HBSS) control group (23 samples per group). Immediately after extraction, the teeth were stored dry for 20 min and then immersed for 30 min in one of the storage media. The teeth were then subjected to collagenase-dispase assay and labeled with 0.5% trypan blue staining solution for determination of cell viability. The number of viable cells was counted under a light microscope and statistically analyzed using anova and post hoc Tukey test (P ≤ 0.05). Statistical analysis demonstrated there was a significant difference (P coconut milk and probiotic milk as well as HBSS in maintaining cell viability. However, there was no significant difference between probiotic milk and HBSS in ability to maintain PDL cell viability (P > 0.05). Coconut milk may not be suitable as an interim transport media due to poor maintenance of cell viability. However, probiotic milk was able to maintain PDL cell viability as well as HBSS. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Corneal edema and permanent blue discoloration of a silicone intraocular lens by methylene blue.

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Stevens, Scott; Werner, Liliana; Mamalis, Nick

2007-01-01

To report a silicone intraocular lens (IOL) stained blue by inadvertent intraoperative use of methylene blue instead of trypan blue and the results of experimental staining of various lens materials with different concentrations of the same dye. A "blue dye" was used to enhance visualization during capsulorhexis in a patient undergoing phacoemulsification with implantation of a three-piece silicone lens. Postoperatively, the patient presented with corneal edema and a discolored IOL. Various IOL materials were experimentally stained using methylene blue. Sixteen lenses (4 silicone, 4 hydrophobic acrylic, 4 hydrophilic acrylic, and 4 polymethylmethacrylate) were immersed in 0.5 mL of methylene blue at concentrations of 1%, 0.1%, 0.01%, and 0.001%. These lenses were grossly and microscopically evaluated for discoloration 6 and 24 hours after immersion. The corneal edema resolved within 1 month after the initial surgical procedure. After explantation, gross and microscopic analyses of the explanted silicone lens revealed that its surface and internal substance had been permanently stained blue. In the experimental study, all of the lenses except the polymethylmethacrylate lenses were permanently stained by methylene blue. The hydrophilic acrylic lenses showed the most intense blue staining in all dye concentrations. This is the first clinicopathological report of IOL discoloration due to intraocular use of methylene blue. This and other tissue dyes may be commonly found among surgical supplies in the operating room and due diligence is necessary to avoid mistaking these dyes for those commonly used during ocular surgery.

Increased viability and resilience of haemolymph cells in blue mussels following pre-treatment with acute high-dose gamma irradiation

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Jaeschke, B. [Stockholm University (Sweden)

2014-07-01

In an initial experiment, blue mussels (Mytilus edulis) were exposed to a range of acute high doses of gamma radiation in the laboratory. Haemolymph was extracted and the haemocytes (blood cells) were scored for cell viability (% living cells) under a microscope, directly after irradiation (0.04, 0.4 or 4 Gy) and again after a subsequent treatment with hydrogen peroxide in vitro (final H{sub 2}O{sub 2} conc.: 0.2 μM). Cell viability in controls (0 Gy) was approximately 100% and no cell death was observable from radiation exposure alone. When treated with H{sub 2}O{sub 2} a decrease in cell viability was seen across all treatments, however this decrease in viability was reduced with increasing radiation pre-treatment (0 Gy = 53%; 0.04 Gy = 66%; 0.4 Gy = 75%; 4 Gy = 83%). To investigate the mechanism for this therapeutic effect observed, the experiment was repeated. Using mussels from a different location, the same, but more extensive method of irradiation (0[control], 0.04, 0.4 Gy, 5 or 40 Gy) and H{sub 2}O{sub 2} treatment was used. Additional haemolymph sub-samples were taken for analysis of catalase concentration. In this second experiment, viability of cells from controls was only 62%, indicating the mussels were in a poorer condition than those of the previous experiment. The lowest level of radiation exposure (0.04 Gy) further decreased the viability (56%). However, at higher doses the viability was increased compared to control, which then gradually declined with increasing dose (0.4 Gy = 75%; 5 Gy = 72%; 40 Gy = 65%). Catalase analysis demonstrated a complimentary pattern of activity of the antioxidant in the haemolymph, directly correlating with radiation dose (0 Gy = 0.2 U; 0.04 Gy = 0.1 U; 0.4 Gy = 1.3 U; 5 Gy = 0.9 U; 40 Gy = 0.1 Gy). Treatment with H{sub 2}O{sub 2} decreased cell viability across all treatments, but no pattern between radiation treatments was discernable. The results indicate that an acute dose of radiation not only has negligible

Resin phantoms as skin simulating layers

CSIR Research Space (South Africa)

Karsten, AE

2011-07-01

Full Text Available is considered to be reactive oxygen species (ROS) that are lethal to the cells. The treatment is a localized treatment due to the short lifetimes of the singlet and triplet oxygen. Photosense?, which is a mixture of sulfonated aluminium(III) phthalocyanines....3.4. Changes in cell viability Change in morphology and viability of cells (controls and experimental) was assessed using an inverted microscope. A trypan blue dye, cell viability assay reagent, based on the principle that live cells possess intact cell...

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

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Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

Toxoplasma gondii: effects of 60 Co ionizing radiation in the viability and infectivity, detected in vitro in LLC-MK2 cells and in vivo in C57BL/6J mice

International Nuclear Information System (INIS)

Hiramoto, Roberto M.; Almeida, Beatriz S.V.; Cardoso, Roselaine P.A.; Andrade Junior, Heitor F.

1997-01-01

Toxoplasmosis, caused by Toxoplasma gondii, promotes devasting disease in fetus and AIDS patients. The longlife immunity of natural infection is inefficient in eliminate tissue infective cysts. Few immunization programs were tested, mostly with attenuated strains. Ionizing radiation were used with successful in vaccine production, without reproductive ability with a relatively normal physiology until reproduction. Here, we tested several schedules of 60 Co irradiation of tachzoites from RH strain of T. gondii, from peritoneal exudate or suspensions of LLC-MK2 infected cells, to optimize the viability and sterility of the irradiated agents. The tachzoites were exposed to 50, 100 and 200 Gy in a GammaCell 220 at 366 Gy/h. The viability was tested by motility, integrity and Trypan Blue dye exclusion. All irradiation schedules maintained a high (>90%) viability of the parasites. Dilutions were injected in C57Bl/6j mice with induction of specific antibodies, no clinical disease but uncertain sterility. Infection of LLC--MK2 cells showed that viable and reproductive parasites were often found in 50 Gy irradiated cells, rarely found in 100 Gy irradiated cells, rarely found in 100 Gy irradied cells, with no growth occuring with 200 Gy irradiated tachzoites. Our data show that 200 Gy 60 Co irradiation blocks the reproductive capacity without affecting the short term viability of tachzoites of T. gondii. (author). 11 refs., 1 fig

Preparation of pre-confluent retinal cells increases graft viability in vitro and in vivo: a mouse model.

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Kevin P Kennelly

Full Text Available PURPOSE: Graft failure remains an obstacle to experimental subretinal cell transplantation. A key step is preparing a viable graft, as high levels of necrosis and apoptosis increase the risk of graft failure. Retinal grafts are commonly harvested from cell cultures. We termed the graft preparation procedure "transplant conditions" (TC. We hypothesized that culture conditions influenced graft viability, and investigated whether viability decreased following TC using a mouse retinal pigment epithelial (RPE cell line, DH01. METHODS: Cell viability was assessed by trypan blue exclusion. Levels of apoptosis and necrosis in vitro were determined by flow cytometry for annexin V and propidium iodide and Western blot analysis for the pro- and cleaved forms of caspases 3 and 7. Graft viability in vivo was established by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL and cleaved caspase 3 immunolabeling of subretinal allografts. RESULTS: Pre-confluent cultures had significantly less nonviable cells than post-confluent cultures (6.6%±0.8% vs. 13.1%±0.9%, p<0.01. Cell viability in either group was not altered significantly following TC. Caspases 3 and 7 were not altered by levels of confluence or following TC. Pre-confluent cultures had low levels of apoptosis/necrosis (5.6%±1.1% that did not increase following TC (4.8%±0.5%. However, culturing beyond confluence led to progressively increasing levels of apoptosis and necrosis (up to 16.5%±0.9%. Allografts prepared from post-confluent cultures had significantly more TUNEL-positive cells 3 hours post-operatively than grafts of pre-confluent cells (12.7%±3.1% vs. 4.5%±1.4%, p<0.001. Subretinal grafts of post-confluent cells also had significantly higher rates of cleaved caspase 3 than pre-confluent grafts (20.2%±4.3% vs. 7.8%±1.8%, p<0.001. CONCLUSION: Pre-confluent cells should be used to maximize graft cell viability.

Nanoparticles containing allotropes of carbon have genotoxic effects on glioblastoma multiforme cells

DEFF Research Database (Denmark)

Hinzmann, Mateusz; Jaworski, Sławomir; Kutwin, Marta

2014-01-01

of the U87 cancer cells. However, incubation with pristine graphene and reduced graphene oxide led to a significant decrease in cell viability, whereas incubation with graphene oxide, graphite, and ultradispersed detonation diamond led to a smaller decrease in cell viability. The results of a comet assay...... viability by Trypan blue assay and level of DNA fragmentation of U87 cells after 24 hours of incubation with 50 μg/mL carbon nanoparticles. DNA fragmentation was studied using single-cell gel electrophoresis. Incubation with nanoparticles containing the allotropes of carbon did not alter the morphology...

Effect of Formaldehyde on Human Middle Ear Epithelial Cells

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Shin Hye Kim

2018-01-01

Full Text Available Formaldehyde (FA is a familiar indoor air pollutant found in everything from cosmetics to clothing, but its impact on the middle ear is unknown. This study investigated whether FA causes cytotoxicity, inflammation, or induction of apoptosis in human middle ear epithelial cells (HMEECs. Cell viability was investigated using the trypan blue assay and a cell counting kit (CCK-8 in HMEECs treated with FA for 4 or 24 h. The expression of genes encoding the inflammatory cytokine tumor necrosis factor alpha (TNF-α and mucin (MUC5AC was analyzed using RT-PCR. Activation of the apoptosis pathway was determined by measuring mitochondrial membrane potential (MMP, cytochrome oxidase, caspase-9/Mch6/Apaf 3, and Caspase-Glo® 3/7 activities. The CCK-8 assay and trypan blue assay results showed a reduction in cell viability in FA-treated HMEECs. FA also increased the cellular expression of TNF-α and MUC5AC and reduced the activities of MMP and cytochrome oxidase. Caspase-9 activity increased in cells stimulated for 4 h, as well as caspase-3/7 activity in cells stimulated for 24 h. The decreased cell viability, the induction of inflammation and mucin gene expression, and the activation of the apoptosis pathway together indicate a link between environmental FA exposure and the development of otitis media.

CSN1S2 protein of goat milk inhibits the decrease of viability and increases the proliferation of MC3T3E1 pre-osteoblast cell in methyl glyoxal exposure

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Choirunil Chotimah

2015-03-01

Full Text Available Objective: To investigate whether the CNS1S2 protein of goat milk is able to inhibit the toxicity of methyl glyoxal (MG towards MC3T3E1 pre-osteoblast cells. Methods: At confluency, pre-osteoblast cells were divided into five groups which included control (untreated, pre-osteoblast cells exposed to 5 µmol/L MG, pre-osteoblast cells exposed to MG in the presence of CSN1S2 protein at doses of 0.025, 0.050, and 0.100 mg/L, respectively. Analysis of reactive oxygen species was done with 2,7-dichlorodihydrofluorescein diacetate fluorochrome. The proliferation and viability of MC3T3E1 cells were measured by trypan blue staining. Malondialdehyde analysis was done colorimetrically. Results: Cell's viabilities were significantly lower in MG+0.050 mg/L CSN1S2 protein of goat milk compared to MG group (P<0.05. MG+0.100 mg/L CSN1S2 protein of goat milk significantly increased the cells viability compared to MG group (P<0.05. The levels of proliferation were significantly higher in MG+0.100 mg/L CSN1S2 protein of goat milk compared to control group and all treatment groups, respectively (P<0.05. Conclusions: High dose of CSN1S2 protein of goat milk (0.100 mg/L in high MG environment inhibits the decrease of viability due to the increases of the proliferation of MC3T3E1 preosteoblast cell.

Blue light-mediated inactivation of Enterococcus faecalis in vitro.

Science.gov (United States)

Pileggi, Giorgio; Wataha, John C; Girard, Myriam; Grad, Iwona; Schrenzel, Jacques; Lange, Norbert; Bouillaguet, Serge

2013-05-01

In dentistry, residual infection remains a major cause of failure after endodontic treatment; many of these infections involve Enterococcus faecalis. In the current study, we explored the possibility that blue light activated photosensitizers could be used, in principle, to inactivate this microbe as an adjunct disinfection strategy for endodontic therapy. Three blue light absorbing photosensitizers, eosin-Y, rose bengal, and curcumin, were tested on E. faecalis grown in planktonic suspensions or biofilms. Photosensitizers were incubated for 30 min with bacteria then exposed to blue light (450-500 nm) for 240 s. Sodium hypochlorite (3%) was used as a control. After 48 h, the viability of E. faecalis was estimated by measuring colony-forming units post-exposure vs. untreated controls (CFU/mL). Blue light irradiation alone did not alter E. faecalis viability. For planktonic cultures, blue light activated eosin-Y (5 μM), rose bengal (1 μM), or curcumin (5 μM) significantly (pcurcumin of 100, 10, and 10 μM respectively, completely suppressed E. faecalis viability (p<0.05). Although the current results are limited to an in vitro model, they support further exploration of blue light activated antimicrobials as an adjunct therapy in endodontic treatment. Copyright © 2012 Elsevier B.V. All rights reserved.

Blue light induced free radicals from riboflavin in degradation of crystal violet by microbial viability evaluation.

Science.gov (United States)

Liang, Ji-Yuan; Yuann, Jeu-Ming P; Hsie, Zong-Jhe; Huang, Shiuh-Tsuen; Chen, Chiing-Chang

2017-09-01

Crystal violet (CV) is applied in daily use mainly as a commercial dye and antimicrobial agent. Waste water containing CV may affect aquatic ecosystems. Riboflavin, also known as vitamin B 2 , is non-toxic and an essential vitamin required for the functions of the human body. Riboflavin is photosensitive to UV and visible light in terms of generating reactive oxygen species. This study investigated the potential application of blue light on riboflavin, so as to come up with an effective way of degrading CV during its treatment. Photosensitivity of CV leading to degradation in the presence of riboflavin was investigated by light intensity, exposure time, and irradiation dosage. The degradation of CV during riboflavin photolysis treatment was studied by a UV/vis spectrometry and chromatography. The effects of CV degradation on microbial viability are relevant when considering the influences on the ecosystem. This study proved that riboflavin photochemical treatment with blue light degrades CV dye by ROS formation. The riboflavin photolysis-treated CV solution appeared to be transparent during conformational transformations of the CV that was rearranged by free radical species generated from riboflavin photolysis. After riboflavin photolysis, colony-forming units (CFUs) were determined for each CV solution. CFU preservation was 85.2% for the CV dissolved riboflavin solution treated with blue light irradiation at 2.0mW/cm 2 for 120min. Degradation of CV by riboflavin photochemical procedures can greatly reduce antimicrobial ability and serve as an environmental friendly waste water treatment method. Our results presented here concerning riboflavin photolysis in degradation of CV provide a novel technique, and a simple and safe practice for environmental decontamination processes. Copyright © 2017 Elsevier B.V. All rights reserved.

Synthetic Beta-Lactam Antibiotics as a Selective Breast Cancer Cell Apoptosis Inducer: Significance in Breast Cancer Prevention and Treatment

Science.gov (United States)

2008-03-01

Hoechst 33258, cremophore, dimethylsulfoxide ( DMSO ) and trypan blue were purchased from Sigma-Aldrich (St. Louis, MO). Apoptag peroxidase in situ...treated with solvent ( DMSO ), 50 or 100 μM of each indicated compound for 24 h, followed by trypan blue exclusion assay. (C) HL-60 leukemic cells were...treated with solvent ( DMSO ), 50 or 100 μM of each indicated analog for 24 h, followed by trypan blue exclusion assay. The numbers given are

Microprocessor-controlled vs. "dump-freezing" platelet and lymphocyte cryopreservation: A quantitative and qualitative comparative study

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Balint Bela

2006-01-01

Full Text Available Background/Aim. Thermodynamical and cryobiological parameters responsible for cell damages during cryopreservation (cryoinjuries have not yet been completely explained. Thus, freezing procedures should be revised, exactly optimized to obtain an enhanced structural and functional recovery of frozen- thawed cells. The aim of this study was to compare microprocessor- controlled (controlled-rate with the compensation of the released fusion heat and “dump-freezing” (uncontrolled- rate of the platelet and lymphocyte cryopreservation efficacy. Methods. Platelet quantitative recovery (post-thaw vs. unfrozen cell count, viability (using hypotonic shock response - HSR, morphological score (PMS, ultrastructural (electron microscopy properties and expression of different surface antigens were investigated. In lymphocyte setting, cell recovery and viability (using trypan blue exclusion test as well as functionality (by plant mitogens were determined. Controlled- rate freezing and uncontrolled-rate cryopreservation were combined with 6% (platelets and 10% (lymphocytes dimethyl sulfoxide (DMSO. Results. Platelet recovery and functionality were superior in the controlled-rate system. The majority of surface antigen expression was reduced in both freezing groups vs. unfrozen cells, but GP140/CD62p was significantly higher in controlled-rate vs. uncontrolled-rate setting. Controlled- rate freezing resulted with better lymphocyte recovery and viability (trypan blue-negative cell percentage. In mitogen-induced lymphocyte proliferative response no significant intergroup difference (controlled-rate vs. uncontrolled-rate were found. Conclusion. The data obtained in this study showned the dependence of cell response on the cryopreservation type. Controlled-rate freezing provided a superior platelet quantitative and functional recovery. Lymphocyte recovery and viability were better in the controlled-rate group, although only a minor intergroup difference for cell

LncRNA-LET inhibits cell viability, migration and EMT while induces apoptosis by up-regulation of TIMP2 in human granulosa-like tumor cell line KGN.

Science.gov (United States)

Han, Qingfang; Zhang, Wenke; Meng, Jinlai; Ma, Li; Li, Aihua

2018-04-01

Polycystic ovary syndrome (PCOS) is a common endocrine disease characterized by hyperandrogenism, irregular menses, and polycystic ovaries. Several long non-coding RNAs (lncRNAs) are aberrantly expressed in PCOS patients; however, little is known about the effects of the lncRNA-low expression in tumor (lncRNA-LET) on PCOS. We aimed to explore the effects of lncRNA-LET on human granulosa-like tumor cell line, KGN. Expression of lncRNA-LET in normal IOSE80 cells and granulosa cells was determined by qRT-PCR. KGN cell viability, apoptosis and migration were measured by trypan blue exclusion method, flow cytometry assay and wound healing assay, respectively. TGF-β1 was used to induce epithelial-mesenchymal transition (EMT) process. LncRNA-LET expression and mRNA expressions of TIMP2 and EMT-related proteins were measured by qRT-PCR. Western blot analysis was used to measure the protein expression of apoptosis-related proteins, EMT-related proteins, TIMP2, and the proteins in the Wnt/β-catenin and Notch signaling pathways. lncRNA-LET was down-regulated in KGN cells, and its overexpression inhibited cell viability and migration, and promoted apoptosis in KGN cells. Overexpression of lncRNA-LET increased the expression of E-cadherin and decreased the expressions of N-cadherin and vimentin in KGN cells. These effects of lncRNA-LET on KGN cells were reversed by TIMP2 suppression. Overexpression of TIMP2 inhibited cell viability, migration and EMT process, and increased apoptosis by activating the Wnt/β-catenin and Notch pathways. Overexpression of lncRNA-LET inhibits cell viability, migration and EMT process, and increases apoptosis in KGN cells by up-regulating the expression of TIMP2 and activating the Wnt/β-catenin and notch signaling pathways. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Corneal epithelial cell viability of an ex vivo porcine eye model.

Science.gov (United States)

Chan, Ka Yin; Cho, Pauline; Boost, Maureen

2014-07-01

The aim was to assess the consistency of corneal epithelial cell viability of an ex vivo porcine eye model. Six porcine eye models (four test and two control) were prepared for each experiment. The model has a computer-controlled mechanical arm, which could move the eyelid of the porcine eye and apply phosphate buffered saline to simulate blinking and lacrimation. The four test eyes were set up to simulate evaporative dry eyes with simulated lacrimation and blinking (one blink and one drop of buffered saline per minute) over three hours. Control A models were set up to collect pre-experimental baseline data, while those of control B were the same as the test eyes but without lacrimation and blinking simulation. All porcine eyes were kept in a closed chamber with temperature and humidity well controlled. After three hours, the cells of all eyes (except control A, which were assessed immediately before commencement of the experiment) were assessed. The eyes were first dipped into 0.4 per cent trypan blue solution. Following the dissection and separation of the cells, the number of dead cells were then counted under the microscope with a field size of 0.25 mm(2). The experiment was repeated 11 times. No significant differences were found in the number of dead cells among the four test eyes in both the central and peripheral cornea. There were significantly more dead cells in the test eyes compared to control A but significantly less when compared to control B. More dead cells were found in the central cornea than the peripheral cornea in the test eyes but the difference was not observed in controls A and B. Epithelial cell viabilities among the four porcine eye models with simulated lacrimation and blinking were consistent. The majority of cells were viable before the experiment and simulated lacrimation and blinking maintained more viable cells over time. © 2014 The Authors. Clinical and Experimental Optometry © 2014 Optometrists Association Australia.

Effects of dipotassium-trioxohydroxytetrafluorotriborate, K2[B3O3F4OH], on cell viability and gene expression of common human cancer drug targets in a melanoma cell line.

Science.gov (United States)

Pojskic, Lejla; Haveric, Sanin; Lojo-Kadric, Naida; Hadzic, Maida; Haveric, Anja; Galic, Zoran; Galic, Borivoj; Vullo, Daniela; Supuran, Claudiu T; Milos, Mladen

2016-12-01

Recently it was found that dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH), is a potent and highly specific inhibitor of precancerous cell processes. We conducted gene expression profiling of human melanoma cells before and after treatment with two concentrations (0.1 and 1 mM) of this boron inorganic derivative in order to assess its effects on deregulation of genes associated with tumor pathways. Parallel trypan blue exclusion assay was performed to assess the cytotoxicity effects of this chemical. Treatment with K2(B3O3F4OH) induced a significant decrease of cell viability in melanoma cellline at both tested concentrations. Furthermore, these treatments caused deregulation of more than 30 genes known as common anti-tumor drug targets. IGF-1 and hTERT were found to be significantly downregulated and this result may imply potential use of K2(B3O3F4OH) as an inhibitor or human telomerase and insulin-like growth factor 1, both of which are associated with various tumor pathways.

Amazonian açai and food dyes for staining arbuscular- micorrhizal fungi

Directory of Open Access Journals (Sweden)

Aline Lourdes Martins Silva

2015-12-01

Full Text Available Arbuscular mycorrhizae microscopy requires differential staining of typical structures. Dyes employed, such as trypan blue, pose risks to health and environment. Alternative dyes such as pen ink and aniline have variable coloring efficiency. In this work, Brachiaria decumbens roots, discolored with caustic soda (NaOH, were stained with açai, annatto, saffron, trypan blue and pen inks. There were significant differences among dyes regarding stained mycorrhizal structures and pictures quality. Acai was considered the best alternative dye, with similar results to trypan blue.

OBSERVATIONS REGARDING OOCYTES STORAGE POST MENDING FROM SLAUGHTER FEMALES

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CARABA V.

2008-01-01

Full Text Available The oocytes viability must be taken as an important selection parameter for successful in vitrocultivation. The ovaries were collected from the slaughterhouse and maintained at 4°C for 7days. Fallowing cumulus -oocytes complexes recovery the viability was tested by two stainingmethods. For the first experiment we used 27 cumulus - oocytes complexes, stained withNeutral red and for the second experiment we used 11 cumulus - oocytes complexes stainedwith Trypan blue. Fallowing staining with Neutral red 23 cumulus - oocytes complexes wereassessed as viable (were stained in red – enzymatic activity within the cells and for the Trypanblue staining 11 cumulus - oocytes complexes were assessed as viable (remained unstained –integers cellular membranes.

Viability of the Fricke dosemeter doped with methylene blue

International Nuclear Information System (INIS)

Souza, V.L.B.; Santos, C.D.A.; Rodrigues, K.R.G.; Cunha, M.S.; Figueiredo, M.D.C.; Melo, R.T.

2009-01-01

This work aims to find the possible utilization of the Fricke dosemeter doped with methylene blue (FMB) for the dosimetry of photodynamic therapy. The FMB was irradiated wit X rays and light emitted diodes demonstrating positive answers to the stimulus, being probably to be used for dosimetric objectives

The effects of beryllium metal particles on the viability and function of cultured rat alveolar macrophages

International Nuclear Information System (INIS)

Finch, G.L.; Lowther, W.T.; Hoover, M.D.; Brooks, A.L.

1988-01-01

Rat pulmonary alveolar macrophages (PAM) were exposed in vitro to beryllium metal particles. The particles used were relatively large (Be-II) and small (Be-V) size fractions of beryllium metal obtained from an aerosol cyclone, and a beryllium metal aerosol generated by laser vaporization of beryllium metal in an argon atmosphere (Be-L). Glass beads (GB) were used as a negative control particle. The endpoints examined included cell killing (trypan blue dye exclusion) and phagocytic ability (sheep red blood cell uptake). Phagocytic ability was inhibited by beryllium particles at concentrations that did not cause appreciable cell killing. Results based on the mass concentration of particles in culture medium were transformed by the amount of specific surface area of the particles to permit expression of toxicity on the basis of amount of surface area of particles per unit volume of culture medium. On a mass concentration basis, the order of cytotoxicity was Be-L > Be-V ∼ Be-II > GB; for inhibition of phagacytosis, the cytotoxicity order was Be-L ∼ Be-V > Be-II > GB. On a surface area concentration basis, the order of toxicity for viability was altered to Be-II > Be-L ∼ Be-V (with GB indeterminant) and to Be-V > Be-II ∼ Be-L > GB for inhibition of phagocytosis. We conclude that there are factors in addition to specific surface area that influence the expression of toxic effects in cultured PAM. (author)

Anti-adhesive and pro-apoptotic effects of 2-hydroxyethyl methacrylate on human gingival fibroblasts co-cultured with Streptococcus mitis strains

Science.gov (United States)

Zara, S; Di Giulio, M; D’Ercole, S; Cellini, L; Cataldi, A

2011-01-01

Aim To evaluate and observe the cellular reactions that occur during the interaction/integration between 2-hydroxyethyl methacrylate/host tissue/microbial environment, in a co-culture of human gingival fibroblasts (HGF) and Streptococcus mitis strains. Methodology Streptococcus mitis were cultured with strains in the presence of 3 mmol L−1 HEMA for 48 h and 72 h. Cytotoxicity was evaluated by the trypan blue dye exclusion test. Apoptosis was evaluated by TUNEL analysis. Adhesion was evaluated by immunofluorescence and western blot analyses. Quantitative analyses of the results were acquired by Qwin Plus 3.5 and QuantityOne I-D analysis software, respectively. The statistical significance of the results was evaluated using t-tests and linear regression tests. Results The trypan blue dye test revealed 47.3% and 46.5% of dead fibroblasts after 48 and 72 h HEMA treatment, respectively, while bacterial viability was not influenced by the presence of HEMA and fibroblasts. The expression of pro-collagen I, involved in fibroblast adhesion, in untreated samples ranged from 12.49% to 6.91% of the positive area after 48 and 72 h, respectively, dropping to below 2% of the positive area in the other experimental conditions. Unlike the trypan blue test, co-cultured samples treated with HEMA showed 20% and 25% versus 17% and 21% (after 48 and 72 h, respectively) of apoptotic cells. Conclusions The evidence for HEMA toxicity and anti-adhesive effects against eukaryotic cells was reduced in the presence of bacteria, suggesting that dental resins should be well polymerized to avoid the spread of toxic monomers within the mouth. PMID:21902700

Photodynamic Effects of Pterin on HeLa Cells

DEFF Research Database (Denmark)

Denofrio, M. Paula; Lorente, Carolina; Breitenbach, Thomas

2011-01-01

cells (HeLa) and that these cells die upon UV-A irradiation of Ptr. Cell death was assessed using two tests: (1) the Rhodamine 123 fluorescence assay for mitochondrial viability and (2) the Trypan Blue assay for membrane integrity. The data suggest that, for Ptr-dependent photoinitiated cell death......Pterins, heterocyclic compounds widespread in biological systems, participate in relevant biological processes and are able to act as photosensitizers. In the present study, we ascertained that 2-aminopteridin-4(3H)-one, abbreviated as Ptr, is readily incorporated into and ⁄ or onto cervical cancer...

Ultrasound-microbubble mediated cavitation of plant cells: effects on morphology and viability.

Science.gov (United States)

Qin, Peng; Xu, Lin; Zhong, Wenjing; Yu, Alfred C H

2012-06-01

The interaction between ultrasound pulses and microbubbles is known to generate acoustic cavitation that may puncture biological cells. This work presents new experimental findings on the bioeffects of ultrasound-microbubble mediated cavitation in plant cells with emphasis on direct observations of morphological impact and analysis of viability trends in tobacco BY-2 cells that are widely studied in higher plant physiology. The tobacco cell suspensions were exposed to 1 MHz ultrasound pulses in the presence of 1% v/v microbubbles (10% duty cycle; 1 kHz pulse repetition frequency; 70 mm between probe and cells; 1-min exposure time). Few bioeffects were observed at low peak negative pressures (cavitation presumably occurred. In contrast, at 0.9 MPa peak negative pressure (with more inertial cavitation activities according to our passive cavitation detection results), random pores were found on tobacco cell wall (observed via scanning electron microscopy) and enhanced exogenous uptake into the cytoplasm was evident (noted in our fluorescein isothiocyanate dextran uptake analysis). Also, instant lysis was observed in 23.4% of cells (found using trypan blue staining) and programmed cell death was seen in 23.3% of population after 12 h (determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling [TUNEL]). These bioeffects generally correspond in trend with those for mammalian cells. This raises the possibility of developing ultrasound-microbubble mediated cavitation into a targeted gene transfection paradigm for plant cells and, conversely, adopting plant cells as experimental test-beds for sonoporation-based gene therapy in mammalian cells. Copyright © 2012 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

Energy Technology Data Exchange (ETDEWEB)

Oliveira, R.J. [Programa de Pós-Graduação em Biologia Celular e Molecular, Instituto de Biociências de Rio Claro, Universidade Estadual Paulista, Rio Claro, SP (Brazil); Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Pós-Graduação em Saúde em Desenvolvimento na Região Centro-Oeste, Faculdade de Medicina “Dr. Hélio Mandetta”, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Mestrado em Farmácia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Mantovani, M.S.; Silva, A.F. da [Departamento de Biologia Geral, Universidade Estadual de Londrina, Londrina, PR (Brazil); Pesarini, J.R. [Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Pós-Graduação em Saúde em Desenvolvimento na Região Centro-Oeste, Faculdade de Medicina “Dr. Hélio Mandetta”, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Mauro, M.O. [Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Doutorado em Biotecnologia e Biodiversidade - Rede Pró Centro-Oeste, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Ribeiro, L.R. [Programa de Pós-Graduação em Biologia Celular e Molecular, Instituto de Biociências de Rio Claro, Universidade Estadual Paulista, Rio Claro, SP (Brazil); Programa de Pós-Graduação em Patologia, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista, Botucatu, SP (Brazil)

2014-03-28

The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.

Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

Directory of Open Access Journals (Sweden)

R.J. Oliveira

2014-04-01

Full Text Available The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.

Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

International Nuclear Information System (INIS)

Oliveira, R.J.; Mantovani, M.S.; Silva, A.F. da; Pesarini, J.R.; Mauro, M.O.; Ribeiro, L.R.

2014-01-01

The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero

on Lymphoblastic Leukemia Jurkat Cells

African Journals Online (AJOL)

human tumor cell line (Hela) by using MTT assay. [13]. In the present study, we have observed the cytotoxic effect of ethanolic extract of C. arvensis against Jurkat cells, a human lymphoblastic leukemia cell line, by using Trypan blue, MTS assay and FACS analysis. It was shown from the trypan blue exclusion assay that ...

Biostimulative effects of Nd:YAG Q-switch dye on normal human fibroblast cultures: study of a new chemosensitizing agent for the Nd:YAG laser

International Nuclear Information System (INIS)

Castro, D.J.; Saxton, R.E.; Fetterman, H.R.; Castro, D.J.; Ward, P.H.

1987-01-01

Kodak Q-switch II is a new chemical with an absorption maxima at 1051 nm, designed to be used as an Nd:YAG dye laser. The potential for this dye as a new chemosensitizing agent in the treatment of connective tissue diseases and wound healing with low energy Nd:YAG laser was examined. Two normal fibroblast cell lines were tested for sensitivity to various levels of this dye in vitro. These cells were exposed to Q-switch II dye at concentrations of 0.01, 0.1, 1, 10, 50, and 100 micrograms/ml for 1 and 24 hours. Cell viability was assessed by the trypan blue exclusion test. Cell duplication and DNA synthesis were measured by the incorporation of [ 3 H]-thymidine at 6 and 24 hours postexposure to Q-switch II dye. At concentrations up to 10 micrograms/ml, both cell lines tested showed no changes in cell viability. However, at concentrations equal or higher than 50 micrograms/ml, more than 40% of the fibroblasts incorporated trypan blue after 24 hours of exposure to this dye, indicating significant cell destruction. The results indicate that Q-switch II dye is nontoxic to normal human fibroblast cultures and showed significant biostimulative effects on cell duplication at concentrations equal to or lower than 10 micrograms/ml. Further studies will be required to determine the usefulness of Q-switch II dye as a new photochemosensitizing agent for potential biostimulation of wound healing and/or treatment of connective tissue diseases with the Nd:YAG laser (near infrared, 1060 nm) at nonthermal levels of energies

Use of fluorescent redox indicators to evaluate cell proliferation and viability

DEFF Research Database (Denmark)

Rasmussen, E.S.

1999-01-01

The performance of two cell viability test kits based on the use of redox indicators yielding fluorescent products, the AlamarBlue assay and a resazurin-based in vitro toxicology assay kit from Sigma, was compared in the present study. Cultures of human neonatal foreskin fibroblasts were exposed...... for 168 h of continuous exposure, but showed equal levels of cytostatic effects in cultures with a low initial cell density after 72 h of exposure. Similar characteristics of the dye solutions were observed by high-performance Liquid chromatography (HPLC) separation and UV spectroscopy, and the major...... components were tentatively identified as resazurin and resorufin. The AlamarBlue assay has gained wide application as a cell viability indicator that allows continuous monitoring of cell proliferation or cytotoxicity in human and animal cells, bacteria, and fungi, but no studies with the deliberate use...

Magnetic labeling and in vitro MR imaging of rat bone marrow mesenchymal stem cells

International Nuclear Information System (INIS)

Cai Jinhua; Feng Gansheng; Wu Hanping; Wang Xin; Li Chuan; Zhao Jiannong; Guo Daqin; Yu Guorong; Liu Guanxing; Wang Shiyi

2006-01-01

Objective: To label rat bone marrow mesenchymal stem cells with feridex combined with poly-l-lysine (PLL), and to determine the feasibility of detection of magnetically labeled stem cells with MR imaging. Methods: Feridex were incubated with PLL for 1 hour to obtain a complex of feridex-PLL. Mesenchymal stem cells isolated from the bone marrows of Wistar rats were cultured and expanded. By the 4th passage, cells were co-incubated overnight with the feridex-PLL complex. Prussian blue staining for demonstrating intracytoplastic nanoparticles and trypan-blue exclusion test for cell viability were performed respectively at 24 h, 1 w, 2 w, 3 w after labeling. MR imaging of cell suspensions was performed by using T 1 WI, T 2 WI and T 2 * WI sequences at a clinical 1.5 T MR system. Results: Numerous intracytoplastic iron particles were stained with Prussian blue. With division of stern cells, the stained particles were seen decreased gradually. Trypan blue exclusion test at 24 h, 1 w, 2 w and 3 w showed that the viability of the labeled cells was 91.00%, 93.00%, 91.75%, and 92.50%, not significantly different with that of nonlabeled cells (P>0.05). For 10 3 , 10 4 and l0 5 cells, T 2 signal intensity decreased by 63.75%, 82.31% and 91.92% respectively, T 2 * signal intensity decreased by 68.24%, 83.01%, and 93.94% respectively. For 10 5 labeled cells, T 2 * signal intensity decreased by 93.75%, 75.92%, 41.75% and 8.83 % respectively at 24 h, 1 w, 2 w and 3 w after labeling. Conclusion: Magnetic labeling of rat bone marrow stem cells with feridex-PLL complex is feasible, efficient and safe. T 2 * WI is the most sensitive sequence to detect the labeled cells. The degree of T 2 signal decreasing may be related to the cell count and division phase. (authors)

Effect of methyl butyrate aroma on the survival and viability of human breast cancer cells in vitro

International Nuclear Information System (INIS)

Khan, M.A.; Rumana Ahmad, R.; Srivastava, A.N.

2016-01-01

Background: Aroma can have far reaching effects on mind, body and soul. Pleasant aromas are known to have a soothing effect on the mind and are known to relieve stress and enhance concentration. Recently, it has been demonstrated that aroma may also have some curative effects as well as benefits and can be used both for prophylaxis and therapy of diseases. Our aim was to test our hypothesis whether aroma can cure or prevent cancer. Methyl butyrate (MB) is the methyl ester of butyric acid having a characteristic sweet and fruity odor like that of apples and pineapples. It occurs in many plant products in minute quantities and in pineapple oil. Methods: In the present study, the effect of aroma of MB has been evaluated on human breast cancer cell line MDA-MB-231 in vitro . The percentage viability of the cell line was determined by using Trypan blue dye exclusion assay. Results: It was found that MB at a concentration of 0.01 M was effective in causing considerable cytotoxicity (40%) in breast cancer cells (without even coming in contact with cells) while at 0.02 M, % cytotoxicity was found to be 50%. Mechanism of action of MB on cancer cells was investigated by acridine orange–ethidium bromide assay using fluorescence microscopy and DNA fragmentation assay. MB aroma appeared to induce necrosis in cancer cells exposed to it. Conclusion: No study involving the effect of aroma/smell on cancer cells has ever been reported before and warrants further investigation on other cancer and normal cell lines.

The effect of hydrostatic pressure on staurosporine-induced neural differentiation in mouse bone marrow‑derived mesenchymal stem cells.

Science.gov (United States)

Javanmard, F; Azadbakht, M; Pourmoradi, M

2016-01-01

In this study, the role of hydrostatic pressure on staurosporine-induced neural differentiation in mouse bone marrow mesenchymal stem cells were investigated. The cells were cultured in treatment medium containing 100 nM of staurosporine for 4 hours; then the cells were affected by hydrostatic pressure (0, 25,50, 100 mmHg). The percentage of cell viability by trypan blue staining and the percentage of cell death by Hoechst/PI differential staining were assessed. We obtained the total neurite length. Expression of β-tubulin III and GFAP (Glial fibrillary acidic protein) proteins were also analyzed by immunocytochemistry. The percentage of cell viability in treatments decreased relative to the increase in hydrostatic pressure and time (p Keywords: bone marrow mesenchymal stem cell, hydrostatic pressure, immunocytochemistry, neural differentiation, neurite length, cell differentiation.

Human amniotic fluid stem cells (hAFSCs expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide DMSO

Directory of Open Access Journals (Sweden)

Shiva Gholizadeh-Ghaleh Aziz

2018-04-01

Full Text Available Human amniotic fluid stem cells (hAFSCs have features intermediate between embryonic and adult SCs, can differentiate into lineages of all three germ layers, and do not develop into tumors in vivo. Moreover, hAFSCs can be easily obtained in routine procedures and there is no ethical or legal limitations regarding their use for clinical and experimental applications. The aim of this study was to assess the effect of slow freezing/thawing and two different concentrations of DMSO (10% DMSO + 90% fetal bovine serum [FBS] and 5% DMSO + 95% FBS on the survival of hAFSCs. hAFSCs were obtained from 5 pregnant women during amniocentesis at 16–22 weeks of gestation. The expression of pluripotency markers (Octamer-binding transcription factor 4 [Oct4] and NANOG by reverse transcription polymerase chain reaction and cell surface markers (cluster of differentiation [CD31], CD44, CD45, and CD90 by flow cytometry was analyzed before and after the slow-freezing. Cell viability was assessed by trypan blue exclusion or MTT assay. Quantitative mRNA expression of Oct4, NANOG, cyclin D1 and p21 was determined by real-time PCR before and after the slow-freezing. Pluripotency of hAFSCs was confirmed by NANOG and POU5F1 (Oct4 gene expression before and after slow-freezing. All hAFSC cultures were positive for CD44 and CD90. A higher viability of hAFSCs was observed after freezing with 90% FBS + 10% DMSO. There was increased expression of NANOG and decreased expression of POU5F1 gene after freezing, compared to control cells (before freezing. DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs. Overall, our results indicate the applicability of slow-freezing and DMSO in cryopreservation of SCs.

Mechanisms of immunological eradication of a syngeneic guinea pig tumor: participation of a component(s) of recipient origin in the expression of systemic adoptive immunity

International Nuclear Information System (INIS)

Shu, S.; Fonseca, L.S.; Kato, H.; Zbar, B.

1983-01-01

The effects of carrageenan and trypan blue on the expression of adoptive immunity to the syngeneic guinea pig line 10 hepatoma were investigated. Adoptive immunity was assessed by observing dermal tumor growth in recipients of immune cells and by bioassays in which tumor challenge sites were transplanted into secondary hosts. Carrageenan abrogated transferred immunity in treated animals as evidenced by dermal tumor growth and by development of fatal ascites tumors in peritoneal cavities of the secondary hosts. Trypan blue, on the other hand, did not abrogate transferred immunity in treated animals. However, the i.p. bioassay revealed the presence of line 10 cells in the tumor challenge sites 10 days after adoptive transfer. In vitro and in vivo exposure of immune spleen cells to carrageenan or trypan blue had no significant effect on the subsequent adoptive transfer, indicating that the inhibitory activity of these agents cannot be attributed to direct toxicity to immune lymphoid cells. Tumor challenge sites taken from carrageenan or trypan blue-treated animals 5 days after adoptive transfer failed to grow progressively when transplanted s.c. into secondary hosts. This observation suggests the presence of immune cells at tumor challenge sites. Thus, the inhibitory effects were unlikely due to interference with recirculation of the i.v.-transferred immune cells. Adoptive immunity was not influenced in guinea pigs that received a lethal dose of irradiation (500 rads). These results demonstrate that a recipient component(s) sensitive to carrageenan and trypan blue but resistant to radiation is essential to the expression of adoptive immunity

[Octanol preconditioning alleviates mouse cardiomyocyte swelling induced by simulated ischemia/reperfusion challenge in vitro].

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Luo, Yukun; Fang, Jun; Fan, Lin; Lin, Chaogui; Chen, Zhaoyang; Chen, Lianglong

2012-10-01

To investigate the role of connexin 43-formed hemichannels in cell volume regulation induced by simulated ischemia/reperfusion (SI/R). Mouse cardiomyocytes isolated on a Langendorff apparatus with enzyme solution were aliquoted into control, SI/R and SI/R +octanol groups. Calcein-AM was used to stain the cells and the cell volume was measured with confocal microscope by stack scanning. Trypan blue was used to measure the cell viability after the treatments. Calcein-AM staining and cofocal microscopy yielded stable and reproducible results for cell volume measurement. Mouse cardiomyocytes subjected to simulated SI/R showed obvious cell swelling as compared with the control cells [(126∓6)% vs 100%, Poctanol preconditioning significantly attenuated the cell swelling [(113∓6)%, Poctanol preconditioning obviously reduced the viability of the cells with SI/R challenge [(31∓2)%, Poctanol can alleviate the cell swelling to enhance the viability of the cardiomyocytes following SI/R.

Cryopreservation of zebrafish (Danio rerio) oocytes by vitrification.

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Guan, M; Rawson, D M; Zhang, T

2010-01-01

Cryopreservation of fish oocytes is challenging because these oocytes have low membrane permeability to water and cryoprotectant and are highly chilling sensitive. Vitrification is considered to be a promising approach for their cryopreservation as it involves rapid freezing and thawing of the oocytes and therefore minimising the chilling injury. In the present study, vitrification properties and the toxicity of a range of vitrification solutions containing different concentrations of Me2SO, methanol, propylene glycol and ethylene glycol were investigated. Two different base media and vitrification methods were compared. The effect of different post-thaw dilution solutions together with incubation periods on oocyte viability were also investigated. Stage III zebrafish oocytes were equilibrated in increasing concentrations of cryoprotectants for 30 min in 3 steps. Oocytes were thawed rapidly in a water bath and cryoprotectants were removed in 4 steps. Oocyte viability was assessed using trypan blue staining. The results showed that vitrification solutions V3 and V4 in KCl buffer had low toxicity and vitrified well. The survivals of oocytes after stepwise dilution using solutions containing permeable cryoprotectants were significant higher than those diluted in 0.5M glucose, and the use of CVA65 vitrification system improved oocyte survival when compared with plastic straws after 30 min at 22 degrees C post-thawing. Cryopreservation of zebrafish oocytes by vitrification is reported here for the first time, although oocyte survivals after cryopreservation assessed by trypan blue staining were relatively high shortly after thawing, they became swollen and translucent after incubation in KCl buffer. Further studies are needed to optimise the post-thaw culturing conditions.

Repeated exposures to blue light-activated eosin Y enhance inactivation of E. faecalis biofilms, in vitro.

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Marinic, Karlo; Manoil, Daniel; Filieri, Anna; Wataha, John C; Schrenzel, Jacques; Lange, Norbert; Bouillaguet, Serge

2015-09-01

In dentistry, antibacterial photodynamic therapy (a-PDT) has shown promising results for inactivating bacterial biofilms causing carious, endodontic and periodontal diseases. In the current study, we assessed the ability of eosin Y exposed to 3 irradiation protocols at inactivating Enterococcus faecalis biofilms, in vitro. E. faecalis biofilms formed on hydroxyapatite disks were incubated with eosin Y (10-80μM), then activated with blue light using different irradiation protocols. Biofilms exposed to continuous exposure were incubated for 40min before being light-activated for 960 s. For the intermittent exposure, biofilms were exposed 4 times to the light/photosensitizer combination (960 s total) without renewing the photosensitizer. For repeated a-PDT, the same light dose was delivered in a series of 4 irradiation periods separated by dark periods; fresh photosensitizer was added between each light irradiation. After treatment, bacteria were immediately labeled with LIVE/DEAD BacLight Bacterial Viability kit and viability was assessed by flow cytometry (FCM). Results were statistically analyzed using one-way ANOVA and Tukey multiple comparison intervals (α=0.05). The viability of E. faecalis biofilms exposed to 10μM eosin Y, was significantly reduced compared to controls (light only-eosin Y only). After a second exposure to blue light-activated eosin Y, viability significantly decreased from 58% to 12% whereas 6.5% of the bacterial biofilm remained live after a third exposure (p<0.05). Only 3.5% of the bacterial population survived after the fourth exposure. The results of this study indicate that blue light-activated eosin Y can photoinactivate E. faecalis biofilms grown on hydroxyapatite disks. Also, repeated exposures to blue light-activated eosin Y were shown to significantly improve efficacy. Further studies seem warranted to optimize the antibacterial activity of blue light-activated eosin Y on major oral pathogens. Copyright © 2015 Elsevier B.V. All

Efeito da nicotina na viabilidade e morfologia de fibroblastos: estudo in vitro Effect of nicotine on the viability and morphology of fibroblasts: in vitro study

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Aurora Esmeralda Traverso Martinez

2002-09-01

incubation period, cell viability was assessed by means of 0.4% trypan blue staining. Cell viability and morphology were assessed in an inverted microscope, by a single examiner, who was blind as to the experimental groups. The experiment was repeated 5 times. Regarding morphology, in the 1-hour conditioned group there was statistically significant difference between the control group and the group with the greatest dose of nicotine. These differences were also observed between the control group and all nicotine groups after 24 hours. The results of the Kruskal-Wallis test revealed that more unviable cells were found in the groups exposed to nicotine, in comparison with the control group, both after 1 and 24 hours of conditioning (p < 0.05. Moreover, with increasing doses of nicotine there was a directly proportional increase in the number of unviable cells, both after 1 and 24 hours of exposure (p = 0.0053 and p = 0.00001, respectively. The conclusion of this study is that nicotine can alter, in vitro, the viability and morphology of fibroblasts in a manner proportional to the dose and time of exposure.

Aspects of nitrogen dioxide toxicity in environmental urban concentrations in human nasal epithelium

International Nuclear Information System (INIS)

Koehler, C.; Ginzkey, C.; Friehs, G.; Hackenberg, S.; Froelich, K.; Scherzed, A.; Burghartz, M.; Kessler, M.; Kleinsasser, N.

2010-01-01

Cytotoxicity and genotoxicity of nitrogen dioxide (NO 2 ) as part of urban exhaust pollution are widely discussed as potential hazards to human health. This study focuses on toxic effects of NO 2 in realistic environmental concentrations with respect to the current limit values in a human target tissue of volatile xenobiotics, the epithelium of the upper aerodigestive tract. Nasal epithelial cells of 10 patients were cultured as an air-liquid interface and exposed to 0.01 ppm NO 2 , 0.1 ppm NO 2 , 1 ppm NO 2 , 10 ppm NO 2 and synthetic air for half an hour. After exposure, genotoxicity was evaluated by the alkaline single-cell microgel electophoresis (Comet) assay and by induction of micronuclei in the micronucleus test. Depression of proliferation and cytotoxic effects were determined using the micronucleus assay and trypan blue exclusion assay, respectively. The experiments revealed genotoxic effects by DNA fragmentation starting at 0.01 ppm NO 2 in the Comet assay, but no micronucleus inductions, no changes in proliferation, no signs of necrosis or apoptosis in the micronucleus assay, nor did the trypan blue exclusion assay show any changes in viability. The present data reveal a possible genotoxicity of NO 2 in urban concentrations in a screening test. However, permanent DNA damage as indicated by the induction of micronuclei was not observed. Further research should elucidate the effects of prolonged exposure.

The effect of HCV Core protein on the expression of miR-150

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Sayad Khanizadeh

2016-09-01

Full Text Available Background : Hepatitis C virus (HCV is considered as one of the major pathogenic agents of chronic liver diseases. Previous studies have shown that HCV proteins can interaction with gene regulatory networks such as microRNAs. The aim of this study was to investigate the effect of HCV core protein on the expression of miR-150 in a cell culture model. Materials and Methods: Plasmids expressing full HCV core protein was transfected into Huh7 cell lines while a GFP expressing plasmid employed as negative control. Subsequently, total RNA extracted and Real-Time PCR performed to measure the expression level of miR-150 expression. Moreover, trypan blue exclusion assay was performed to investigate the effect of core protein on cell viability. Results: The gene expression analysis of miR-150 in Huh7 cells showed that endogenous HCV core protein could significantly down regulation of miR-150 when compared to GFP control plasmid and normal cells (P<0.01. Beside, core protein induced no significant proliferative or cytotoxic effects on hepatic cells as determined by trypan blue exclusion assay (P<0.05. Conclusion: Our study suggests that HCV core protein can led to down regulation of miR-150 expression. This data revealed that HCV protein interactions with cell regulatory machinery may contribute to pathogenesis of chronic liver diseases.

Accurate measurement of peripheral blood mononuclear cell concentration using image cytometry to eliminate RBC-induced counting error.

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Chan, Leo Li-Ying; Laverty, Daniel J; Smith, Tim; Nejad, Parham; Hei, Hillary; Gandhi, Roopali; Kuksin, Dmitry; Qiu, Jean

2013-02-28

Peripheral blood mononuclear cells (PBMCs) have been widely researched in the fields of immunology, infectious disease, oncology, transplantation, hematological malignancy, and vaccine development. Specifically, in immunology research, PBMCs have been utilized to monitor concentration, viability, proliferation, and cytokine production from immune cells, which are critical for both clinical trials and biomedical research. The viability and concentration of isolated PBMCs are traditionally measured by manual counting with trypan blue (TB) using a hemacytometer. One of the common issues of PBMC isolation is red blood cell (RBC) contamination. The RBC contamination can be dependent on the donor sample and/or technical skill level of the operator. RBC contamination in a PBMC sample can introduce error to the measured concentration, which can pass down to future experimental assays performed on these cells. To resolve this issue, RBC lysing protocol can be used to eliminate potential error caused by RBC contamination. In the recent years, a rapid fluorescence-based image cytometry system has been utilized for bright-field and fluorescence imaging analysis of cellular characteristics (Nexcelom Bioscience LLC, Lawrence, MA). The Cellometer image cytometry system has demonstrated the capability of automated concentration and viability detection in disposable counting chambers of unpurified mouse splenocytes and PBMCs stained with acridine orange (AO) and propidium iodide (PI) under fluorescence detection. In this work, we demonstrate the ability of Cellometer image cytometry system to accurately measure PBMC concentration, despite RBC contamination, by comparison of five different total PBMC counting methods: (1) manual counting of trypan blue-stained PBMCs in hemacytometer, (2) manual counting of PBMCs in bright-field images, (3) manual counting of acetic acid lysing of RBCs with TB-stained PBMCs, (4) automated counting of acetic acid lysing of RBCs with PI-stained PBMCs

Nosocomial outbreak of Pseudomonas aeruginosa endophthalmitis.

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Mateos, I; Valencia, R; Torres, M J; Cantos, A; Conde, M; Aznar, J

2006-11-01

We describe an outbreak of nosocomial endophthalmitis due to a common source, which was determined to be trypan blue solution prepared in the hospital's pharmacy service. We assume that viable bacteria probably gained access to the trypan blue stock solution during cooling after autoclaving. The temporal cluster of Pseudomonas aeruginosa endophthalmitis was readily perceived on the basis of clinical and microbiological findings, and an exogenous source of contamination was unequivocally identified by means of DNA fingerprinting.

Effect of sirolimus on urinary bladder cancer T24 cell line

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Oliveira Paula A

2009-01-01

Full Text Available Abstract Background Sirolimus is recently reported to have antitumour effects on a large variety of cancers. The present study was performed to investigate sirolimus's ability to inhibit growth in T24 bladder cancer cells. Methods T24 bladder cancer cells were treated with various concentrations of sirolimus. MTT assay was used to evaluate the proliferation inhibitory effect on T24 cell line. The viability of T24 cell line was determined by Trypan blue exclusion analysis. Results Sirolimus inhibits the growth of bladder carcinoma cells and decreases their viability. Significant correlations were found between cell proliferation and sirolimus concentration (r = 0.830; p Conclusion Sirolimus has an anti-proliferation effect on the T24 bladder carcinoma cell line. The information from our results is useful for a better understanding sirolimus's anti-proliferative activity in the T24 bladder cancer cell line.

N-Acetyl-L-Cysteine Affords Protection against Lead-Induced Cytotoxicity and Oxidative Stress in Human Liver Carcinoma (HepG2 Cells

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Paul B. Tchounwou

2007-06-01

Full Text Available Although lead exposure has declined in recent years as a result of change to lead-free gasoline, several epidemiological have pointed out that it represents a medical and public health emergency, especially in young children consuming high amounts of lead-contaminated flake paints. A previous study in our laboratory indicated that lead exposure induces cytotoxicity in human liver carcinoma cells. In the present study, we evaluated the role of oxidative stress in lead-induced toxicity, and the protective effect of the anti-oxidant n-acetyl-l-cysteine (NAC. We hypothesized that oxidative stress plays a role in lead-induced cytotoxicity, and that NAC affords protection against this adverse effect. To test this hypothesis, we performed the MTT [3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide] assay and the trypan blue exclusion test for cell viability. We also performed the thiobarbituric acid test for lipid peroxidation. Data obtained from the MTT assay indicated that NAC significantly increased the viability of HepG2 cells in a dosedependent manner upon 48 hours of exposure. Similar trend was obtained with the trypan blue exclusion test. Data generated from the thiobarbituric acid test showed a significant (p ≤ 0.05 increase of MDA levels in lead nitrate-treated HepG2 cells compared to control cells. Interestingly, the addition of NAC to lead nitrate-treated HepG2 cells significantly decreased cellular content of reactive oxygen species (ROS, as evidenced by the decrease in lipid peroxidation byproducts. Overall, findings from this study suggest that NAC inhibits lead nitrate-induced cytotoxicity and oxidative stress in HepG2 cells. Hence, NAC may be used as a salvage therapy for lead-induced toxicity in exposed persons.

Bovine ocular squamous cell carcinoma: UV sensitivity in lymphocytes

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Lavin, M.F.; Jennings, P.A.; Hughes, D.J. (Queensland Univ., Brisbane (Australia))

1982-05-01

Increased sensitivity to UV light has been demonstrated in Phytohaemagglutinin stimulated lymphocytes from normal and tumour-bearing Hereford cattle when compared to lymphocytes from other breeds. Trypan blue exclusion and inhibition of DNA synthesis were used to determine cell viability. The results obtained from time course and radiation dose experiments demonstrate biphasic survival kinetics. This is indicative of at least two separate cell populations, exhibiting differential sensitivity to UV. The increased sensitivity to UV observed in Herefords may reflect a general sensitivity to UV or alternatively a different cellular constitution in the mitogen stimulated cultures. DNA repair synthesis, measured in the presence of hydroxyurea, was of similar levels in cell cultures from Herefords and one of the control breeds.

Coloración de azul de metileno como alternativa para determinar la viabilidad de larvas libres de Trichinella spiralis Methylene blue test for the determination of viability of free larvae of Trichinella spiralis

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V.R. Randazzo

2010-06-01

Full Text Available El objetivo de la presente investigación fue determinar si la coloración con azul de metileno, de probada utilidad para demostrar la viabilidad de protoescólices de Echinococcus granulosus, puede evidenciar también la viabilidad de larvas libres de Trichinella spiralis. Para ello se utilizaron tres suspensiones de larvas de T. spiralis (M1, M2 y M3, las que fueron expuestas a diferentes condiciones y observadas a distintos tiempos: M1 se expuso a -30 °C y se observó a los 70 días; M2 a 80 °C durante 5 minutos y se observó inmediatamente, y M3 se mantuvo a 4 °C durante todo el experimento, como testigo del 100% de vitalidad. Cada suspensión contenía 500 estadios larvarios libres. Se emplearon 100 μl de solución de azul de metileno 1:10 000 en agua destilada, agregados a igual volumen de suspensión de larvas, y las muestras fueron observadas al microscopio óptico para evaluar la motilidad. Los resultados evidenciaron que cuando las larvas de T. spiralis estaban muertas (M1 y M2, el 100% se coloreaban totalmente de azul en su interior, y las estructuras internas, en relación con la capa quitinosa, aparecían retraídas y algunas fragmentadas. En la suspensión testigo M3 las larvas no se coloreaban, y como prueba adicional de viabilidad se pudo observar su típico movimiento en espiral en el 100% de ellas.The present research was carried out with the purpose of determining whether or not the methylene blue test could also evidence viability of free larvae of Trichinella spiralis, taking into account its usefulness in the determination of viability of protoscolices in Echinococcus granulosus. To this end, three T. spiralis larval suspensions (M1, M2 and M3 were used, each containing 500 free larval stages. A hundred μl of methylene blue solution 1:10000 in distilled water were added to 100 μl of the larval suspension. Larvae were observed under optical microscopy to evaluate motility. M1 was exposed to -30 °C and could be

Safety of intracameral injection of gatifloxacin, levofloxacin on corneal endothelial structure and viability.

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Choi, Jin A; Chung, Sung Kun

2009-10-01

To investigate the safety of intracameral injection of gatifloxacin, levofloxacin in a rabbit model as prophylaxis against endophthalmitis. Twenty-four eyes of New Zealand white rabbits were randomly divided into 3 treatment groups: levofloxacin, gatifloxacin, and balanced salt solution (BSS) control groups. After 100 microL of each was injected into the anterior chamber, endothelial toxicity was evaluated by measuring the central corneal thicknesses and the clinical toxicity scores using a slit-lamp at post-procedure days 3 and 7. The percent of dead cells was determined by vital staining with alizarin red and trypan blue at 7 days after injection. Finally, in each group, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were performed for the evaluation of structural integrity. The toxicity scores were increased at post-procedure days 3 and 7, but the difference among the groups was not statistically significant (P = 0.661, 0.216, respectively). With regard to baseline corneal thickness, only the levofloxacin group exhibited a significant increase from baseline (P = 0.028), whereas the other treatment groups showed no difference from baseline (P = 0.128 in gatifloxacin, 0.161 in BSS group). The mean corneal endothelial damage was 0.81 +/- 0.31% in the levofloxacin group, 0.56 +/- 0.47% in the gatifloxacin group, and 0.53 +/- 0.52% in the BSS group, with no statistically significant difference noted among the groups (P = 0.582). SEM revealed a well-preserved hexagonal endothelial cell mosaic and normal microvilli on the endothelial cell surface in the gatifloxacin and control groups. However, the levofloxacin group showed slightly disintegrated cellular borders. TEM revealed that each group maintained normal intracellular organization, whereas the levofloxacin group exhibited slightly flat cell configuration with irregular folds on the apical cell surface. Intracameral injection of gatifloxacin and levofloxacin was nontoxic in terms of

The effect of various antibiotics on the labelling efficiency of human white blood cells with 111In-oxine

International Nuclear Information System (INIS)

Sinzinger, Helmut; Granegger, Susanne

1988-01-01

Earlier clinical studies revealed that in patients suffering from chronic osteomyelitis undergoing antibiotic therapy the white blood cell scanning missed the right diagnosis in 40% of cases, whereas all the acute untreated cases were imaged correctly. Thus, it was suspected that an impaired labelling efficiency and white blood cell function might have been causative. Retrospective analysis of labelling efficiency exhibited no difference between patients on antibiotics and those not on antibiotics. Prospective cellular viability testing in 81 patients, 71 of whom were on various antibiotics, using latex particles (phagocytosis) and the Trypan blue exclusion test, did not reveal any different function behaviour either. Examining the labelling efficiency (after 111 In-oxine and 111 In-oxine-sulphate labelling), recovery, half-life and viability of white blood cells of 107 patients undergoing therapy with various antibiotics as compared to controls, it becomes evident that the antibiotic therapy is not causative of the clinical difference observed. (author)

Comparison of 111In-oxine and 111In-acetylacetone for the labeling of cells: in vivo and in vitro biological testing

International Nuclear Information System (INIS)

Mathias, C.J.; Heaton, W.A.; Welch, M.J.

1981-01-01

Several complexes of indium were compared as cell labels: indium-111-acetylacetone, indium-111-oxine, and indium-111-chloride complexed with 8-hydroxyquinoline (oxine) immediately prior to use. In labeling with acetylacetone, it was shown that the labeling efficiency is directly proportional to the amount of acetylacetone present, but the cell viability (as measured by in vitro aggregation studies), is inversely proportional to the amount of acetylacetone present. Biological studies were carried out in dogs using indium-111-labeled platelets; survival times and recovery values obtained with platelets labeled using all three techniques were similar. The same solutions were also used to label white blood cells; labeling efficiencies of greater than 80% were obtained in all cases, and the viability (as measured by trypan blue exclusion) was high in all cases. Chemotactic ability of the white cells labeled with indium-111-oxine is higher than that of unlabeled control cells; however, cells labeled with indium-111-acetylacetone were the same as the unlabeled control cells. (author)

Bovine ocular squamous cell carcinoma: UV sensitivity in lymphocytes

International Nuclear Information System (INIS)

Lavin, M.F.; Jennings, P.A.; Hughes, D.J.

1982-01-01

Increased sensitivity to UV light has been demonstrated in Phytohaemagglutinin stimulated lymphocytes from normal and tumour-bearing Hereford cattle when compared to lymphocytes from other breeds. Trypan blue exclusion and inhibition of DNA synthesis were used to determine cell viability. The results obtained from time course and radiation dose experiments demonstrate biphasic survival kinetics. This is indicative of at least two separate cell populations, exhibiting differential sensitivity to UV. The increased sensitivity to UV observed in Herefords may reflect a general sensitivity to UV or alternatively a different cellular constitution in the mitogen stimulated cultures. DNA repair synthesis, measured in the presence of hydroxyurea, was of similar levels in cell cultures from Herefords and one of the control breeds. (author)

Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma

International Nuclear Information System (INIS)

Dyberg, Cecilia; Papachristou, Panagiotis; Haug, Bjørn Helge; Lagercrantz, Hugo; Kogner, Per; Ringstedt, Thomas; Wickström, Malin; Johnsen, John Inge

2016-01-01

The non-canonical Wnt/Planar cell polarity (PCP) signaling pathway is a major player in cell migration during embryonal development and has recently been implicated in tumorigenesis. Transfections with cDNA plasmids or siRNA were used to increase and suppress Prickle1 and Vangl2 expression in neuroblastoma cells and in non-tumorigenic cells. Cell viability was measured by trypan blue exclusion and protein expression was determined with western blotting. Transcriptional activity was studied with luciferase reporter assay and mRNA expression with real-time RT-PCR. Immunofluorescence stainings were used to study the effects of Vangl2 overexpression in non-tumorigenic embryonic cells. Statistical significance was tested with t-test or one-way ANOVA. Here we show that high expression of the PCP core genes Prickle1 and Vangl2 is associated with low-risk neuroblastoma, suppression of neuroblastoma cell growth and decreased Wnt/β-catenin signaling. Inhibition of Rho-associated kinases (ROCKs) that are important in mediating non-canonical Wnt signaling resulted in increased expression of Prickle1 and inhibition of β-catenin activity in neuroblastoma cells. In contrast, overexpression of Vangl2 in MYC immortalized neural stem cells induced accumulation of active β-catenin and decreased the neural differentiation marker Tuj1. Similarly, genetically modified mice with forced overexpression of Vangl2 in nestin-positive cells showed decreased Tuj1 differentiation marker during embryonal development. Our experimental data demonstrate that high expression of Prickle1 and Vangl2 reduce the growth of neuroblastoma cells and indicate different roles of PCP proteins in tumorigenic cells compared to normal cells. These results suggest that the activity of the non-canonical Wnt/PCP signaling pathway is important for neuroblastoma development and that manipulation of the Wnt/PCP pathway provides a possible therapy for neuroblastoma. The online version of this article (doi:10.1186/s

Comparison of Cell Viability and Embryoid Body Size of Two Embryonic Stem Cell Lines After Different Exposure Times to Bone Morphogenetic Protein 4

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Nehleh Zarei Fard

2015-03-01

Full Text Available Background: Activation of bone morphogenetic protein 4 (BMP4 signaling pathway in embryonic stem (ES cells plays an important role in controlling cell proliferation, differentiation, and apoptosis. Adverse effects of BMP4 occur in a time dependent manner; however, little is known about the effect of different time exposure of this growth factor on cell number in culture media. In this study, we investigated the role of two different exposure times to BMP4 in cell viability, embryoid body (EB, size, and cavitation of ES cells. Methods: Embryonic stem cells (R1 and B1 lines were released from the feeder cell layers and were cultured using EBs protocol by using the hanging drop method and monolayer culture system. The cells were cultured for 5 days with 100 ng/mL BMP4 from the beginning (++BMP4 or after 48 h (+BMP4 of culture and their cell number were counted by trypan blue staining. The data were analyzed using non-parametric two-tailed Mann-Whitney test. P<0.05 was considered as significant. Results: In EB culture protocol, cell number significantly decreased in +BMP4 culture condition with greater cavity size compared to the ++BMP4 condition at day 5 (P=0.009. In contrast, in monolayer culture system, there was no significant difference in the cell number between all groups (P=0.91. Conclusion: The results suggest that short-term exposure of BMP4 is required to promote cavitation in EBs according to lower cell number in +BMP4 condition. Different cell lines showed different behavior in cavitation formation.

Effect of temperature on immunocompetence of the blue mussel (Mytilus edulis

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Alexandre Beaudry

2016-08-01

Full Text Available The blue mussel is a filter-feeding bivalve commonly used in ecotoxicological monitoring as a sentinel species. Due to climate change and the increase of temperature expected in marine environment, it is important to anticipate potential impacts on this species. The aim of this study was to investigate the immunocompetence of blue mussels acclimated to different temperatures and on the effects of increasing temperatures (5, 10 and 20°C. Different indices and gonad maturation stages were also determined throughout the experiments. Cell viability, phagocytosis, serum lysozyme activity and cyclooxygenase (COX activity were evaluated as immune parameters. The cellular immunity was also evaluated after hemocytes exposure to various cadmium concentrations in vitro. The results obtained demonstrate modulation of hemocyte viability and the ability of these cells to phagocytize in absence of contaminants. After the exposure to cadmium, hemocytes showed greater viability at 5°C while maintaining a higher phagocytic competence. In addition, the lysozyme activity stayed stable at all tested temperatures, contrary to that of COX, which increased when the mussels were maintained at 20°C. The evaluation of indices demonstrated no reduction of general conditions during all the experiment despite the increase of temperature and the reduction of the digestive gland weight. Moreover, the lack of food does not affect gonad maturation and the spawning process.

Thermochemical ablation therapy of VX2 tumor using a permeable oil-packed liquid alkali metal.

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Ziyi Guo

Full Text Available Alkali metal appears to be a promising tool in thermochemical ablation, but, it requires additional data on safety is required. The objective of this study was to explore the effectiveness of permeable oil-packed liquid alkali metal in the thermochemical ablation of tumors.Permeable oil-packed sodium-potassium (NaK was prepared using ultrasonic mixing of different ratios of metal to oil. The thermal effect of the mixture during ablation of muscle tissue ex vivo was evaluated using the Fluke Ti400 Thermal Imager. The thermochemical effect of the NaK-oil mixture on VX2 tumors was evaluated by performing perfusion CT scans both before and after treatment in 10 VX2 rabbit model tumors. VX2 tumors were harvested from two rabbits immediately after treatment to assess their viability using trypan blue and hematoxylin and eosin (H.E. staining.The injection of the NaK-oil mixture resulted in significantly higher heat in the ablation areas. The permeable oil controlled the rate of heat released during the NaK reaction with water in the living tissue. Perfusion computed tomography and its parameter map confirmed that the NaK-oil mixture had curative effects on VX2 tumors. Both trypan blue and H.E. staining showed partial necrosis of the VX2 tumors.The NaK-oil mixture may be used successfully to ablate tumor tissue in vivo. With reference to the controlled thermal and chemical lethal injury to tumors, using a liquid alkali in ablation is potentially an effective and safe method to treat malignant tumors.

Mechanism of H₂O₂-induced oxidative stress regulating viability and biocontrol ability of Rhodotorula glutinis.

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Chen, Jian; Li, Boqiang; Qin, Guozheng; Tian, Shiping

2015-01-16

The use of antagonistic yeasts to control postharvest pathogens is a promising alternative to fungicides. The effectiveness of the antagonists against fungal pathogens is greatly dependent on their viability, which is usually mediated by reactive oxygen species (ROS). Here, we investigated the effects of H₂O₂-induced oxidative stress on the viability and biocontrol efficacy of Rhodotorula glutinis and, using flow cytometric analysis, observed the changes of ROS accumulation and apoptosis in the yeast cells with or without H₂O₂ treatment. We found that the viability of R. glutinis decreased in a time- and dose-dependent manner under H₂O₂-induced oxidative stress. Compared to the control, yeast cells exposed to oxidative stress exhibited more accumulation of ROS and higher levels of protein oxidative damage, but showed lower efficacy for biocontrol of Penicillium expansum causing blue mold rot on peach fruit. The results indicate that apoptosis is a main cause of the cell viability loss in R. glutinis, which is attributed to ROS accumulation under oxidative stress. These findings offer a plausible explanation that oxidative stress affects biocontrol efficacy of R. glutinis via regulating its viability and cell apoptosis. Copyright © 2014 Elsevier B.V. All rights reserved.

Electron Microscopic Changes of Rabbit Retina after Chromovitrectomy Using Combined Dyes (Experimental Study

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B. M. Aznabaev

2018-01-01

Full Text Available Purpose: to evaluate on experimental model electron-microscopic changes of rabbit retina after staining of the posterior eye segmentwith combined dyes based on Trypan blue and Brilliant blue G for the assessment of their safety. Methods. The study was performed onChinchilla breed rabbits. Combined dyes based on Trypan blue and Brilliant blue were used: MembraneBlue-Dual (DORC, Netherlandsand “Staining solution for ophthalmic surgery” (JCS “Optimedservis”, Russia. Standard three-port vitrectomy technique was used. After vitreous removal dyes were injected in vitreous cavity and exposed for 10 seconds and then removed. The vitreous cavity was filled by a balanced salt solution. An electron-microscopic evaluation was performed on 5, 14 and 30 days after surgery. Eyes were enucleated in 20 minutes after animal was killed by air embolization. Intact eyes were used as a control, all samples were prepared in same сonditions. The damage of the retina architectonics and the presence of intracellular inclusions were evaluated. Results. The staged character of pathomorphological changes was revealed. On the 5th day moderate edema and hydropic dystrophy of neurons were registered. On the 14th day, there was no negative dynamics. On day 30, the signs of edema and dystrophy of neurons practically disappeared, which may indicate a fundamental reversibility of the registered changes. Conclusion. Investigated dyes for staining intraocular structures based on Trypan blue and Brilliant blue did not cause significant histomorphological changes and toxic effects on retinal cell structures. Detected electron microscopic changes were insignificant, had reversible character and could be mostly caused by a surgical injury.

Cell Uptake and Validation of Novel PECs for Biomedical Applications.

Science.gov (United States)

Palamà, Ilaria E; Musarò, Mariarosaria; Coluccia, Addolorata M L; D'Amone, Stefania; Gigli, Giuseppe

2011-01-01

This pilot study provides the proof of principle for biomedical application of novel polyelectrolyte complexes (PECs) obtained via electrostatic interactions between dextran sulphate (DXS) and poly(allylamine hydrochloride) (PAH). Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that DXS/PAH polyelectrolyte complexes were Monodispersed with regular rounded-shape features and average diameters of 250 nm at 2 : 1 weight ratios of DXS/PAH. Fluorescently labelled DXS and fluorescein-isothiocyanate- (FITC-)conjugate DXS were used to follow cell uptake efficiency of PECs and biodegradability of their enzymatically degradable DXS-layers by using confocal laser scanning microscopy (CLSM). Moreover, quantitative MTT and Trypan Blue assays were employed to validate PECs as feasible and safe nanoscaled carriers at single-cell level without adverse effects on metabolism and viability.

Cell Uptake and Validation of Novel PECs for Biomedical Applications

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Ilaria E. Palamà

2011-01-01

Full Text Available This pilot study provides the proof of principle for biomedical application of novel polyelectrolyte complexes (PECs obtained via electrostatic interactions between dextran sulphate (DXS and poly(allylamine hydrochloride (PAH. Scanning electron microscopy (SEM and atomic force microscopy (AFM showed that DXS/PAH polyelectrolyte complexes were Monodispersed with regular rounded-shape features and average diameters of 250 nm at 2 : 1 weight ratios of DXS/PAH. Fluorescently labelled DXS and fluorescein-isothiocyanate- (FITC-conjugate DXS were used to follow cell uptake efficiency of PECs and biodegradability of their enzymatically degradable DXS-layers by using confocal laser scanning microscopy (CLSM. Moreover, quantitative MTT and Trypan Blue assays were employed to validate PECs as feasible and safe nanoscaled carriers at single-cell level without adverse effects on metabolism and viability.

The effects of monosodium urate monohydrate crystals on chondrocyte viability and function: implications for development of cartilage damage in gout.

Science.gov (United States)

Chhana, Ashika; Callon, Karen E; Pool, Bregina; Naot, Dorit; Gamble, Gregory D; Dray, Michael; Pitto, Rocco; Bentley, Jarome; McQueen, Fiona M; Cornish, Jillian; Dalbeth, Nicola

2013-12-01

Cartilage damage is frequently observed in advanced destructive gout. The aim of our study was to investigate the effects of monosodium urate monohydrate (MSU) crystals on chondrocyte viability and function. The alamarBlue assay and flow cytometry were used to assess the viability of primary human chondrocytes and cartilage explants following culture with MSU crystals. The number of dead chondrocytes in cartilage explants cultured with MSU crystals was quantified. Real-time PCR was used to determine changes in the relative mRNA expression levels of chondrocytic genes. The histological appearance of cartilage in joints affected by gout was also examined. MSU crystals rapidly reduced primary human chondrocyte and cartilage explant viability in a dose-dependent manner (p gout, normal cartilage architecture was lost, with empty chondrocyte lacunae observed. MSU crystals have profound inhibitory effects on chondrocyte viability and function. Interactions between MSU crystals and chondrocytes may contribute to cartilage damage in gout through reduction of chondrocyte viability and promotion of a catabolic state.

Immobilization of Electroporated Cells for Fabrication of Cellular Biosensors: Physiological Effects of the Shape of Calcium Alginate Matrices and Foetal Calf Serum

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Nikos Katsanakis

2009-01-01

Full Text Available In order to investigate the physiological effect of transfected cell immobilization in calcium alginate gels, we immobilized electroporated Vero cells in gels shaped either as spherical beads or as thin membrane layers. In addition, we investigated whether serum addition had a positive effect on cell proliferation and viability in either gel configuration. The gels were stored for four weeks in a medium supplemented or not with 20% (v/v foetal calf serum. Throughout a culture period of four weeks, cell proliferation and cell viability were assayed by optical microscopy after provision of Trypan Blue. Non-elaborate culture conditions (room temperature, non-CO2 enriched culture atmosphere were applied throughout the experimental period in order to evaluate cell viability under less than optimal storage conditions. Immobilization of electroporated cells was associated with an initially reduced cell viability, which was gradually increased. Immobilization was associated with maintenance of cell growth for the duration of the experimental period, whereas electroporated cells essentially died after a week in suspension culture. Considerable proliferation of immobilized cells was observed in spherical alginate beads. In both gel configurations, addition of serum was associated with increased cell proliferation. The results of the present study could contribute to an improvement of the storability of biosensors based on electroporated, genetically or membrane-engineered cells.

Changes in the corneal Na-K ATPase levels in eyes stored in moist chamber at 4°C

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Devi B

1996-01-01

Full Text Available This report deals with a chronological measurement of Na-K ATPase enzyme activity in human and bovine corneas stored in a moist chamber at 4°C. Paired human and bovine eyes were sterilized by the standard eye bank procedure and stored up to 6 days. At the desired time, the corneal endothelium was assayed for Na-K ATPase activity. The protein content of each tissue sample was also determined. In a parallel set of experiments, the viability of identical stored corneas was determined by trypan blue and alizarin red staining technique, and morphometric analysis was done to quantify the extent of the corneal endothelial damage. The human corneas showed that there was a significant progressive decrease in the Na-K ATPase activity as the storage time increased. The decrease was related to morphological endothelial damage.

Effects of hydrostatic pressure and supercritical carbon dioxide on the viability of Botryococcus braunii algae cells.

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Yildiz-Ozturk, Ece; Ilhan-Ayisigi, Esra; Togtema, Arnoud; Gouveia, Joao; Yesil-Celiktas, Ozlem

2018-05-01

In bio-based industries, Botryococcus braunii is identified as a potential resource for production of hydrocarbons having a wide range of applications in chemical and biopolymer industries. For a sustainable production platform, the algae cultivation should be integrated with downstream processes. Ideally the algae are not harvested, but the product is isolated while cultivation and growth is continued especially if the doubling time is slow. Consequently, hydrocarbons can be extracted while keeping the algae viable. In this study, the effects of pressure on the viability of B. braunii cells were tested hydrostatically and under supercritical CO 2 conditions. Viability was determined by light microscopy, methylene blue uptake and by re-cultivation of the algae after treatments to follow the growth. It was concluded that supercritical CO 2 was lethal to the algae, whereas hydrostatic pressure treatments up to 150 bar have not affected cell viability and recultivation was successful. Copyright © 2018 Elsevier Ltd. All rights reserved.

Rabbit model of intervertebral disc degeneration by external compression device characterized by X-ray, MRI, histology, and cell viability

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Ismail Ismail

2006-12-01

Full Text Available Appropriate experimental animal models, which mimic the degenerative process occurring in human intervertebral disc (IVD breakdown and can be used for new treatment studies such as tissue engineering or disc distraction are lacking. We studied the external compression device that used by Kroeber et al to create intervertebral disc degeneration in rabbit model characterized by X-ray, MRI, Histology, and Cell Viability. Ten NZW rabbit were randomly assigned to one of five groups. Intervertebral disc VL4-L5 are compressed using an external loading device, 1.9 MPa. First group rabbit are loaded for 14 days, second loaded for 28 days, thirth group are loaded for 14 days, and unloaded for 14 days, fourth group loaded for 28 days and unloaded for 28 days. The fifth group, rabbits underwent a sham operation. Additional, rabbits were used as sample for cell viability study. In disc height : sample in group one have biggest decreasing of disc height, that is 23.9 unit. In MRI assessment, the worst grade is grade 3. In histological score, the worst group is group three (58.69, and the best is group 4 (45.69. Group one have the largest dead cell, that are 403.5, and the smallest is group four (124.75. Trypan blue staining showed that group four have better viable cell (91.1 compare than group three (86.4. The study conclude disc degeneration can be created by external axial loading for 14 days in rabbit intervertebral disc. Duration of 28 days unloading gave better result for cells to recover. (Med J Indones 2006; 15:199-207  Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Keywords: Rabbit model –intervertebral disc degeneration- external compression device-X-ray, MRI, Histology, and Cell viabilty /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso

Viability Theory

CERN Document Server

Aubin, Jean-Pierre; Saint-Pierre, Patrick

2011-01-01

Viability theory designs and develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty that are found in many domains involving living beings, from biological evolution to economics, from environmental sciences to financial markets, from control theory and robotics to cognitive sciences. It involves interdisciplinary investigations spanning fields that have traditionally developed in isolation. The purpose of this book is to present an initiation to applications of viability theory, explai

Hyaluronic acid effect on adipose-derived stem cells. Biological in vitro evaluation.

Science.gov (United States)

Moreno, A; Martínez, A; Olmedillas, S; Bello, S; de Miguel, F

2015-01-01

To evaluate the in vitro effects of hyaluronic acid (HA) on adipose-derived stem cells (ASC) in order to consider the possibility of their combined used in the treatment of knee arthrosis. The ASC cells were grown both in the presence and absence of AH, and several studies were carried out: proliferation (WST8) and cell viability studies (Alamar Blue® and Trypan Blue), possible chondrogenic differentiation (collagen type 2 expression) by RT-PCR, AH receptor expression (CD44) by flow cytometry and RT-QPCR, and expression of inflammatory and anti-inflammatory factors (IL-6, TGFß, IL-10) by RT-QPCR. The number of ASC significantly increased after 7 days with HA (158±39%, p <0.05). Additionally, the cell viability of the ASC treated with HA after 1, 3, 5 and 7 days was similar to that of the control cells, being considered non-toxic. There were no changes observed in the expression of CD44 and chondrogenic differentiation. TGFß expression was not modified after AH treatment, but there was a 4-fold decrease in IL-6 expression and IL-10 expression increased up to 2-fold compared to control cells. Hyaluronic acid favours ASC proliferation without causing cellular toxicity, and inducing an anti-inflammatory profile in these cells. Hyaluronic acid appears to be a suitable vehicle for the intra-articular administration of mesenchymal stem cells. Copyright © 2014 SECOT. Published by Elsevier Espana. All rights reserved.

Viability, Apoptosis, Proliferation, Activation, and Cytokine Secretion of Human Keratoconus Keratocytes after Cross-Linking

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Xuefei Song

2015-01-01

Full Text Available Purpose. The purpose of this study was to determine the impact of cross-linking (CXL on viability, apoptosis, proliferation, activation, and cytokine secretion of human keratoconus (KC keratocytes, in vitro. Methods. Primary KC keratocytes were cultured in DMEM/Ham’s F12 medium supplemented with 10% FCS and underwent UVA illumination (370 nm, 2 J/cm2 during exposure to 0.1% riboflavin and 20% Dextran in PBS. Twenty-four hours after CXL, viability was assessed using Alamar blue assay; apoptosis using APO-DIRECT Kit; proliferation using ELISA-BrdU kit; and CD34 and alpha-smooth muscle actin (α-SMA expression using flow cytometry. Five and 24 hours after CXL, FGFb, HGF, TGFβ1, VEGF, KGF, IL-1β, IL-6, and IL-8 secretion was measured using enzyme-linked-immunoabsorbent assay (ELISA. Results. Following CXL, cell viability and proliferation decreased (P0.06. Five hours after CXL, FGFb secretion increased significantly (P=0.037; however no other cytokine secretion differed significantly from controls after 5 or 24 hours (P>0.12. Conclusions. Cross-linking decreases viability, triggers apoptosis, and inhibits proliferation, without an impact on multipotent hematopoietic stem cell transformation and myofibroblastic transformation of KC keratocytes. CXL triggers FGFb secretion of KC keratocytes transiently (5 hours, normalizing after 24 hours.

Rapid, high-efficiency labeling of leukocytes with In-111 after hemolytic removal of erythrocytes

International Nuclear Information System (INIS)

Karesh, S.M.; Henkin, R.E.

1985-01-01

During the labeling of leukocytes with Indium-111, conventional methodology involves separation and washing to remove red cells. This technique results in the loss of a significant number of leukocytes. Citrated whole blood of ten normal volunteers was studied for an alternate labeling method following sedimentation for 30 to 45 minutes and low speed centrifugation of the leukocyte-rich plasma. The average labeling for these ten volunteers by Indium-111 was 90% versus 60% by the older technique. Viability as measured by the trypan blue exclusion test was greater than 95%, WBC losses were essentially zero, and no WBC clumping was observed. Eighteen patients referred for leukocyte imaging were studied by this method. In this patient population, there was 91% labeling with viability greater than 95% and no evidence of clumping. Less than 5% RBC's were noted in any lot. Indium-111 WBC activity 20 minutes post injection averaged 79% of whole blood activity. This modification results in decreased losses of white cells, reduces preparation time to less than 2 hours, and significantly improves the labeling efficiency of the final product. Liver/spleen ratios and image quality were unchanged from the original method

Accelerated stem cell labeling with ferucarbotran and protamine

Energy Technology Data Exchange (ETDEWEB)

Golovko, Daniel M.; Henning, Tobias; Bauer, Jan S. [Department of Radiology and Biomedical Imaging, University of California San Francisco, San Francisco, CA (United States); Settles, Marcus; Rummeny, Ernst J. [Technical University Munich, Department of Radiology, Munich (Germany); Frenzel, Thomas [Bayer Schering Pharma AG, Berlin (Germany); Mayerhofer, Artur [Ludwig-Maximilians-Universitaet, Institute of Cell Biology, Munich (Germany); Daldrup-Link, Heike E. [Department of Radiology and Biomedical Imaging, University of California San Francisco, San Francisco, CA (United States); UCSF Medical Center, Contrast Agent Research Group, Department of Radiology, San Francisco, CA (United States)

2010-03-15

To develop and characterize a clinically applicable, fast and efficient method for stem cell labeling with ferucarbotran and protamine for depiction with clinical MRI. The hydrodynamic diameter, zeta potential and relaxivities of ferucarbotran and varying concentrations of protamine were measured. Once the optimized ratio was found, human mesenchymal stem cells (MSCs) were labeled at varying incubation times (1-24 h). Viability was assessed via Trypan blue exclusion testing. 150,000 labeled cells in Ficoll solution were imaged with T1-, T2- and T2*-weighted sequences at 3 T, and relaxation rates were calculated. Varying the concentrations of protamine allows for easy modification of the physicochemical properties. Simple incubation with ferucarbotran alone resulted in efficient labeling after 24 h of incubation while assisted labeling with protamine resulted in similar results after only 1 h. Cell viability remained unaffected. R2 and R2* relaxation rates were drastically increased. Electron microscopy confirmed intracellular iron oxide uptake in lysosomes. Relaxation times correlated with results from ICP-AES. Our results show internalization of ferucarbotran can be accelerated in MSCs with protamine, an approved heparin antagonist and potentially clinically applicable uptake-enhancing agent. (orig.)

Yeast viability and concentration analysis using lens-free computational microscopy and machine learning

Science.gov (United States)

Feizi, Alborz; Zhang, Yibo; Greenbaum, Alon; Guziak, Alex; Luong, Michelle; Chan, Raymond Yan Lok; Berg, Brandon; Ozkan, Haydar; Luo, Wei; Wu, Michael; Wu, Yichen; Ozcan, Aydogan

2017-03-01

Research laboratories and the industry rely on yeast viability and concentration measurements to adjust fermentation parameters such as pH, temperature, and pressure. Beer-brewing processes as well as biofuel production can especially utilize a cost-effective and portable way of obtaining data on cell viability and concentration. However, current methods of analysis are relatively costly and tedious. Here, we demonstrate a rapid, portable, and cost-effective platform for imaging and measuring viability and concentration of yeast cells. Our platform features a lens-free microscope that weighs 70 g and has dimensions of 12 × 4 × 4 cm. A partially-coherent illumination source (a light-emitting-diode), a band-pass optical filter, and a multimode optical fiber are used to illuminate the sample. The yeast sample is directly placed on a complementary metal-oxide semiconductor (CMOS) image sensor chip, which captures an in-line hologram of the sample over a large field-of-view of >20 mm2. The hologram is transferred to a touch-screen interface, where a trained Support Vector Machine model classifies yeast cells stained with methylene blue as live or dead and measures cell viability as well as concentration. We tested the accuracy of our platform against manual counting of live and dead cells using fluorescent exclusion staining and a bench-top fluorescence microscope. Our regression analysis showed no significant difference between the two methods within a concentration range of 1.4 × 105 to 1.4 × 106 cells/mL. This compact and cost-effective yeast analysis platform will enable automatic quantification of yeast viability and concentration in field settings and resource-limited environments.

In vitro anti-inflammatory and phytochemical properties of crude ...

African Journals Online (AJOL)

pc owner

2013-09-25

Sep 25, 2013 ... RPMI 1640 medium, fetal bovine serum (FBS), trypan blue,. Histopaque-1077, penicillin G ..... biochemical parameters of male guinea pigs. J. Appl. Sci. Environ. ... Rheumatoid arthritis: Inflammation and bone loss. Wien. Med.

Differential responses of cells from human skin keratinocyte and bovine mammary epithelium to attack by pore-forming Staphylococcus aureus alpha-toxin.

Science.gov (United States)

Suriyaphol, Gunnaporn; Sarikaputi, Meena; Suriyaphol, Prapat

2009-11-01

Human skin keratinocytes HaCat attacked by Staphylococcus aureus alpha-toxin showed a transient drop of cellular ATP levels whereas in toxin-perforated bovine mammary epithelial cells (BMEC), the ATP levels dropped more slowly. Morphologically, during the ATP level depletion, HaCat cell developed a spacious intracellular vacuole together with the transient influx of trypan blue. WST-1 signal, which tested the function of mitochondrial enzyme in viable cells, also decreased concomitantly. On the other hand, BMEC excluded trypan blue and vacuolation was not observed throughout the experiment. We conclude that mammary epithelial cells resist the toxin better than keratinocytes. This is the first report showing that alpha-toxin enhances transient membrane permeability to large molecules, temporary vacuole formation and the transient defect of mitochondrial enzyme in viable cells without cell lysis.

Effect of albumin and polyvinyl alcohol on the vitality, motility and acrosomal integrity of canine spermatozoa incubated in vitro.

Science.gov (United States)

Risopatrón, J; Catalán, S; Miska, W; Schill, W-B; Sánchez, R

2002-12-01

Sperm culture media used for in vitro fertilization (IVF) procedures are important factors concerning the viability, motility and acrosomal integrity of spermatozoa. The aim of this study was to investigate the effects of three different sperm diluting media, tissue culture medium (TCM-199), sperm culture medium (Sp-TALP) and human tubular fluid (HTF) supplemented with varying concentrations of bovine serum albumin (1, 4 and 6%) or polyvinyl alcohol (0.8%) on the acrosomal integrity, motility and viability of canine spermatozoa. Ejaculates collected from four dogs were diluted in all media and spermatozoa were separated from seminal plasma by the swim-up technique. Sperm progressive motility was assessed using a phase contrast microscope. Viability and acrosomal integrity were evaluated using a dual stain technique (Giemsa-Trypan blue). The results demonstrated that the number of live canine spermatozoa was similar in culture media supplemented or not supplemented with macromolecules. A minimal concentration of albumin (1%) in the three media showed similar effects on vitality, motility and acrosomal integrity, as had higher concentrations (4 and 6%). The percentage of acrosome-intact spermatozoa was markedly higher after HTF (94.1%) than after TCM-199 (70.1%) or Sp-TALP (71.0%) without supplementation. It is concluded that serum bovine albumin, irrespective of the concentration, preserved sperm viability and function, and HTF is the most suitable medium for preserving the acrosome in canine spermatozoa prepared for in vitro manipulation through short incubation.

Effects of in vitro Brevetoxin Exposure on Apoptosis and Cellular Metabolism in a Leukemic T Cell Line (Jurkat

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John W. Sleasman

2008-06-01

Full Text Available Harmful algal blooms (HABs of the toxic dinoflagellate, Karenia brevis, produce red tide toxins, or brevetoxins. Significant health effects associated with red tide toxin exposure have been reported in sea life and in humans, with brevetoxins documented within immune cells from many species. The objective of this research was to investigate potential immunotoxic effects of brevetoxins using a leukemic T cell line (Jurkat as an in vitro model system. Viability, cell proliferation, and apoptosis assays were conducted using brevetoxin congeners PbTx-2, PbTx-3, and PbTx-6. The effects of in vitro brevetoxin exposure on cell viability and cellular metabolism or proliferation were determined using trypan blue and MTT (1-(4,5-dimethylthiazol-2-yl-3,5- diphenylformazan, respectively. Using MTT, cellular metabolic activity was decreased in Jurkat cells exposed to 5 - 10 μg/ml PbTx-2 or PbTx-6. After 3 h, no significant effects on cell viability were observed with any toxin congener in concentrations up to 10 μg/ml. Viability decreased dramatically after 24 h in cells treated with PbTx-2 or -6. Apoptosis, as measured by caspase-3 activity, was significantly increased in cells exposed to PbTx-2 or PbTx-6. In summary, brevetoxin congeners varied in effects on Jurkat cells, with PbTx-2 and PbTx-6 eliciting greater cellular effects compared to PbTx-3.

Assessment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage.

Science.gov (United States)

Huo, Li-Jun; Ma, Xing-Hong; Yang, Zeng-Ming

2002-10-15

The purpose of this study was to assess sperm quality in extended boar semen during in vitro storage in order to determine which extender should be used and how long boar semen can be stored. Freshly ejaculated boar semen was diluted with equal volumes of Beltsville thaw solution (BTS), Androhep, KIEV or Zorlesco extenders and stored at 17 degrees C for up to 15 days. Sperm quality was evaluated by examining viability using SYBR-14/PI and Hoechst 33258 staining, mitochondrial activity using 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) staining, acrosome intactness by Coomassie blue staining, and capacitation status by chlortetracycline (CTC) staining. There were over 50% viable spermatozoa in boar semen extended with Zorlesco and Androhep extenders on Day 13 of storage. The percentage of JC-1-stained spermatozoa was 53.8 +/- 2.1% for Zorlesco and 57.7 +/- 1.60% for Androhep extenders on Day 13 of storage. The percentage of acrosome-intact spermatozoa detected by Coomassie blue staining was higher than that in the SYBR-14PI-, Hoechst 33258-, and JC-1-stained samples in our study. The results from SYBR-14/PI, Hoechst 33258, JC-1, and Coomassie blue staining were highly correlated (r > or = 0.9461). There were less than 15% capacitated spermatozoa in the semen extended with BTS, Androhep and Zorlesco extenders during 9 days of storage. However, most viable boar spermatozoa became capacitated by Day 13 of storage. The rank order of four extenders for maintaining sperm viability and mitochondrial activity was as follows: Androhep, Zorlesco, BTS, KIEV.

TiO{sub 2}/PCL hybrid materials synthesized via sol–gel technique for biomedical applications

Energy Technology Data Exchange (ETDEWEB)

Catauro, M., E-mail: michelina.catauro@unina2.it [Department of Industrial and Information Engineering, Second University of Naples, Via Roma 29, 81031 Aversa (Italy); Bollino, F.; Papale, F. [Department of Industrial and Information Engineering, Second University of Naples, Via Roma 29, 81031 Aversa (Italy); Marciano, S.; Pacifico, S. [Department Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Via Vivaldi 43, 81100 Caserta (Italy)

2015-02-01

The aim of the present work has been the synthesis of organic/inorganic hybrid materials based on titanium dioxide and poly(ε-caprolactone) (PCL) to be used in the biomedical field. Several materials have been synthesized using sol–gel methods by adding different amounts of polymer to the inorganic sol. The obtained gels have been characterized using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). The FT-IR data allowed us to hypothesize that the structure formed was that of an interpenetrating network, realized by hydrogen bonds between Ti-OH groups in the sol–gel intermediate species and carbonyl groups in the polymer repeating units. SEM and AFM analyses highlighted that the obtained materials were nanostructurated hybrids. To evaluate the biological properties of the hybrids, their bioactivity and cytotoxicity were investigated as a function of the PCL amount. The bioactivity of the synthesized systems was proven by the formation of a hydroxyapatite layer on the surface of samples soaked in a fluid simulating human blood plasma (SBF). MTT cytotoxicity tests and Trypan Blue dye exclusion tests were carried out exposing NIH-3T3 mouse embryonic fibroblasts for 24 and 48 h to extracts from the investigated hybrid materials. The results showed that all the hybrids had a non-cytotoxic effect on target cells. - Highlights: • TiO{sub 2}/PCL hybrids were obtained by the sol–gel process for biomedical applications. • Synthesized materials were found to be first-class hybrid nanocomposites. • Hybrids appear to be bioactive, a fundamental characteristic for osseointegration. • MTT and Trypan Blue viability test show that the materials are biocompatible. • The organic phase is able to modulate the biocompatibility of the materials.

Effects of SO/sub 2/ or NOx on toxic and genotoxic properties of chemical carcinogens. II. Short term in vivo studies

Energy Technology Data Exchange (ETDEWEB)

Pool, B L; Brendler, S; Klein, R G; Monarca, S; Pasquini, R; Schmezer, P; Zeller, W J

1988-07-01

Short term in vivo studies were performed to study biological effects of the common air pollutants SO2 or NOx and their influence on the genotoxic activities of nitrosamines. Hepatocytes and lung cells were isolated from Sprague-Dawley rats which had inhaled 50 p.p.m. of SO2 or NOx for 2 weeks. After incubating the cells for 1 h, genotoxicity was determined in hepatocytes by measuring DNA single-strand breaks induced by N-nitroso-acetoxymethylmethylamine, N-nitrosodimethylamine and N-nitrosomethylbenzylamine. Parameters of toxicity (trypan blue exclusion and leakage of serum enzymes) were determined in both liver and lung cells also following 1 h incubation. The activities of aryl hydrocarbon hydroxylase (AHH), nitrosodimethylamine demethylase (NDMA-D) and glutathione-S-transferase (GST) were determined in subcellular microsomal fractions isolated from lung and liver tissues. Finally, as a measure of overall toxicity, the activities of various serum enzymes were determined in the blood serum of the rats. It was found that the induction of DNA single-strand breaks by three nitrosamines was decreased in hepatocytes from SO2-treated animals. The viability of rat hepatocytes and of rat lung cells, as determined by trypan blue exclusion, was similar in all three treatment groups immediately after isolation, as well as after 1 h incubation with DMSO or with the nitrosamines. In contrast, the leakage of enzymes was different in hepatocytes of SO2-treated rats, since lactate dehydrogenase activity was decreased. Leakage of enzymes from the lung cells did not differ from group to group, but was lower than from hepatocytes. Foreign compound metabolizing enzymes were mainly decreased in NOx-treated animals, namely AHH, NDMA-D and GST in liver and GST in the lung. For SO2-treated animals NDMA-D was increased in liver and GST was decreased in lung.

TiO2/PCL hybrid materials synthesized via sol–gel technique for biomedical applications

International Nuclear Information System (INIS)

Catauro, M.; Bollino, F.; Papale, F.; Marciano, S.; Pacifico, S.

2015-01-01

The aim of the present work has been the synthesis of organic/inorganic hybrid materials based on titanium dioxide and poly(ε-caprolactone) (PCL) to be used in the biomedical field. Several materials have been synthesized using sol–gel methods by adding different amounts of polymer to the inorganic sol. The obtained gels have been characterized using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). The FT-IR data allowed us to hypothesize that the structure formed was that of an interpenetrating network, realized by hydrogen bonds between Ti-OH groups in the sol–gel intermediate species and carbonyl groups in the polymer repeating units. SEM and AFM analyses highlighted that the obtained materials were nanostructurated hybrids. To evaluate the biological properties of the hybrids, their bioactivity and cytotoxicity were investigated as a function of the PCL amount. The bioactivity of the synthesized systems was proven by the formation of a hydroxyapatite layer on the surface of samples soaked in a fluid simulating human blood plasma (SBF). MTT cytotoxicity tests and Trypan Blue dye exclusion tests were carried out exposing NIH-3T3 mouse embryonic fibroblasts for 24 and 48 h to extracts from the investigated hybrid materials. The results showed that all the hybrids had a non-cytotoxic effect on target cells. - Highlights: • TiO 2 /PCL hybrids were obtained by the sol–gel process for biomedical applications. • Synthesized materials were found to be first-class hybrid nanocomposites. • Hybrids appear to be bioactive, a fundamental characteristic for osseointegration. • MTT and Trypan Blue viability test show that the materials are biocompatible. • The organic phase is able to modulate the biocompatibility of the materials

Cytotoxic effects of cyanoacrylates used as retrograde filling materials: an in vitro analysis

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Azevedo Cledson Lima de

2003-01-01

Full Text Available Cyanoacrylate has been used in medicine and dentistry for many years. It has been used as a postextraction dressing and retrograde filling material in endodontic surgery. The aim of this study was to evaluate the cytotoxic effects of Histoacryl and other two homologue ethyl cyanoacrylates, Super Bonder and Ultrabond, on cultured fibroblasts, using the Trypan blue dye exclusion assay. The cyanoacrylates were applied to round glass coverslips, which were placed in contact with NIH 3T3 cells. After 0, 6, 12 and 24 h (short-term assay; viability and 1, 3, 5 and 7 days (long-term assay; survival, the cells were examined under phase light microscopy and counted. The data were compared by the Kruskal-Wallis test. In the short-term experiments, only the cultures of the Ultrabond group (GIV presented significant smaller percentages of cell viability than the cultures of the other groups (GI: control; GII: Super Bonder; GIII: Histoacryl. Although the cultures of the Super Bonder group (GII presented smaller percentages of cell viability than cultures of the other groups (GI, GIII, GIV at the long-term assay, this group was the only experimental group presenting a continuous and progressive cell growth. Our results have shown an in vitro biocompatibility of Histoacryl and ethyl cyanoacrylate homologues. These cyanoacrylates could therefore be of importance for endodontic purposes.

The in vitro viability and growth of fibroblasts cultured in the presence of different bone grafting materials (NanoBone and Straumann Bone Ceramic).

Science.gov (United States)

Kauschke, E; Rumpel, E; Fanghänel, J; Bayerlein, T; Gedrange, T; Proff, P

2006-02-01

Different clinical applications, including dentistry, are making increasing demands on bone grafting material. In the present study we have analysed the viability, proliferation and growth characteristics of fibroblasts cultured in vitro together with two different bone grafting materials, NanoBone and Straumann Bone Ceramic, over a period of 24 and 28 days respectively. Viability was measured at least every 72 hours by using the alamarBlue assay, a test that measures quantitatively cell proliferation and viability but does not require cell fixation or extraction. After one week of culture fibroblast viability was as high as in controls for both grafting materials and remained high (> 90%) for the duration of the experiment. Cell growth was evaluated microscopically. Scanning electron microscopy revealed a dense fibroblast growth at the surface of both bone grafting materials after three weeks of in vitro culture. Generally, our in vitro analyses contribute to further insights into cell - scaffold interactions.

Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

Science.gov (United States)

Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

2015-06-01

Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

The rights and wrongs of blood-brain barrier permeability studies

DEFF Research Database (Denmark)

Saunders, Norman R; Dreifuss, Jean-Jacques; Dziegielewska, Katarzyna M

2014-01-01

Careful examination of relevant literature shows that many of the most cherished concepts of the blood-brain barrier are incorrect. These include an almost mythological belief in its immaturity that is unfortunately often equated with absence or at least leakiness in the embryo and fetus....... The original concept of a blood-brain barrier is often attributed to Ehrlich; however, he did not accept that permeability of cerebral vessels was different from other organs. Goldmann is often credited with the first experiments showing dye (trypan blue) exclusion from the brain when injected systemically......, but not when injected directly into it. Rarely cited are earlier experiments of Bouffard and of Franke who showed methylene blue and trypan red stained all tissues except the brain. The term "blood-brain barrier" "Blut-Hirnschranke" is often attributed to Lewandowsky, but it does not appear in his papers...

Trophic transfer and in vivo immunotoxicological effects of tributyltin (TBT) in polar seastar Leptasterias polaris

International Nuclear Information System (INIS)

Bekri, Khalida; Pelletier, Emilien

2004-01-01

This study investigated the potential in vivo immunotoxic effects of tributyltin (TBT) on amoebocytes of 6-armed seastar Leptasterias polaris. Tested animals were contaminated by trophic transfer via alive contaminated prey consisting of blue mussels (3 μg TBT g -1 wet weight (WW) tissue) exposed to seawater containing dissolved TBT. Four biomarkers of immunotoxicological effects were monitored over 45 days at different sampling times (9, 24, 48 and 72 h, 11, 18, 25, 32 and 45 days): amoebocytes count (AC), cell viability using Trypan blue exclusion test, phagocytic activity (PA) using a suspension of dead bacteria labelled with fluorescein isothiocyanate (FITC) and injected directly in the coelomic fluid of the animals, and lysosomal integrity (LI) using the neutral red (NR) retention test. Data showed that TBT and its metabolites (DBT and MBT) bioaccumulated preferentially in pyloric caeca, whereas gonads contained only small quantities. Despite the differences in exposure periods to the contaminated diet and in burdens of butyltins (BTs) ingested by the various contaminated groups, there were no significant differences in body burdens of BTs. Only 6.2±2.0% of total ingested BTs were retained in soft tissues of seastars. Even if butyltins were not detected in the coelomic fluid (CF), their detrimental effects have been detected in the phagocytic activity of amoebocytes and their lysosomal retention of neutral red, but no effects were observed on amoebocytes count and their viability. These results show that seastar L. polaris possesses adequate mechanisms to depurate ingested TBT without supporting major disturbances of its immune defence system. By their ability to digest whole contaminated prey and eliminate only dissolved metabolites, L. polaris and other seastars with the same preying mode could play a role of 'recycling organisms' in coastal environments where toxicants, such as butyltins and other metallic species are accumulated by bivalves and

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African Journals Online (AJOL)

ebutamanya

2016-07-07

Jul 7, 2016 ... Cataract surgery with intraocular lens implantation in children aged 5-15 in .... logMAR 2.00 intact light perception = logMAR 2.30, defect light ... under blue trypan). .... and contact lenses) are less suitable in these settings.

Statistical optimization of harvesting Chlorella vulgaris using a novel bio-source, Strychnos potatorum

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Sirajunnisa Abdul Razack

2015-09-01

Full Text Available The present study was aimed at harvesting microalga, Chlorella vulgaris, by bioflocculation using seed powder of clearing nut, Strychnos potatorum. The research was essentially the prime step to yield a large biomass for utilising the cells in biodiesel production. Optimization of the parameters influencing bioflocculation was carried out statistically using RSM. The optimized conditions were 100 mg L−1 bioflocculant concentration, 35 °C temperature, 150 rpm agitation speed and 30 min incubation time and resulted in a maximum efficiency of 99.68%. Through cell viability test, using Trypan blue stain, it was found that cells were completely intact when treated with bioflocculant, but destroyed when exposed to chemical flocculant, alum. The overall study represented that S. potatorum could potentially be a bioflocculant of microalgal cells and a promising substitute for expensive and hazardous chemical flocculants. Moreover, this bioflocculant demonstrated their utility to harvest microalgal cells by economically, effectively and in an ecofriendly way.

Development of Taenia pisiformis in golden hamster (Mesocricetus auratus

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Maravilla Pablo

2011-07-01

Full Text Available Abstract The life cycle of Taenia pisiformis includes canines as definitive hosts and rabbits as intermediate hosts. Golden hamster (Mesocricetus auratus is a rodent that has been successfully used as experimental model of Taenia solium taeniosis. In the present study we describe the course of T. pisiformis infection in experimentally infected golden hamsters. Ten females, treated with methyl-prednisolone acetate were infected with three T. pisiformis cysticerci each one excised from one rabbit. Proglottids released in faeces and adults recovered during necropsy showed that all animals were infected. Eggs obtained from the hamsters' tapeworms, were assessed for viability using trypan blue or propidium iodide stains. Afterwards, some rabbits were inoculated with eggs, necropsy was performed after seven weeks and viable cysticerci were obtained. Our results demonstrate that the experimental model of adult Taenia pisiformis in golden hamster can replace the use of canines in order to study this parasite and to provide eggs and adult tapeworms to be used in different types of experiments.

Assessment of swim-up and discontinuous density gradient in sperm sex preselection for bovine embryo production

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A.C Lucio

2012-06-01

Full Text Available The purpose of this work was to associate the modified swim-up method with centrifugation in density gradient for the separation of X-bearing spermatozoa. Sperm viability and integrity were evaluated through the Trypan Blue/Giemsa staining method. Quality control of centrifuged spermatozoa was performed in in vitro produced embryos. The results were validated by the sex ratio of in vitro produced embryos using PCR by Y- specific sequences present in bovine male genomic DNA. After determining genetic sex of in vitro produced embryos, the results showed difference (P<0.05 in deviation of sex ratio when comparing the control group (45.2% females with the other spermatozoa selection procedures (60.6% females (P<0.05. The sperm selection methods are capable of selecting X-bearing spermatozoa without compromising the spermatozoa fertility (cleavage and blastocyst rates, 70% and 26%, respectively and were considered relevant methods to be introduced in bovine in vitro produced embryo programs.

Exposure to brominated flame retardant PBDE-99 affects cytoskeletal protein expression in the neonatal mouse cerebral cortex

DEFF Research Database (Denmark)

Alm, Henrik; Kultima, Kim; Scholz, Birger

2008-01-01

, and the cytotoxic and apoptotic effects of PBDE-99 in primary cultures of fetal rat cortical cells. We used two-dimensional difference gel electrophoresis (2D-DIGE) to analyze protein samples isolated from the cortex of NMRI mice 24h after exposure to a single oral dose of 12 mg/kg PBDE-99 on post-natal day 10....... Protein resolution was enhanced by sample pre-fractionation. In the cell model, we determined cell viability using the trypan blue exclusion assay, and apoptosis using immunocytochemical detection of cleaved caspase-3. We determined the identity of 111 differentially expressed proteins, 32 (29%) of which...... are known to be cytoskeleton-related. Similar to previous findings in the striatum, we found elevated levels of the neuron growth-associated protein Gap43 in the cortex. In cultured cortical cells, a high concentration of PBDE-99 (30 microM) induced cell death without any apparent increase in caspase-3...

Viability of the Fricke dosemeter doped with methylene blue; Viabilidade do dosimetro Fricke dopado com azul de metileno

Energy Technology Data Exchange (ETDEWEB)

Souza, V.L.B.; Santos, C.D.A.; Rodrigues, K.R.G.; Cunha, M.S.; Figueiredo, M.D.C.; Melo, R.T. [Centro Regional de Ciencias Nucleares do Nordeste (CRCN-NE/CNEN-PE), Recife, PE (Brazil)

2009-07-01

This work aims to find the possible utilization of the Fricke dosemeter doped with methylene blue (FMB) for the dosimetry of photodynamic therapy. The FMB was irradiated wit X rays and light emitted diodes demonstrating positive answers to the stimulus, being probably to be used for dosimetric objectives

Comparison of GSM Modulated and CW Radiofrequency Radiation on Cells

International Nuclear Information System (INIS)

Pavicic, I.; Marjanovic, A.M.; Trosic, I.

2011-01-01

The aim of our study was to evaluate and compare effect of global system of mobile (GSM) modulation and continuous wave (CW) radiofrequency radiation (RF) on proliferation ability and viability of V79 Chinese hamster lung cells. Previously prepared samples of cells in culture were exposed for 1, 2 and 3 hours both to 915 MHz GSM modulated and to 935 MHz CW RF field in gigahertz transversal electromagnetic mode cell (GTEM-cell). Electric field strength for cells exposed to GSM modulation was set at 10 V/m and for CW exposed cells was 8.2 V/m. Average specific absorption rate (SAR) was calculated to be for GSM 0.23 W/kg and for CW 0.12 W/kg. V79 samples were plated in concentration of 1x10 4 cells/mL. Cell proliferation was determined by cell counts for each hour of exposure during five post-exposure days. Trypan blue exclusion test was used to determine cell viability. In comparison to control cell samples, proliferation of GSM irradiated cells showed significant decrease after 3 hours of exposure on the second and third post-exposure day. CW exposed cell samples showed significant decrease after 3 hours of exposure on the third post-exposure day. Viability of GSM and CW exposed cells did not significantly differ from matched control cell samples. Both applied RF fields have shown similar effect on cell culture growth, and cell viability of V79 cell line. In addition, applied GSM modulated RF radiation demonstrate bigger influence on proliferation of cells. (author)

Assessment of organ culture for the conservation of human skin allografts.

Science.gov (United States)

Hautier, A; Sabatier, F; Stellmann, P; Andrac, L; Nouaille De Gorce, Y; Dignat-George, F; Magalon, G

2008-03-01

Human skin allografts are used in the treatment of severe burns and their preservation is therefore critical for optimal clinical benefit. Current preservation methods, such as 4 degrees C storage or cryopreservation, cannot prevent the decrease of tissue viability. The aim of this study was to assess viability and function of skin allografts in a new skin organ culture model, allowing conservation parameters as close as possible to physiological conditions: 32 degrees C, air-liquid interface and physiological skin tension. Twelve skin samples, harvested from 6 living surgical donors, were conserved 35 days in two conditions: conservation at 4 degrees C and organ culture. Viability and function of skin samples were investigated at Day 0, 7, 14, 21, 28 and 35 using cell culture methods (trypan blue exclusion, Colony Forming Efficiency and Growth Rate), histopathological and histoenzymological studies (Ki67 immunostaining). In the two conditions, fibroblast and keratinocyte viability was progressively affected by storage, with a significant decrease observed after 35 days. No statistical difference could be observed between the two conditions. The two methods were also comparable regarding alterations of fibroblast and keratinocyte culture parameters, which were respectively significantly reduced at Day 7 and 21, compared to fresh skin. By contrast, histopathological and histoenzymological studies revealed a better preservation of skin architecture and proliferative potential at 4 degrees C, as compared to organ culture. These results indicate that skin organ culture does not provide significant advantages for skin allograft preservation. However, its potential use as an experimental model to study skin physiology and wound healing should be further evaluated.

Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK.

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Maninder Bhogal

Full Text Available To establish a method for assessing graft viability, in-vivo, following corneal transplantation.Optimization of calcein AM fluorescence and toxicity assessment was performed in cultured human corneal endothelial cells and ex-vivo corneal tissue. Descemet membrane endothelial keratoplasty grafts were incubated with calcein AM and imaged pre and post preparation, and in-situ after insertion and unfolding in a pig eye model. Global, macroscopic images of the entire graft and individual cell resolution could be attained by altering the magnification of a clinical confocal scanning laser microscope. Patterns of cell loss observed in situ were compared to those seen using standard ex-vivo techniques.Calcein AM showed a positive dose-fluorescence relationship. A dose of 2.67μmol was sufficient to allow clear discrimination between viable and non-viable areas (sensitivity of 96.6% with a specificity of 96.1% and was not toxic to cultured endothelial cells or ex-vivo corneal tissue. Patterns of cell loss seen in-situ closely matched those seen on ex-vivo assessment with fluorescence viability imaging, trypan blue/alizarin red staining or scanning electron microscopy. Iatrogenic graft damage from preparation and insertion varied between 7-35% and incarceration of the graft tissue within surgical wounds was identified as a significant cause of endothelial damage.In-situ graft viability assessment using clinical imaging devices provides comparable information to ex-vivo methods. This method shows high sensitivity and specificity, is non-toxic and can be used to evaluate immediate cell viability in new grafting techniques in-vivo.

Optimization of Neutral Comet Assay for studying DNA double-strand breaks in pea and wheat

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Ivelina Nikolova

2013-01-01

Full Text Available This study describes an adaptation of the Comet assay under neutral conditions for mono- and dicotyledonous plants pea (Pisum sativum L. and wheat (Triticum aestivum L.. Modifications concern lysis and electrophoresis steps, respectively. Electrophoresis was carried out varying the intensity of the electric field. A linear relationship between the percentages of DNA in the tail from control background with alteration of intensity was found. Trypan blue dye exclusion test was used in order to determine the intactness of nuclear membrane of the isolated nuclei from both plant model systems. Assessment was conducted on non-irradiated and irradiated nuclei on a monolayer with three doses of UVC. It was found that the share of intact nuclei (trypan blue negative ones is about 95% in controls. Gradual dose-related increase of damaged nuclei was observed in both species, reaching statistical significance only at the higher dose applied.

Ultrastructural Analysis of Human Breast Cancer Cells during Their Overtime Interaction with Cerium Oxide Nanoparticles

KAUST Repository

AlAbbadi, Shatha H.

2016-12-01

Cerium oxide nanoparticles have been proposed as an anticancer agent, thanks to their ability of tuning the redox activity in accordance to different conditions, which lead to selective roles on healthy and cancer cells. Recent evidence suggested the ability of these nanoparticles to be toxic against cancer cells, while confer protection from oxidative stress, toward healthy cells. The main focus of this study was to determine the ultrastructural effects of cerium oxide nanoparticles over multiple incubation time of 1, 3, and 7 days on breast healthy and cancer cells. Cellular characterizations were carried out using electron microscopes, both transmission and scanning electron microscopes, while the viability assessments were performed by propidium iodide and trypan blue viability assays. The obtained results of the viability assays and electron microscopy suggested higher toxic effects on the cancer cell line viability by using a nanoceria dose of 300 μg/mL after 1 day of treatment. Such effects were shown to be preserved at 3 days, and in a longer time point of 7 days. On the contrary, the healthy cells underwent less effects on their viability at time point of 1 and 7 days. The 3 days treatment demonstrated a reduction on the number of cells that did not correlate with an increase of the dead cells, which suggested a possible initial decrease of the cell growth rate, which could be due to the high intracellular loading of nanoparticles. To conclude, the overall result of this experiment suggested that 300 μg/mL of CeO2 nanoparticles is the most suitable dose, within the range and the time point tested, which induces long-lasting cytotoxic effects in breast cancer cells, without harming the normal cells, as highlighted by the viability assays and ultrastructural characterization of electron microscopy analysis.

Low-power laser irradiation did not stimulate breast cancer cells following ionizing radiation

Science.gov (United States)

Silva, C. R.; Camargo, C. F. M.; Cabral, F. V.; Ribeiro, M. S.

2016-03-01

Cancer has become a public health problem worldwide. Radiotherapy may be a treatment to a number of types of cancer, frequently using gamma-radiation with sources such as 137Cs and 60Co, with varying doses, dose rates, and exposure times to obtain a better as a stimulant for cell proliferation and tissue healing process. However, its effects on cancer cells are not yet well elucidated. The purpose of this work was to evaluate the effects of the LPL on breast cancer cultures after ionizing radiation. The breast cancer-MDA-MB-231 cells were gamma irradiated by a 60Co source, with dose of 2.5 Gy. After 24h, cells were submitted to LPL irradiation using a red laser emitting at λ= 660 nm, with output power of 40 mW and exposure time of 30 s and 60 s. The plates were uniformly irradiated, with energy of 1.2 J and 2.4 J, respectively. Cell viability was analyzed using the exclusion method with trypan blue. Our results show that breast cancer cells submitted to LPL after ionizing radiation remained 95 % viable. No statistically significant differences were observed between laser and control untreated cells, (P > 0.05). These findings suggest that LPL did not influenced cancer cells viability.

Cytotoxicity evaluation of a copaiba oil-based root canal sealer compared to three commonly used sealers in endodontics

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Angela Delfina Bittencourt Garrido

2015-01-01

Full Text Available Background: The constant development of new root canal sealers has allowed the solution of a large number of clinical cases in endodontics, however, cytotoxicity of such sealers must be tested before their validation as filling materials. The aim of this study was to evaluate the cytotoxic effect of a new Copaiba oil-based root canal sealer (Biosealer [BS] on osteoblast-like Osteo-1 cells. Materials and Methods: The experimental groups were formed according to the culture medium conditioned with the tested sealers, as follows: Control group (CG (culture medium without conditioning; Sealer 26 (S26 - culture medium + S26; Endofill (EF - culture medium + EF; AH Plus (AHP - culture medium + AHP; and BS - culture medium + BS (Copaiba oil-based sealer. The conditioned culture medium was placed in contact with 2 × 10 4 cells cultivated on 60 mm diameter Petri dishes for 24 h. Then, hemocytometer count was performed to evaluate cellular viability, using Trypan Blue assay. The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for cellular viability were statistically analyzed (1-way ANOVA, Tukey′s test - P 0.05. Conclusion: The Copaiba oil-based root canal sealer presented promising results in terms of cytotoxicity which indicated its usefulness as a root canal sealer.

Cytotoxicity evaluation of a copaiba oil-based root canal sealer compared to three commonly used sealers in endodontics

Science.gov (United States)

Garrido, Angela Delfina Bittencourt; de Cara, Sueli Patricia Harumi Miyagi; Marques, Marcia Martins; Sponchiado, Emílio Carlos; Garcia, Lucas da Fonseca Roberti; de Sousa-Neto, Manoel Damião

2015-01-01

Background: The constant development of new root canal sealers has allowed the solution of a large number of clinical cases in endodontics, however, cytotoxicity of such sealers must be tested before their validation as filling materials. The aim of this study was to evaluate the cytotoxic effect of a new Copaiba oil-based root canal sealer (Biosealer [BS]) on osteoblast-like Osteo-1 cells. Materials and Methods: The experimental groups were formed according to the culture medium conditioned with the tested sealers, as follows: Control group (CG) (culture medium without conditioning); Sealer 26 (S26) - culture medium + S26; Endofill (EF) - culture medium + EF; AH Plus (AHP) - culture medium + AHP; and BS - culture medium + BS (Copaiba oil-based sealer). The conditioned culture medium was placed in contact with 2 × 104 cells cultivated on 60 mm diameter Petri dishes for 24 h. Then, hemocytometer count was performed to evaluate cellular viability, using Trypan Blue assay. The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for cellular viability were statistically analyzed (1-way ANOVA, Tukey's test - P 0.05). Conclusion: The Copaiba oil-based root canal sealer presented promising results in terms of cytotoxicity which indicated its usefulness as a root canal sealer. PMID:25878676

Protective effect of infrared-A radiation against damage induced by UVB radiation in the melan-a cell line.

Science.gov (United States)

Portantiolo Lettnin, Aline; Teixeira Santos Figueiredo Salgado, Mariana; Gonsalez Cruz, Camila; Manoel Rodrigues da Silva-Júnior, Flávio; Cunha Gonzalez, Vinícius; de Souza Votto, Ana Paula; Santos Trindade, Gilma; de Moraes Vaz Batista Filgueira, Daza

2016-10-01

The present work evaluated the infrared-A (IR-A) protective effect using a light-emitting diode (LED) lamp against the cytotoxic effects of ultraviolet B radiation (UVB). Effects on cell viability (Trypan blue assay), DNA damage (comet assay), lipid peroxidation (FOX method), reactive oxygen species production and antioxidant capacity were analyzed in melan-a, a non-tumoral murine melanocytic cell line. To define the doses used in the interaction experiments between IR-A+UVB, dose/response curves were made after exposure to IR-A or UVB. The IR-A dose chosen was 0.8J/cm(2) because this dose caused no significant inhibition of proliferation effects and viability decreased. For UVB exposure, a dose of 0.015J/cm(2), which showed a decrease in viable cell number by approximately 50% in relation to control until 72h, was selected. For IR-A+UVB, cell proliferation recovery was showed, decreasing DNA damage and lipid peroxide content when compared to UVB alone. Besides, the results obtained for ROS and antioxidant capacity showed that the protection observed was probably not related to decreased oxidative stress. In conclusion, non-thermal IR-A was capable of protecting the melan-a cells from UVB induced damage. Copyright © 2016 Elsevier B.V. All rights reserved.

Anti-tumor activity of tetrodotoxin extracted from the Masked Puffer ...

African Journals Online (AJOL)

as demonstrated by trypan blue stain (d and f) and after treatment with Annexin V stain (e & g) (X1000). Notice ... hair erection and loss of appetite. ..... effect of vitamin A against carcinogenic effect of 7, 12- DMBA on the liver of albino rat. Egypt.

Chitosan scaffold modified with D-(+) raffinose and enriched with thiol-modified gelatin for improved osteoblast adhesion

International Nuclear Information System (INIS)

Galli, C; Parisi, L; Smerieri, A; Lumetti, S; Manfredi, E; Macaluso, G M; Elviri, L; Bianchera, A; Bettini, R; Lagonegro, P

2016-01-01

The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml −1 BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties. (paper)

Preliminary viability studies of fibroblastic cells cultured on microcrystalline and nanocrystalline diamonds produced by chemical vapour deposition method

Directory of Open Access Journals (Sweden)

Ana Amélia Rodrigues

2013-02-01

Full Text Available Implant materials used in orthopedics surgery have demonstrated some disadvantages, such as metallic corrosion processes, generation of wear particles, inflammation reactions and bone reabsorption in the implant region. The diamond produced through hot-filament chemical vapour deposition method is a new potential biomedical material due to its chemical inertness, extreme hardness and low coefficient of friction. In the present study we analysis two samples: the microcrystalline diamond and the nanocrystalline diamond. The aim of this study was to evaluate the surface properties of the diamond samples by scanning electron microscopy, Raman spectroscopy and atomic force microscopy. Cell viability and morphology were assessed using thiazolyl blue tetrazolium bromide, cytochemical assay and scanning electron microscopy, respectively. The results revealed that the two samples did not interfere in the cell viability, however the proliferation of fibroblasts cells observed was comparatively higher with the nanocrystalline diamond.

Assembled microneedle arrays enhance the transport of compounds varying over a large range of molecular weight across human dermatomed skin

NARCIS (Netherlands)

Verbaan, F.J.; Bal, S.M.; van den Berg, D.J.; Groenink, W.H.H.; Verpoorten, H.; Lüttge, Regina; Bouwstra, J.A.

2007-01-01

In this study, we demonstrate the feasibility to use microneedle arrays manufactured from commercially available 30G hypodermal needles to enhance the transport of compounds up to a molecular weight of 72 kDa. Piercing of human dermatomed skin with microneedle arrays was studied by Trypan Blue

Quantifying fungal viability in air and water samples using quantitative PCR after treatment with propidium monoazide (PMA)

International Nuclear Information System (INIS)

Vesper, Stephen; McKinstry, Craig A.; Hartmann, Chris; Neace, Michelle; Yoder, Stephanie; Vesper, Alex

2007-01-01

A method is described to discriminate between live and dead cells of the infectious fungi Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Mucor racemosus, Rhizopus stolonifer and Paecilomyces variotii. To test the method, conidial suspensions were heat inactivated at 85 C or held at 5 C (controls) for 1 h. Polycarbonate filters (25 mm diameter, 0.8 (micro)m pore size) were placed on 'welled' slides (14 mm diameter) and the filters treated with either PBS or PMA. Propidium monoazide (PMA), which enters dead cells but not live cells, was incubated with cell suspensions, exposed to blue wavelength light-emitting diodes (LED) to inactivate remaining PMA and secure intercalation of PMA with DNA of dead cells. Treated cells were extracted and the live and dead cells evaluated with quantitative PCR (QPCR). After heat treatment and DNA modification with PMA, all fungal species tested showed an approximate 100- to 1000-fold difference in cell viability estimated by QPCR analysis which was consistent with estimates of viability based on culturing.

Risco de contaminação do corante azul de tripano após primeira utilização Risk of contamination of trypan blue dye after first use

Directory of Open Access Journals (Sweden)

João Baptista Nigro Santiago Malta

2006-02-01

Full Text Available OBJETIVOS: Determinar o potencial risco de contaminação do frasco de azul de tripano (AT depois de utilizado pela primeira vez e estocado em diferentes condições de temperatura e umidade, assim como identificar os possíveis fatores de contaminação, microrganismos mais freqüentemente envolvidos e simultaneamente avaliar as propriedades bacteriostáticas e bactericidas do corante. MÉTODOS: Realizado estudo experimental, prospectivo, em que 30 frascos de AT foram divididos em três grupos (A: controle, B: armazenamento em geladeira e C: armazenamento em armário. O corante era aspirado e semeado em placas de ágar sangue e tubo de ágar Sabouraud. No grupo A o AT foi semeado apenas logo após a abertura dos frascos (tempo zero - T0, nos grupos B e C ocorreu semeadura nos T0, T1 (1 dia, T2 (2 dias, T7 (7 dias e T10 (10 dias após abertura dos frascos. No 10º dia os frascos dos grupos B e C também foram submetidos a um raspado do lado interno do frasco após abertura. Concomitantemente foi realizado teste de ação inibitória do corante AT para estudo da atividade bacteriostática e bactericida. RESULTADOS: As semeaduras realizadas no T0 não apresentaram contaminação. Entre os T1 e T10 mais o raspado houve apenas 1 frasco contaminado armazenado em geladeira. O microrganismo encontrado foi o Aspergillus niger. Foi comprovado que o corante não apresenta ação bactericida e bacteriostática para as bactérias testadas. CONCLUSÕES: Nas condições do estudo não houve contaminação dos frascos armazenados em armário e 1 frasco (10% armazenado em geladeira apresentou contaminação após abertura e uso inicial. A fonte de contaminação talvez seja o lado externo do produto. O AT não apresenta propriedades bactericidas e bacteriostáticas para as bactérias testadas e na concentração utilizada.PURPOSE: To determine the potential risk of contamination of a trypan blue bottle (TB after first use and after being stored under different

Neuroprotective effects of riluzole: an electrophysiological and histological analysis in an in vitro model of ischemia.

Science.gov (United States)

Siniscalchi, A; Zona, C; Sancesario, G; D'Angelo, E; Zeng, Y C; Mercuri, N B; Bernardi, G

1999-06-01

The protective effects of riluzole against the neuronal damage caused by O2 and glucose deprivation (ischemia) was investigated in rat cortical slices by recording electrophysiologically the cortico-cortical field potential and by evaluating histologically the severity of neuronal death. Five minutes of ischemia determined an irreversible depression of the amplitude of the field potential. In addition, this insult caused a clear enhancement of the number of death cells that were specifically colored with trypan blue (a vital colorant which stains altered cells). We found that riluzole, which by itself depressed the synaptic transmission, neuroprotected when perfused 15-20 min before and during ischemia. In fact, due to the treatment with riluzole, the ischemia-induced irreversible depression of the field potential recovered and less cells were stained with trypan blue. These findings demonstrate that riluzole prevents neuronal death in an in vitro model of ischemia and suggest a therapeutic use of this drug in order to reduce the pathophysiological outcomes of stroke.

Release kinetics and cell viability of ibuprofen nanocrystals produced by melt-emulsification.

Science.gov (United States)

Fernandes, A R; Dias-Ferreira, J; Cabral, C; Garcia, M L; Souto, E B

2018-06-01

The clinical use of poorly water-soluble drugs has become a big challenge in pharmaceutical development due to the compromised bioavailability of the drugs in vivo. Nanocrystals have been proposed as a formulation strategy to improve the dissolution properties of these drugs. The benefits of using nanocrystals in drug delivery, when compared to other nanoparticles, are related to their production facilities, simple structure, and suitability for a variety of administration routes. High pressure homogenization (HPH) is the most promising production process, which can be employed at low or high temperatures. Ibuprofen nanocrystals with a mean size below 175 nm, and polydispersity below 0.18, have been produced by melt-emulsification, followed by HPH. Two nanocrystal formulations, differing on the surfactant composition, have been produced, their in vitro ibuprofen release tested in Franz diffusion cells and adjusted to several kinetic models (zero order, first order, Higuchi, Hixson-Crowell, Korsmeyer-Peppas, Baker-Lonsdale and Weibull model). Cell viability was assessed at 3, 6 and 24 h of incubation on human epithelial colorectal cells (Caco-2) by AlamarBlue ® colorimetric assay. For both formulations, Caco-2 cells viability was dependent on the drug concentration and time of exposure. Copyright © 2018 Elsevier B.V. All rights reserved.

Study of internalization and viability of multimodal nanoparticles for labeling of human umbilical cord mesenchymal stem cells

International Nuclear Information System (INIS)

Miyaki, Liza Aya Mabuchi; Sibov, Tatiana Tais; Pavon, Lorena Favaro; Mamani, Javier Bustamante; Gamarra, Lionel Fernel

2012-01-01

Objective: To analyze multimodal magnetic nanoparticles-Rhodamine B in culture media for cell labeling, and to establish a study of multimodal magnetic nanoparticles-Rhodamine B detection at labeled cells evaluating they viability at concentrations of 10 μg Fe/mL and 100μg Fe/mL. Methods: We performed the analysis of stability of multimodal magnetic nanoparticles-Rhodamine B in different culture media; the mesenchymal stem cells labeling with multimodal magnetic nanoparticles-Rhodamine B; the intracellular detection of multimodal magnetic nanoparticles-Rhodamine B in mesenchymal stem cells, and assessment of the viability of labeled cells by kinetic proliferation. Results: The stability analysis showed that multimodal magnetic nanoparticles-Rhodamine B had good stability in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium. The mesenchymal stem cell with multimodal magnetic nanoparticles-Rhodamine B described location of intracellular nanoparticles, which were shown as blue granules co-localized in fluorescent clusters, thus characterizing magnetic and fluorescent properties of multimodal magnetic nanoparticles Rhodamine B. Conclusion: The stability of multimodal magnetic nanoparticles-Rhodamine B found in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium assured intracellular mesenchymal stem cells labeling. This cell labeling did not affect viability of labeled mesenchymal stem cells since they continued to proliferate for five days. (author)

Rapid, portable and cost-effective yeast cell viability and concentration analysis using lensfree on-chip microscopy and machine learning

KAUST Repository

Feizi, Alborz

2016-09-24

Monitoring yeast cell viability and concentration is important in brewing, baking and biofuel production. However, existing methods of measuring viability and concentration are relatively bulky, tedious and expensive. Here we demonstrate a compact and cost-effective automatic yeast analysis platform (AYAP), which can rapidly measure cell concentration and viability. AYAP is based on digital in-line holography and on-chip microscopy and rapidly images a large field-of-view of 22.5 mm2. This lens-free microscope weighs 70 g and utilizes a partially-coherent illumination source and an opto-electronic image sensor chip. A touch-screen user interface based on a tablet-PC is developed to reconstruct the holographic shadows captured by the image sensor chip and use a support vector machine (SVM) model to automatically classify live and dead cells in a yeast sample stained with methylene blue. In order to quantify its accuracy, we varied the viability and concentration of the cells and compared AYAP\\'s performance with a fluorescence exclusion staining based gold-standard using regression analysis. The results agree very well with this gold-standard method and no significant difference was observed between the two methods within a concentration range of 1.4 × 105 to 1.4 × 106 cells per mL, providing a dynamic range suitable for various applications. This lensfree computational imaging technology that is coupled with machine learning algorithms would be useful for cost-effective and rapid quantification of cell viability and density even in field and resource-poor settings.

Rapid, portable and cost-effective yeast cell viability and concentration analysis using lensfree on-chip microscopy and machine learning.

Science.gov (United States)

Feizi, Alborz; Zhang, Yibo; Greenbaum, Alon; Guziak, Alex; Luong, Michelle; Chan, Raymond Yan Lok; Berg, Brandon; Ozkan, Haydar; Luo, Wei; Wu, Michael; Wu, Yichen; Ozcan, Aydogan

2016-11-01

Monitoring yeast cell viability and concentration is important in brewing, baking and biofuel production. However, existing methods of measuring viability and concentration are relatively bulky, tedious and expensive. Here we demonstrate a compact and cost-effective automatic yeast analysis platform (AYAP), which can rapidly measure cell concentration and viability. AYAP is based on digital in-line holography and on-chip microscopy and rapidly images a large field-of-view of 22.5 mm 2 . This lens-free microscope weighs 70 g and utilizes a partially-coherent illumination source and an opto-electronic image sensor chip. A touch-screen user interface based on a tablet-PC is developed to reconstruct the holographic shadows captured by the image sensor chip and use a support vector machine (SVM) model to automatically classify live and dead cells in a yeast sample stained with methylene blue. In order to quantify its accuracy, we varied the viability and concentration of the cells and compared AYAP's performance with a fluorescence exclusion staining based gold-standard using regression analysis. The results agree very well with this gold-standard method and no significant difference was observed between the two methods within a concentration range of 1.4 × 10 5 to 1.4 × 10 6 cells per mL, providing a dynamic range suitable for various applications. This lensfree computational imaging technology that is coupled with machine learning algorithms would be useful for cost-effective and rapid quantification of cell viability and density even in field and resource-poor settings.

Viability, invariance and applications

CERN Document Server

Carja, Ovidiu; Vrabie, Ioan I

2007-01-01

The book is an almost self-contained presentation of the most important concepts and results in viability and invariance. The viability of a set K with respect to a given function (or multi-function) F, defined on it, describes the property that, for each initial data in K, the differential equation (or inclusion) driven by that function or multi-function) to have at least one solution. The invariance of a set K with respect to a function (or multi-function) F, defined on a larger set D, is that property which says that each solution of the differential equation (or inclusion) driven by F and issuing in K remains in K, at least for a short time.The book includes the most important necessary and sufficient conditions for viability starting with Nagumo's Viability Theorem for ordinary differential equations with continuous right-hand sides and continuing with the corresponding extensions either to differential inclusions or to semilinear or even fully nonlinear evolution equations, systems and inclusions. In th...

Use of lymphoblastoid cell lines to evaluate the hypersensitivity to ultraviolet radiation in Cockayne syndrome

International Nuclear Information System (INIS)

Otsuka, F.; Tarone, R.E.; Cayeux, S.; Robbins, J.H.

1984-01-01

Cockayne syndrome (CS) is a rare autosomal recessive disease characterized by acute sun sensitivity, cachectic dwarfism, and neurologic and skeletal abnormalities. Cultured skin fibroblasts from patients with this disease are known to be hypersensitive to the lethal effects of 254-nm UV radiation. The authors have studied the sensitivity of 254-nm UV radiation of lymphoblastoid lines derived from 3 typical CS patients, 1 atypical CS patient who had a very late age of onset of clinical manifestations, 2 patients who had both xeroderma pigmentosum (XP) and typical CS, and 3 heterozygous parents of these patients. Post-UV survival was determined by the trypan-blue dye-exclusion method. The lymphoblastoid lines from the 3 typical CS patients, the atypical CS patient, and the 2 patients with both CS and XP had decreased post-UV viability in comparison with lines from normal donors. Lines from the heterozygous parents had normal post-UV viability. The post-UV viability of the typical CS lines was similar to that of a XP complementation group C line. The relative post-UV viability of lymphoblastoid lines from the typical CS patients was similar to the relative post-UV survival of their fibroblast lines. The lymphoblastoid line from the atypical CS patient had a post-UV viability similar to that of the typical CS patients. Thus, the relative hypersensitivity of CS patients cells in vitro does not reflect the severity or age of onset of the patients clinical manifestations. The lymphoblastoid lines from the 2 patients who had both CS and XP were significantly more sensitive to the UV radiation than those from patients with only CS. Our studies demonstrate that lymphoblastoid lines from patients with CS are appropriate and useful cell lines for the study of the inherited hypersensitivity to UV radiation

Direct effect of curcumin on porcine ovarian cell functions.

Science.gov (United States)

Kádasi, Attila; Maruniaková, Nora; Štochmaľová, Aneta; Bauer, Miroslav; Grossmann, Roland; Harrath, Abdel Halim; Kolesárová, Adriana; Sirotkin, Alexander V

2017-07-01

Curcuma longa Linn (L.) is a plant widely used in cooking (in curry powder a.o.) and in folk medicine, but its action on reproductive processes and its possible mechanisms of action remain to be investigated. The objective of this study was to examine the direct effects of curcumin, the major Curcuma longa L. molecule, on basic ovarian cell functions such as proliferation, apoptosis, viability and steroidogenesis. Porcine ovarian granulosa cells were cultured with and without curcumin (at doses of 0, 1, 10 and 100μg/ml of medium). Markers of proliferation (accumulation of PCNA) and apoptosis (accumulation of bax) were analyzed by immunocytochemistry. The expression of mRNA for PCNA and bax was detected by RT-PCR. Cell viability was detected by trypan blue exclusion test. Release of steroid hormones (progesterone and testosterone) was measured by enzyme immunoassay (EIA). It was observed that addition of curcumin reduced ovarian cell proliferation (expression of both PCNA and its mRNA), promoted apoptosis (accumulation of both bax and its mRNA), reduced cell viability, and stimulated both progesterone and testosterone release. These observations demonstrate the direct suppressive effect of Curcuma longa L./curcumin on female gonads via multiple mechanisms of action - suppression of ovarian cell proliferation and viability, promotion of their apoptosis (at the level of mRNA transcription and subsequent accumulation of promoters of genes regulating these activities) and release of anti-proliferative and pro-apoptotic progesterone and androgen. The potential anti-gonadal action of curcumin should be taken into account by consumers of Curcuma longa L.-containing products. Copyright © 2017 Elsevier B.V. All rights reserved.

Osteoarticular cells tolerate short-term exposure to nitisinone-implications in alkaptonuria.

Science.gov (United States)

Mistry, J B; Jackson, D J; Bukhari, M; Taylor, A M

2016-02-01

Alkaptonuria (AKU) is a rare genetic disease resulting in severe, rapidly progressing, early onset multi-joint osteoarthropathy. A potential therapy, nitisinone, is being trialled that reduces the causative agent; homogentisic acid (HGA) and in a murine model has shown to prevent ochronosis. Little is currently known about the effect nitisinone has on osteoarticular cells; these cells suffer most from the presence of HGA and its polymeric derivatives. This led us to investigate nitisinone's effect on chondrocytes and osteoblast-like cells in an in vitro model. Human C20/A4 immortalized chondrocytes, and osteosarcoma cells MG63 cultured in DMEM, as previously described. Confluent cells were then plated into 24-well plates at 4 × 10(4) cells per well in varying concentrations of nitisinone. Cells were cultured for 7 days with medium changes every third day. Trypan blue assay was used to determine viability and the effect of nitisinone concentration on cells. Statistical analysis was performed using analysis of variance, and differences between groups were determined by Newman-Keuls post-test. Analysis of C20/A4 chondrocyte and MG63 osteoblast-like cell viability when cultured in different concentrations of nitisinone demonstrates that there is no statistically significant difference in cell viability compared to control cultures. There is currently no literature surrounding the use of nitisinone in human in vitro models, or its effect on chondrocytes or osteoblast like cells. Our results show that nitisinone does not appear detrimental to cell viability of chondrocytes or osteoblast-like cells, which adds to the evidence that this therapy could be useful in treating AKU.

MORPHOLOGICAL CHARACTERISTIC OF SPERMATOGONIA AND TESTES DISSOCIATION : A Preliminary Study for the Germ Cell Transplantation in Giant Gouramy (Osphronemus gouramy

Directory of Open Access Journals (Sweden)

Irma Andriani

2010-12-01

Full Text Available The recent study were attempting to develop spermatogonial germ cell transplantation as a tool to preserve and propagate male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Spermatogonia identification and testes dissociation were the first necessary steps to obtain highly amount and viable population of spermatogonia as donor cells for transplantation. Using giant gouramy testes as a model, spermatogonia was histological characterized and two methods of testes dissociations were compared (i.e. medium A contained 0.5% trypsin in PBS and medium B contained 0.5% trypsin and DNase 10 IU/μL in PBS complemented with CaCl2, Hepes and FCS. Optimal incubation times (1, 2, 3, 4 and 5 hours in dissociation medium were also determined. Freshly isolated testes of immature giant gouramy were minced in dissociation medium and then incubated to get monodisperce cell suspension. Parameters observed were number and viability of spermatogonia (ø > 10 μm. The viability was analyzed using trypan blue exclusion dye. The results showed that the average number of spermatogonia observed in medium B was higher than in medium A (P0.05. The viability of spermatogonia decreased by the increasing duration time of dissociation. The viability of spermatogonia started to decrease significantly in 2 hours incubation time in medium A and 4 hours incubation time in medium B (P<0.05. In conclusion, application of dissociation medium B yielded higher number of viable spermatogonia than dissociation medium A.

High-luminosity blue and blue-green gallium nitride light-emitting diodes.

Science.gov (United States)

Morkoç, H; Mohammad, S N

1995-01-06

Compact and efficient sources of blue light for full color display applications and lighting eluded and tantalized researchers for many years. Semiconductor light sources are attractive owing to their reliability and amenability to mass manufacture. However, large band gaps are required to achieve blue color. A class of compound semiconductors formed by metal nitrides, GaN and its allied compounds AIGaN and InGaN, exhibits properties well suited for not only blue and blue-green emitters, but also for ultraviolet emitters and detectors. What thwarted engineers and scientists from fabricating useful devices from these materials in the past was the poor quality of material and lack of p-type doping. Both of these obstacles have recently been overcome to the point where highluminosity blue and blue-green light-emitting diodes are now available in the marketplace.

Blue ocean strategy.

Science.gov (United States)

Kim, W Chan; Mauborgne, Renée

2004-10-01

Despite a long-term decline in the circus industry, Cirque du Soleil profitably increased revenue 22-fold over the last ten years by reinventing the circus. Rather than competing within the confines of the existing industry or trying to steal customers from rivals, Cirque developed uncontested market space that made the competition irrelevant. Cirque created what the authors call a blue ocean, a previously unknown market space. In blue oceans, demand is created rather than fought over. There is ample opportunity for growth that is both profitable and rapid. In red oceans--that is, in all the industries already existing--companies compete by grabbing for a greater share of limited demand. As the market space gets more crowded, prospects for profits and growth decline. Products turn into commodities, and increasing competition turns the water bloody. There are two ways to create blue oceans. One is to launch completely new industries, as eBay did with online auctions. But it's much more common for a blue ocean to be created from within a red ocean when a company expands the boundaries of an existing industry. In studying more than 150 blue ocean creations in over 30 industries, the authors observed that the traditional units of strategic analysis--company and industry--are of limited use in explaining how and why blue oceans are created. The most appropriate unit of analysis is the strategic move, the set of managerial actions and decisions involved in making a major market-creating business offering. Creating blue oceans builds brands. So powerful is blue ocean strategy, in fact, that a blue ocean strategic move can create brand equity that lasts for decades.

Electronic properties of blue phosphorene/graphene and blue phosphorene/graphene-like gallium nitride heterostructures.

Science.gov (United States)

Sun, Minglei; Chou, Jyh-Pin; Yu, Jin; Tang, Wencheng

2017-07-05

Blue phosphorene (BlueP) is a graphene-like phosphorus nanosheet which was synthesized very recently for the first time [Nano Lett., 2016, 16, 4903-4908]. The combination of electronic properties of two different two-dimensional materials in an ultrathin van der Waals (vdW) vertical heterostructure has been proved to be an effective approach to the design of novel electronic and optoelectronic devices. Therefore, we used density functional theory to investigate the structural and electronic properties of two BlueP-based heterostructures - BlueP/graphene (BlueP/G) and BlueP/graphene-like gallium nitride (BlueP/g-GaN). Our results showed that the semiconducting nature of BlueP and the Dirac cone of G are well preserved in the BlueP/G vdW heterostructure. Moreover, by applying a perpendicular electric field, it is possible to tune the position of the Dirac cone of G with respect to the band edge of BlueP, resulting in the ability to control the Schottky barrier height. For the BlueP/g-GaN vdW heterostructure, BlueP forms an interface with g-GaN with a type-II band alignment, which is a promising feature for unipolar electronic device applications. Furthermore, we discovered that both G and g-GaN can be used as an active layer for BlueP to facilitate charge injection and enhance the device performance.

Quantifying fungal viability in air and water samples using quantitative PCR after treatment with propidium monoazide (PMA)

Energy Technology Data Exchange (ETDEWEB)

Vesper, Stephen; McKinstry, Craig A.; Hartmann, Chris; Neace, Michelle; Yoder, Stephanie; Vesper, Alex

2007-11-28

A method is described to discriminate between live and dead cells of the infectious fungi Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Mucor racemosus, Rhizopus stolonifer and Paecilomyces variotii. To test the method, conidial suspensions were heat inactivated at 85 °C or held at 5 °C (controls) for 1 h. Polycarbonate filters (25 mm diameter, 0.8 μm pore size) were placed on "welled" slides (14 mm diameter) and the filters treated with either PBS or PMA. Propidium monoazide (PMA), which enters dead cells but not live cells, was incubated with cell suspensions, exposed to blue wavelength light-emitting diodes (LED) to inactivate remaining PMA and secure intercalation of PMAwith DNA of dead cells. Treated cells were extracted and the live and dead cells evaluated with quantitative PCR (QPCR). After heat treatment and DNA modification with PMA, all fungal species tested showed an approximate 100- to 1000-fold difference in cell viability estimated by QPCR analysis which was consistent with estimates of viability based on culturing.

Cytotoxicity and fluorescence studies of silica-coated CdSe quantum dots for bioimaging applications

International Nuclear Information System (INIS)

Vibin, Muthunayagam; Vinayakan, Ramachandran; John, Annie; Raji, Vijayamma; Rejiya, Chellappan S.; Vinesh, Naresh S.; Abraham, Annie

2011-01-01

The toxicological effects of silica-coated CdSe quantum dots (QDs) were investigated systematically on human cervical cancer cell line. Trioctylphosphine oxide capped CdSe QDs were synthesized and rendered water soluble by overcoating with silica, using aminopropyl silane as silica precursor. The cytotoxicity studies were conducted by exposing cells to freshly synthesized QDs as a function of time (0–72 h) and concentration up to micromolar level by Lactate dehydrogenase assay, MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay, Neutral red cell viability assay, Trypan blue dye exclusion method and morphological examination of cells using phase contrast microscope. The in vitro analysis results showed that the silica-coated CdSe QDs were nontoxic even at higher loadings. Subsequently the in vivo fluorescence was also demonstrated by intravenous administration of the QDs in Swiss albino mice. The fluorescence images in the cryosections of tissues depicted strong luminescence property of silica-coated QDs under biological conditions. These results confirmed the role of these luminescent materials in biological labeling and imaging applications.

Cytotoxicity and fluorescence studies of silica-coated CdSe quantum dots for bioimaging applications

Energy Technology Data Exchange (ETDEWEB)

Vibin, Muthunayagam [University of Kerala, Department of Biochemistry (India); Vinayakan, Ramachandran [National Institute for Interdisciplinary Science and Technology (CSIR), Photosciences and Photonics (India); John, Annie [Sree Chitra Tirunal Institute of Medical Sciences and Technology, Biomedical Technology Wing (India); Raji, Vijayamma; Rejiya, Chellappan S.; Vinesh, Naresh S.; Abraham, Annie, E-mail: annieab2@yahoo.co.in [University of Kerala, Department of Biochemistry (India)

2011-06-15

The toxicological effects of silica-coated CdSe quantum dots (QDs) were investigated systematically on human cervical cancer cell line. Trioctylphosphine oxide capped CdSe QDs were synthesized and rendered water soluble by overcoating with silica, using aminopropyl silane as silica precursor. The cytotoxicity studies were conducted by exposing cells to freshly synthesized QDs as a function of time (0-72 h) and concentration up to micromolar level by Lactate dehydrogenase assay, MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay, Neutral red cell viability assay, Trypan blue dye exclusion method and morphological examination of cells using phase contrast microscope. The in vitro analysis results showed that the silica-coated CdSe QDs were nontoxic even at higher loadings. Subsequently the in vivo fluorescence was also demonstrated by intravenous administration of the QDs in Swiss albino mice. The fluorescence images in the cryosections of tissues depicted strong luminescence property of silica-coated QDs under biological conditions. These results confirmed the role of these luminescent materials in biological labeling and imaging applications.

Early cell death detection with digital holographic microscopy.

Directory of Open Access Journals (Sweden)

Nicolas Pavillon

Full Text Available BACKGROUND: Digital holography provides a non-invasive measurement of the quantitative phase shifts induced by cells in culture, which can be related to cell volume changes. It has been shown previously that regulation of cell volume, in particular as it relates to ionic homeostasis, is crucially involved in the activation/inactivation of the cell death processes. We thus present here an application of digital holographic microscopy (DHM dedicated to early and label-free detection of cell death. METHODS AND FINDINGS: We provide quantitative measurements of phase signal obtained on mouse cortical neurons, and caused by early neuronal cell volume regulation triggered by excitotoxic concentrations of L-glutamate. We show that the efficiency of this early regulation of cell volume detected by DHM, is correlated with the occurrence of subsequent neuronal death assessed with the widely accepted trypan blue method for detection of cell viability. CONCLUSIONS: The determination of the phase signal by DHM provides a simple and rapid optical method for the early detection of cell death.

Growth Inhibition of Osteosarcoma Cell Lines in 3D Cultures: Role of Nitrosative and Oxidative Stress.

Science.gov (United States)

Gorska, Magdalena; Krzywiec, Pawel Bieniasz; Kuban-Jankowska, Alicja; Zmijewski, Michal; Wozniak, Michal; Wierzbicka, Justyna; Piotrowska, Anna; Siwicka, Karolina

2016-01-01

3D cell cultures have revolutionized the understanding of cell behavior, allowing culture of cells with the possibility of resembling in vivo intercellular signaling and cell-extracellular matrix interaction. The effect of limited oxygen penetration into 3D culture of highly metastatic osteosarcoma 143B cells in terms of expression of nitro-oxidative stress markers was investigated and compared to standard 2D cell culture. Human osteosarcoma (143B cell line) cells were cultured as monolayers, in collagen and Matrigel. Cell viability, gene expression of nitro-oxidative stress markers, and vascular endothelial growth factor were determined using Trypan blue assay, quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Three-dimensional environments modify nitro-oxidative stress and influence gene expression and cell proliferation of OS 143B cells. Commercial cell lines might not constitute a good model of 3D cultures for bone tissue engineering, as they are highly sensitive to hypoxia, and hypoxic conditions can induce oxidation of the cellular environment. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Control of proliferation rate of N27 dopaminergic neurons using Transcranial Magnetic Stimulation orientation

Science.gov (United States)

Meng, Yiwen; Hadimani, Ravi; Anantharam, Vellareddy; Kanthasamy, Anumantha; Jiles, David

2015-03-01

Transcranial magnetic stimulation (TMS) has been used to investigate possible treatments for a variety of neurological disorders. However, the effect that magnetic fields have on neurons has not been well documented in the literature. We have investigated the effect of different orientation of magnetic field generated by TMS coils with a monophasic stimulator on the proliferation rate of N27 neuronal cells cultured in flasks and multi-well plates. The proliferation rate of neurons would increase by exposed horizontally adherent N27 cells to a magnetic field pointing upward through the neuronal proliferation layer compared with the control group. On the other hand, proliferation rate would decrease in cells exposed to a magnetic field pointing downward through the neuronal growth layer compared with the control group. We confirmed results obtained from the Trypan-blue and automatic cell counting methods with those from the CyQuant and MTS cell viability assays. Our findings could have important implications for the preclinical development of TMS treatments of neurological disorders and represents a new method to control the proliferation rate of neuronal cells.

IN VITRO CYTOTOXICITY STUDY OF AGAVE AMERICANA, STRYCHNOS NUX-VOMICA AND ARECA CATECHU EXTRACTS USING MCF-7 CELL LINE

Directory of Open Access Journals (Sweden)

Anajwala Chetan C.

2010-06-01

Full Text Available Research is focusing on the search for new types of natural chemotherapeutic agent that is plant based medicines which are proving to be excellent sources of new compounds. In present research study, an attempt was made to prove cytotoxicity activity of various parts of medicinal plants such as Agave americana, Strychnos nux-vomica and Areca catechu using MCF-7 and Vero cell line. Various parts of the medicinal plants were extracted by soxhlet apparatus using solvents likes methanol and water. By trypan blue dye exclusion method, Viability of MCF-7 and Vero cell lines were 85.50 and 81.13%, respectively. IC50 value of methanol extract of Agave americana leaves and aqueous extract of Areca catechu fruits were found to be 545.9 & 826.1 µg/ml by SRB assay and 775.1 & 1461µg/ml by MTT assay, respectively, against MCF-7 cell line. From cytotoxicity study data by SRB and MTT assay, it revealed that methanol extract of Agave americana and aqueous extract of Areca catechu are potent cytotoxic.

Antitumor activity of colloidal silver on MCF-7 human breast cancer cells

Directory of Open Access Journals (Sweden)

Franco-Molina Moisés A

2010-11-01

Full Text Available Abstract Background Colloidal silver has been used as an antimicrobial and disinfectant agent. However, there is scarce information on its antitumor potential. The aim of this study was to determine if colloidal silver had cytotoxic effects on MCF-7 breast cancer cells and its mechanism of cell death. Methods MCF-7 breast cancer cells were treated with colloidal silver (ranged from 1.75 to 17.5 ng/mL for 5 h at 37°C and 5% CO2 atmosphere. Cell Viability was evaluated by trypan blue exclusion method and the mechanism of cell death through detection of mono-oligonucleosomes using an ELISA kit and TUNEL assay. The production of NO, LDH, and Gpx, SOD, CAT, and Total antioxidant activities were evaluated by colorimetric assays. Results Colloidal silver had dose-dependent cytotoxic effect in MCF-7 breast cancer cells through induction of apoptosis, shown an LD50 (3.5 ng/mL and LD100 (14 ng/mL (*P Conclusions The present results showed that colloidal silver might be a potential alternative agent for human breast cancer therapy.

Antitumor activity of colloidal silver on MCF-7 human breast cancer cells.

Science.gov (United States)

Franco-Molina, Moisés A; Mendoza-Gamboa, Edgar; Sierra-Rivera, Crystel A; Gómez-Flores, Ricardo A; Zapata-Benavides, Pablo; Castillo-Tello, Paloma; Alcocer-González, Juan Manuel; Miranda-Hernández, Diana F; Tamez-Guerra, Reyes S; Rodríguez-Padilla, Cristina

2010-11-16

Colloidal silver has been used as an antimicrobial and disinfectant agent. However, there is scarce information on its antitumor potential. The aim of this study was to determine if colloidal silver had cytotoxic effects on MCF-7 breast cancer cells and its mechanism of cell death. MCF-7 breast cancer cells were treated with colloidal silver (ranged from 1.75 to 17.5 ng/mL) for 5 h at 37°C and 5% CO2 atmosphere. Cell Viability was evaluated by trypan blue exclusion method and the mechanism of cell death through detection of mono-oligonucleosomes using an ELISA kit and TUNEL assay. The production of NO, LDH, and Gpx, SOD, CAT, and Total antioxidant activities were evaluated by colorimetric assays. Colloidal silver had dose-dependent cytotoxic effect in MCF-7 breast cancer cells through induction of apoptosis, shown an LD50 (3.5 ng/mL) and LD100 (14 ng/mL) (*P colloidal silver. The present results showed that colloidal silver might be a potential alternative agent for human breast cancer therapy.

Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells

International Nuclear Information System (INIS)

Barboza, Carlos Augusto Galvão; Ginani, Fernanda; Soares, Diego Moura; Henriques, Águida Cristina Gomes; Freitas, Roseana de Almeida

2014-01-01

To evaluate the effect of low-level laser irradiation on the proliferation and possible nuclear morphological changes of mouse mesenchymal stem cells. Mesenchymal stem cells derived from bone marrow and adipose tissue were submitted to two applications (T0 and T48 hours) of low-level laser irradiation (660nm; doses of 0.5 and 1.0J/cm"2). The trypan blue assay was used to evaluate cell viability, and growth curves were used to analyze proliferation at zero, 24, 48, and 72 hours. Nuclear alterations were evaluated by staining with DAPI (4'-6-diamidino-2-phenylindole) at 72 hours. Bone marrow-derived mesenchymal stem cells responded to laser therapy in a dose-dependent manner. Higher cell growth was observed when the cells were irradiated with a dose of 1.0J/cm"2, especially after 24 hours (p<0.01). Adipose-derived mesenchymal stem cells responded better to a dose of 1.0J/cm"2, but higher cell proliferation was observed after 48 hours (p<0.05) and 72 hours (p<0.01). Neither nuclear alterations nor a significant change in cell viability was detected in the studied groups. Low-level laser irradiation stimulated the proliferation of mouse mesenchymal stem cells without causing nuclear alterations. The biostimulation of mesenchymal stem cells using laser therapy might be an important tool for regenerative therapy and tissue engineering

Procaine Induces Epigenetic Changes in HCT116 Colon Cancer Cells

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Hussein Sabit

2016-01-01

Full Text Available Colon cancer is the third most commonly diagnosed cancer in the world, and it is the major cause of morbidity and mortality throughout the world. The present study aimed at treating colon cancer cell line (HCT116 with different chemotherapeutic drug/drug combinations (procaine, vorinostat “SAHA,” sodium phenylbutyrate, erlotinib, and carboplatin. Two different final concentrations were applied: 3 μM and 5 μM. Trypan blue test was performed to assess the viability of the cell before and after being treated with the drugs. The data obtained showed that there was a significant decrease in the viability of cells after applying the chemotherapeutic drugs/drug combinations. Also, DNA fragmentation assay was carried out to study the effect of these drugs on the activation of apoptosis-mediated DNA degradation process. The results indicated that all the drugs/drug combinations had a severe effect on inducing DNA fragmentation. Global DNA methylation quantification was performed to identify the role of these drugs individually or in combination in hypo- or hypermethylating the CpG dinucleotide all over the genome of the HCT116 colon cancer cell line. Data obtained indicated that different combinations had different effects in reducing or increasing the level of methylation, which might indicate the effectiveness of combining drugs in treating colon cancer cells.

Rocuronium is more hepatotoxic than succinylcholine in vitro.

Science.gov (United States)

Sauer, Martin; Piel, Ines; Haubner, Cristof; Richter, Georg; Mann, Miriam; Nöldge-Schomburg, Gabriele; Mencke, Thomas

2017-09-01

The development of liver failure is a major problem in critically ill patients. The hepatotoxicity of many drugs, as one important reason for liver failure, is poorly screened for in human models. Rocuronium and succinylcholine are neuromuscular blocking agents used for tracheal intubation and for rapid-sequence induction. We used an in-vitro test with a permanent cell line and compared rocuronium and succinylcholine for hepatotoxicity. In-vitro study. A basic science laboratory, University Hospital Rostock, Germany. The basic test compound is the permanent human liver cell line HepG2/C3A. In a standardised microtitre plate assay the toxicity of different concentrations of rocuronium, succinylcholine and plasma control was tested. After two incubation periods of 3 days, the viability of cells (XTT test, lactate dehydrogenase release and trypan blue staining), micro-albumin synthesis and the cytochrome 1A2 activity (metabolism of ethoxyresorufin) were measured. Differences between rocuronium and succinylcholine were assessed using the Kruskal-Wallis one-way test and two-tailed Mann-Whitney U test. Rocuronium, but not succinylcholine, led to a significant dose-dependent decrease of viability, albumin synthesis and cytochrome 1A2 activity of test cells. An in-vitro test with a cell line showed hepatotoxicity of rocuronium that was dose-dependent. Further studies are needed to investigate the underlying mechanisms of the effects of rocuronium on hepatic cellular integrity. Not suitable.

Involvement of ERK in NMDA receptor-independent cortical neurotoxicity of hydrogen sulfide

International Nuclear Information System (INIS)

Kurokawa, Yuko; Sekiguchi, Fumiko; Kubo, Satoko; Yamasaki, Yoshiko; Matsuda, Sachi; Okamoto, Yukari; Sekimoto, Teruki; Fukatsu, Anna; Nishikawa, Hiroyuki; Kume, Toshiaki; Fukushima, Nobuyuki; Akaike, Akinori; Kawabata, Atsufumi

2011-01-01

Highlights: ► Hydrogen sulfide causes NMDA receptor-independent neurotoxicity in mouse fetal cortical neurons. ► Activation of ERK mediates the toxicity of hydrogen sulfide. ► Apoptotic mechanisms are involved in the hydrogen-induced cell death. -- Abstract: Hydrogen sulfide (H 2 S), a gasotransmitter, exerts both neurotoxicity and neuroprotection, and targets multiple molecules including NMDA receptors, T-type calcium channels and NO synthase (NOS) that might affect neuronal viability. Here, we determined and characterized effects of NaHS, an H 2 S donor, on cell viability in the primary cultures of mouse fetal cortical neurons. NaHS caused neuronal death, as assessed by LDH release and trypan blue staining, but did not significantly reduce the glutamate toxicity. The neurotoxicity of NaHS was resistant to inhibitors of NMDA receptors, T-type calcium channels and NOS, and was blocked by inhibitors of MEK, but not JNK, p38 MAP kinase, PKC and Src. NaHS caused prompt phosphorylation of ERK and upregulation of Bad, followed by translocation of Bax to mitochondria and release of mitochondrial cytochrome c, leading to the nuclear condensation/fragmentation. These effects of NaHS were suppressed by the MEK inhibitor. Our data suggest that the NMDA receptor-independent neurotoxicity of H 2 S involves activation of the MEK/ERK pathway and some apoptotic mechanisms.

EFEITO DOS MEIOS DILUÍDORES MMC (MÍNIMA CONTAMINAÇÃO E LGM (LACTOSE-GEMA MODIFICADO NA VIABILIDADE DO SÊMEN DE CÃES CONSERVADO POR 24 HORAS EM EQUITAINER®.

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Tania Goes de Pinho

2011-03-01

Full Text Available The aim of this study was to compare the viability of canine semen after dilution in Minimum Contamination Medium (MMC and Modified Lactose-Egg yolk extender (LGM and incubation at 4 ºC for 12 and 24 hours. Thirteen ejaculates were collected from 5 adult dogs by digital manipulation. Macroscopic and microscopic characteristics were assessed right after collection. Semen was divided to be diluted (1:3 in MMC and LGM and subsequently chilled in Equitainer®. Seminal parameters of motility and spermatic morphology, membrane integrity (hyposmotic test, spermatic viability and spontaneous acrosome reaction (Trypan-blue Giemsa stain were evaluated in fresh and chilled semen after 12 and 24 hours of incubation at 4 ºC. No difference between extenders was identified in semen conservation after 12 hours. After 24 hours just spermatic motility was different (p>0.05. But, after both periods of conservation, semen diluted in LGM maintained most of the characteristics verified in the fresh semen. The mean of true and false acrosome reaction did not exceed 2% in both semen extenders, which demonstrates no influence of media component and incubation period on these phenomena at 4 ºC. In conclusion, these results indicate that both extenders can be used in canine semen conservation at 4 ºC in contêiner during 12 hours without significant changes in semen characteristics.

The antiproliferative and apoptotic effects of apigenin on glioblastoma cells.

Science.gov (United States)

Stump, Trevor A; Santee, Brittany N; Williams, Lauren P; Kunze, Rachel A; Heinze, Chelsae E; Huseman, Eric D; Gryka, Rebecca J; Simpson, Denise S; Amos, Samson

2017-07-01

Glioblastoma (GBM) is highly proliferative, infiltrative, malignant and the most deadly form of brain tumour. The epidermal growth factor receptor (EGFR) is overexpressed, amplified and mutated in GBM and has been shown to play key and important roles in the proliferation, growth and survival of this tumour. The goal of our study was to investigate the antiproliferative, apoptotic and molecular effects of apigenin in GBM. Proliferation and viability tests were carried out using the trypan blue exclusion, MTT and lactate dehydrogenase (LDH) assays. Flow cytometry was used to examine the effects of apigenin on the cell cycle check-points. In addition, we determined the effects of apigenin on EGFR-mediated signalling pathways by Western blot analyses. Our results showed that apigenin reduced cell viability and proliferation in a dose- and time-dependent manner while increasing cytotoxicity in GBM cells. Treatment with apigenin-induced is poly ADP-ribose polymerase (PARP) cleavage and caused cell cycle arrest at the G2M checkpoint. Furthermore, our data revealed that apigenin inhibited EGFR-mediated phosphorylation of mitogen-activated protein kinase (MAPK), AKT and mammalian target of rapamycin (mTOR) signalling pathways and attenuated the expression of Bcl-xL. Our results demonstrated that apigenin has potent inhibitory effects on pathways involved in GBM proliferation and survival and could potentially be used as a therapeutic agent for GBM. © 2017 Royal Pharmaceutical Society.

Dental pulp stem cells immobilized in alginate microspheres for applications in bone tissue engineering.

Science.gov (United States)

Kanafi, M M; Ramesh, A; Gupta, P K; Bhonde, R R

2014-07-01

To immobilize dental pulp stem cells (DPSC) in alginate microspheres and to determine cell viability, proliferation, stem cell characteristics and osteogenic potential of the immobilized DPSCs. Human DPSCs isolated from the dental pulp were immobilized in 1% w/v alginate microspheres. Viability and proliferation of immobilized DPSCs were determined by trypan blue and MTT assay, respectively. Stem cell characteristics of DPSCs post immobilization were verified by labelling the cells with CD73 and CD90. Osteogenic potential of immobilized DPSCs was assessed by the presence of osteocalcin. Alizarin red staining and O-cresolphthalein complexone method confirmed and quantified calcium deposition. A final reverse transcriptase PCR evaluated the expression of osteogenic markers - ALP, Runx-2 and OCN. More than 80% of immobilized DPSCs were viable throughout the 3-week study. Proliferation appeared controlled and consistent unlike DPSCs in the control group. Presence of CD73 and CD90 markers confirmed the stem cell nature of immobilized DPSCs. The presence of osteocalcin, an osteoblastic marker, was confirmed in the microspheres on day 21. Mineralization assays showed high calcium deposition indicating elevated osteogenic potential of immobilized DPSCs. Osteogenic genes- ALP, Runx-2 and OCN were also upregulated in immobilized DPSCs. Surprisingly, immobilized DPSCs in the control group cultured in conventional stem cell media showed upregulation of osteogenic genes and expressed osteocalcin. Dental pulp stem cells immobilized in alginate hydrogels exhibit enhanced osteogenic potential while maintaining high cell viability both of which are fundamental for bone tissue regeneration. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Effects of Ranolazine on Astrocytes and Neurons in Primary Culture.

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Martin Aldasoro

Full Text Available Ranolazine (Rn is an antianginal agent used for the treatment of chronic angina pectoris when angina is not adequately controlled by other drugs. Rn also acts in the central nervous system and it has been proposed for the treatment of pain and epileptic disorders. Under the hypothesis that ranolazine could act as a neuroprotective drug, we studied its effects on astrocytes and neurons in primary culture. We incubated rat astrocytes and neurons in primary cultures for 24 hours with Rn (10-7, 10-6 and 10-5 M. Cell viability and proliferation were measured using trypan blue exclusion assay, MTT conversion assay and LDH release assay. Apoptosis was determined by Caspase 3 activity assay. The effects of Rn on pro-inflammatory mediators IL-β and TNF-α was determined by ELISA technique, and protein expression levels of Smac/Diablo, PPAR-γ, Mn-SOD and Cu/Zn-SOD by western blot technique. In cultured astrocytes, Rn significantly increased cell viability and proliferation at any concentration tested, and decreased LDH leakage, Smac/Diablo expression and Caspase 3 activity indicating less cell death. Rn also increased anti-inflammatory PPAR-γ protein expression and reduced pro-inflammatory proteins IL-1 β and TNFα levels. Furthermore, antioxidant proteins Cu/Zn-SOD and Mn-SOD significantly increased after Rn addition in cultured astrocytes. Conversely, Rn did not exert any effect on cultured neurons. In conclusion, Rn could act as a neuroprotective drug in the central nervous system by promoting astrocyte viability, preventing necrosis and apoptosis, inhibiting inflammatory phenomena and inducing anti-inflammatory and antioxidant agents.

Evaluation of royal jelly as an alternative to fetal bovine serum in cell culture using cell proliferation assays and live cell imaging.

Science.gov (United States)

Musa, Marahaini; Nasir, Nurul Fatihah Mohamad; Thirumulu, Kannan Ponnuraj

2014-01-01

Royal jelly is a nutritious substance produced by the young nurse bees and contains significant amounts of proteins which are important for cell growth and proliferation. The aim of this study was to evaluate the effect of royal jelly as an alternative to fetal bovine serum (FBS) in cell culture using cell proliferation assays and live cell imaging. MRC-5 cells were treated with various concentrations of royal jelly extract in MTT assay. The control groups were comprised of Alpha-Minimal Essential Medium (α-MEM) alone and α-MEM with 10% FBS. Subsequently, the cell proliferation was studied for 10 days using Alamar Blue assay and live cell imaging from 48 to 72 h. The population doubling time (PDT) was determined using trypan blue assay after live cell imaging. In MTT assay, 0.156 and 0.078 mg/ml of royal jelly produced higher cell viability compared to positive control group but were not significantly different (P > 0.05). In the Alamar Blue assay, 0.156 and 0.078 mg/ml of royal jelly produced greater percentage of reduction at day 3 even though no significant difference was found (P > 0.05). Based on live cell imaging, the PDT for positive, negative, 0.156 and 0.078 mg/ml of royal jelly groups were 29.09, 62.50, 41.67 and 41.67 h respectively. No significant difference was found in the PDT between all the groups (P > 0.05). Royal jelly does not exhibit similar ability like FBS to facilitate cell growth under the present test conditions.

THE EFFECT OF X-RAY ON THE SKIN OF VITALLY STAINED WHITE MICE

Science.gov (United States)

Cori, Gerty T.

1924-01-01

1. The time interval between radiation and the occurrence of epilation is shorter in mice injected with trypan blue than in normal animals. 2. An x-ray unit defined as that causing total spontaneous epilation on the skin of the mouse is suggested. It corresponds to four to five human erythema doses. PMID:19868874

Paris polyphylla extract inhibits proliferation and promotes apoptosis ...

African Journals Online (AJOL)

Purpose: To investigate the effect of Paris polyphylla extract (PPE) on proliferation and apoptosis in A549 human lung cancer cells. Methods: Morphological changes were examined by microscopy in A549 cells after exposure to PPE. Trypan blue staining of living cells was used to aid the construction of the cell growth curve ...

Dermatoscopy of blue vitiligo.

Science.gov (United States)

Chandrashekar, L

2009-07-01

Blue vitiligo is a distinct variant of vitiligo characterized by a blue-grey appearance of the skin, which corresponds histologically with absence of epidermal melanocytes and presence of numerous dermal melanophages. A 23-year-old woman of Indian origin with Fitzpatrick skin type V presented with a 1-month history of normoaesthetic depigmented macules over the right forearm, dorsa of the hands and right areola. The macule over the right forearm had a bluish tinge. A clinical diagnosis of vitiligo vulgaris with blue vitiligo was made. Dermatoscopy of the interface between the blue macule and the hypopigmented macule revealed a linear depigmented macule in the centre with multiple blue dots and absence of epidermal melanin on the side of the blue macule, and reticular pigmentation with a few depigmented macules and scattered blue dots over the side of the hypopigmented macule. Blue vitiligo was described previously in a patient seropositive for human immunodeficiency virus, and believed to represent postinflammatory hyperpigmentation in areas bordering the vitiliginous patches as a result of psoralen ultraviolet A treatment. This case is unusual because of its rarity and the description of the associated dermatoscopical findings.

Studies on acute toxic effects to keratinocytes induced by hematoporphyrin derivatives and laser light.

Science.gov (United States)

Artuc, M; Ramshad, M; Kappus, H

1989-01-01

Human epidermal keratinocytes were grown in culture and the uptake of hematoporphyrin derivatives (HPDs) used in photodynamic therapy was estimated. Keratinocytes loaded with HPDs were irradiated with laser light of 632 nm generated by a helium-neon laser and cell toxicity was determined by the trypan blue exclusion test and the measurement of enzyme release. With increasing intracellular concentration of HPDs and with increasing intensity of the laser light, an increasing number of cells took up trypan blue and released the cytosolic enzyme lactate dehydrogenase and the lysosomal enzyme acid phosphatase after 1 h incubation of the irradiated cells at 37 degrees C. Cytotoxicity was less pronounced when the irradiated cells were incubated at 0 degree C indicating the involvement of enzyme reactions in cell death. No lipid peroxidation as measured by malondialdehyde and ethane formation was detectable. Our results suggest that during photodynamic therapy with HPDs and laser light epidermal keratinocytes may be seriously damaged. The data indicate that not lipid peroxidation but rather the activation of lysosomal enzymes is responsible for the cytotoxicity observed.

Multifractal resilience and viability

Science.gov (United States)

Tchiguirinskaia, I.; Schertzer, D. J. M.

2017-12-01

The term resilience has become extremely fashionable and there had been many attempts to provide operational definition and in fact metrics going beyond a set of more or less ad-hoc indicators. The viability theory (Aubin and Saint-Pierre, 2011) have been used to give a rather precise mathematical definition of resilience (Deffuant and Gilbert, 2011). However, it does not grasp the multiscale nature of resilience that is rather fundamental as particularly stressed by Folke et al (2010). In this communication, we first recall a preliminary attempt (Tchiguirinskaia et al., 2014) to define multifractal resilience with the help of the maximal probable singularity. Then we extend this multifractal approach to the capture basin of the viability, therefore the resilient basin. Aubin, J P, A. Bayen, and P Saint-Pierre (2011). Viability Theory. New Directions. Springer, Berlin,. Deffuant, G. and Gilbert, N. (eds) (2011) Viability and Resilience of Complex Systems. Springer Berlin.Folke, C., S R Carpenter, B Walker, M Sheffer, T Chapin, and J Rockstroem (2010). Resilience thinking: integrating re- silience, adaptability and transformability. Ecology and So- ciety, 14(4):20, Tchiguirinskaia,I., D. Schertzer, , A. Giangola-Murzyn and T. C. Hoang (2014). Multiscale resilience metrics to assess flood. Proceedings of ICCSA 2014, Normandie University, Le Havre, France -.

A cell transportation solution that preserves live circulating tumor cells in patient blood samples

International Nuclear Information System (INIS)

Stefansson, Steingrimur; Adams, Daniel L.; Ershler, William B.; Le, Huyen; Ho, David H.

2016-01-01

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90 % viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs

A cell transportation solution that preserves live circulating tumor cells in patient blood samples.

Science.gov (United States)

Stefansson, Steingrimur; Adams, Daniel L; Ershler, William B; Le, Huyen; Ho, David H

2016-05-06

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90% viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs after

Origin of colour stability in blue/orange/blue stacked phosphorescent white organic light-emitting diodes

International Nuclear Information System (INIS)

Kim, Sung Hyun; Jang, Jyongsik; Yook, Kyoung Soo; Lee, Jun Yeob

2009-01-01

The origin of colour stability in phosphorescent white organic light-emitting diodes (PHWOLEDs) with a blue/orange/blue stacked emitting structure was studied by monitoring the change in a recombination zone. A balanced recombination zone shift between the blue and the orange light-emitting layers was found to be responsible for the colour stability in the blue/orange/blue stacked PHWOLEDs.

Comparison of the Viability of Cryopreserved Fat Tissue in Accordance with the Thawing Temperature

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So-Min Hwang

2015-03-01

Full Text Available BackgroundAdipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at -20℃ by using thawing methods used in clinics.MethodsThe survival rates of adipocytes in the following thawing groups were measured: natural thawing at 25℃ for 15 minutes; natural thawing at 25℃ for 5 minutes, followed by rapid thawing at 37℃ in a water bath for 5 minutes; and rapid thawing at 37℃ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes.ResultsIn the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05. The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant.ConclusionsIt was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at 37℃. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

Blue-Green Algae

Science.gov (United States)

... that taking a specific blue-green algae product (Super Blue-Green Algae, Cell Tech, Klamath Falls, OR) ... system. Premenstrual syndrome (PMS). Depression. Digestion. Heart disease. Memory. Wound healing. Other conditions. More evidence is needed ...

Blue gods, blue oil, and blue people.

Science.gov (United States)

Fairbanks, V F

1994-09-01

Studies of the composition of coal tar, which began in Prussia in 1834, profoundly affected the economies of Germany, Great Britain, India, and the rest of the world, as well as medicine and surgery. Such effects include the collapse of the profits of the British indigo monopoly, the growth in economic power of Germany based on coal tar chemistry, and an economic crisis in India that led to more humane tax laws and, ultimately, the independence of India and the end of the British Empire. Additional consequences were the development of antiseptic surgery and the synthesis of a wide variety of useful drugs that have eradicated infections and alleviated pain. Many of these drugs, particularly the commonly used analgesics, sulfonamides, sulfones, and local anesthetics, are derivatives of aniline, originally called "blue oil" or "kyanol." Some of these aniline derivatives, however, have also caused aplastic anemia, agranulocytosis, and methemoglobinemia (that is, "blue people"). Exposure to aniline drugs, particularly when two or three aniline drugs are taken concurrently, seems to be the commonest cause of methemoglobinemia today.

[Acute blue urticaria following subcutaneous injection of patent blue dye].

Science.gov (United States)

Hamelin, A; Vial-Dupuy, A; Lebrun-Vignes, B; Francès, C; Soria, A; Barete, S

2015-11-01

Patent blue (PB) is a lymphatic vessel dye commonly used in France for sentinel lymph node detection in breast cancer, and less frequently in melanoma, and which may induce hypersensitivity reactions. We report a case of acute blue urticaria occurring within minutes of PB injection. Ten minutes after PB injection for sentinel lymph node detection during breast cancer surgery, a 49-year-old woman developed generalised acute blue urticaria and eyelid angioedema without bronchospasm or haemodynamic disturbance, but requiring discontinuation of surgery. Skin testing using PB and the anaesthetics given were run 6 weeks after the episode and confirmed PB allergy. PB was formally contra-indicated. Immediate hypersensitivity reactions to PB have been reported for between 0.24 and 2.2% of procedures. Such reactions are on occasion severe, chiefly involving anaphylactic shock. Two mechanisms are probably associated: non-specific histamine release and/or an IgE-mediated mechanism. Skin tests are helpful in confirming the diagnosis of PB allergy. Blue acute urticaria is one of the clinical manifestations of immediate hypersensitivity reactions to patent blue dye. Skin tests must be performed 6 weeks after the reaction in order to confirm the diagnosis and formally contra-indicate this substance. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Photothermal effects of multi-walled carbon nanotubes on the viability of BT-474 cancer cells

International Nuclear Information System (INIS)

Chou, Hung-Tao; Wang, Tsung-Pao; Lee, Chi-Young; Tai, Nyan-Hwa; Chang, Hwan-You

2013-01-01

Functionalized multi-walled carbon nanotubes (f-MWCNTs) were conjugated to an antibody of BT-474 cancer cells (f-MWCNTs-ab), and the photothermal effect of the f-MWCNTs-ab for BT-474 cancer cell destruction was demonstrated. After near-infrared irradiation, the f-MWCNTs-ab were more capable of killing cancer cells and possessed higher cell specificity than f-MWCNTs. Quantitative results showed that the viability of the cancer cells was affected by the concentration of the f-MWCNTs-ab solution, irradiation time, and settling time after irradiation. The membrane impermeable fluorescence dye ethidium bromide was used to detect cell viability after near-infrared irradiation, and the results agreed with those obtained from the Alamar Blue cell viability assay. The EtBr fluorescence results suggest that the cell membrane, attached to f-MWCNTs-ab, was damaged after irradiation, which led to cell death and necrosis. Using confocal microscopy, a few f-MWCNTs-ab were detected in the cell, indicating the endocytosis effect. The results not only explain the improved efficiency of thermotherapy but also indicate that necrosis may result from protein denaturation attributing to the heated f-MWCNTs-ab in the cell. Highlights: ► f-MWCNTs conjugated with anti-HER2 antibody by chemical method. ► Kill breast cancer cells by using low dose f-MWCNTs-ab due to photothermal effect. ► Use EtBr fluorescent to prove that the cell membrane was broken by heated f-MWCNTs. ► Few f-MWCNTs-ab were detected in the cell indicating the endocytosis effect. ► Necrosis may result from protein denaturation due to contact with the heated CNTs.

Jagua blue derived from Genipa americana L. fruit: A natural alternative to commonly used blue food colorants?

Science.gov (United States)

Brauch, J E; Zapata-Porras, S P; Buchweitz, M; Aschoff, J K; Carle, R

2016-11-01

Due to consumers' increasing health awareness, food industry aims at replacing synthetic dyes by natural counterparts. The substitution of blue synthetic dyes is particularly challenging since current natural alternatives such as phycocyanin (Spirulina) suffer from poor stability. Jagua blue (produced from Genipa americana L. fruit) might represent a potential novel blue pigment source. However, only little is known about its color properties, and application in food systems. Therefore, the blue color and the stability of Jagua blue were assessed for the first time and compared to commonly used colorants, namely, Spirulina, brilliant blue FCF (Blue no. 1), and indigo carmine (Blue no. 2). The reaction rate of Jagua blue was independent of its concentration, confirming thermal degradation to follow first-order kinetics. Between pH 3.6 and 5.0, the color hue of Jagua blue solutions was similar to that of Blue no. 2. However, Jagua blue revealed markedly higher storage stabilities (t 1/2 =86-105days) than Blue no. 2 (t 1 /2 ≤9days) and was less susceptible to acidic pH of 3.6 (t 1 /2 =86days) than Spirulina (t 1 /2 =70days). High negative b* values (blueness) of colored gelatin gels were only obtained for Jagua blue and Spirulina, and the former exhibited higher light stabilities (t 1 /2 =15days) than Spirulina gels (t 1 /2 =4days). Our findings indicate Jagua blue to be a most promising alternative to synthetic dyes, providing relevant information regarding potential food applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Fictional Black Blues Figure: Blues Music and the Art of Narrative Self-Invention

OpenAIRE

Mack, Kimberly

2015-01-01

The Fictional Black Blues Figure: Blues Music and the Art of Narrative Self-Invention, Kimberly MackMy dissertation examines representations of black American blues musicians in contemporary American fiction, drama, and popular music, and it argues that blues music can be examined as a narrative art rooted in the tradition of fictionalized autobiographical self-fashioning. I contend that the contemporary, multi-racial, literary and musical characters in my project who participate in so-called...

Coomassie Brilliant Blue G is a more potent antagonist of P2 purinergic responses than Reactive Blue 2 (Cibacron Blue 3GA) in rat parotid acinar cells

International Nuclear Information System (INIS)

Soltoff, S.P.; McMillian, M.K.; Talamo, B.R.

1989-01-01

The ability of Brilliant Blue G (Coomassie Brilliant Blue G) and Reactive Blue 2 (Cibacron Blue 3GA) to block the effects of extracellular ATP on rat parotid acinar cells was examined by evaluating their effects on ATP-stimulated 45Ca 2+ entry and the elevation of [Ca 2+ ]i (Fura 2 fluorescence). ATP (300 microM) increased the rate of Ca 2+ entry to more than 25-times the basal rate and elevated [Ca 2+ ]i to levels more than three times the basal value. Brilliant Blue G and Reactive Blue 2 greatly reduced the entry of 45 Ca 2+ into parotid cells, but the potency of Brilliant Blue G (IC50 approximately 0.4 microM) was about 100-times that of Reactive Blue 2. Fura 2 studies demonstrated that inhibitory concentrations of these compounds did not block the cholinergic response of these cells, thus demonstrating the selectivity of the dye compounds for purinergic receptors. Unlike Reactive Blue 2, effective concentrations of Brilliant Blue G did not substantially quench Fura 2 fluorescence. The greater potency of Brilliant Blue G suggests that it may be very useful in identifying P2-type purinergic receptors, especially in studies which utilize fluorescent probes

Measuring Blue Space Visibility and 'Blue Recreation' in the Everyday Lives of Children in a Capital City.

Science.gov (United States)

Pearson, Amber L; Bottomley, Ross; Chambers, Tim; Thornton, Lukar; Stanley, James; Smith, Moira; Barr, Michelle; Signal, Louise

2017-05-26

Blue spaces (water bodies) may promote positive mental and physical health through opportunities for relaxation, recreation, and social connections. However, we know little about the nature and extent of everyday exposure to blue spaces, particularly in settings outside the home or among children, nor whether exposure varies by individual or household characteristics. Wearable cameras offer a novel, reliable method for blue space exposure measurement. In this study, we used images from cameras worn over two days by 166 children in Wellington, New Zealand, and conducted content and blue space quantification analysis on each image ( n = 749,389). Blue space was identified in 24,721 images (3.6%), with a total of 23 blue recreation events. Visual exposure and participation in blue recreation did not differ by ethnicity, weight status, household deprivation, or residential proximity to the coastline. Significant differences in both visual exposure to blue space and participation in blue recreation were observed, whereby children from the most deprived schools had significantly higher rates of blue space exposure than children from low deprivation schools. Schools may be important settings to promote equitable blue space exposures. Childhood exposures to blue space may not follow the expected income inequality trends observed among adults.

Stimulation of human gingival fibroblasts viability and growth by roots treated with high intensity lasers, photodynamic therapy and citric acid.

Science.gov (United States)

Karam, Paula Stephania Brandão Hage; Ferreira, Rafael; Oliveira, Rodrigo Cardoso; Greghi, Sebastião Luiz Aguiar; de Rezende, Maria Lúcia Rubo; Sant'Ana, Adriana Campos Passanezi; Zangrando, Mariana Schutzer Ragghianti; Damante, Carla Andreotti

2017-09-01

The aim of this study was to compare the effect of root biomodification by lasers, citric acid and antimicrobial photodynamic therapy (aPDT) on viability and proliferation of human gingival fibroblasts (FGH). Groups were divided in control (CC - only cells), and root fragments treated by: scaling and root planing (positice control - SC), Er:YAG (ER-60mJ,10pps,10Hz,10s,2940nm), Nd:YAG (ND-0.5W,15Hz,10s,1640nm), antimicrobial photodynamic therapy (PDT-InGaAIP,30mW,45J/cm 2 ,30s,660nm,toluidine blue O), citric acid plus tetracycline (CA). Fibroblasts (6th passage, 2×10 3 ) were cultivated in a 24-h conditioned medium by the treated root fragments. Cell viability was measured by MTT test at 24, 48, 72 and 96h. In a second experiment, FGH cells (10 4 ) were cultivated on root fragments which received the same treatments. After 24, 48, 72h the number of cells was counted in SEM pictures. In addition, chemical elements were analyzed by energy dispersive spectroscopy (EDS). Data was analyzed by two-way ANOVA (first experiment), repeated measures ANOVA (second experiment) and ANOVA (EDS experiment) tests complemented by Tukey's test (pplaning stimulated fibroblast viability while Er:YAG and Nd:YAG treated root surfaces presented higher number of cells. Copyright © 2017. Published by Elsevier Ltd.

75 FR 65525 - Anthem Blue Cross Blue Shield, Claim Management Services, Inc. Operations, a Division of...

Science.gov (United States)

2010-10-25

... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-74,327] Anthem Blue Cross Blue Shield, Claim Management Services, Inc. Operations, a Division of Wellpoint, Inc., Green Bay, WI; Notice... former workers of Anthem Blue Cross Blue Shield, Claim Management Services, Inc. Operations, a Division...

Dyes adsorption blue vegetable and blue watercolor by natural zeolites modified with surfactants

International Nuclear Information System (INIS)

Jardon S, C. C.; Olguin G, M. T.; Diaz N, M. C.

2009-01-01

In this work was carried out the dyes removal blue vegetable and blue watercolor of aqueous solutions, to 20 C, at different times and using a zeolite mineral of Parral (Chihuahua, Mexico) modified with hexadecyl trimethyl ammonium bromide or dodecyl trimethyl ammonium bromide. The zeolite was characterized before and after of its adaptation with NaCl and later with HDTMABr and DTMABr. For the materials characterization were used the scanning electron microscopy of high vacuum; elementary microanalysis by X-ray spectroscopy of dispersed energy and X-ray diffraction techniques. It was found that the surfactant type absorbed in the zeolite material influences on the adsorption process of the blue dye. Likewise, the chemical structure between the vegetable blue dye and the blue watercolor, determines the efficiency of the color removal of the water, by the zeolites modified with the surfactants. (Author)

Spontaneous and radiation induced cell death in HeLa S3 human carcinoma

International Nuclear Information System (INIS)

Zaric, B.; Milosavljevic, B.; Radojcic, M.

2001-01-01

Radiation biologists have classified radiation-induced cell death based on cell proliferative capacity to either mitotic or interphase death. Cytologists have revealed two morphologically and biochemically diverse forms of cell death, apoptosis and necrosis. While the knowledge of the former is already well exploited by radiologists, cell susceptibility to apoptosis and necrosis is still under investigation. We studied characteristics of spontaneous cell death, and dose dependence and time course of radiation-induced cell death of human uterine cervix epitheloid carcinoma HeLaS 3 in culture. Cells were irradiated with 2-40 Gy of γ-rays. The effect on growth, viability, morphology and genomic DNA structure were followed 24-72 h after irradiation. Cell viability was evaluated by trypan-blue exclusion assay and cell morphology by in situ DNA staining with propidium iodide. Cell genomic DNA fragmentation pattern was determined by electrophoresis on 2% agarose gels. At all cell densities 25-35% cells were PI positive and their DNA was fragmented to a high molecular size (≥20 kbp), but the internucleosomal ladder was not observed. A significant decrease in viability to 33% was observed 72 h post 40 Gy irradiation. It corresponded to 55% of PI positive cells. A smear of smaller DNA fragments (0.1-1 kbp), 24 h after 10-20 Gy irradiation was considered as proof that the dominant form of radiation-induced cell death was necrosis. It was concluded that the dominant form of radiation-induced cell death in HeLaS 3 population was necrosis and the radiation dose which caused 50% of cell death after 72 h (termed ND 50 ) was between 30-40 Gy. (author)

Pollen viability and membrane lipid composition

NARCIS (Netherlands)

Bilsen, van D.G.J.L.

1993-01-01

In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

Synthesis of oleic acid functionalized Fe3O4 magnetic nanoparticles and studying their interaction with tumor cells for potential hyperthermia applications.

Science.gov (United States)

Jadhav, Neena V; Prasad, Amresh I; Kumar, Amit; Mishra, R; Dhara, Sangita; Babu, K R; Prajapat, C L; Misra, N L; Ningthoujam, R S; Pandey, B N; Vatsa, R K

2013-08-01

In the present study, oleic acid (OA) functionalized Fe3O4 magnetic nanoparticles (MN) were synthesized following modified wet method of MN synthesis. The optimum amount of OA required for capping of MN and the amount of bound and unbound/free OA was determined by thermogravimetric analysis (TGA). Further, we have studied the effect of water molecules, associated with MN, on the variation in their induction heating ability under alternating current (AC) magnetic field conditions. We have employed a new approach to achieve dispersion of OA functionalized MN (MN-OA) in aqueous medium using sodium carbonate, which improves their biological applicability. Interactions amongst MN, OA and sodium carbonate were studied by Fourier transform infrared spectroscopy (FT-IR). Intracellular localization of MN-OA was studied in mouse fibrosarcoma cells (WEHI-164) by prussian blue staining and confocal laser scanning microscopy (CLSM) using nile blue A as a fluorescent probe. Results showed MN-OA to be interacting mainly with the cell membrane. Their hyperthermic killing ability was evaluated in WEHI-164 cells by trypan blue method. Cells treated with MN-OA in combination with induction heating showed decreased viability as compared to respective induction heating controls. These results were supported by altered cellular morphology after treatment of MN-OA in combination with induction heating. Further, the magnitude of apoptosis was found to be ~5 folds higher in cells treated with MN-OA in combination with induction heating as compared to untreated control. These results suggest the efficacy of MN-OA in killing of tumor cells by cellular hyperthermia. Copyright © 2013 Elsevier B.V. All rights reserved.

shRNA-mediated EMMPRIN silencing inhibits human leukemic monocyte lymphoma U937 cell proliferation and increases chemosensitivity to adriamycin.

Science.gov (United States)

Gao, Hui; Jiang, Qixiao; Han, Yantao; Peng, Jianjun; Wang, Chunbo

2015-03-01

EMMPRIN is a widely distributed cell surface glycoprotein, which plays an important role in tumor progression and confers resistance to some chemotherapeutic drugs. Recent studies have shown that EMMPRIN overexpression indicates poor prognosis in acute myeloid leukemia (AML). However, little was known on the role of EMMPRIN in leukemia. Human leukemia cell line U937 was stably transfected with a EMMPRIN-targeted shRNA-containing vector to investigate the effect of EMMPRIN on cellular functions. EMMPRIN expression was monitored by qRT-PCR and Western blotting. Cell viability and proliferation were determined by trypan blue exclusion and BrdU labeling, respectively. Cell cycle and apoptosis were analyzed by flow cytometry. Cytotoxicity of chemotherapeutic agent adriamycin on cells was assessed by MTT assay. Knockdown of EMMPRIN gene significantly inhibited cell viability and decreased cell proliferation. Fluorescence-activated cell-sorting analysis revealed that the reduced EMMPRIN expression resulted in cell cycle arrest at G1 phase and induced apoptosis. Meanwhile, western blotting analysis showed that EMMPRIN knockdown was associated with downregulation of cell cycle- and apoptosis-related molecules including cyclin D1, cyclin E, as well as increase in cleavage of caspase-3 and PARP. This study also showed that silencing of EMMPRIN sensitized U937 cells to Adriamycin. EMMPRIN is involved in proliferation, growth, and chemosensitivity of human AML line U937, indicating that EMMPRIN may be a promising therapeutic target for AML.

Modified titanium surface with gelatin nano gold composite increases osteoblast cell biocompatibility

International Nuclear Information System (INIS)

Lee, Young-Hee; Bhattarai, Govinda; Aryal, Santosh; Lee, Nan-Hee; Lee, Min-Ho; Kim, Tae-Gun; Jhee, Eun-Chung; Kim, Hak-Yong; Yi, Ho-Keun

2010-01-01

This study examined the gelatin nano gold (GnG) composite for surface modification of titanium in addition to insure biocompatibility on dental implants or biomaterials. The GnG composite was constructed by gelatin and hydrogen tetrachloroaurate in presence of reducing agent, sodium borohydrate (NabH 4 ). The GnG composite was confirmed by UV-VIS spectroscopy and transmission electron microscopy (TEM). A dipping method was used to modify the titanium surface by GnG composite. Surface was characterized by scanning electron microscopy (SEM) and energy dispersive X-ray (EDX). The MC-3T3 E1 cell viability was assessed by trypan blue and the expression of proteins to biocompatibility were analyzed by Western blotting. The GnG composite showed well dispersed character, the strong absorption at 530 nm, roughness, regular crystal and clear C, Na, Cl, P, and Au signals onto titanium. Further, this composite allowed MC-3T3 E1 growth and viability compared to gelatin and pure titanium. It induced ERK activation and the expression of cell adherent molecules, FAK and SPARC, and growth factor, VEGF. However, GnG decreased the level of SAPK/JNK. This shows that GnG composite coated titanium surfaces have a good biocompatibility for osteoblast growth and attachment than in intact by simple and versatile dipping method. Furthermore, it offers good communication between cell and implant surfaces by regulating cell signaling and adherent molecules, which are useful to enhance the biocompatibility of titanium surfaces.

Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells

Energy Technology Data Exchange (ETDEWEB)

Barboza, Carlos Augusto Galvão; Ginani, Fernanda [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil); Soares, Diego Moura [Universidade Federal de Pernambuco, Recife, PE (Brazil); Henriques, Águida Cristina Gomes; Freitas, Roseana de Almeida [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil)

2014-07-01

To evaluate the effect of low-level laser irradiation on the proliferation and possible nuclear morphological changes of mouse mesenchymal stem cells. Mesenchymal stem cells derived from bone marrow and adipose tissue were submitted to two applications (T0 and T48 hours) of low-level laser irradiation (660nm; doses of 0.5 and 1.0J/cm{sup 2}). The trypan blue assay was used to evaluate cell viability, and growth curves were used to analyze proliferation at zero, 24, 48, and 72 hours. Nuclear alterations were evaluated by staining with DAPI (4'-6-diamidino-2-phenylindole) at 72 hours. Bone marrow-derived mesenchymal stem cells responded to laser therapy in a dose-dependent manner. Higher cell growth was observed when the cells were irradiated with a dose of 1.0J/cm{sup 2}, especially after 24 hours (p<0.01). Adipose-derived mesenchymal stem cells responded better to a dose of 1.0J/cm{sup 2}, but higher cell proliferation was observed after 48 hours (p<0.05) and 72 hours (p<0.01). Neither nuclear alterations nor a significant change in cell viability was detected in the studied groups. Low-level laser irradiation stimulated the proliferation of mouse mesenchymal stem cells without causing nuclear alterations. The biostimulation of mesenchymal stem cells using laser therapy might be an important tool for regenerative therapy and tissue engineering.

Optimization of the isolation and cultivation of Cyprinus carpio primary hepatocytes

Science.gov (United States)

Yanhong, Fan; Chenghua, He; Guofang, Liu

2008-01-01

The aquatic environment is affected by numerous chemical contaminants. There is an increasing need to identify these chemicals and to evaluate their potential toxicity towards aquatic life. In this research we optimized techniques for primary cell culture of Cyprinus carpio hepatocytes as one adjunct model for ecotoxicological evaluation of the potential hazards of xenobiotics in the aquatic environment. In this study, Cyprinus carpio hepatocytes were isolated by mechanical separation, two-step collagenase perfusion, and pancreatin digestion. The hepatocytes or parenchymal cells could be separated from cell debris and from non-parenchymal cells by low-speed centrifugation (Percoll gradient centrifugation). The harvested hepatocytes were suspended in DMEM, M199 (cultured in 5% CO2), or L-15 (cultured without 5% CO2) medium then cultured at 17, 27, or 37 °C. Cell yield was counted by use of a hemocytometer, and the viability of the cells was assessed by use of the Trypan blue exclusion test. Results from these studies showed that the best method of isolation was pancreatin digestion (the cell yield was 2.7 × 108 per g (liver weight) and the viability was 98.4%) and the best medium was M199 (cultured in 5% CO2) or L-15 (cultured without 5% CO2). The optimum culture temperature was 27 °C. The primary hepatocytes culture of Cyprimus carpio grew well and satisfied requirements for most toxicological experiments in this condition. PMID:19002769

Optimization of the isolation and cultivation of Cyprinus carpio primary hepatocytes.

Science.gov (United States)

Yanhong, Fan; Chenghua, He; Guofang, Liu; Haibin, Zhang

2008-10-01

The aquatic environment is affected by numerous chemical contaminants. There is an increasing need to identify these chemicals and to evaluate their potential toxicity towards aquatic life. In this research we optimized techniques for primary cell culture of Cyprinus carpio hepatocytes as one adjunct model for ecotoxicological evaluation of the potential hazards of xenobiotics in the aquatic environment. In this study, Cyprinus carpio hepatocytes were isolated by mechanical separation, two-step collagenase perfusion, and pancreatin digestion. The hepatocytes or parenchymal cells could be separated from cell debris and from non-parenchymal cells by low-speed centrifugation (Percoll gradient centrifugation). The harvested hepatocytes were suspended in DMEM, M199 (cultured in 5% CO(2)), or L-15 (cultured without 5% CO(2)) medium then cultured at 17, 27, or 37 degrees C. Cell yield was counted by use of a hemocytometer, and the viability of the cells was assessed by use of the Trypan blue exclusion test. Results from these studies showed that the best method of isolation was pancreatin digestion (the cell yield was 2.7 x 10(8) per g (liver weight) and the viability was 98.4%) and the best medium was M199 (cultured in 5% CO(2)) or L-15 (cultured without 5% CO(2)). The optimum culture temperature was 27 degrees C. The primary hepatocytes culture of Cyprimus carpio grew well and satisfied requirements for most toxicological experiments in this condition.

N-Methyl-D aspartate receptor-mediated effect on glucose transporter-3 levels of high glucose exposed-SH-SY5Y dopaminergic neurons.

Science.gov (United States)

Engin, Ayse Basak; Engin, Evren Doruk; Karakus, Resul; Aral, Arzu; Gulbahar, Ozlem; Engin, Atilla

2017-11-01

High glucose and insulin lead to neuronal insulin resistance. Glucose transport into the neurons is achieved by regulatory induction of surface glucose transporter-3 (GLUT3) instead of the insulin. N-methyl-D aspartate (NMDA) receptor activity increases GLUT3 expression. This study explored whether an endogenous NMDA receptor antagonist, kynurenic acid (KynA) affects the neuronal cell viability at high glucose concentrations. SH-SY5Y neuroblastoma cells were exposed to 150-250 mg/dL glucose and 40 μU/mL insulin. In KynA and N-nitro-l-arginine methyl ester (L-NAME) supplemented cultures, oxidative stress, mitochondrial metabolic activity (MTT), nitric oxide as nitrite+nitrate (NOx) and GLUT3 were determined at the end of 24 and 48-h incubation periods. Viable cells were counted by trypan blue dye. High glucose-exposed SH-SY5Y cells showed two-times more GLUT3 expression at second 24-h period. While GLUT3-stimulated glucose transport and oxidative stress was increased, total mitochondrial metabolic activity was significantly reduced. Insulin supplementation to high glucose decreased NOx synthesis and GLUT3 levels, in contrast oxidative stress increased three-fold. KynA significantly reduced oxidative stress, and increased MTT by regulating NOx production and GLUT3 expression. KynA is a noteworthy compound, as an endogenous, specific NMDA receptor antagonist; it significantly reduces oxidative stress, while increasing cell viability at high glucose and insulin concentrations. Copyright © 2017 Elsevier Ltd. All rights reserved.

CIRRHOSIS INDUCES APOPTOSIS IN RENAL TISSUE THROUGH INTRACELLULAR OXIDATIVE STRESS

Directory of Open Access Journals (Sweden)

Keli Cristina Simões da SILVEIRA

2015-03-01

Full Text Available Background Renal failure is a frequent and serious complication in patients with decompensated cirrhosis. Objectives We aimed to evaluate the renal oxidative stress, cell damage and impaired cell function in animal model of cirrhosis. Methods Secondary biliary cirrhosis was induced in rats by ligation of the common bile duct. We measured TBARS, ROS and mitochondrial membrane potential in kidney as markers of oxidative stress, and activities of the antioxidant enzymes. Relative cell viability was determined by trypan blue dye-exclusion assay. Annexin V-PE was used with a vital dye, 7-AAD, to distinguish apoptotic from necrotic cells and comet assay was used for determined DNA integrity in single cells. Results In bile duct ligation animals there was significant increase in the kidney lipoperoxidation and an increase of the level of intracellular ROS. There was too an increase in the activity of all antioxidant enzymes evaluated in the kidney. The percentage viability was above 90% in the control group and in bile duct ligation was 64.66% and the dominant cell death type was apoptosis. DNA damage was observed in the bile duct ligation. There was a decreased in the mitochondrial membrane potential from 71.40% ± 6.35% to 34.48% ± 11.40% in bile duct ligation. Conclusions These results indicate that intracellular increase of ROS cause damage in the DNA and apoptosis getting worse the renal function in cirrhosis.

Investigating Ceria Nanocrystals Uptake by Glioblastoma Multiforme Cells and its Related Effects: An Electron Microscopy Study

KAUST Repository

Aloufi, Bader

2017-01-22

Cerium oxide nanoparticles have been utilized widely nowadays in cancer research. It has been suggested by many studies that these nanoparticles are capable of having dual antioxidant behavior in healthy and cancer microenvironment; where in physiological condition, they act as antioxidant and do not affect the healthy cells, while in tumor-like condition; they act as an oxidase, and result in a selective killing for the cancer cells. In this experiment, the interaction of nanoceria with glioblastoma and healthy astrocyte cells was examined, and further correlated with the in vitro cytotoxic effects of various nanoceria concentrations (100 and 300 µg/ml) and exposure times (12, 24, and 48 hours). Electron microscopes were used to investigate the cellular-NPs interactions, and to examine the related cytotoxic effects in combination with trypan blue and propidium iodide viability assays. Our data suggest the following results. First, the two cell lines demonstrated capability of taken up the ceria through endocytosis pathway, where the NPs were recognized engulfed by double membrane vesicles at various regions over the cellular cytoplasm. Secondly, cerium oxide nanoparticles were found to affect the glioblastoma cells, but not so severely the corresponding healthy astrocytes at the various concentrations and incubation times, as revealed by the viability assays and the electron microscopy analysis. Thirdly, the viability of the glioblastoma cells after the treatment displayed a declined trend when increasing the ceria concentrations, but did not show such dependency with regard to the different time points. In all cases, the healthy astrocyte cells showed slight alterations in mitochondrial shape which did not influence their viability. Among the various nanoceria concentrations and exposure times, the most efficient dose of treatment was found to be with a concentration of 300 µg/ml at a time point of 24-hour, where higher reduction on the viability of

Algae viability over time in a ballast water sample

Science.gov (United States)

Gollasch, Stephan; David, Matej

2018-03-01

The biology of vessels' ballast water needs to be analysed for several reasons, one of these being performance tests of ballast water management systems. This analysis includes a viability assessment of phytoplankton. To overcome logistical problems to get algae sample processing gear on board of a vessel to document algae viability, samples may be transported to land-based laboratories. Concerns were raised how the storage conditions of the sample may impact algae viability over time and what the most appropriate storage conditions were. Here we answer these questions with a long-term algae viability study with daily sample analysis using Pulse-Amplitude Modulated (PAM) fluorometry. The sample was analysed over 79 days. We tested different storage conditions: fridge and room temperature with and without light. It seems that during the first two weeks of the experiment the viability remains almost unchanged with a slight downwards trend. In the continuing period, before the sample was split, a slightly stronger downwards viability trend was observed, which occurred at a similar rate towards the end of the experiment. After the sample was split, the strongest viability reduction was measured for the sample stored without light at room temperature. We concluded that the storage conditions, especially regarding temperature and light exposure, have a stronger impact on algae viability compared to the storage duration and that inappropriate storage conditions reduce algal viability. A sample storage time of up to two weeks in a dark and cool environment has little influence on the organism viability. This indicates that a two week time duration between sample taking on board a vessel and the viability measurement in a land-based laboratory may not be very critical.

Blue Ocean Thinking

Science.gov (United States)

Orem, Donna

2016-01-01

This article describes a concept called the "blue ocean thinking strategy," developed by W. Chan Kim and Renée Mauborgne, professors at INSEAD, an international graduate school of business in France. The "blue ocean" thinking strategy considers opportunities to create new markets for services, rather than focusing solely on…

Blue Emission in Proteins

OpenAIRE

Sarkar, Sohini; Sengupta, Abhigyan; Hazra, Partha; Mandal, Pankaj

2014-01-01

Recent literatures reported blue-green emission from amyloid fibril as exclusive signature of fibril formation. This unusual visible luminescence is regularly used to monitor fibril growth. Blue-green emission has also been observed in crystalline protein and in solution. However, the origin of this emission is not known exactly. Our spectroscopic study of serum proteins reveals that the blue-green emission is a property of protein monomer. Evidences suggest that semiconductor-like band struc...

Photothermal effects of multi-walled carbon nanotubes on the viability of BT-474 cancer cells

Energy Technology Data Exchange (ETDEWEB)

Chou, Hung-Tao [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Wang, Tsung-Pao [Department of Medical Science, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Lee, Chi-Young [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Tai, Nyan-Hwa, E-mail: nhtai@mx.nthu.edu.tw [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Chang, Hwan-You, E-mail: hychang@mx.nthu.edu.tw [Department of Medical Science, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China)

2013-03-01

Functionalized multi-walled carbon nanotubes (f-MWCNTs) were conjugated to an antibody of BT-474 cancer cells (f-MWCNTs-ab), and the photothermal effect of the f-MWCNTs-ab for BT-474 cancer cell destruction was demonstrated. After near-infrared irradiation, the f-MWCNTs-ab were more capable of killing cancer cells and possessed higher cell specificity than f-MWCNTs. Quantitative results showed that the viability of the cancer cells was affected by the concentration of the f-MWCNTs-ab solution, irradiation time, and settling time after irradiation. The membrane impermeable fluorescence dye ethidium bromide was used to detect cell viability after near-infrared irradiation, and the results agreed with those obtained from the Alamar Blue cell viability assay. The EtBr fluorescence results suggest that the cell membrane, attached to f-MWCNTs-ab, was damaged after irradiation, which led to cell death and necrosis. Using confocal microscopy, a few f-MWCNTs-ab were detected in the cell, indicating the endocytosis effect. The results not only explain the improved efficiency of thermotherapy but also indicate that necrosis may result from protein denaturation attributing to the heated f-MWCNTs-ab in the cell. Highlights: Black-Right-Pointing-Pointer f-MWCNTs conjugated with anti-HER2 antibody by chemical method. Black-Right-Pointing-Pointer Kill breast cancer cells by using low dose f-MWCNTs-ab due to photothermal effect. Black-Right-Pointing-Pointer Use EtBr fluorescent to prove that the cell membrane was broken by heated f-MWCNTs. Black-Right-Pointing-Pointer Few f-MWCNTs-ab were detected in the cell indicating the endocytosis effect. Black-Right-Pointing-Pointer Necrosis may result from protein denaturation due to contact with the heated CNTs.

Growth and viability of Aedes albopictus cell line in vitro after cesium-137 gamma irradiation

International Nuclear Information System (INIS)

Blakely, E.A.

1975-01-01

The radiosensitivity of the cultured mosquito cell line Aedes albopictus (Skuse) was investigated. Population growth was followed by total cell counts and by viable cell counts on aliquots of cultures exposed to various doses of gamma radiation during exponential growth. Viable cell determinations were based on the cellular exclusion of the dye, alcian blue, in a procedure adapted to the insect cells in culture. Viability determinations in the irradiated exponential cultures indicated that initially there was some increase in the gestation, suggesting that gonadal steroids may have unusual effects on uterine physiology and biochemistry in this species. Consequently, studies were undertaken to elucidate some of the basic responses of hamster uteri to estradiol benzoate and progesterone under conditions of protein malnutrition, actinomycin D administration and corticosterone injection. Furthermore, the effects of gonadal steroids on uteri of pregnant ovariectomized hamsters were studied

Electronic cigarettes induce DNA strand breaks and cell death independently of nicotine in cell lines.

Science.gov (United States)

Yu, Vicky; Rahimy, Mehran; Korrapati, Avinaash; Xuan, Yinan; Zou, Angela E; Krishnan, Aswini R; Tsui, Tzuhan; Aguilera, Joseph A; Advani, Sunil; Crotty Alexander, Laura E; Brumund, Kevin T; Wang-Rodriguez, Jessica; Ongkeko, Weg M

2016-01-01

Evaluate the cytotoxicity and genotoxicity of short- and long-term e-cigarette vapor exposure on a panel of normal epithelial and head and neck squamous cell carcinoma (HNSCC) cell lines. HaCaT, UMSCC10B, and HN30 were treated with nicotine-containing and nicotine-free vapor extract from two popular e-cigarette brands for periods ranging from 48 h to 8 weeks. Cytotoxicity was assessed using Annexin V flow cytometric analysis, trypan blue exclusion, and clonogenic assays. Genotoxicity in the form of DNA strand breaks was quantified using the neutral comet assay and γ-H2AX immunostaining. E-cigarette-exposed cells showed significantly reduced cell viability and clonogenic survival, along with increased rates of apoptosis and necrosis, regardless of e-cigarette vapor nicotine content. They also exhibited significantly increased comet tail length and accumulation of γ-H2AX foci, demonstrating increased DNA strand breaks. E-cigarette vapor, both with and without nicotine, is cytotoxic to epithelial cell lines and is a DNA strand break-inducing agent. Further assessment of the potential carcinogenic effects of e-cigarette vapor is urgently needed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mammalian cells exposed to ionizing radiation: structural and biochemical aspects

International Nuclear Information System (INIS)

Sabanero, M.; Flores V, L. L.; Azorin V, J. C.; Vallejo, M. A.; Cordova F, T.; Sosa A, M.; Castruita D, J. P.; Barbosa S, G.

2015-10-01

Acute or chronic exposure to ionizing radiation is a factor that may be hazardous to health. It has been reported that exposure to low doses of radiation (less than 50 mSv / year) and subsequently exposure to high doses have greater effects in people. However, it is unknown molecular and biochemical level alteration. This study, analyzes the susceptibility of a biological system (HeLa Atcc CCL-2 human cervix cancer cell line) to ionizing radiation (6 and 60 mSv/ 90). Our evaluate multiple variables such as: total protein profile, mitochondrial metabolic activity (XTT assay), cell viability (Trypan blue exclusion assay), cytoskeleton (actin micro filaments), nuclei (D API), genomic DNA. The results indicate, that cells exposed to ionizing radiation structurally show alterations in nuclear phenotype and aneuploidy, further disruption in the tight junctions and consequently on the distribution of actin micro filaments. Similar alterations were observed in cells treated with a genotoxic agent (200μM H 2 O 2 /1 h). In conclusion, this multi-criteria assessment enables precise comparisons of the effects of radiation between any biological systems. However, it is necessary to determine stress markers for integration of the effects of ionizing radiation. (Author)

Piperlongumine induces apoptosis and autophagy in leukemic cells through targeting the PI3K/Akt/mTOR and p38 signaling pathways.

Science.gov (United States)

Wang, Hongfei; Wang, Yongqiang; Gao, Hongmei; Wang, Bing; Dou, Lin; Li, Yin

2018-02-01

Piperlongumine is an alkaloid compound extracted from Piper longum L. It is a chemical substance with various pharmacological effects and medicinal value, including anti-tumor, lipid metabolism regulatory, antiplatelet aggregation and analgesic properties. The present study aimed to understand whether piperlongumine induces the apoptosis and autophagy of leukemic cells, and to identify the mechanism involved. Cell viability and autophagy were detected using MTT, phenazine methyl sulfate and trypan blue exclusion assays. The apoptosis rate was calculated using flow cytometry. The protein expression levels of microtubule-associated protein 1A/1B-light chain 3, Akt and mechanistic target of rapamycin (mTOR) were measured using western blotting. The cell growth of leukemic cells was completely inhibited following treatment with piperlongumine, and marked apoptosis was also induced. Dead cells as a result of autophagy were stained using immunofluorescence and observed under a light microscope. Phosphoinositide 3-kinase (PI3K)/Akt/mTOR signaling was suppressed by treatment with piperlongumine, while p38 signaling and caspase-3 activity were induced by treatment with piperlongumine. It was concluded that piperlongumine induces apoptosis and autophagy in leukemic cells through targeting the PI3K/Akt/mTOR and p38 signaling pathways.

Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG2 cells

International Nuclear Information System (INIS)

Guo Dongping; Li Xiaoyu; Sun, Ping; Tang Yibo; Chen Xiuying; Chen Qi; Fan Leming; Zang Bin; Shao Lizheng; Li Xiaorong

2006-01-01

Ultrasound-targeted microbubble destruction had been employed in gene delivery and promised great potential. Liver has unique features that make it attractive for gene therapy. However, it poses formidable obstacles to hepatocyte-specific gene delivery. This study was designed to test the efficiency of therapeutic gene transfer and expression mediated by ultrasound/microbubble strategy in HepG 2 cell line. Air-filled albumin microbubbles were prepared and mixed with plasmid DNA encoding low density lipoprotein receptor (LDLR) and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured HepG 2 cells under variable ultrasound conditions. Transfection rate of the transferred gene and cell viability were assessed by FACS analysis, confocal laser scanning microscopy, Western blot analysis and Trypan blue staining. The result demonstrated that microbubbles with ultrasound irradiation can significantly elevate exogenous LDLR gene expression and the expressed LDLRs were functional and active to uptake their ligands. We conclude that ultrasound-targeted microbubble destruction has the potential to promote safe and efficient LDLR gene transfer into hepatocytes. With further refinement, it may represent an effective nonviral avenue of gene therapy for liver-involved genetic diseases

A rapid, efficient, and economic device and method for the isolation and purification of mouse islet cells.

Directory of Open Access Journals (Sweden)

Yin Zongyi

Full Text Available Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods. Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P 1000 islets. In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.

Effect of light polarization on the efficiency of photodynamic therapy of basal cell carcinomas: an in vitro cellular study.

Science.gov (United States)

JalalKamali, M; Nematollahi-Mahani, S N; Shojaei, M; Shamsoddini, A; Arabpour, N

2018-02-01

In an in vitro study, the effect of light polarization on the efficiency of 5-aminolaevulinic acid (ALA) photodynamic therapy (PDT) of basal cell carcinoma (BCC) was investigated. Three states of light polarization (non-polarized, linearly polarized, and circularly polarized) were considered. Cells were exposed to green (532 pm 20 nm) irradiation from light emitting diodes. Cell survival was measured by the colorimetric assay (WST-1) and Trypan blue staining. The colorimetric assay showed a pronounced decrease in the cell viability (up to 30%) using polarized light compared to the non-polarized one in the wavelength region used. Similar results were obtained by the cell counting method (20-30% increase in cell death). The observed effect was dependent on the concentration of photosensitizer. The effect is more expressed in the case of linearly polarized light compared to the circularly polarized one. Results show that the use of polarized light increases the efficiency of in vitro ALA-PDT of BCC. Utilizing polarized light, it is possible to obtain the same effect from PDT by lower concentrations of photosensitizer. Additionally, the concentration dependency of PDT response and photo-bleaching is also reduced.

Mammalian cells exposed to ionizing radiation: structural and biochemical aspects

Energy Technology Data Exchange (ETDEWEB)

Sabanero, M.; Flores V, L. L. [Universidad de Guanajuato, Departamento de Biologia, DCNE, Noria Alta s/n, 36250 Guanajuato, Gto. (Mexico); Azorin V, J. C.; Vallejo, M. A.; Cordova F, T.; Sosa A, M. [Universidad de Guanajuato, Departamento de Ingenieria Fisica, DCI, Loma del Bosque 103, Col. Lomas del Campestre, 37150 Leon, Guanajuato (Mexico); Castruita D, J. P. [Universidad de Guadalajara, Departamento de Ecologia, CUCBA, Las Agujas, 45100 Zapopan, Jalisco (Mexico); Barbosa S, G., E-mail: myrna.sabanero@gmail.com [Universidad de Guanajuato, Departamento de Ciencias Medicas, DCS, 20 de Enero No. 929, Col. Obregon, 37000 Leon, Guanajuato (Mexico)

2015-10-15

Acute or chronic exposure to ionizing radiation is a factor that may be hazardous to health. It has been reported that exposure to low doses of radiation (less than 50 mSv / year) and subsequently exposure to high doses have greater effects in people. However, it is unknown molecular and biochemical level alteration. This study, analyzes the susceptibility of a biological system (HeLa Atcc CCL-2 human cervix cancer cell line) to ionizing radiation (6 and 60 mSv/ 90). Our evaluate multiple variables such as: total protein profile, mitochondrial metabolic activity (XTT assay), cell viability (Trypan blue exclusion assay), cytoskeleton (actin micro filaments), nuclei (D API), genomic DNA. The results indicate, that cells exposed to ionizing radiation structurally show alterations in nuclear phenotype and aneuploidy, further disruption in the tight junctions and consequently on the distribution of actin micro filaments. Similar alterations were observed in cells treated with a genotoxic agent (200μM H{sub 2}O{sub 2}/1 h). In conclusion, this multi-criteria assessment enables precise comparisons of the effects of radiation between any biological systems. However, it is necessary to determine stress markers for integration of the effects of ionizing radiation. (Author)

Synthesis and characterization of chitosan-alginate scaffolds for seeding human umbilical cord derived mesenchymal stem cells.

Science.gov (United States)

Kumbhar, Sneha G; Pawar, S H

2016-01-01

Chitosan and alginate are two natural and accessible polymers that are known to be biocompatible, biodegradable and possesses good antimicrobial activity. When combined, they exhibit desirable characteristics and can be created into a scaffold for cell culture. In this study interaction of chitosan-alginate scaffolds with mesenchymal stem cells are studied. Mesenchymal stem cells were derived from human umbilical cord tissues, characterized by flow cytometry and other growth parameters studied as well. Proliferation and viability of cultured cells were studied by MTT Assay and Trypan Blue dye exclusion assay. Besides chitosan-alginate scaffold was prepared by freeze-drying method and characterized by FTIR, SEM and Rheological properties. The obtained 3D porous structure allowed very efficient seeding of hUMSCs that are able to inhabit the whole volume of the scaffold, showing good adhesion and proliferation. These materials showed desirable rheological properties for facile injection as tissue scaffolds. The results of this study demonstrated that chitosan-alginate scaffold may be promising biomaterial in the field of tissue engineering, which is currently under a great deal of examination for the development and/or restoration of tissue and organs. It combines the stem cell therapy and biomaterials.

Anti-proliferative, Cytotoxic and NF-ĸB Inhibitory Properties of Spiro(Lactone-Cyclohexanone) Compounds in Human Leukemia.

Science.gov (United States)

Bouhenna, Mustapha M; Orlikova, Barbora; Talhi, Oualid; Schram, Ben; Pinto, Diana C G A; Taibi, Nadia; Bachari, Khaldoun; Diederich, Marc; Silva, Artur M S; Mameri, Nabil

2017-09-01

NF-ĸB affects most aspects of cellular physiology. Deregulation of NF-ĸB signaling is associated with inflammatory diseases and cancer. In this study, we evaluated the cytotoxic and NF-ĸB inhibition potential of new spiro(lactone-cyclohexanone) compounds in two different human leukemia cell lines (U937 and K562). The anti-proliferative effects of the spiro(lactone-cyclohexanone) compounds on human K562 and U937 cell lines was evaluated by trypan blue staining, as well as their involvement in NF-kB regulation were analyzed by luciferase reporter gene assay, Caspase-3/7 activities were evaluated to analyze apoptosis induction. Both spiro(coumarin-cyclohexanone) 4 and spiro(6- methyllactone-cyclohexanone) 9 down-regulated cancer cell viability and proliferation. Compound 4 inhibited TNF-α-induced NF-ĸB activation in a dose-dependent manner and induced caspase-dependent apoptosis in both leukemia cell lines. Results show that compound 4 and compound 9 have potential as anti-cancer agents. In addition, compound 4 exerted NF-kB inhibition activity in leukemia cancer cells. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Evaluation of Code Blue Implementation Outcomes

Directory of Open Access Journals (Sweden)

Bengü Özütürk

2015-09-01

Full Text Available Aim: In this study, we aimed to emphasize the importance of Code Blue implementation and to determine deficiencies in this regard. Methods: After obtaining the ethics committee approval, 225 patient’s code blue call data between 2012 and 2014 January were retrospectively analyzed. Age and gender of the patients, date and time of the call and the clinics giving Code Blue, the time needed for the Code Blue team to arrive, the rates of false Code Blue calls, reasons for Code Blue calls and patient outcomes were investigated. Results: A total of 225 patients (149 male, 76 female were evaluated in the study. The mean age of the patients was 54.1 years. 142 (67.2% Code Blue calls occurred after hours and by emergency unit. The mean time for the Code Blue team to arrive was 1.10 minutes. Spontaneous circulation was provided in 137 patients (60.8%; 88 (39.1% died. The most commonly identified possible causes were of cardiac origin. Conclusion: This study showed that Code Blue implementation with a professional team within an efficient and targeted time increase the survival rate. Therefore, we conclude that the application of Code Blue carried out by a trained team is an essential standard in hospitals. (The Medical Bulletin of Haseki 2015; 53:204-8

Terminology for pregnancy loss prior to viability

DEFF Research Database (Denmark)

Kolte, A M; Bernardi, L A; Christiansen, O B

2015-01-01

Pregnancy loss prior to viability is common and research in the field is extensive. Unfortunately, terminology in the literature is inconsistent. The lack of consensus regarding nomenclature and classification of pregnancy loss prior to viability makes it difficult to compare study results from...... different centres. In our opinion, terminology and definitions should be based on clinical findings, and when possible, transvaginal ultrasound. With this Early Pregnancy Consensus Statement, it is our goal to provide clear and consistent terminology for pregnancy loss prior to viability....

Tunable photonic crystals with partial bandgaps from blue phase colloidal crystals and dielectric-doped blue phases.

Science.gov (United States)

Stimulak, Mitja; Ravnik, Miha

2014-09-07

Blue phase colloidal crystals and dielectric nanoparticle/polymer doped blue phases are demonstrated to combine multiple components with different symmetries in one photonic material, creating a photonic crystal with variable and micro-controllable photonic band structure. In this composite photonic material, one contribution to the band structure is determined by the 3D periodic birefringent orientational profile of the blue phases, whereas the second contribution emerges from the regular array of the colloidal particles or from the dielectric/nanoparticle-doped defect network. Using the planewave expansion method, optical photonic bands of the blue phase I and II colloidal crystals and related nanoparticle/polymer doped blue phases are calculated, and then compared to blue phases with no particles and to face-centred-cubic and body-centred-cubic colloidal crystals in isotropic background. We find opening of local band gaps at particular points of Brillouin zone for blue phase colloidal crystals, where there were none in blue phases without particles or dopants. Particle size and filling fraction of the blue phase defect network are demonstrated as parameters that can directly tune the optical bands and local band gaps. In the blue phase I colloidal crystal with an additionally doped defect network, interestingly, we find an indirect total band gap (with the exception of one point) at the entire edge of SC irreducible zone. Finally, this work demonstrates the role of combining multiple - by symmetry - differently organised components in one photonic crystal material, which offers a novel approach towards tunable soft matter photonic materials.

Prodrug-activating Gene Therapy with Rabbit Cytochrome P450 4B1/4-Ipomeanol or 2-Aminoanthracene System in Glioma Cells

Energy Technology Data Exchange (ETDEWEB)

Jang, Su Jin; Kang, Joo Hyun; Lee, Tae Sup; Kim, Sung Joo; Kim, Kwang Il; Lee, Yong Jin; Cheon, Gi Jeong; Choi, Chang Woon; Lim, Sang Moo [Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul (Korea, Republic of)

2010-09-15

We determined the cytotoxic properties of cytochrome P450 4B1 (CYP4B1) activated 4-ipomeanol (4-ipo) and 2-aminoanthracene (2-AA) in rat glioma to verify the CYP4B1/4-ipo or 2-AA system for prodrug-activating gene therapy. The cyp4B1 cDNA was cloned into pcDNA3.1/ Hygro from rabbit lung total RNA (pcDAN-cyp4B1). Lentiviral vector encoding firefly luciferase (fLuc) was infected into C6 (rat glioma), and the fLuc-expressing cell was selected (C6-L). After transfection with pcDNA-cyp4B1 vector into C6-L, the single clone expressing cyp4B1 gene was selected (C6-CL). Prodrug for various concentrations of 4-ipo or 2-AA was treated for 72 h and 96 h. The cell survival rate of C6-CL was determined using MTT assay and trypan-blue dye exclusion methods. By RT-PCR analysis, fLuc and CYP4B1 expression was detected in C6-CL, but not in C6. MTT assay and trypan-blue dye exclusion showed that IC'5'0 of C6-CL was 0.3 mM and <0.01 mM after 4-ipo or 2-AA treatment at 96 h or 72 h exposure, respectively. Cell survivals of C6-CL were more rapidly reduced after treatment with 4-ipo or 2-AA than those of C6-L cells. The cell survival rate with MTT and trypan-blue dye exclusion assay was well correlated with fLuc activity in C6-CL cells. Conclusions CYP4B1-based prodrug-activating gene therapy may have the potential to treat glioma and the cytotoxic effects of CYP4B1 enzyme activated 4-ipo or 2-AA in C6, and could be clearly determined by bioluminescent activity in C6-CL.

Abundance, viability and culturability of Antarctic bacteria

Digital Repository Service at National Institute of Oceanography (India)

LokaBharathi, P.A.; DeSouza, M.J.B.D.; Nair, S.; Chandramohan, D.

The viability of total number of bacteria decide the mineralisation rate in any ecosystem and ultimately the fertility of the region. This study aims at establishing the extent of viability in the standing stock of the Antarctic bacterial population...

Assessment of myocardial viability by MR imaging

International Nuclear Information System (INIS)

Sandstede, Joern J.W.

2003-01-01

Diagnosis of myocardial viability after infarction focuses on the prediction of functional improvement of dysfunctional myocardium after revascularization therapy. Magnetic resonance imaging provides different approaches for the detection of myocardial viability. Measurement of end-diastolic wall thickness is easy to perform and has a high sensitivity, but a low specificity, and can only be used 4 months after myocardial infarction due to infarct healing processes. Low-dose dobutamine stress has a good sensitivity with a high specificity for the prediction of wall motion improvement, but this is only true for patients with a singular dysfunctional area and only slightly depressed cardiac function. Late enhancement allows for direct visualization of necrotic or scarred tissue. By measuring the transmural extent of late enhancement, the probability of mechanical improvement can precisely be given. Imaging of microvascular obstruction by first-pass perfusion or late enhancement gives additional information on viability and patient prognosis. Metabolic imaging techniques, such as 31 P-MR spectroscopy and 23 Na-MR imaging, provide further insights into the mechanisms of myocardial infarction and viability. In conclusion, cardiac MRI offers several clinically usable approaches for the assessment of myocardial viability and will probably become the method of choice in the near future. (orig.)

Effect of low-power red light laser irradiation on the viability of human skin fibroblast

Energy Technology Data Exchange (ETDEWEB)

Bednarska, K.; Rozga, B.; Leyko, W.; Bryszewska, M. [Institute of Biophysics, University of Lodz (Poland); Kolodziejczyk, K.; Szosland, D. [Diabetological Clinic, Medical Academy of Lodz (Poland)

1998-10-01

Human skin fibroblast monolayers (S-126 cell line) were exposed to laser radiation (wavelength 670 nm, power density 40 mW/cm{sup 2}). The energy densities were 2 J/cm{sup 2} and 12 J/cm{sup 2}, respectively, and the irradiation was carried out at a temperature of 22 C. For fibroblast viability evaluation, the colorimetric assay (conversion of thiazolyl blue to formazan) was used. The experiments were carried out at 37 C, in the presence of 5% CO{sub 2}, and at different time periods of incubation after irradiation (2, 4, 8 h and 1, 2, 3, 4, 5 days). The results indicated that there was a certain stimulating effect on the long-term proliferation of skin fibroblasts and that the stimulation proceeded in two stages, the first one 2 h and the second one 3 days post-irradiation. (orig.) With 4 figs., 2 tabs., 13 refs.

"Blue-Collar Blues" uurib töösuhteid uutes oludes / Janar Ala

Index Scriptorium Estoniae

Ala, Janar, 1979-

2009-01-01

Tööproblemaatikat käsitlev näitus "Blue-Collar Blues" Tallinna Kunstihoones ja Tallinna Kunstihoone galeriis 31. jaanuarini 2010, kuraator Anders Härm. Lähemalt belgia-mehhiko kunstniku Francis Alys'e videost, austria kunstniku Oliver Ressleri ning venetsueela-saksa politoloogi Dario Azzelini videost "Viis tehast. Tööliste kontroll Venezuelas"

Annonaceous acetogenin mimic AA005 induces cancer cell death via apoptosis inducing factor through a caspase-3-independent mechanism

OpenAIRE

Han, Bing; Wang, Tong-Dan; Shen, Shao-Ming; Yu, Yun; Mao, Chan; Yao, Zhu-Jun; Wang, Li-Shun

2015-01-01

Background Annonaceous acetogenins are a family of natural products with antitumor activities. Annonaceous acetogenin mimic AA005 reportedly inhibits mammalian mitochondrial NADH-ubiquinone reductase (Complex I) and induces gastric cancer cell death. However, the mechanisms underlying its cell-death-inducing activity are unclear. Methods We used SW620 colorectal adenocarcinoma cells to study AA005 cytotoxic activity. Cell deaths were determined by Trypan blue assay and flow cytometry, and rel...

The spectrum of dermatoscopic patterns in blue nevi.

Science.gov (United States)

Di Cesare, Antonella; Sera, Francesco; Gulia, Andrea; Coletti, Gino; Micantonio, Tamara; Fargnoli, Maria Concetta; Peris, Ketty

2012-08-01

Blue nevi are congenital or acquired, dermal dendritic melanocytic proliferations that can simulate melanocytic and nonmelanocytic lesions including melanoma, cutaneous metastasis of melanoma, Spitz/Reed nevi, and basal cell carcinoma. We sought to investigate global and local dermatoscopic patterns of blue nevi compared with melanomas and basal cell carcinomas. We retrospectively analyzed global and local features in 95 dermatoscopic images of blue nevi and in 190 melanomas and basal cell carcinomas that were selected as control lesions on the basis of similar pigmentation. Lesion pigmentation was classified as monochromatic, dichromatic, or multichromatic. A global pattern characterized by homogeneous pigmentation was observed in all of 95 (100%) blue nevi. Eighty of 95 (84.2%) blue nevi presented a homogeneous pattern consisting of one color (blue, black, or brown) or two colors (blue-brown, blue-gray, or blue-black). Fifteen of 95 (15.8%) blue nevi had a multichromatic (blue, gray, black, brown, and/or red) pigmentation. In all, 47 of 95 (49.5%) blue nevi were characterized by pigmentation in the absence of pigment network or any other local dermatoscopic features. And 48 of 95 (50.5%) blue nevi showed local dermatoscopic patterns including whitish scarlike depigmentation, dots/globules, vascular pattern, streaks, and networklike pattern. The study was retrospective and involved only Caucasian people of Italian origin. The characteristic feature of blue nevi is a homogeneous pigmentation that is blue, blue-gray, blue-brown, or blue-black. We showed that a wide spectrum of local dermatoscopic features (whitish scarlike depigmentation, dots/globules, peripheral streaks or vessels) may also be present. In such cases, clinical and dermatoscopic distinction from melanoma or nonmelanocytic lesions may be difficult or impossible, and surgical excision is necessary. Copyright © 2011 American Academy of Dermatology, Inc. Published by Mosby, Inc. All rights reserved.

Formation of highly toxic hydrogen cyanide upon ruby laser irradiation of the tattoo pigment phthalocyanine blue

Science.gov (United States)

Schreiver, Ines; Hutzler, Christoph; Laux, Peter; Berlien, Hans-Peter; Luch, Andreas

2015-08-01

Since laser treatment of tattoos is the favored method for the removing of no longer wanted permanent skin paintings, analytical, biokinetics and toxicological data on the fragmentation pattern of commonly used pigments are urgently required for health safety reasons. Applying dynamic headspace—gas chromatography with mass spectrometric detection (DHS—GC/MS) and comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GCxGC—ToF-MS), we identified 1,2-benzene dicarbonitrile, benzonitrile, benzene, and the poisonous gas hydrogen cyanide (HCN) as main fragmentation products emerging dose-dependently upon ruby laser irradiation of the popular blue pigment copper phthalocyanine in suspension. Skin cell viability was found to be significantly compromised at cyanide levels of ≥1 mM liberated during ruby laser irradiation of >1.5 mg/ml phthalocyanine blue. Further, for the first time we introduce pyrolysis-GC/MS as method suitable to simulate pigment fragmentation that may occur spontaneously or during laser removal of organic pigments in the living skin of tattooed people. According to the literature such regular tattoos hold up to 9 mg pigment/cm2 skin.

The Blue Coma: The Role of Methylene Blue in Unexplained Coma After Cardiac Surgery.

Science.gov (United States)

Martino, Enrico Antonio; Winterton, Dario; Nardelli, Pasquale; Pasin, Laura; Calabrò, Maria Grazia; Bove, Tiziana; Fanelli, Giovanna; Zangrillo, Alberto; Landoni, Giovanni

2016-04-01

Methylene blue commonly is used as a dye or an antidote, but also can be used off label as a vasopressor. Serotonin toxicity is a potentially lethal and often misdiagnosed condition that can result from drug interaction. Mild serotonin toxicity previously was reported in settings in which methylene blue was used as a dye. The authors report 3 cases of life-threatening serotonin toxicity in patients undergoing chronic selective serotonin reuptake inhibitor (SSRI) therapy who also underwent cardiac surgery and received methylene blue to treat vasoplegic syndrome. An observational study. A cardiothoracic intensive care unit (ICU) in a teaching hospital. Three patients who received methylene blue after cardiac surgery, later discovered to be undergoing chronic SSRI therapy. None. All 3 patients received high doses of fentanyl during general anesthesia. They all developed vasoplegic syndrome and consequently were given methylene blue in the ICU. All 3 patients developed serotonin toxicity, including coma, after this administration and diagnostic tests were negative for acute intracranial pathology. Coma lasted between 1 and 5 days. Two patients were discharged from the ICU shortly after awakening, whereas the third patient experienced a complicated postoperative course for concomitant refractory low-cardiac-output syndrome. Patients undergoing chronic SSRI therapy should not be administered methylene blue to treat vasoplegic syndrome. Copyright © 2016 Elsevier Inc. All rights reserved.

Metabolic Abnormalities Detected in Phase II Evaluation of Doxycycline in Dogs with Multicentric B-Cell Lymphoma.

Science.gov (United States)

Hume, Kelly R; Sylvester, Skylar R; Borlle, Lucia; Balkman, Cheryl E; McCleary-Wheeler, Angela L; Pulvino, Mary; Casulo, Carla; Zhao, Jiyong

2018-01-01

Doxycycline has antiproliferative effects in human lymphoma cells and in murine xenografts. We hypothesized that doxycycline would decrease canine lymphoma cell viability and prospectively evaluated its clinical tolerability in client-owned dogs with spontaneous, nodal, multicentric, substage a, B-cell lymphoma, not previously treated with chemotherapy. Treatment duration ranged from 1 to 8 weeks (median and mean, 3 weeks). Dogs were treated with either 10 ( n  = 6) or 7.5 ( n  = 7) mg/kg by mouth twice daily. One dog had a stable disease for 6 weeks. No complete or partial tumor responses were observed. Five dogs developed grade 3 and/or 4 metabolic abnormalities suggestive of hepatopathy with elevations in bilirubin, ALT, ALP, and/or AST. To evaluate the absorption of oral doxycycline in our study population, serum concentrations in 10 treated dogs were determined using liquid chromatography tandem mass spectrometry. Serum levels were variable and ranged from 3.6 to 16.6 µg/ml (median, 7.6 µg/ml; mean, 8.8 µg/ml). To evaluate the effect of doxycycline on canine lymphoma cell viability in vitro , trypan blue exclusion assay was performed on canine B-cell lymphoma cell lines (17-71 and CLBL) and primary B-cell lymphoma cells from the nodal tissue of four dogs. A doxycycline concentration of 6 µg/ml decreased canine lymphoma cell viability by 80%, compared to matched, untreated, control cells (mixed model analysis, p  canine lymphoma, combination therapy may be worthwhile if future research determines that doxycycline can alter cell survival pathways in canine lymphoma cells. Due to the potential for metabolic abnormalities, close monitoring is recommended with the use of this drug in tumor-bearing dogs. Additional research is needed to assess the tolerability of chronic doxycycline therapy.

The pyruvic acid analog 3-bromopyruvate interferes with the tetrazolium reagent MTS in the evaluation of cytotoxicity.

Science.gov (United States)

Ganapathy-Kanniappan, Shanmugasundaram; Geschwind, Jean-Francois H; Kunjithapatham, Rani; Buijs, Manon; Syed, Labiq H; Rao, Pramod P; Ota, Shinichi; Vali, Mustafa

2010-04-01

3-Bromopyruvate (3BrPA) is a pyruvate analog known for its alkylating property. Recently, several reports have documented the antiglycolytic and anticancer effects of 3BrPA and its potential for therapeutic applications. 3BrPA-mediated cytotoxicity has been evaluated in vitro by various methods including tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)-based assays such as MTT, MTS, and so on. However, growing body of evidences has shown that tetrazolium reagent may interfere with the test compounds. In this study, we investigated whether the tetrazolium reagent interferes with the assessment of 3BrPA cytotoxicity. The results of the tetrazolium-based MTS assay were compared with 3 distinct cell viability detection methods, that is, Trypan Blue staining, ATP depletion, and Annexin V staining in 2 different cell lines, Vx-2 and HepG2. The MTS assay data showed false positive results by indicating increased cell viability at 1 mM and 2 mM 3BrPA whereas the other cell viability assays demonstrated that both Vx-2 and HepG2 cells are not viable at the same treatment conditions. In order to validate the direct interaction of 3BrPA with MTS reagent, we tested cell-free media incubated with different concentrations of 3BrPA. The results of cell-free media showed an increase in absorbance in a dose-dependent manner confirming the interaction of MTS with 3BrPA. Thus, our data clearly demonstrate that 3BrPA interferes with the accuracy of MTS-based cytotoxicity evaluation. Hence, we suggest that employing multiple methods of biochemical as well as morphological cytotoxicity assays is critical to evaluate 3BrPA-mediated cell death.

Simvastatin induces apoptosis by a Rho-dependent mechanism in cultured cardiac fibroblasts and myofibroblasts

International Nuclear Information System (INIS)

Copaja, Miguel; Venegas, Daniel; Aranguiz, Pablo; Canales, Jimena; Vivar, Raul; Catalan, Mabel; Olmedo, Ivonne; Rodriguez, Andrea E.; Chiong, Mario; Leyton, Lisette; Lavandero, Sergio; Diaz-Araya, Guillermo

2011-01-01

Several clinical trials have shown the beneficial effects of statins in the prevention of coronary heart disease. Additionally, statins promote apoptosis in vascular smooth muscle cells, in renal tubular epithelial cells and also in a variety of cell lines; yet, the effects of statins on cardiac fibroblast and myofibroblast, primarily responsible for cardiac tissue healing are almost unknown. Here, we investigated the effects of simvastatin on cardiac fibroblast and myofibroblast viability and studied the molecular cell death mechanism triggered by simvastatin in both cell types. Methods: Rat neonatal cardiac fibroblasts and myofibroblasts were treated with simvastatin (0.1-10 μM) up to 72 h. Cell viability and apoptosis were evaluated by trypan blue exclusion method and by flow cytometry, respectively. Caspase-3 activation and Rho protein levels and activity were also determined by Western blot and pull-down assay, respectively. Results: Simvastatin induces caspase-dependent apoptosis of cardiac fibroblasts and myofibroblasts in a concentration- and time-dependent manner, with greater effects on fibroblasts than myofibroblasts. These effects were prevented by mevalonate, farnesylpyrophosphate and geranylgeranylpyrophosphate, but not squalene. These last results suggest that apoptosis was dependent on small GTPases of the Rho family rather than Ras. Conclusion: Simvastatin triggered apoptosis of cardiac fibroblasts and myofibroblasts by a mechanism independent of cholesterol synthesis, but dependent of isoprenilation of Rho protein. Additionally, cardiac fibroblasts were more susceptible to simvastatin-induced apoptosis than cardiac myofibroblasts. Thus simvastatin could avoid adverse cardiac remodeling leading to a less fibrotic repair of the damaged tissues. - Research Highlights: → Simvastatin decreases CF and CMF viability independent of cholesterol synthesis. → Simvastatin induces CF and CMF apoptosis in a caspase-dependent manner being CMF more resistant

Nanoparticles containing allotropes of carbon have genotoxic effects on glioblastomamultiforme cells

Directory of Open Access Journals (Sweden)

Hinzmann M

2014-05-01

Full Text Available Mateusz Hinzmann,1 Slawomir Jaworski,1 Marta Kutwin,1 Joanna Jagiello,2 Rafal Kozinski,2 Mateusz Wierzbicki,1 Marta Grodzik,1 Ludwika Lipinska,2 Ewa Sawosz,1 Andrè Chwalibog31Division of Nanobiotechnology, Warsaw University of Life Sciences, 2Institute of Electronic Materials Technology, Warsaw, Poland; 3Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Copenhagen, DenmarkAbstract: The carbon-based nanomaterial family consists of nanoparticles containing allotropes of carbon, which may have a number of interactions with biological systems. The objective of this study was to evaluate the toxicity of nanoparticles comprised of pristine graphene, reduced graphene oxide, graphene oxide, graphite, and ultradispersed detonation diamond in a U87 cell line. The scope of the work consisted of structural analysis of the nanoparticles using transmission electron microscopy, evaluation of cell morphology, and assessment of cell viability by Trypan blue assay and level of DNA fragmentation of U87 cells after 24 hours of incubation with 50 µg/mL carbon nanoparticles. DNA fragmentation was studied using single-cell gel electrophoresis. Incubation with nanoparticles containing the allotropes of carbon did not alter the morphology of the U87 cancer cells. However, incubation with pristine graphene and reduced graphene oxide led to a significant decrease in cell viability, whereas incubation with graphene oxide, graphite, and ultradispersed detonation diamond led to a smaller decrease in cell viability. The results of a comet assay demonstrated that pristine graphene, reduced graphene oxide, graphite, and ultradispersed detonation diamond caused DNA damage and were therefore genotoxic in U87 cells, whereas graphene oxide was not.Keywords: nanostructures, graphene, graphite, diamond, glioblastoma multiforme, geno toxicity

FOXP3 expression is modulated by TGF-β1/NOTCH1 pathway in human melanoma

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Skarmoutsou, Eva; Bevelacqua, Valentina; D'Amico, Fabio; Russo, Angela; Spandidos, Demetrios A.; Scalisi, Aurora

2018-01-01

Forkhead box protein 3 (FOXP3) transcription factor is expressed by immune cells and several human cancers and is associated with tumor aggressiveness and unfavorable clinical outcomes. NOTCH and transforming growth factor-β (TGF-β) protumorigenic effects are mediated by FOXP3 expression in several cancer models; however, their interaction and role in melanoma is unknown. We investigated TGF-β-induced FOXP3 gene expression during NOTCH1 signaling inactivation. Primary (WM35) and metastatic melanoma (A375 and A2058) cell lines and normal melanocytes (NHEM) were used. FOXP3 subcellular distribution was evaluated by immuno cytochemical analysis. Gene expression levels were assessed by reverse transcription-quantitative polymerase chain reaction. Protein levels were assessed by western blot analysis. The γ-secretase inhibitor (GSI) was used for NOTCH1 inhibition and recombinant human (rh)TGF-β was used for melanoma cell stimulation. Cell proliferation and viability were respectively assessed by MTT and Trypan blue dye assays. FOXP3 mRNA and protein levels were progressively higher in WM35, A375 and A2058 cell lines compared to NHEM and their levels were further increased after stimulation with rh-TGF-β. TGF-β-mediated FOXP3 expression was mediated by NOTCH1 signaling. Inhibition of NOTCH1 with concomitant rh-TGF-β stimulation determined the reduction in gene expression and protein level of FOXP3. Finally, melanoma cell line proliferation and viability were reduced by NOTCH1 inhibition. The results show that nn increase in FOXP3 expression in metastatic melanoma cell lines is a potential marker of tumor aggressiveness and metastasis. NOTCH1 is a central mediator of TGF-β-mediated FOXP3 expression and NOTCH1 inhibition produces a significant reduction of melanoma cell proliferation and viability. PMID:29620159

Capsaicin induces cell cycle arrest and apoptosis in human KB cancer cells.

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Lin, Chia-Han; Lu, Wei-Cheng; Wang, Che-Wei; Chan, Ya-Chi; Chen, Mu-Kuan

2013-02-25

Capsaicin, a pungent phytochemical in a variety of red peppers of the genus Capsicum, has shown an anti-proliferative effect on various human cancer cell lines. In contrast, capsaicin has also been considered to promote the growth of cancer cells. Thus, the effects of capsaicin on various cell types need to be explored. The anti-proliferative effects of capsaicin on human KB cancer cells are still unknown. Therefore, we examined the viability, cell cycle progression, and factors associated with apoptosis in KB cells treated with capsaicin. The cell proliferation/viability and cytotoxicity of KB cells exposed to capsaicin were determined by a sulforhodamine B colorimetric assay and trypan blue exclusion. Apoptosis was detected by Hoechst staining and confirmed by western blot analysis of poly-(ADP-ribose) polymerase cleavage. Cell cycle distribution and changes of the mitochondrial membrane potential were analyzed by flow cytometry. Furthermore, the expression of caspase 3, 8 and 9 was evaluated by immunoblotting. We found that treatment of KB cells with capsaicin significantly reduced cell proliferation/viability and induced cell death in a dose-dependent manner compared with that in the untreated control. Cell cycle analysis indicated that exposure of KB cells to capsaicin resulted in cell cycle arrest at G2/M phase. Capsaicin-induced growth inhibition of KB cells appeared to be associated with induction of apoptosis. Moreover, capsaicin induced disruption of the mitochondrial membrane potential as well as activation of caspase 9, 3 and poly-(ADP-ribose) polymerase in KB cells. Our data demonstrate that capsaicin modulates cell cycle progression and induces apoptosis in human KB cancer cells through mitochondrial membrane permeabilization and caspase activation. These observations suggest an anti-cancer activity of capsaicin.

Corantes vitais em cromovitrectomia Vital dyes in chromovitrectomy

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Eduardo Dib

2009-12-01

Full Text Available O objetivo do artigo é apresentar os dados atuais da aplicação de corantes vitais durante cirurgia vitreorretiniana, "cromovitrectomia", bem como uma revisão da literatura atual sobre o assunto no tocante às técnicas de aplicação, indicações e complicações em cromovitrectomia. Um grande número de publicações tem abordado o perfil tóxico da indocianina verde na cromovitrectomia. Dados experimentais mostram uma toxicidade dose-dependente da mesma em várias populações de células retinianas. Novas gerações de corantes incluem: azul tripan, azul patente, acetato de triancinolona, infracianina verde, fluoresceína sódica, azul de bromofenol, acetato de fluorometolona e azul brilhante. Novos instrumentos podem permitir um corar seletivo de estruturas durante a vitrectomia. Este artigo mostra que o campo da cromovitrectomia está em plena expansão de pesquisas. Os corantes de primeira linha são a indocianina verde, infracianina verde e o azul brilhante. Azul patente, azul de bromofenol e azul tripan surgem como novos adjuvantes para melhor observação da membrana epirretiniana. Demais corantes que surgiram merecem maior investigação.The aim of this article is to present the current data with regard to the application of vital dyes during vitreoretinal surgery, "chromovitrectomy", as well as to overview the current literature regarding the properties of dyes, techniques of application, indications and complications in chromovitrectomy. A large body of published research has recently addressed the toxicity profile of indocyanine green for chromovitrectomy. Experimental data demonstrate dose-dependent toxicity of indocyanine green to various retinal cells. Newer generation vital dyes for chromovitrectomy include trypan blue, patent blue, triamcinolone acetonide, infracyanine green, sodium fluorescein, bromophenol blue, fluorometholone acetate and brilliant blue. Novel instruments may enable a selective painting of preretinal

Why Blue-Collar Blacks Help Less

OpenAIRE

Smith, Sandra Susan; Young, Kara Alexis

2013-01-01

Why are blue-collar blacks less likely to help jobseekers than jobholders from other ethnoracial groups or even than more affluent blacks? Drawing from in-depth, semi-structured interviews with 97 black and Latino workers at one large, public sector employer, we find that blue-collar black workers both helped less proactively and rejected more requests for assistance than did blue-collar Latino and white-collar black workers. We attribute blue-collar blacks’ more passive engagement to their...

Doctors' perspectives on the viability of rural practice.

Science.gov (United States)

Jones, J A; Humphreys, J S; Adena, M A

2004-01-01

Private practitioners play a vital role in meeting the health needs of rural communities. However, the prospect of operating a private practice business in rural Australia seems to be increasingly unattractive, because many communities are forced to recruit salaried or overseas-trained doctors. This study focuses on rural practices as businesses whose viability influences their attractiveness for the recruitment and retention of practitioners. The specific objectives are to ascertain which factors contribute to or threaten practice viability in rural areas, and whether they vary according to the degree of rurality or geographical remoteness. This study is based on data collected from a national study into the viability of rural general practice undertaken jointly by the Rural Doctors Association of Australia and Monash University School of Rural Health Bendigo. The Rural Remote and Metropolitan Area (RRMA) classification was used as the indicator of rurality. The study surveyed all general practitioners practising in rural or remote regions of Australia (RRMAs 3 to 7). Only practitioners with some financial interest in the practice were selected for this analysis. Free-text responses to the two questions 'What are the key factors contributing to the viability of your practice?' and 'What factors would put the viability of your practice at risk?' were analysed using qualitative content analysis. Factors were derived iteratively through higher-level aggregation of responses. Chi-square tests were used to make comparisons across the RRMA categories. The national survey achieved a response rate of 35% of the entire population of GPs practising in RRMA 3 to 7 regions. Of these, 1050 respondents were relevant to this analysis. Seven major factors were identified by practitioners as the main contributors to practice viability. 'Practice characteristics' was nominated by 59% of respondents, followed by 'Income' (31%), 'Personal circumstances', 'Workforce' and 'Community

Link overlap, viability, and mutual percolation in multiplex networks

International Nuclear Information System (INIS)

Min, Byungjoon; Lee, Sangchul; Lee, Kyu-Min; Goh, K.-I.

2015-01-01

Many real-world complex systems are best modeled by multiplex networks. The multiplexity has proved to have broad impact on the system’s structure and function. Most theoretical studies on multiplex networks to date, however, have largely ignored the effect of the link overlap across layers despite strong empirical evidences for its significance. In this article, we investigate the effect of the link overlap in the viability of multiplex networks, both analytically and numerically. After a short recap of the original multiplex viability study, the distinctive role of overlapping links in viability and mutual connectivity is emphasized and exploited for setting up a proper analytic framework. A rich phase diagram for viability is obtained and greatly diversified patterns of hysteretic behavior in viability are observed in the presence of link overlap. Mutual percolation with link overlap is revisited as a limit of multiplex viability problem, and the controversy between existing results is clarified. The distinctive role of overlapping links is further demonstrated by the different responses of networks under random removals of overlapping and non-overlapping links, respectively, as well as under several link-removal strategies. Our results show that the link overlap facilitates the viability and mutual percolation; at the same time, the presence of link overlap poses a challenge in analytical approaches to the problem

Raman analysis of cobalt blue pigment in blue and white porcelain: A reassessment

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Jiang, Xiaochenyang; Ma, Yanying; Chen, Yue; Li, Yuanqiu; Ma, Qinglin; Zhang, Zhaoxia; Wang, Changsui; Yang, Yimin

2018-02-01

Cobalt blue is a famous pigment in human history. In the past decade it is widely reported that the cobalt aluminate has been detected in ancient ceramics as blue colorant in glaze, yet the acquired Raman spectra are incredibly different from that of synthesised references, necessitating a reassessment of such contradictory scenario with more accurate analytic strategies. In this study, micro-Raman spectroscopy (MRS) and scanning electron microscopy (SEM) in association with energy dispersive spectrometry (EDS) were performed on under-glaze cobalt pigments from one submerged blue and white porcelain shard dated from Wanli reign (1573-1620 CE) of Ming dynasty (1365-1644 CE) excavated at Nan'ao I shipwreck off the southern coast of China. The micro-structural inspection reveals that the pigment particles have characteristics of small account, tiny size, heterogeneously distribution, and more importantly, been completely enwrapped by well-developed anorthite crystals in the glaze, indicating that the signals recorded in previous publications are probably not from cobalt pigments themselves but from outside thickset anorthite shell. The further spectromicroscopic analyses confirm this presumption when the accurate spectra of cobalt aluminate pigment and surrounding anorthite were obtained separately with precise optical positioning. Accordingly, we reassess and clarify the previous Raman studies dedicated to cobalt blue pigment in ancient ceramics, e.g. cobalt blue in celadon glaze, and in turn demonstrate the superiority and necessity of coupling spectroscopic analysis with corresponding structure observation, especially in the characterization of pigments from complicated physico-chemical environment like antiquities. Thus, this study promotes a better understanding of Raman spectroscopy study of cobalt blue pigments in art and archaeology field.

A comparative study of three cytotoxicity test methods for nanomaterials using sodium lauryl sulfate.

Science.gov (United States)

Kwon, Jae-Sung; Kim, Kwang-Mahn; Kim, Kyoung-Nam

2014-10-01

The biocompatibility evaluation of nanomaterials is essential for their medical diagnostic and therapeutic usage, where a cytotoxicity test is the simplest form of biocompatibility evaluation. Three methods have been commonly used in previous studies for the cytotoxicity testing of nanomaterials: trypan blue exclusion, colorimetric assay using water soluble tetrazolium (WST), and imaging under a microscope following calcein AM/ethidium homodimer-1 staining. However, there has yet to be a study to compare each method. Therefore, in this study three methods were compared using the standard reference material of sodium lauryl sulfate (SLS). Each method of the cytotoxicity test was carried out using mouse fibroblasts of L-929 exposed to different concentrations of SLS. Compared to the gold standard trypan blue exclusion test, both colorimetric assay using water soluble tetrazolium (WST) and imaging under microscope with calcein AM/ethidium homodimer-1 staining showed results that were not statistically different. Also, each method exhibited various advantages and disadvantages, which included the need of equipment, time taken for the experiment, and provision of additional information such as cell morphology. Therefore, this study concludes that all three methods of cytotoxicity testing may be valid, though careful consideration will be needed when selecting tests with regard to time, finances, and the amount of information required by the researcher(s).

Novel Threadlike Structures May Be Present on the Large Animal Organ Surface: Evidence in Swine Model

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Kyoung-Hee Bae

2013-01-01

Full Text Available Background. The types of embryonic development probably provoke different paths of novel threadlike structure (NTS development. The authors hypothesized that NTS may be easily observed on the surface of swine intestines by using trypan blue staining method and visualization under an optical microscope. Methods. General anesthesia was administered to 2 Yorkshire pigs. The abdominal walls of the pigs were carefully dissected along the medial alba. NTSs were identified on organ surfaces under a stereoscopic microscope after trypan blue staining. Isolated NTS specimens obtained from the large intestine were subjected to 4′,6-diamidino-2-phenylindole (DAPI staining and observed using the polarized light microscopy to confirm whether the obtained structure fits the definition of NTS. Results. We found elastic, semitransparent threadlike structures (forming a network structure that had a milky-white color in situ and in vivo in swine large intestines. The samples showed distinct extinction of polarized light at every 90 degrees, and nucleus was shown to be rod shaped by DAPI staining, indicating that they meet the criteria of NTS. Conclusion. We used a swine model to demonstrate that NTS may be present on large animal organ surfaces. Our results may permit similar studies by using human specimens.

A viability analysis for a stock/price model

Science.gov (United States)

Jerry, Chakib; Raissi, Nadia

2012-09-01

We examine the conditions for the sustainability of a stock/price system based on the use of a marine renewable resource. Instead of studying the environmental and economic interactions in terms of optimal control, we focus on the viability of the system. These viability/crisis situations are defined by a set of economic state constraints. This constraints combine a guaranteed consumption and a minimum income for fishermen. Using the mathematical concept of viability kernel, we reveal that with only economics constraints we guarantee a perennial stock/price system.

Low power laser effects in cancer cells and fibroblasts submitted the ionizing radiation; Efeitos do laser de baixa potencia em celulas de linhagem tumoral e fibroblastos submetidos a radiacao ionizante

Energy Technology Data Exchange (ETDEWEB)

Silva, Camila Ramos

2015-07-01

Cancer is considered a public health problem worldwide. According to Brazil's the National Cancer Institute (INCA), 576,000 new cases of cancer were estimated for 2015 in Brazil, representing the second leading cause of death. Radiotherapy may be a treatment to several of types of cancer, frequently using ionizing radiation to eradicate or prevent the proliferation of tumor cells. This treatment, however, can lead to death of non-tumor cells around in irradiated tissue. Given this, adjuvant therapies that can minimize the side effects of ionizing radiation are of extremely importance. In this context, low power laser (LPL) may be an alternative to modulate the response of healthy cells to ionizing radiation. In this study, cells of human gingival fibroblasts (FMM1) and breast cancer (MDAMB- 231) were exposed to gamma radiation at doses of 2.5 and 10 Gy. After twenty-four hours, cell were irradiated with LPL ( λ= 660 nm, 40 mW and total area of 0.04 cm²) with energy densities of 30, 60, 90, 120 and 150 J/cm². The cell viability was measured during four days, using the trypan blue technique. The influence of LPL on the cell cycle and on expression of the nuclear antigen of cellular proliferation (PCNA) was evaluated by flow cytometry. The expression of β-Galactosidase was the chosen method to assess cell senescence. Considering our adopted parameters, and focusing on the non-tumor cells, we have observed an increase in: 1) cell viability; 2) cell population in phases S and G{sub 2}/M cell cycle; 3) PCNA expression with decrease in senescence. No alterations were observed in the cell viability, with greater population in phases S and G{sub 2}/M cell cycle, while the number of senescent cells and the expression of PCNA were decreased. Therefore, we have concluded that the LPL promoted effects on both cell lineages, with increased cell viability on FMM1 cells, whether cancer cells maintained a decreased proliferation. (author)

Assessing the Cytotoxicity of Black Carbon As A Model for Ultrafine Anthropogenic Aerosol Across Human and Murine Cells: A Chronic Exposure Model of Nanosized Particulate Matter

Science.gov (United States)

Salinas, E.

2015-12-01

BC: 100μM, 300μM, 600μM, 1,250μM, 2,500μM, and 5,000μM conducting the Trypan Blue (TB) Exclusion Assay, Calcein-AM Viability Assay, and CellTiter-Glo Viability Assay.

Low power laser effects in cancer cells and fibroblasts submitted the ionizing radiation

International Nuclear Information System (INIS)

Silva, Camila Ramos

2015-01-01

Cancer is considered a public health problem worldwide. According to Brazil's the National Cancer Institute (INCA), 576,000 new cases of cancer were estimated for 2015 in Brazil, representing the second leading cause of death. Radiotherapy may be a treatment to several of types of cancer, frequently using ionizing radiation to eradicate or prevent the proliferation of tumor cells. This treatment, however, can lead to death of non-tumor cells around in irradiated tissue. Given this, adjuvant therapies that can minimize the side effects of ionizing radiation are of extremely importance. In this context, low power laser (LPL) may be an alternative to modulate the response of healthy cells to ionizing radiation. In this study, cells of human gingival fibroblasts (FMM1) and breast cancer (MDAMB- 231) were exposed to gamma radiation at doses of 2.5 and 10 Gy. After twenty-four hours, cell were irradiated with LPL ( λ= 660 nm, 40 mW and total area of 0.04 cm²) with energy densities of 30, 60, 90, 120 and 150 J/cm². The cell viability was measured during four days, using the trypan blue technique. The influence of LPL on the cell cycle and on expression of the nuclear antigen of cellular proliferation (PCNA) was evaluated by flow cytometry. The expression of β-Galactosidase was the chosen method to assess cell senescence. Considering our adopted parameters, and focusing on the non-tumor cells, we have observed an increase in: 1) cell viability; 2) cell population in phases S and G 2 /M cell cycle; 3) PCNA expression with decrease in senescence. No alterations were observed in the cell viability, with greater population in phases S and G 2 /M cell cycle, while the number of senescent cells and the expression of PCNA were decreased. Therefore, we have concluded that the LPL promoted effects on both cell lineages, with increased cell viability on FMM1 cells, whether cancer cells maintained a decreased proliferation. (author)

Comparison of 864 and 935 MHz microwave radiation effects on cell culture

International Nuclear Information System (INIS)

Pavicic, I.; Trosic, I.; Sarolic, A.

2005-01-01

The aim of our study was to evaluate and compare the effect of 864 and 935 MHz microwave radiation on proliferation, colony forming and viability of Chinese hamster lung cells, cell line V79. Cell cultures were exposed both to the 864 MHz microwave field in transversal electromagnetic mode cell (TEM-cell) and to the 935 MHz field in Gigahertz transversal electromagnetic mode cell (GTEM-cell) for 1, 2 and 3 hours. Philips PM 5508 generator connected with a signal amplifier generated the frequency of 864 MHz, whereas Hewlett Packard HP8657A signal generator was used to generate the frequency of 935 MHz. The average specific absorption rate (SAR) was 0.08 W/kg for 864 MHz and 0.12 W/kg for 935 MHz. To determine the cell growth, V79 cells were plated in the concentration of 1x10 4 cells per milliliter of nutrient medium. Cells were cultured in a humidified atmosphere at 37 degrees of C in 5% CO 2 . Cell proliferation was determined by cell counts for each hour of exposure during the five post-exposure days. To identify colony-forming ability, cells were cultivated in the concentration of 40 cells/mL of medium and incubated as described above. Colony-forming ability was assessed for each exposure time by colony count on post-exposure day 7. Trypan blue exclusion test was used to determine cell viability. On post-exposure day 3, the growth curve of 864 MHz irradiated cells showed a significant decrease (p less than 0.05) after 2 and 3 hours of exposure in comparison with control cells. Cells exposed to 935 MHz radiation showed a significant decrease (p less than 0.05) after 3 hours of exposure on post-exposure day 3. Both the colony-forming ability and viability of 864 MHz and 935 MHz exposed cells did not significantly differ from matched control cells. In conclusion, both applied RF/MW fields have shown similar effects on cell culture growth, colony forming and cell viability of the V79 cell line.(author)

Unraveling the Mystery of the Blue Fog: Structure, Properties, and Applications of Amorphous Blue Phase III.

Science.gov (United States)

Gandhi, Sahil Sandesh; Chien, Liang-Chy

2017-12-01

The amorphous blue phase III of cholesteric liquid crystals, also known as the "blue fog," are among the rising stars in materials science that can potentially be used to develop next-generation displays with the ability to compete toe-to-toe with disruptive technologies like organic light-emitting diodes. The structure and properties of the practically unobservable blue phase III have eluded scientists for more than a century since it was discovered. This progress report reviews the developments in this field from both fundamental and applied research perspectives. The first part of this progress report gives an overview of the 130-years-long scientific tour-de-force that very recently resulted in the revelation of the mysterious structure of blue phase III. The second part reviews progress made in the past decade in developing electrooptical, optical, and photonic devices based on blue phase III. The strong and weak aspects of the development of these devices are underlined and criticized, respectively. The third- and-final part proposes ideas for further improvement in blue phase III technology to make it feasible for commercialization and widespread use. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cytotoxicity and Bioactivity of Calcium Silicate Cements Combined with Niobium Oxide in Different Cell Lines.

Science.gov (United States)

Mestieri, Leticia Boldrin; Gomes-Cornélio, Ana Lívia; Rodrigues, Elisandra Márcia; Faria, Gisele; Guerreiro-Tanomaru, Juliane Maria; Tanomaru-Filho, Mário

2017-01-01

The aim of this study was to evaluate the cytotoxicity and bioactivity of calcium silicate-based cements combined with niobium oxide (Nb2O5) micro and nanoparticles, comparing the response in different cell lines. This evaluation used four cell lines: two primary cultures (human dental pulp cells - hDPCs and human dental follicle cells - hDFCs) and two immortalized cultures (human osteoblast-like cells - Saos-2 and mouse periodontal ligament cells - mPDL). The tested materials were: White Portland Cement (PC), mineral trioxide aggregate (MTA), white Portland cement combined with microparticles (PC/Nb2O5µ) or nanoparticles (PC/Nb2O5n) of niobium oxide (Nb2O5). Cytotoxicity was evaluated by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) and trypan blue exclusion assays and bioactivity by alkaline phosphatase (ALP) enzyme activity. Results were analyzed by ANOVA and Tukey test (a=0.05). PC/Nb2O5n presented similar or higher cell viability than PC/Nb2O5µ in all cell lines. Moreover, the materials presented similar or higher cell viability than MTA. Saos-2 exhibited high ALP activity, highlighting PC/Nb2O5µ material at 7 days of exposure. In conclusion, calcium silicate cements combined with micro and nanoparticles of Nb2O5 presented cytocompatibility and bioactivity, demonstrating the potential of Nb2O5 as an alternative radiopacifier agent for these cements. The different cell lines had similar response to cytotoxicity evaluation of calcium silicate cements. However, bioactivity was more accurately detected in human osteoblast-like cell line, Saos-2.

Telmisartan Exerts Anti-Tumor Effects by Activating Peroxisome Proliferator-Activated Receptor-γ in Human Lung Adenocarcinoma A549 Cells

Directory of Open Access Journals (Sweden)

Juan Li

2014-03-01

Full Text Available Telmisartan, a member of the angiotensin II type 1 receptor blockers, is usually used for cardiovascular diseases. Recent studies have showed that telmisartan has the property of PPARγ activation. Meanwhile, PPARγ is essential for tumor proliferation, invasion and metastasis. In this work we explore whether telmisartan could exert anti-tumor effects through PPARγ activation in A549 cells. MTT and trypan blue exclusion assays were included to determine the survival rates and cell viabilities. RT-PCR and western blotting were used to analyze the expression of ICAM-1, MMP-9 and PPARγ. DNA binding activity of PPARγ was evaluated by EMSA. Our data showed that the survival rates and cell viabilities of A549 cells were all reduced by telmisartan in a time- and concentration-dependent manner. Meanwhile, our results also demonstrated that telmisartan dose-dependently inhibited the expression of ICAM-1 and MMP-9. Moreover, the cytotoxic and anti-proliferative effects, ICAM-1 and MMP-9 inhibitive properties of telmisartan were totally blunted by the PPARγ antagonist GW9662. Our findings also showed that the expression of PPARγ was up-regulated by telmisartan in a dose dependent manner. And, the EMSA results also figured out that DNA binding activity of PPARγ was dose-dependently increased by telmisartan. Additionally, our data also revealed that telmisartan-induced PPARγ activation was abrogated by GW9662. Taken together, our results indicated that telmisartan inhibited the expression of ICAM-1 and MMP-9 in A549 cells, very likely through the up-regulation of PPARγ synthesis.

Effect of dental materials on gluconeogenesis in rat kidney tubules

Energy Technology Data Exchange (ETDEWEB)

Reichl, F.X.; Durner, J.; Mueckter, H.; Elsenhans, B.; Forth, W. [Muenchen Univ. (Germany). Walter-Straub-Institut fuer Pharmakologie und Toxikologie; Kunzelmann, K.H.; Hickel, R. [Department of Operative/Restorative Dentistry, Periodontology and Pedodontics, Ludwig-Maximilians-University of Munich (Germany); Spahl, W. [Institute of Organic Chemistry, Ludwig-Maximilians-University of Munich (Germany); Hume, W.R. [Dental Research Institute, Univ. of California, Los Angeles, CA (United States); Moes, G.W. [TNO Prins-Maurits-Laboratorium, Rijswijk (Netherlands)

1999-09-01

The effect of dental composite components triethyleneglycoldimethacrylate (TEGDMA) and hydroxyethylmethacrylate (HEMA) as well as mercuric chloride (HgCl{sub 2}) and methylmercury chloride (MeHgCl) on gluconeogenesis was investigated in isolated rat kidney tubules. From starved rats kidney tubules were prepared and isolated by digestion with collagenase. Every 10 min up to 60 min 1-ml samples were drawn from the cell suspension for quantitating the glucose content. Glucose formation in controls was 3.3 {+-} 0.2 nmol/mg . per min (mean {+-} SEM, n=21). Relative rates of glucose formation were obtained by expressing individual rates as a percentage of the corresponding control. X-Y concentration curves (effective concentration, EC) of the substances were calculated by fitting a four-parametric sigmoid function to the relative rates of glucose formation at various test concentrations. At the end of the incubation period cell viability was assessed by trypan blue exclusion. Cell viability decreased within the 60 min interval from 90 to approx. 80% (controls), <25 (HEMA), <20 (TEGDMA), <10 (MeHgCl), and <10% (HgCl{sub 2}). Values of 50% effective concentration (EC{sub 50}) were calculated from fitted curves. EC{sub 50} values were (mmol; mean {+-} SEM; n=4): HEMA, 17.7 {+-} 2.9; TEGDMA, 1.8 {+-} 0.2; MeHgCl, 0.018 {+-} 0.0005; and HgCl{sub 2}, 0.0016 {+-} 0.0005. The toxic effect of HgCl{sub 2} was {proportional{underscore}to}1000 or 10 000 higher than that of the dental composite components TEGDMA or HEMA, respectively. (orig.)

Auranofin, an Anti-Rheumatic Gold Compound, Modulates Apoptosis by Elevating the Intracellular Calcium Concentration ([Ca{sup 2+}]{sub i}) in MCF-7 Breast Cancer Cells

Energy Technology Data Exchange (ETDEWEB)

Varghese, Elizabeth; Büsselberg, Dietrich, E-mail: dib2015@qatar-med.cornell.edu [Weil Cornell Medical College in Qatar, Qatar Foundation-Education City, P.O. Box 24144 Doha (Qatar)

2014-11-06

Auranofin, a transition metal complex is used for the treatment of rheumatoid arthritis but is also an effective anti-cancer drug. We investigate the effects of Auranofin in inducing cell death by apoptosis and whether these changes are correlated to changes of intracellular calcium concentration ([Ca{sup 2+}]{sub i}) in breast cancer cells (MCF-7). Cytotoxicity of Auranofin was evaluated using MTS assay and the Trypan blue dye exclusion method. With fluorescent dyes SR-FLICA and 7-AAD apoptotic death and necrotic death were differentiated by Flow cytometry. A concentration dependent decrease in the viability occurred and cells were shifted to the apoptotic phase. Intracellular calcium ([Ca{sup 2+}]{sub i}) was recorded using florescence microscopy and a calcium sensitive dye (Fluo-4 AM) with a strong negative correlation (r = −0.713) to viability. Pharmacological modulators 2-APB (50 μM), Nimodipine (10 μM), Caffeine (10 mM), SKF 96365(20 μM) were used to modify calcium entry and release. Auranofin induced a sustained increase of [Ca{sup 2+}]{sub i} in a concentration and time dependent manner. The use of different blockers of calcium channels did not reveal the source for the rise of [Ca{sup 2+}]{sub i}. Overall, elevation of [Ca{sup 2+}]{sub i} by Auranofin might be crucial for triggering Ca{sup 2+}-dependent apoptotic pathways. Therefore, in anti-cancer therapy, modulating [Ca{sup 2+}]{sub i} should be considered as a crucial factor for the induction of cell death in cancer cells.

Effects of ultraviolet-visible irradiation in the presence of melanin isolated from human black or red hair upon Ehrlich ascites carcinoma cells

Energy Technology Data Exchange (ETDEWEB)

Menon, I.A.; Persad, S.; Ranadive, N.S.; Haberman, H.F.

1983-07-01

The present study is an attempt to investigate the possibility that ultraviolet irradiation in the presence of pheomelanin may be more harmful to cells than the irradiation in the presence of eumelanin. The effects of UV-visible irradiation upon Ehrlich ascites carcinoma cells in the presence of the melanin isolated from human black hair (eumelanin) or from red hair (pheomelanin) were investigated. Irradiation of these cells was found to produce cell lysis, as observed by leakage of 51Cr from labeled cells and intracellular lactic dehydrogenase from the cells and decrease in cell viability demonstrated by the trypan blue exclusion test. The three parameters were quantitatively parallel to one another under various experimental conditions, namely different periods of irradiation and irradiation in the presence of different concentrations of melanin. The above effects were more pronounced when the irradiation was carried out in the presence of melanin from red hair than in the presence of black-hair melanin. In the absence of either melanin, the irradiation did not produce any significant effect in cell viability or cell lysis. Irradiation of the cells in the presence of red-hair melanin also decreased the transplantability of these cells. These observations clearly show that irradiation of cells in the presence of pheomelanin could produce cytotoxic effects. The present experimental design may have application in the development of in vitro models for the study of UV radiation-induced cutaneous carcinogenesis. The reactions of pheomelanin may be related to the susceptibility of ''Celtic'' skin to UV radiation-induced skin damage and carcinogenesis.

"Blue-Collar Blues" : kõigi maade töötud, ühinege! / Ants Juske

Index Scriptorium Estoniae

Juske, Ants, 1956-2016

2009-01-01

Rahvusvaheline näitus "Blue-Collar Blues" Tallinna Kunstihoones ja Kunstihoone galeriis 31. jaanuarini 2010. Kuraator Anders Härm. Näituse ajendiks on 1. juulist 2009 Eestis kehtima hakanud töölepinguseadus, näituse fookus on töösuhetel

Population-specific life histories contribute to metapopulation viability

Science.gov (United States)

Halsey, Samniqueka J.; Bell, Timothy J.; McEachern, A. Kathryn; Pavlovic, Noel B.

2016-01-01

Restoration efforts can be improved by understanding how variations in life-history traits occur within populations of the same species living in different environments. This can be done by first understanding the demographic responses of natural occurring populations. Population viability analysis continues to be useful to species management and conservation with sensitivity analysis aiding in the understanding of population dynamics. In this study, using life-table response experiments and elasticity analyses, we investigated how population-specific life-history demographic responses contributed to the metapopulation viability of the Federally threatened Pitcher's thistle (Cirsium pitcheri). Specifically, we tested the following hypotheses: (1) Subpopulations occupying different environments within a metapopulation have independent demographic responses and (2) advancing succession results in a shift from a demographic response focused on growth and fecundity to one dominated by stasis. Our results showed that reintroductions had a positive contribution to the metapopulation growth rate as compared to native populations which had a negative contribution. We found no difference in succession on the contribution to metapopulation viability. In addition, we identified distinct population-specific contributions to metapopulation viability and were able to associate specific life-history demographic responses. For example, the positive impact of Miller High Dunes population on the metapopulation growth rate resulted from high growth contributions, whereas increased time of plant in stasis for the State Park Big Blowout population resulted in negative contributions. A greater understanding of how separate populations respond in their corresponding environment may ultimately lead to more effective management strategies aimed at reducing extinction risk. We propose the continued use of sensitivity analyses to evaluate population-specific demographic influences on

Neuroprotective effect of the natural iron chelator, phytic acid in a cell culture model of Parkinson's disease

International Nuclear Information System (INIS)

Xu Qi; Kanthasamy, Anumantha G.; Reddy, Manju B.

2008-01-01

Disrupted iron metabolism and excess iron accumulation has been reported in the brains of Parkinson's disease (PD) patients. Because excessive iron can induce oxidative stress subsequently causing degradation of nigral dopaminergic neurons in PD, we determined the protective effect of a naturally occurring iron chelator, phytic acid (IP6), on 1-methyl-4-phenylpyridinium (MPP + )-induced cell death in immortalized rat mesencephalic/dopaminergic cells. Cell death was induced with MPP + in normal and iron-excess conditions and cytotoxicity was measured by thiazolyl blue tetrazolium bromide (MTT assay) and trypan blue staining. Apoptotic cell death was also measured with caspase-3 activity, DNA fragmentation, and Hoechst nuclear staining. Compared to MPP + treatment, IP6 (30 μmol/L) increased cell viability by 19% (P + treatment was decreased by 55% (P < 0.01) and 52% (P < 0.05), respectively with IP6. Cell survival was increased by 18% (P < 0.05) and 42% (P < 0.001) with 30 and 100 μmol/L of IP6, respectively in iron-excess conditions. A 40% and 52% (P < 0.001) protection was observed in caspase-3 activity with 30 and 100 μmol/L IP6, respectively in iron-excess condition. Similarly, a 45% reduction (P < 0.001) in DNA fragmentation was found with 100 μmol/L IP6. In addition, Hoechst nuclear staining results confirmed the protective effect of IP6 against apoptosis. Similar protection was also observed with the differentiated cells. Collectively, our results demonstrate a significant neuroprotective effect of phytate in a cell culture model of PD

Live cell imaging reveals different modes of cytotoxic action of extracts derived from commonly used luting cements.

Science.gov (United States)

Trumpaitė-Vanagienė, Rita; Čebatariūnienė, Alina; Tunaitis, Virginijus; Pūrienė, Alina; Pivoriūnas, Augustas

2018-02-01

To compare cytotoxicity of extracts derived from commonly used luting cements: Hoffmann's Zinc Phosphate (ZPC), GC Fuji Plus Resin Modified Glass Ionomer (RMGIC) and 3M ESPE RelyX Unicem Resin Cement (RC) on primary human gingival fibroblasts (HGFs). HGFs were exposed to different concentrations of the ZPC, RMGIC and RC extracts. The cytotoxicity was assessed with the PrestoBlue Cell Viability Reagent and viable cells were counted by a haemocytometer using the trypan blue exclusion test. In order to determine the primary mechanism of the cell death induced by extracts from different luting cements, the real-time monitoring of caspase-3/-7 activity and membrane integrity of cells was employed. The extracts from the RMGIC and ZPC decreased the metabolic activity and numbers of viable cells. Unexpectedly, the extracts from the RC evoked only small effects on the metabolic activity of HGFs with a decreasing number of viable cells in a dose-and time-dependent manner. The live cell imaging revealed that the apoptosis was the primary mechanism of a cell death induced by the extracts derived from the RMGIC, whereas the extracts from the RC and ZPC induced a cell death through a necrotic and caspase-independent pathway. The apoptosis was the primary mechanism of the cell death induced by the extracts derived from the RMGIC, whereas the extracts from the RC and ZPC induced a cell death via a necrotic pathway. We suggest that metabolic assays commonly used to assess the cytotoxicity of luting cements should be validated by alternative methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

Poporodní blues – česká adaptace dotazníku „Maternity blues questionnaire“

Czech Academy of Sciences Publication Activity Database

Takács, L.; Smolík, Filip; Mlíková Seidlerová, J.; Čepický, P.; Hoskovcová, S.

2016-01-01

Roč. 81, č. 5 (2016), s. 355-368 ISSN 1210-7832 Institutional support: RVO:68081740 Keywords : EPDS postpartum mood * Maternity Blues Questionnaire * postnatal depression * postpartum blues * postpartum depression Subject RIV: AN - Psychology

Studies on the Mechanism and Prevention of Decompression Sickness.

Science.gov (United States)

1982-07-12

for trypan blue extraction from tissues or blood) con- sisted of four volumes of 95% ethanol and one volume of 17% benzalkonium chloride (Zephi- ran...sacrificed (in the striking reduction in mortality and absence or de- the experimental group the animals that did not succumb crease in severity of the...smooth muscle stimulating activity. The nature of the smooth muscle stimulating agent (s) involved has not yet been determined. Partial reduction in the

Petiveria alliacea extracts uses multiple mechanisms to inhibit growth of human and mouse tumoral cells

OpenAIRE

Urue?a, Claudia; Cifuentes, Claudia; Casta?eda, Diana; Arango, Amparo; Kaur, Punit; Asea, Alexzander; Fiorentino, Susana

2008-01-01

Abstract Background There is ethnopharmacological evidence that Petiveria alliacea can have antitumor activity; however, the mechanism of its cytotoxic activity is not well understood. We assessed multiple in vitro biological activities of an ethyl acetate soluble plant fraction over several tumor cell lines. Methods Tumor cell lines were evaluated using the following tests: trypan blue exclusion test, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], flow cytometry, cytosk...

Effect of a solution containing citrate/Methylene Blue/parabens on Staphylococcus aureus bacteria and biofilm, and comparison with various heparin solutions.

Science.gov (United States)

Sauer, Karin; Steczko, Janusz; Ash, Stephen R

2009-05-01

Some antibiotic solutions increase bacterial resistance and may cause toxic side effects. Heparin, frequently used as an anticoagulant in catheter lock solutions, may cause bleeding and stimulate biofilm formation. The aim of this study was to investigate the effect of a new antibacterial/antithrombotic solution, citrate/Methylene Blue/parabens (C/MB/P), versus various heparin solutions on the viability and the structure of preformed mature biofilms of Staphylococcus aureus bacteria. The degree of eradication of both planktonic and sessile microorganisms was evaluated. The changes in the structure of biofilms after exposure to C/MB/P and several concentrations of heparin were analysed by means of confocal laser scanning microscopy. COMSTAT image analysis was utilized to compare biofilm biomass, average and maximum height, surface coverage and roughness coefficient. Viability studies were performed on both biofilms and supernatant solutions. C/MB/P, in contrast to heparin solutions, significantly reduced biofilm biomass and thickness and reduced viability by 5 log when compared with saline treatment. No viable planktonic bacteria were detected and the few remaining biofilm cells appeared to be lysed. In contrast, most heparin solutions only reduced viability up to 1.0 log and failed to eradicate planktonic bacteria. C/MB/P has a rapid bactericidal effect on the preformed, mature biofilm of S. aureus. The structural changes of biofilms treated with C/MB/P, together with the observed log reduction of viable biofilm cells, confirmed the high potential of this solution to eliminate sessile bacteria. Furthermore, the tested solution entirely eliminated planktonic bacteria detached from the biofilm.

Experimental exposure of blue mussels (Mytilus galloprovincialis) to high levels of benzo[a]pyrene and possible implications for human health.

Science.gov (United States)

Speciale, A; Zena, R; Calabrò, C; Bertuccio, C; Aragona, M; Saija, A; Trombetta, D; Cimino, F; Lo Cascio, P

2018-04-15

Polycyclic aromatic hydrocarbons (PAHs) are lipophilic compounds able to accumulate in the food chain. Mussels showed to bioaccumulate contaminants, such as PAHs, so that recurrent consumption of such contaminated food represents a risk for human health. This study was aimed to elucidate if acute exposure of Mediterranean blue mussel (Mytilus galloprovincialis), a bivalve of great economic importance in several countries, to a PAH, benzo[a]pyrene (B[a]P), at doses able to induce cytochrome P450 1A (CYP1A) and pathological changes in mussel gills, can produce accumulation in soft tissue. We explored the cytotoxic effects (cell viability, DNA laddering, and glutathione levels) of in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to organic extracts obtained from blue mussels previously exposed for 12 and 72h via water to B[a]P (0.5-1mg/L). In our experimental conditions, B[a]P induced CYP1A induction and morphological changes in mussel gills and a significant B[a]P accumulation in soft tissue. Conversely, exposing PBMCs to organic extracts obtained from contaminated mussels, resulted in a significant reduction of cell viability and cell glutathione content, and in an increase in DNA laddering. This confirms that consumption of mussels from B[a]P polluted waters might affect human health. Our data lead us to suggest that CYP1A activity in mussel gills may be useful (more than the amount of detected PAHs in the mussel edible tissue) as a marker in assessment of risk for health of consumers exposed to PAHs through ingestion of shellfish. Copyright © 2017 Elsevier Inc. All rights reserved.

76 FR 22923 - Wellpoint, Inc. D/B/A/Anthem Blue Cross & Blue Shield Enterprise Provider Data Management Team...

Science.gov (United States)

2011-04-25

.../B/A/Anthem Blue Cross & Blue Shield Enterprise Provider Data Management Team Including On-Site... & Blue Shield, Enterprise Provider Data Management Team, Including On-Site Leased Workers From Kelly... Of Kentucky, Enterprise Provider Data Management Team, Louisville, Kentucky TA-W-74,895B Wellpoint...

Importance of Donor Chondrocyte Viability for Osteochondral Allografts.

Science.gov (United States)

Cook, James L; Stannard, James P; Stoker, Aaron M; Bozynski, Chantelle C; Kuroki, Keiichi; Cook, Cristi R; Pfeiffer, Ferris M

2016-05-01

Osteochondral allograft (OCA) transplantation provides a biological treatment option for functional restoration of large articular cartilage defects in multiple joints. While successful outcomes after OCA transplantation have been linked to viable donor chondrocytes, the importance of donor cell viability has not been comprehensively validated. To use a canine model to determine the importance of donor chondrocyte viability at the time of implantation with respect to functional success of femoral condylar OCAs based on radiographic, gross, cell viability, histologic, biochemical, and biomechanical outcome measures. Controlled laboratory study. After approval was obtained from the institutional animal care and use committee, adult female dogs (N = 16) were implanted with 8-mm cylindrical OCAs from male dogs in the lateral and medial femoral condyles of 1 knee. OCAs were preserved for 28 or 60 days after procurement, and chondrocyte viability was quantified before implantation. Two different storage media, temperatures, and time points were used to obtain a spectrum of percentage chondrocyte viability at the time of implantation. A successful outcome was defined as an OCA that was associated with graft integration, maintenance of hyaline cartilage, lack of associated cartilage disorder, and lack of fibrillation, fissuring, or fibrous tissue infiltration of the allograft based on subjective radiographic, gross, and histologic assessments at 6 months after implantation. Chondrocyte viability ranged from 23% to 99% at the time of implantation. All successful grafts had >70% chondrocyte viability at the time of implantation, and no graft with chondrocyte viability <70% was associated with a successful outcome. Live-dead stained sections and histologic findings with respect to cell morphological features suggested that successful grafts were consistently composed of viable chondrocytes in lacunae, while grafts that were not successful were composed of nonviable

Fast Blue RR—Siloxane Derivatized Materials Indicate Wound Infection Due to a Deep Blue Color Development

Directory of Open Access Journals (Sweden)

Doris Schiffer

2015-09-01

Full Text Available There is a strong need for simple and fast methods for wound infection determination. Myeloperoxidase, an immune system-derived enzyme was found to be a suitable biomarker for wound infection. Hence, alkoxysilane-derivatized Fast Blue RR was immobilized via simple hydrolytic polymerization. The resulting enzyme-responsive siloxane layers were incubated with myeloperoxidase, wound fluid or hemoglobin. The reaction was monitored via HPLC measurements and the color development quantified spectrophotometrically. Myeloperoxidase was indeed able to oxidize immobilized Fast Blue RR leading to a blue colored product. No conversion was detected in non-infected wound fluids. The visible color changes of these novel materials towards blue enable an easy distinction between infected and non-infected wound fluids.

Genomic copy number analysis of a spectrum of blue nevi identifies recurrent aberrations of entire chromosomal arms in melanoma ex blue nevus.

Science.gov (United States)

Chan, May P; Andea, Aleodor A; Harms, Paul W; Durham, Alison B; Patel, Rajiv M; Wang, Min; Robichaud, Patrick; Fisher, Gary J; Johnson, Timothy M; Fullen, Douglas R

2016-03-01

Blue nevi may display significant atypia or undergo malignant transformation. Morphologic diagnosis of this spectrum of lesions is notoriously difficult, and molecular tools are increasingly used to improve diagnostic accuracy. We studied copy number aberrations in a cohort of cellular blue nevi, atypical cellular blue nevi, and melanomas ex blue nevi using Affymetrix's OncoScan platform. Cases with sufficient DNA were analyzed for GNAQ, GNA11, and HRAS mutations. Copy number aberrations were detected in 0 of 5 (0%) cellular blue nevi, 3 of 12 (25%) atypical cellular blue nevi, and 6 of 9 (67%) melanomas ex blue nevi. None of the atypical cellular blue nevi displayed more than one aberration, whereas complex aberrations involving four or more regions were seen exclusively in melanomas ex blue nevi. Gains and losses of entire chromosomal arms were identified in four of five melanomas ex blue nevi with copy number aberrations. In particular, gains of 1q, 4p, 6p, and 8q, and losses of 1p and 4q were each found in at least two melanomas. Whole chromosome aberrations were also common, and represented the sole finding in one atypical cellular blue nevus. When seen in melanomas, however, whole chromosome aberrations were invariably accompanied by partial aberrations of other chromosomes. Three melanomas ex blue nevi harbored aberrations, which were absent or negligible in their precursor components, suggesting progression in tumor biology. Gene mutations involving GNAQ and GNA11 were each detected in two of eight melanomas ex blue nevi. In conclusion, copy number aberrations are more common and often complex in melanomas ex blue nevi compared with cellular and atypical cellular blue nevi. Identification of recurrent gains and losses of entire chromosomal arms in melanomas ex blue nevi suggests that development of new probes targeting these regions may improve detection and risk stratification of these lesions.

Assessment of bacterial endospore viability with fluorescent dyes.

Science.gov (United States)

Laflamme, C; Lavigne, S; Ho, J; Duchaine, C

2004-01-01

To validate three fluorescence viability assays designed primarily for vegetative cells on pure Bacillus endospores. Purified fresh and gamma-irradiated Bacillus endospores (Bacillus cereus, B. coagulans and two strains of B. subtilis) were used. The viability assays were: 5-cyano-2,3-diotolyl tetrazolium chloride (CTC) to test respiratory activity and early germination, DiBAC4(3) and Live/Dead BacLight to measure membrane energization and permeabilization, respectively. Gamma irradiation treatment completely eliminated spore culturability and was used as negative control. The untreated spores showed respiratory activity after 1 h of incubation and this was characteristic of almost 100% of spores after 24 h. The membrane potential assessment gave no answer about spore viability. A lower proportion of untreated spores had permeabilized membrane compared with gamma-irradiated spores using Live/Dead BacLight (P plate count. This study shows that fluorescence tests could be applied to assess viability in potentially pathogenic Bacillus spore preparations within 1 h.

Pollen Viability and Autogamy Fitness in Bauhinia forficata Link (Fabaceae

Directory of Open Access Journals (Sweden)

Luana Camila Capitani

2018-05-01

Full Text Available ABSTRACT Bauhinia forficata (Fabaceae occurs in many phytophysiognomies of southern Brazil, however its ecological relevance is not well understood. The present study was developed in the Central Depression of Rio Grande do Sul and aimed to determine variations in pollen viability along flowering, ability to perform autogamy and dye efficiency for the viability test. Pollen viability was determined by colorimetry as well as the ability to perform autogamy by isolating floral buds, being evaluated in eleven matrices. Average pollen viability was 81.43%, with the highest average value obtained with the dye 2,3,5- Triphenyltetrazolium Chloride (TTC (84.11%. Safranin was not a good indicator at the tested concentration. No correlation was found between pollen viability and flowering time. The species demonstrated an inability to perform autogamy.

The lithium abundance of M67 blue stragglers - A constraint on the blue straggler phenomenon

International Nuclear Information System (INIS)

Pritchet, C.J.; Glaspey, J.W.

1991-01-01

Upper limits have been placed on the line strength of the 6707 A Li I resonance doublet in seven blue stragglers in M67. The corresponding upper limits on abundances range from log N(Li) less than about 1.3 to less than about 2.3. This result is significantly below the level of log N(Li) about 3.1 + or - 0.1 found in field main-sequence stars of comparable temperature. It is concluded that some form of mixing has affected the outer envelopes of blue stragglers. (Such mixing has been proposed as the mechanism needed to prolong the lifetimes of blue stragglers relative to normal main-sequence stars at the same luminosity). Virtually all mechanisms for the production of blue stragglers other than mixing, binary mass transfer, or binary coalescence appear to be ruled out by the present observations. 45 refs

Ecology of blue straggler stars

CERN Document Server

Carraro, Giovanni; Beccari, Giacomo

2015-01-01

The existence of blue straggler stars, which appear younger, hotter, and more massive than their siblings, is at odds with a simple picture of stellar evolution. Such stars should have exhausted their nuclear fuel and evolved long ago to become cooling white dwarfs. They are found to exist in globular clusters, open clusters, dwarf spheroidal galaxies of the Local Group, OB associations and as field stars. This book summarises the many advances in observational and theoretical work dedicated to blue straggler stars. Carefully edited extended contributions by well-known experts in the field cover all the relevant aspects of blue straggler stars research: Observations of blue straggler stars in their various environments; Binary stars and formation channels; Dynamics of globular clusters; Interpretation of observational data and comparison with models. The book also offers an introductory chapter on stellar evolution written by the editors of the book.

Myocardial viability assessment using nuclear imaging

International Nuclear Information System (INIS)

Matsunari, Ichiro; Hisada, Kinichi; Taki, Junichi; Nakajima, Kenichi; Tonami, Norihisa

2003-01-01

Myocardial assessment continues to be an issue in patients with coronary artery disease and left ventricular dysfunction. Nuclear imaging has long played an important role in this field. In particular, PET imaging using 18 F-fluorodeoxyglucose is regarded as the metabolic gold standard of tissue viability, which has been supported by a wide clinical experience. Viability assessment using SPECT techniques has gained more wide-spread clinical acceptance than PET, because it is more widely available at lower cost. Moreover, technical advances in SPECT technology such as gated-SPECT further improve the diagnostic accuracy of the test. However, other imaging techniques such as dobutamine echocardiography have recently emerged as competitors to nuclear imaging. It is also important to note that they sometimes may work in a complementary fashion to nuclear imaging, indicating that an appropriate use of these techniques may significantly improve their overall accuracy. In keeping these circumstances in mind, further efforts are necessary to further improve the diagnostic performance of nuclear imaging as a reliable viability test. (author) 107 refs

Natural Blue Food Colour

DEFF Research Database (Denmark)

Roda-Serrat, Maria Cinta

In recent years, there has been a growing tendency to avoid the use of artificial colorants and additives in food products, especially after some studies linked their consumption with behavioural changes in children. However, the incorporation of colorants from natural origin remains a challenge...... for food technologists, as these are typically less vivid and less stable than their synthetic alternatives. Regarding blue colorants, phycocyanins from cyanobacteria are currently in the spotlight as promising new natural blue colorants. Phycocyanins are proteins which blue colour results from...... the presence of the chromophore phycocyanobilin (PCB), a covalently attached linear tetrapyrrole. The applications of phycocyanins as food colorants are however limited, as they show poor stability in certain conditions of pH, light and temperature. Cleavage of PCB from the protein followed by careful product...

Can greening of aquaculture sequester blue carbon?

Science.gov (United States)

Ahmed, Nesar; Bunting, Stuart W; Glaser, Marion; Flaherty, Mark S; Diana, James S

2017-05-01

Globally, blue carbon (i.e., carbon in coastal and marine ecosystems) emissions have been seriously augmented due to the devastating effects of anthropogenic pressures on coastal ecosystems including mangrove swamps, salt marshes, and seagrass meadows. The greening of aquaculture, however, including an ecosystem approach to Integrated Aquaculture-Agriculture (IAA) and Integrated Multi-Trophic Aquaculture (IMTA) could play a significant role in reversing this trend, enhancing coastal ecosystems, and sequestering blue carbon. Ponds within IAA farming systems sequester more carbon per unit area than conventional fish ponds, natural lakes, and inland seas. The translocation of shrimp culture from mangrove swamps to offshore IMTA could reduce mangrove loss, reverse blue carbon emissions, and in turn increase storage of blue carbon through restoration of mangroves. Moreover, offshore IMTA may create a barrier to trawl fishing which in turn could help restore seagrasses and further enhance blue carbon sequestration. Seaweed and shellfish culture within IMTA could also help to sequester more blue carbon. The greening of aquaculture could face several challenges that need to be addressed in order to realize substantial benefits from enhanced blue carbon sequestration and eventually contribute to global climate change mitigation.

The fading of irradiated blue-colored pearls

International Nuclear Information System (INIS)

Okamoto, Shinichi

1982-01-01

The fading of irradiated and natural blue-colored pearls was investigated in this experiment. Thirty natural blue-colored pearls and sixty irradiated blue-colored pearls were used. Some of them were placed at a light position of RT. Another pearls were placed at a dark position of 50 0 C. The irradiated pearls placed at a light position of RT didn't show remarkable fading in their color in 294 days. But the natural blue-colored pearls showed a little recovery from 4% to 8% in reflection factors in 223 days at RT. The irradiated pearls placed at a dark position of 50 0 C showed the recovery from 9% to 14% in 264 days independently of irradiation times. The natural blue-colored pearls also showed the bleaching from 5% to 10% in reflection factor in 86 days at 50 0 C. Both irradiated and natural blue-colored pearls hardly showed their remarkable changes in their chromaticities independently of temperatures. (author)

Blue space geographies: Enabling health in place.

Science.gov (United States)

Foley, Ronan; Kistemann, Thomas

2015-09-01

Drawing from research on therapeutic landscapes and relationships between environment, health and wellbeing, we propose the idea of 'healthy blue space' as an important new development Complementing research on healthy green space, blue space is defined as; 'health-enabling places and spaces, where water is at the centre of a range of environments with identifiable potential for the promotion of human wellbeing'. Using theoretical ideas from emotional and relational geographies and critical understandings of salutogenesis, the value of blue space to health and wellbeing is recognised and evaluated. Six individual papers from five different countries consider how health can be enabled in mixed blue space settings. Four sub-themes; embodiment, inter-subjectivity, activity and meaning, document multiple experiences within a range of healthy blue spaces. Finally, we suggest a considerable research agenda - theoretical, methodological and applied - for future work within different forms of blue space. All are suggested as having public health policy relevance in social and public space. Copyright © 2015 Elsevier Ltd. All rights reserved.

In vitro ovicidal and cestocidal effects of toxins from Bacillus thuringiensis on the canine and human parasite Dipylidium caninum.

Science.gov (United States)

Peña, Guadalupe; Aguilar Jiménez, Fortino Agustín; Hallal-Calleros, Claudia; Morales-Montor, Jorge; Hernández-Velázquez, Víctor Manuel; Flores-Pérez, Fernando Iván

2013-01-01

Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum). Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μ g/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μ g/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

In Vitro Ovicidal and Cestocidal Effects of Toxins from Bacillus thuringiensis on the Canine and Human Parasite Dipylidium caninum

Directory of Open Access Journals (Sweden)

Guadalupe Peña

2013-01-01

Full Text Available Bacillus thuringiensis is a gram-positive soil-dwelling bacterium that is commonly used as a biological pesticide. This bacterium may also be used for biological control of helminth parasites in domestic animals. In this study, we evaluated the possible ovicidal and cestocidal effects of a total protein extract of B. thuringiensis native strains on the zoonotic cestode parasite of dogs, Dipylidium caninum (D. caninum. Dose and time response curves were determined by coincubating B. thuringiensis proteins at concentration ranging from 100 to 1000 μg/mL along with 4000 egg capsules of D. caninum. Egg viability was evaluated using the trypan blue exclusion test. The lethal concentration of toxins on eggs was 600 μg/ml, and the best incubation time to produce this effect was 3 h. In the adult stage, the motility and the thickness of the tegument were used as indicators of damage. The motility was inhibited by 100% after 8 hours of culture compared to the control group, while the thickness of the cestode was reduced by 34%. Conclusively, proteins of the strain GP526 of B. thuringiensis directly act upon D. caninum showing ovicidal and cestocidal effects. Thus, B. thuringiensis is proposed as a potential biological control agent against this zoonosis.

Assessment of Cytotoxic Activity of Rosemary (Rosmarinus officinalis L.), Turmeric (Curcuma longa L.), and Ginger (Zingiber officinale R.) Essential Oils in Cervical Cancer Cells (HeLa)

Science.gov (United States)

Santos, P. A. S. R.; Avanço, G. B.; Nerilo, S. B.; Marcelino, R. I. A.; Janeiro, V.; Valadares, M. C.

2016-01-01

The objective of this study was to evaluate the cytotoxic activity of rosemary (REO, Rosmarinus officinalis L.), turmeric (CEO, Curcuma longa L.), and ginger (GEO, Zingiber officinale R.) essential oils in HeLa cells. Cytotoxicity tests were performed in vitro, using tetrazolium (MTT) and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC50 obtained was 36.6 μg/mL for CEO and 129.9 μg/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs), and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81 μg/mL of CEO and 32.12 μg/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activity in vitro for CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells. PMID:28042599

A novel protein from edible fungi Cordyceps militaris that induces apoptosis

Directory of Open Access Journals (Sweden)

Ke-Chun Bai

2018-01-01

Full Text Available Cordyceps militaris is a dietary therapeutic fungus that is an important model species in Cordyceps research. In this study, we purified a novel protein from the fruit bodies of C. militaris and designated it as Cordyceps militaris protein (CMP. CMP has a molecular mass of 18.0 kDa and is not glycosylated. Interestingly, CMP inhibited cell viability in murine primary cells and other cell lines in a time- and dose-dependent manner. Using trypan blue staining and a lactate dehydrogenase release assay, we showed that CMP caused cell death in the murine hepatoma cell line BNL 1MEA.7R.1. Furthermore, the frequency of BNL 1MEA.7R.1 cells at the sub-G1 stage was increased by CMP. Apoptosis, as determined by Annexin V and propidium iodide analysis, indicated that CMP could mediate BNL 1MEA.7R.1 apoptosis, but not necrosis. After coincubation with CMP, a decrease in mitochondria potential was detected using 3,3′-dihexyloxacarbocyanine iodide. These results suggest that CMP is a harmful protein that induces apoptosis through a mitochondrion-dependent pathway. Stability experiments demonstrated that heat treatment and alkalization degraded CMP and further destroyed its cell-death-inducing ability, implying that cooking is necessary for food containing C. militaris.

Markers Of Apoptosis In HEP-2 Cells In Vitro Damaged By PHOTOFRIN And He:Ne Laser

International Nuclear Information System (INIS)

KORRAA, S.; ELMAGHRABY, T.K.; HELMY, H.M.

2009-01-01

Photodynamic therapy (PDT), which is a treatment for cancer and certain non-cancerous conditions, requires exposure of cells or tissue to a photosensitizing drug followed by irradiation with visible light of the appropriate wavelength. Although PDT can produce apoptosis or necrosis or a combination of the two mechanisms, PDT is a highly efficient if it induces apoptosis. The aim of the present study is to investigate the effect of combination between photofrin and He:Ne laser on killing Hep-2 cells in vitro, 2 and 24 hours post-laser irradiation. Also, to determine the markers of apoptosis in terms of levels of Bcl-2 protein and Bax mRNA relative expression in PDT treated cells six hours post laser irradiation. Cells viability was measured by trypan blue exclusion test (by light microscope). The results showed that photofrin in combination with He:Ne laser was efficient in decreasing the number of viable cells. It caused 81.47± 2.0 % cell death 2 hours post laser irradiation then reached 52.37± 2.24 % 24 hours post laser irradiation. PDT induced an increase in Bax mRNA relative expression and a decrease in Bcl-2 protein as measured 6 hours post laser irradiation.

Assessment of Cytotoxic Activity of Rosemary (Rosmarinus officinalis L., Turmeric (Curcuma longa L., and Ginger (Zingiber officinale R. Essential Oils in Cervical Cancer Cells (HeLa

Directory of Open Access Journals (Sweden)

P. A. S. R. Santos

2016-01-01

Full Text Available The objective of this study was to evaluate the cytotoxic activity of rosemary (REO, Rosmarinus officinalis L., turmeric (CEO, Curcuma longa L., and ginger (GEO, Zingiber officinale R. essential oils in HeLa cells. Cytotoxicity tests were performed in vitro, using tetrazolium (MTT and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC50 obtained was 36.6 μg/mL for CEO and 129.9 μg/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs, and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81 μg/mL of CEO and 32.12 μg/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activity in vitro for CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells.

Increased radiosensitivity and radiothermosensitivity of human pancreatic MIA PaCa-2 and U251 glioblastoma cell lines treated with the novel Hsp90 inhibitor NVP-HSP990

International Nuclear Information System (INIS)

Milanović, Dušan; Firat, Elke; Grosu, Anca Ligia; Niedermann, Gabriele

2013-01-01

Heat shock Protein 90 (Hsp90) is a molecular chaperone that folds, stabilizes, and functionally regulates many cellular proteins involved in oncogenic signaling and in the regulation of radiosensitivity. It is upregulated in response to stress such a heat. Hyperthermia is a potent radiosensitizer, but induction of Hsp90 may potentially limit its efficacy. Our aim was to investigate whether the new Hsp90 inhibitor NVP-HSP990 increases radiosensitivity, thermosensitivity and radiothermosensitivity of human tumor cell lines. U251 glioblastoma and MIA PaCa-2 pancreatic carcinoma cells were used. To determine clonogenic survival, colony forming assays were performed. Cell viability and proliferation were assesed by Trypan blue staining. Cell cycle and apoptosis analyses were performed by flow cytometry. DAPI staining was used to detect mitotic catastrophe. NVP-HSP990 increased the thermosensitivity, radiosensitivity and radio-thermosensitivity of both cell lines in clonogenic assays. 72 hours after irradiation with 4 Gy, a significant reduction in cell number associated with considerable G2/M acumulation and mitotic catastrophe as well as cell death by apoptosis/necrosis was observed. Treatment with NVP-HSP990 strongly sensitized U251 and MIA PaCa-2 cells to hyperthermia and ionizing radiation or combination thereof through augmentation of G2/M arrest, mitotic catastrophe and associated apoptosis

Paracetamol (acetaminophen) attenuates in vitro mast cell and peripheral blood mononucleocyte cell histamine release induced by N-acetylcysteine.

Science.gov (United States)

Coulson, James; Thompson, John Paul

2010-02-01

The treatment of acute paracetamol (acetaminophen) poisoning with N-acetylcysteine (NAC) is frequently complicated by an anaphylactoid reaction to the antidote. The mechanism that underlies this reaction is unclear. We used the human mast cell line 1 (HMC-1) and human peripheral blood mononucleocytes (PBMCs) to investigate the effects of NAC and paracetamol on histamine secretion in vitro. HMC-1 and human PBMCs were incubated in the presence of increasing concentrations of NAC +/- paracetamol. Cell viability was determined by the Trypan Blue Assay, and histamine secretion was measured by ELISA. NAC was toxic to HMC-1 cells at 100 mg/mL and to PBMCs at 67 mg/mL. NAC increased HMC-1 and PBMC histamine secretion at concentrations of NAC from 20 to 50 mg/mL and 2.5 to 100 mg/mL, respectively. NAC-induced histamine secretion by both cell types was reduced by co-incubation with 2.5 mg/mL of paracetamol. Paracetamol (acetaminophen) is capable of modifying histamine secretion in vitro. This may explain the clinical observation of a lower incidence of adverse reactions to NAC in vivo when higher concentrations of paracetamol are present than when paracetamol concentrations are low. Paracetamol (acetaminophen) attenuates in vitro mast cell and PBMC cell histamine release induced by NAC.

2-ethylpyridine, a cigarette smoke component, causes mitochondrial damage in human retinal pigment epithelial cells in vitro

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S Mansoor

2014-01-01

Full Text Available Purpose: Our goal was to identify the cellular and molecular effects of 2-ethylpyridine (2-EP, a component of cigarette smoke on human retinal pigment epithelial cells (ARPE-19 in vitro. Materials and Methods: ARPE-19 cells were exposed to varying concentrations of 2-EP. Cell viability (CV was measured by a trypan blue dye exclusion assay. Caspase-3/7 and caspase-9 activities were measured by fluorochrome assays. The production of reactive oxygen/nitrogen species (ROS/RNS was detected with a 2′,7′-dichlorodihydrofluorescein diacetate dye assay. The JC-1 assay was used to measure mitochondrial membrane potential (ΔΨm. Mitochondrial redox potential was measured using a RedoxSensor Red kit and mitochondria were evaluated with Mitotracker dye. Results: After 2-EP exposure, ARPE-19 cells showed significantly decreased CV, increased caspase-3/7 and caspase-9 activities, elevated ROS/RNS levels, decreased ΔΨm value and decreased redox fluorescence when compared with control samples. Conclusions: These results show that 2-EP treatment induced cell death by caspase-dependent apoptosis associated with an oxidative stress and mitochondrial dysfunction. These data represent a possible mechanism by which smoking contributes to age-related macular degeneration and other retinal diseases and identify mitochondria as a target for future therapeutic interventions.

Biodistribution studies of {sup 99m}Tc-labeled myoblasts in a murine model of muscular dystrophy

Energy Technology Data Exchange (ETDEWEB)

Colombo, F.R. E-mail: colombof@policlinico.mi.it; Torrente, Y.; Casati, R.; Benti, R.; Corti, S.; Salani, S.; D' Angelo, M.G.; DeLiso, A.; Scarlato, G.; Bresolin, N.; Gerundini, P

2001-11-01

The purpose of this study was twofold: first, to evaluate the myoblast labeling of various {sup 99m}Tc complexes and to select the complex that best accomplishes this labeling, and second to evaluate the biodistribution of myoblasts labeled with this complex using mice with MDX muscular dystrophy (the murine homologue of Duchenne's muscular dystrophy). The following ligands were used to prepare the corresponding {sup 99m}Tc complexes: hexakis-methoxy-isobutyl-isonitrile (MIBI), bis(2-ethoxyethyl)diphosphinoethane (Tf), (RR,SS)-4,8-diaza-3,6,6,9-tetramethyl-undecane-2,10-dione-bisoxime (HM-PAO), bis(N-ethyl)dithiocarbamate (NEt), and bis(N-ethoxy, N-ethyl)dithiocarbamate (NOEt). One million murine myoblasts were incubated for 30-60 minutes with 5 mCi of each of the 99mTc complexes prepared from the above ligands. Viability was assessed by microscopic counting after trypan blue staining, and the radioactivity absorbed in the cells was measured after centrifugation. The compound with the highest uptake in cellular pellets was [{sup 99m}Tc]N-NOEt. The biodistribution of myoblasts labeled with this complex was evaluated after intraaortic injection in dystrophic mice. Such an approach has the potential of effecting widespread gene transfer through the bloodstream to muscles lacking dystrophin.

Application of a non-hazardous vital dye for cell counting with automated cell counters.

Science.gov (United States)

Kim, Soo In; Kim, Hyun Jeong; Lee, Ho-Jae; Lee, Kiwon; Hong, Dongpyo; Lim, Hyunchang; Cho, Keunchang; Jung, Neoncheol; Yi, Yong Weon

2016-01-01

Recent advances in automated cell counters enable us to count cells more easily with consistency. However, the wide use of the traditional vital dye trypan blue (TB) raises environmental and health concerns due to its potential teratogenic effects. To avoid this chemical hazard, it is of importance to introduce an alternative non-hazardous vital dye that is compatible with automated cell counters. Erythrosin B (EB) is a vital dye that is impermeable to biological membranes and is used as a food additive. Similarly to TB, EB stains only nonviable cells with disintegrated membranes. However, EB is less popular than TB and is seldom used with automated cell counters. We found that cell counting accuracy with EB was comparable to that with TB. EB was found to be an effective dye for accurate counting of cells with different viabilities across three different automated cell counters. In contrast to TB, EB was less toxic to cultured HL-60 cells during the cell counting process. These results indicate that replacing TB with EB for use with automated cell counters will significantly reduce the hazardous risk while producing comparable results. Copyright © 2015 Logos Biosystems, Inc. Published by Elsevier Inc. All rights reserved.

Biodistribution studies of 99mTc-labeled myoblasts in a murine model of muscular dystrophy

International Nuclear Information System (INIS)

Colombo, F.R.; Torrente, Y.; Casati, R.; Benti, R.; Corti, S.; Salani, S.; D'Angelo, M.G.; DeLiso, A.; Scarlato, G.; Bresolin, N.; Gerundini, P.

2001-01-01

The purpose of this study was twofold: first, to evaluate the myoblast labeling of various 99m Tc complexes and to select the complex that best accomplishes this labeling, and second to evaluate the biodistribution of myoblasts labeled with this complex using mice with MDX muscular dystrophy (the murine homologue of Duchenne's muscular dystrophy). The following ligands were used to prepare the corresponding 99m Tc complexes: hexakis-methoxy-isobutyl-isonitrile (MIBI), bis(2-ethoxyethyl)diphosphinoethane (Tf), (RR,SS)-4,8-diaza-3,6,6,9-tetramethyl-undecane-2,10-dione-bisoxime (HM-PAO), bis(N-ethyl)dithiocarbamate (NEt), and bis(N-ethoxy, N-ethyl)dithiocarbamate (NOEt). One million murine myoblasts were incubated for 30-60 minutes with 5 mCi of each of the 99mTc complexes prepared from the above ligands. Viability was assessed by microscopic counting after trypan blue staining, and the radioactivity absorbed in the cells was measured after centrifugation. The compound with the highest uptake in cellular pellets was [ 99m Tc]N-NOEt. The biodistribution of myoblasts labeled with this complex was evaluated after intraaortic injection in dystrophic mice. Such an approach has the potential of effecting widespread gene transfer through the bloodstream to muscles lacking dystrophin

Assessment of Cytotoxic Activity of Rosemary (Rosmarinus officinalis L.), Turmeric (Curcuma longa L.), and Ginger (Zingiber officinale R.) Essential Oils in Cervical Cancer Cells (HeLa).

Science.gov (United States)

Santos, P A S R; Avanço, G B; Nerilo, S B; Marcelino, R I A; Janeiro, V; Valadares, M C; Machinski, Miguel

2016-01-01

The objective of this study was to evaluate the cytotoxic activity of rosemary (REO, Rosmarinus officinalis L.), turmeric (CEO, Curcuma longa L.), and ginger (GEO, Zingiber officinale R.) essential oils in HeLa cells. Cytotoxicity tests were performed in vitro , using tetrazolium (MTT) and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC 50 obtained was 36.6  μ g/mL for CEO and 129.9  μ g/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs), and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81  μ g/mL of CEO and 32.12  μ g/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activity in vitro for CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells.

Induction of the acrosome reaction test to in vitro estimate embryo production in Nelore cattle

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M.Z. Costa

2010-08-01

Full Text Available The effectiveness of induction of the acrosome reaction (AR test as a parameter to in vitro estimate embryo production (IVP in Nelore breed and the AR pattern by the Trypan Blue/Giemsa (TB stain were evaluated. Frozen semen samples from ten Nelore bulls were submitted to AR induction and were also evaluated for cleavage and blastocyst rates. The treatments utilized for AR induction were: control (TALP medium, TH (TALP medium + 10μg heparin, TL (TALP medium + 100μg lysophosphatidylcholine and THL (TALP medium + 10μg heparin + 100μg lysophosphatidylcholine. Sperm acrosomal status and viability were evaluated by TB staining at 0 and after 4h incubation at 38°C. The results obtained for AR presented a significant difference (P<0.05 in the percentage of acrosome reacted live sperm after 4h of incubation in the treatments that received heparin. The cleavage and blastocyst rates were 60% and 38% respectively and a significant difference was observed among bulls (P<0.05. It was founded a satisfactory model to estimate the cleavage and blastocyst rates by AR induction test. Therefore, it can be concluded that the induction of the AR test is a valuable tool to predict the IVP in Nelore breed.

New nanostructural biomaterials based on active silicate systems and hydroxyapatite: characterization and genotoxicity in human peripheral blood lymphocytes.

Science.gov (United States)

Opačić-Galić, V; Petrović, V; Zivković, S; Jokanović, V; Nikolić, B; Knežević-Vukčević, J; Mitić-Ćulafić, D

2013-06-01

To characterize and investigate the genotoxic effect of a new endodontic cement based on dicalcium- and tricalcium-silicate (CS) with hydroxyapatite (HA) on human lymphocytes. Hydrothermal treatment was applied for synthesis of CS and HA. The final mixture HA-CS, with potential to be used in endodontic practice, is composed of CS (34%) and HA (66%). Human lymphocytes were incubated with HA, HA-CS and CS for 1 h, at 37 °C and 5% CO2. Cell viability was determined using the trypan blue exclusion assay. To evaluate the level of DNA damage comet assay (single cell gel electrophoresis) was performed. For the statistical analysis anova and Duncan's Post Hoc Test were used. The SEM analysis indicated that CS consisted mostly of agglomerates of several micrometers in size, built up from smaller particles, with dimensions between 117 and 477 nm. This is promising because dimensions of agglomerates are not comparable with channels inside the cell membranes, whereas their nano-elements provide evident activity, important for faster setting of these mixtures compared to MTA. Values of DNA damage obtained in the comet assay indicated low genotoxic risk of the new endodontic materials. The significantly improved setting characteristics and low genotoxic risk of the new material support further research. © 2012 International Endodontic Journal.

Starbursts in Blue compact dwarf galaxies

International Nuclear Information System (INIS)

Thuan, T.X.

1987-01-01

We summarize all the arguments for a bursting mode of star formation in blue compact dwarf galaxies. We show in particular how spectral synthesis of far ultraviolet spectra of Blue compact dwarf galaxy constitutes a powerful way for studying the star formation history in these galaxies. Blue compact dwarf galaxy luminosity functions show jumps and discontinuities. These jumps act like fossil records of the star-forming bursts, helping us to count and date the bursts

Present assessment of myocardial viability by nuclear imaging.

Science.gov (United States)

Saha, G B; MacIntyre, W J; Brunken, R C; Go, R T; Raja, S; Wong, C O; Chen, E Q

1996-10-01

Prospective delineation of viable from nonviable myocardium in patients with coronary artery disease in an important factor in deciding whether a patient should be revascularized or treated medically. Two common techniques--single-photon emission computed tomography (SPECT) and positron-emission computed tomography (PET)--are used in nuclear medicine using various radiopharmaceuticals for the detection of myocardial viability in patients. Thallium-201 (201Tl) and technetium-99m (99mTc)-sestamibi are the common radiopharmaceuticals used in different protocols using SPECT, whereas fluoride-18 (18F)-fluorodeoxyglucose (FDG) and rubidium-82 (82Rb) are most widely used in PET. The SPECT protocols involve stress/redistribution, stress/redistribution/reinjection, and rest/redistribution imaging techniques. Many studies have compared the results of 201Tl and (99mTc)-sestamibi SPECT with those of FDG PET; in some studies, concordant results have been found between delayed thallium and FDG results, indicating that 201Tl, although considered a perfusion agent, shows myocardial viability. Discordant results in a number of studies have been found between sestamibi and FDG, suggesting that the efficacy of sestamibi as a viability marker has yet to be established. Radiolabeled fatty acids such as iodine-123 (123I)-para-iodophenylpentadecanoic acid and carbon-11 (11C)-palmitic acid have been used for the assessment of myocardial viability with limited success. 11C-labeled acetate is a good marker of oxidative metabolism in the heart and has been used to predict the reversibility of wall motion abnormalities. (18F)-FDG is considered the marker of choice for myocardial viability, although variable results are obtained under different physiological conditions. Detection of myocardial viability can be greatly improved by developing new equipment and radiopharmaceuticals of better quality.

Hubble's View of Little Blue Dots

Science.gov (United States)

Kohler, Susanna

2018-02-01

The recent discovery of a new type of tiny, star-forming galaxy is the latest in a zoo of detections shedding light on our early universe. What can we learn from the unique little blue dots found in archival Hubble data?Peas, Berries, and DotsGreen pea galaxies identified by citizen scientists with Galaxy Zoo. [Richard Nowell Carolin Cardamone]As telescope capabilities improve and we develop increasingly deeper large-scale surveys of our universe, we continue to learn more about small, faraway galaxies. In recent years, increasing sensitivity first enabled the detection of green peas luminous, compact, low-mass (10 billion solar masses; compare this to the Milky Ways 1 trillion solar masses!) galaxies with high rates of star formation.Not long thereafter, we discovered galaxies that form stars similarly rapidly, but are even smaller only 330 million solar masses, spanning less than 3,000 light-years in size. These tiny powerhouses were termed blueberries for their distinctive color.Now, scientists Debra and Bruce Elmegreen (of Vassar College and IBM Research Division, respectively) report the discovery of galaxies that have even higher star formation rates and even lower masses: little blue dots.Exploring Tiny Star FactoriesThe Elmegreens discovered these unique galaxies by exploring archival Hubble data. The Hubble Frontier Fields data consist of deep images of six distant galaxy clusters and the parallel fields next to them. It was in the archival data for two Frontier Field Parallels, those for clusters Abell 2744 and MAS J0416.1-2403, that the authors noticed several galaxies that stand out as tiny, bright, blue objects that are nearly point sources.Top: a few examples of the little blue dots recently identified in two Hubble Frontier Field Parallels. Bottom: stacked images for three different groups of little blue dots. [Elmegreen Elmegreen 2017]The authors performed a search through the two Frontier Field Parallels, discovering a total of 55 little blue dots

The effects of storage conditions on the viability of ...

African Journals Online (AJOL)

SARAKA DANIEL

2015-01-07

Jan 7, 2015 ... E-mail: ndsaraka@yahoo.fr, ndsaraka@gmail.com. Tel: 225 ..... sample viability and control of contaminants that may mask the ... viability and composition of the Escherichia coli flora in faecal ... Microbiologie alimentaire, 8e.

A business model design framework for viability : a business ecosystem approach

NARCIS (Netherlands)

D'Souza, Austin; Velthuijsen, Hugo; Wortmann, J.C.; Huitema, George

2015-01-01

Purpose: To facilitate the design of viable business models by proposing a novel business model design framework for viability. Design: A design science research method is adopted to develop a business model design framework for viability. The business model design framework for viability is

Viability and Virulence of Entomopathogenic Nematodes Exposed to Ultraviolet Radiation.

Science.gov (United States)

Shapiro-Ilan, David I; Hazir, Selcuk; Lete, Luis

2015-09-01

Entomopathogenic nematodes (EPNs) can be highly effective biocontrol agents, but their efficacy can be reduced due to exposure to environmental stress such as from ultraviolet (UV) radiation. Our objectives were to 1) compare UV tolerance among a broad array of EPN species, and 2) investigate the relationship between reduced nematode viability (after exposure to UV) and virulence. Nematodes exposed to a UV radiation (254 nm) for 10 or 20 min were assessed separately for viability (survival) and virulence to Galleria mellonella. We compared 9 different EPN species and 15 strains: Heterorhabditis bacteriophora (Baine, fl11, Oswego, and Vs strains), H. floridensis (332), H. georgiana (Kesha), H. indica (HOM1), H. megidis (UK211), Steinernema carpocapsae (All, Cxrd, DD136, and Sal strains), S. feltiae (SN), S. rarum (17C&E), and S. riobrave (355). In viability assessments, steinernematids, particularly strains of S. carpocapsae, generally exhibited superior UV tolerance compared with the heterorhabditids. However, some heterorhabditids tended to be more tolerant than others, e.g., H. megidis and H. bacteriophora (Baine) were most susceptible and H. bacteriophora (Vs) was the only heterorhabditid that did not exhibit a significant effect after 10 min of exposure. All heterorhabditids experienced reduced viability after 20 min exposure though several S. carpocapsae strains did not. In total, after 10 or 20 min exposure, the viability of seven nematode strains did not differ from their non-UV exposed controls. In virulence assays, steinernematids (particularly S. carpocapsae strains) also tended to exhibit higher UV tolerance. However, in contrast to the viability measurements, all nematodes experienced a reduction in virulence relative to their controls. Correlation analysis revealed that viability among nematode strains is not necessarily related to virulence. In conclusion, our results indicate that the impact of UV varies substantially among EPNs, and viability alone

The synthetic inhibitor of Fibroblast Growth Factor Receptor PD166866 controls negatively the growth of tumor cells in culture

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Castelli Mauro

2009-12-01

Full Text Available Abstract Background Many experimental data evidence that over-expression of various growth factors cause disorders in cell proliferation. The role of the Fibroblast Growth Factors (FGF in growth control is indisputable: in particular, FGF1 and its tyrosine kinase receptor (FGFR1 act through a very complex network of mechanisms and pathways. In this work we have evaluated the antiproliferative activity effect of PD166866, a synthetic molecule inhibiting the tyrosin kinase action of FGFR1. Methods Cells were routinely grown in Dulbecco Modified Eagle's medium supplemented with newborn serum and a penicillin-streptomycin mixture. Cell viability was evaluated by Mosmann assay and by trypan blue staining. DNA damage was assessed by in situ fluorescent staining with Terminal Deoxynucleotidyl Transferase dUTP nick end labeling (TUNEL assay. Assessment of oxidative stress at membrane level was measured by quantitative analysis of the intra-cellular formation of malonyl-dialdheyde (MDA deriving from the decomposition of poly-unsaturated fatty acids. The expression of Poly-ADP-Ribose-Polymerase (PARP, consequent to DNA fragmentation, was evidenced by immuno-histochemistry utilizing an antibody directed against an N-terminal fragment of the enzyme. Results The bioactivity of the drug was investigated on Hela cells. Cytoxicity was assessed by the Mosmann assay and by vital staining with trypan blue. The target of the molecule is most likely the cell membrane as shown by the significant increase of the intracellular concentration of malonyl-dihaldheyde. The increase of this compound, as a consequence of the treatment with PD166866, is suggestive of membrane lipoperoxidation. The TUNEL assay gave a qualitative, though clear, indication of DNA damage. Furthermore we demonstrate intracellular accumulation of poly-ADP-ribose polymerase I. This enzyme is a sensor of nicks on the DNA strands and this supports the idea that treatment with the drug induces cell

In vitro biocompatibility tests of two commercial types of mineral trioxide aggregate Testes de biocompatibilidade in vitro de duas formas comerciais do agregado de trióxido mineral

Directory of Open Access Journals (Sweden)

Daniel Araki Ribeiro

2005-09-01

Full Text Available Recently, regular and white mineral trioxide aggregate (MTA are being used in Dentistry as retrofilling materials. Genotoxicity and cytotoxicity tests form an important part of cancer research and risk assessment of potential carcinogens. Thus, the goal of this study was to examine the genotoxicity and cytotoxicity of regular and white MTA in vitro by the single cell gel (comet assay and trypan blue exclusion test, respectively. Mouse lymphoma cells were exposed to two presentation forms of MTA at final concentrations ranging from 1 to 1,000 µg/mL for 3 h at 37°C. The results showed that both compounds tested did not produce genotoxic effects at all concentrations evaluated. Likewise, no statistically significant differences (p > 0.05 were observed in cytotoxicity. Taken together, our results suggest that regular and white MTA are not genotoxins and are not able to interfere in cellular viability as assessed by single cell gel (comet assay and trypan blue assay, respectively.Recentemente, o agregado de trióxido mineral (MTA regular e branco estão sendo utilizados na Odontologia como materiais para obturação retrógrada de canais radiculares. Testes de genotoxicidade e citotoxicidade formam uma importante parte da pesquisa do câncer e da avaliação de risco de carcinógenos potenciais. Assim, o objetivo deste estudo foi examinar a genotoxicidade e citotoxicidade do MTA branco e regular in vitro pelo teste do cometa e teste de exclusão pelo azul de tripan, respectivamente. Células do linfoma murino foram expostas às duas formas de apresentação do MTA nas concentrações finais de 1 a 1.000 µg/mL por 3 horas a 37°C. Os resultados mostraram que ambos os compostos testados não produziram efeito genotóxico em todas as concentrações testadas. Da mesma forma, nenhuma diferença estatisticamente significativa (p > 0,05 foi observada na citotoxicidade. Em suma, nossos resultados sugerem que o MTA regular e branco não são genotoxinas e

Variable blue straggler stars in NGC 5466

International Nuclear Information System (INIS)

Harris, H.C.; Mateo, M.; Olszewski, E.W.; Nemec, J.M.

1990-01-01

Nine variable blue stragglers have been found in the globular cluster NGC 5466. The six dwarf Cepheids in this cluster coexist in the instability strip with other nonvariable stars. The three eclipsing binaries are among the hottest of the blue stragglers. The hypothesis is discussed that all blue stragglers in this cluster have undergone mass transfer in close binaries. Under this hypothesis, rotation and spin-down play important roles in controlling the evolution of blue stragglers in old clusters and in affecting some of their observational properties. 14 refs

In vitro assays for predicting tumor cell response to radiation by apoptotic pathways

International Nuclear Information System (INIS)

Algan, Oe.; Hanks, G.E.; Biade, S.; Chapman, J.D.

1995-01-01

Purpose: We had previously shown that the rate of spontaneous and radiation-induced apoptosis was significantly greater in well-differentiated compared to anaplastic Dunning prostate carcinomas. The goal of this study was to define the most useful assay for quantifying radiation-induced apoptotic cell death and to determine if measured rates of radiation-induced apoptosis in tumor cell populations can predict treatment outcome. Materials and Methods: The time course and extent of radiation-induced apoptosis after single doses of Cesium-137 gamma-rays were measured by five different assays. These included gross DNA degradation, nucleosome ladder formation, labeling of 3'-OH ends in DNA with an immunofluorescence probe, immunofluorescence vital stains (LIVE/DEAD[reg] EUKOLIGHT TM ) and trypan blue. The majority of these studies were performed with DU-145 human prostate cells. Data was analyzed to determine the component of cell inactivation resulting from apoptosis with the modified linear quadratic equation, -1n (SF) = (α a + α p ) D + β p D 2 , were α a represents cell inactivation by radiation-induced apoptosis, α p and β p represent cell death by proliferative mechanisms and D represents radiation dose. Results: These studies indicated that DU-145 cell death after radiation occurs over two distinct time periods. The first phase of death begins shortly after irradiation and plateaus within 16-24 hr. This process of cell death has properties consistent with apoptosis as determined by 3'-OH DNA end-labeling and nucleosome ladder assays. The second phase of cell death (determined by viability staining) begins approximately 48 hr after irradiation and continues until the remainder of inactivated cells express their death. This longer phase of cell inactivation probably represents proliferative cell death and other non-apoptotic mechanisms. The five different assays were performed on DU-145 cells 24 hr after irradiation with 10 Gy. Significant nucleosome ladders

Molecular evidence for the subspecific differentiation of blue sheep (Pseudois nayaur) and polyphyletic origin of dwarf blue sheep (Pseudois schaeferi).

Science.gov (United States)

Tan, Shuai; Zou, Dandan; Tang, Lei; Wang, Gaochao; Peng, Quekun; Zeng, Bo; Zhang, Chen; Zou, Fangdong

2012-06-01

Blue sheep (Pseudois nayaur), a Central Asian ungulate with restricted geographic distribution, exhibits unclear variation in morphology and phylogeographic structure. The composition of species and subspecies in the genus Pseudois is controversial, particularly with respect to the taxonomic designation of geographically restricted populations. Here, 26 specimens including 5 dwarf blue sheep (Pseudois schaeferi), which were collected from a broad geographic region in China, were analyzed for 2 mitochondrial DNA fragments (cytochrome b and control region sequences). In a pattern consistent with geographically defined subspecies, we found three deeply divergent mitochondrial lineages restricted to different geographic regions. The currently designated two subspecies of blue sheep, Pseudois nayaur nayaur and Pseudois nayaur szechuanensis, were recognized in the phylogenetic trees. In addition, the Helan Mountain population showed distinct genetic characteristics from other geographic populations, and thus should be classified as a new subspecies. In contrast, dwarf blue sheep clustered closely with some blue sheep from Sichuan Province in the phylogenetic trees. Therefore, dwarf blue sheep appear to be a subset of Pseudois nayaur szechuanensis. After considering both population genetic information and molecular clock analysis, we obtained some relevant molecular phylogeographic information concerning the historical biogeography of blue sheep. These results also indicate that western Sichuan was a potential refugium for blue sheep during the Quaternary period.

The discrimination of (non-denim) blue cotton.

Science.gov (United States)

Palmer, Ray; Hutchinson, William; Fryer, Verity

2009-03-01

This study was conducted to determine the degree of discrimination obtained between non-denim blue cotton fibres using visible-UV range microspectrophotometry alone. To this end, samples of fibres were taken from 100, nondenim, blue cotton, outer garments, including t-shirts, trousers and jumpers and subjected to analysis by both visible and UV range microspectrophotometry. The results obtained from the samples of each garment were compared to determine if they 'matched' or not. From an initial visual comparison of the garments it was possible to subdivide the samples into two populations consisting of 73 'dark blue' garments and 27 'mid-blue' garments. It was found that of the 73 'dark blue' garments, 22 distinct sub-populations could be distinguished using visible range MSP, this figure being increased to 43 when the analysis was extended into the UVW range. In the case of the 27 'mid-blue' garments, 9 distinct sub-populations were discriminated using visible range MSP, this figure being increased to 17 when the analysis was extended into the UV range. The discriminating power (i.e., the number of discriminated pairs divided by the number of possible pairs) of visible range microspectrophotometry was calculated as 0.89 for 'mid-blue' garments and 0.87 for 'dark blue' garments. Extending microspectrophotometry into the UV range increased discrimination by 7%, giving a discriminating power of 0.96 for both mid and dark blue cotton fibres which was similar to that reported by a previous study where this method was combined with light and fluorescence microscopy. Intra-garment variation was found to be negligible. The implications of this study for casework are discussed and a revised analytical pathway for the comparison of this fibre type/colour combination using microspectrophotometry as a primary screening tool, is proposed.

STUDY ON POLLEN VIABILITY AS BIOINDICATOR OF AIR QUALITY

Directory of Open Access Journals (Sweden)

Florentina ŞTEFLEA

2012-01-01

Full Text Available The aim of this research is to estimate the relationship between pollen viability and atmospheric pollution (in polluted and non-polluted conditions. The study was carried out in the city of Timisoara. Two areas, with different intensity of road traffic (very high and absent but all characterized by the presence of the same plant species, were selected. The pollen of herbaceous spontaneous species, arboreal species and a shrub species was used (Robinia pseudacacia, Aesculus x carnea, Catalpa bignonioides, Albizzia julibrissin, Rosa canina, Sambucus nigra, Malva neglecta, Ranunculus acer, Trifolium repens, Cichorium intybus. The pollen of these species was treated with TTC (2, 3, 5 Tryphenil-Tetrazolium-Chloride staining solution and viability was then estimated by light microscopy. The results of the mean pollen viability percentage of the examined species are reported. Pollen viability of herbaceous plants is significantly different between the two environments.

BLUES function method in computational physics

Science.gov (United States)

Indekeu, Joseph O.; Müller-Nedebock, Kristian K.

2018-04-01

We introduce a computational method in physics that goes ‘beyond linear use of equation superposition’ (BLUES). A BLUES function is defined as a solution of a nonlinear differential equation (DE) with a delta source that is at the same time a Green’s function for a related linear DE. For an arbitrary source, the BLUES function can be used to construct an exact solution to the nonlinear DE with a different, but related source. Alternatively, the BLUES function can be used to construct an approximate piecewise analytical solution to the nonlinear DE with an arbitrary source. For this alternative use the related linear DE need not be known. The method is illustrated in a few examples using analytical calculations and numerical computations. Areas for further applications are suggested.

Viability of dielectrophoretically trapped neuronal cortical cells in culture

NARCIS (Netherlands)

Heida, Tjitske; Vulto, P; Rutten, Wim; Marani, Enrico

2001-01-01

Negative dielectrophoretic trapping of neural cells is an efficient way to position neural cells on the electrode sites of planar micro-electrode arrays. The preservation of viability of the neural cells is essential for this approach. This study investigates the viability of postnatal cortical rat

Flow cytometric assessment of viability of lactic acid bacteria

NARCIS (Netherlands)

Bunthof, C.J.; Bloemen, K.; Breeuwer, P.; Rombouts, F.M.; Abee, T.

2001-01-01

The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA

Sperm viability staining in ecology and evolution: potential pitfalls

DEFF Research Database (Denmark)

Holman, Luke

2009-01-01

The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced a nu...

Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

Directory of Open Access Journals (Sweden)

AK Sarvel

2006-10-01

Full Text Available Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%. When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs; mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

Application of lift and squeeze technique in phacoemulsification of hypermature cataract

Directory of Open Access Journals (Sweden)

Geng-Ying Li

2014-07-01

Full Text Available AIM: To evaluate the effectiveness and security of lift and squeeze technique in phacoemulsification of hypermature cataract.METHODS: From June 2010 to June 2013, totally 156 eyes with hypermature cataract, which received phacoemulsification in our hospital, were enrolled. Lift and squeeze technique was used to chop the nucleus, and 1g/L Trypan blue was used for capsulorhexis. Average time of phaco complication, corneal edema and visual outcome were recorded.RESULTS: The best-corrected visual acuity(BCVA was 0.1-0.4 in 15 eyes(9.6%, 0.5-0.7 in 82(52.6%eyes, and 0.8-1.0 in 59(37.8%eyes at 3mo after surgery. The phaco time was 25-56s(average 42±10s, the maximum phaco power was 30%. Posterior capsular rupture and vitreous loss happened in 2 eyes(1.3%, and the IOLs were implanted in the sulcus. Corneal edema classified at grade Ⅰ were seen in 12 eyes(7.7%, and 5 eyes(3.2%at gradeⅡ, no eye at grade Ⅲ and grade Ⅳ. The mean endothelial cell loss was 8.7% at 3mo.CONCLUSION: Crystalline lens capsule staining with Trypan blue increase the success rate of intact continuous curvilinear capsulorhexis(CCC. The lift and squeeze technique reduces the stress on the zonules and capsule, and decreases the phaco time and phaco power.

New insights into the role of Mn and Fe in coloring origin of blue decorations of blue-and-white porcelains by XANES spectroscopy

International Nuclear Information System (INIS)

Zhu, Jian; Luo, Wugan; Ming, Chaofang; Wang, Changsui; Chen, Dongliang; Xu, Wei; Wang, Lihua

2013-01-01

Blue and white porcelain is one of the most valuable ancient ceramics varieties in ancient China. It is well known for its beautiful blue decorations. However, the origin of its blue color has not been very clear till now. In this research, two blue and white porcelains from Jingdezhen, Jiangxi province were selected and Mn and Fe K-edge XANES spectra were recorded from blue decorations with or without transparent glaze. Results showed that Mn K-edge XANES features were almost identical between different samples while that of iron changed. The above findings indicated the positive role of iron in the variation of blue decorations. As for manganese, although more system researches were need, its negative role on the variations of the tone of blue decorations was obtained. On the other hand, the paper also revealed the XAFS results will be affect by the glaze layer above the pigment. These findings provided us more information to understand the coloring origin of blue decorations of blue-and-white porcelain by means of XANES spectroscopy.

New insights into the role of Mn and Fe in coloring origin of blue decorations of blue-and-white porcelains by XANES spectroscopy

Science.gov (United States)

Zhu, Jian; Luo, Wugan; Chen, Dongliang; Xu, Wei; Ming, Chaofang; Wang, Changsui; Wang, Lihua

2013-04-01

Blue and white porcelain is one of the most valuable ancient ceramics varieties in ancient China. It is well known for its beautiful blue decorations. However, the origin of its blue color has not been very clear till now. In this research, two blue and white porcelains from Jingdezhen, Jiangxi province were selected and Mn and Fe K-edge XANES spectra were recorded from blue decorations with or without transparent glaze. Results showed that Mn K-edge XANES features were almost identical between different samples while that of iron changed. The above findings indicated the positive role of iron in the variation of blue decorations. As for manganese, although more system researches were need, its negative role on the variations of the tone of blue decorations was obtained. On the other hand, the paper also revealed the XAFS results will be affect by the glaze layer above the pigment. These findings provided us more information to understand the coloring origin of blue decorations of blue-and-white porcelain by means of XANES spectroscopy.

Pollen viability and germination in Jatropha ribifolia and Jatropha ...

African Journals Online (AJOL)

The aim of this work is to assess pollen viability using the staining technique and in vitro germination with different concentrations of sucrose in Jatropha ribifolia and Jatropha mollissima, contributing to the knowledge of the reproductive biology and subsidizing their conservation, management and utilization. Pollen viability ...

Nature's palette: the search for natural blue colorants.

Science.gov (United States)

Newsome, Andrew G; Culver, Catherine A; van Breemen, Richard B

2014-07-16

The food and beverage industry is seeking to broaden the palette of naturally derived colorants. Although considerable effort has been devoted to the search for new blue colorants in fruits and vegetables, less attention has been directed toward blue compounds from other sources such as bacteria and fungi. The current work reviews known organic blue compounds from natural plant, animal, fungal, and microbial sources. The scarcity of blue-colored metabolites in the natural world relative to metabolites of other colors is discussed, and structural trends common among natural blue compounds are identified. These compounds are grouped into seven structural classes and evaluated for their potential as new color additives.

Analytical Research to Determine the effects of the Components of ONGABO on the Viability of HepG2 Cancer Cells by Using the Sovereign, Minister, Assistant and Courier Principle ().

Science.gov (United States)

Shin, Jeong-Hun; Jun, Seung-Lyul; Hwang, Sung-Yeoun; Ahn, Seong-Hun

2012-12-01

This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle () to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea) at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen) reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Although Ginseng Radix (Red Ginseng) and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. In the sovereign, minister, assistant and courier principle (), Ginseng Radix (Red Ginseng) corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in cell viability in HepG2. We hope this study provides motivation for advanced research

Phototherapy with blue and green mixed-light is as effective against unconjugated jaundice as blue light and reduces oxidative stress in the Gunn rat model.

Science.gov (United States)

Uchida, Yumiko; Morimoto, Yukihiro; Uchiike, Takao; Kamamoto, Tomoyuki; Hayashi, Tamaki; Arai, Ikuyo; Nishikubo, Toshiya; Takahashi, Yukihiro

2015-07-01

Phototherapy using blue light-emitting diodes (LED) is effective against neonatal jaundice. However, green light phototherapy also reduces unconjugated jaundice. We aimed to determine whether mixed blue and green light can relieve jaundice with minimal oxidative stress as effectively as either blue or green light alone in a rat model. Gunn rats were exposed to phototherapy with blue (420-520 nm), filtered blue (FB; 440-520 nm without 1.00), respectively. Blue plus green phototherapy is as effective as blue phototherapy and it attenuates irradiation-induced oxidative stress. Combined blue and green spectra might be effective against neonatal hyperbilirubinemia. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

A Role for MEK-Interacting Protein 1 In Hormone Responsiveness of ER Positive Breast Cancer Cells

Science.gov (United States)

2011-10-01

48 hours, ER- positiv e cell lines tran sfected with MP1siRNA (but not control siR NA) rounded up and detached fr om the plate, and trypan blue...phenotype to MCF-7. To quantitate the effect of MP1 knockdown, attached and detached cells were collected at 48 h following siRNA transfection, stained...Immunoblot from a representative experiment. Lower panel: Quantitation of MP1/Actin ratios in three independent experiments (mean ± SD, *pɘ.05). Figure

Cytopathogenic effects in enterocytelike Caco-2 cells differentiate virulent from avirulent Listeria strains.

OpenAIRE

Pine, L; Kathariou, S; Quinn, F; George, V; Wenger, J D; Weaver, R E

1991-01-01

We have developed a simple test that differentiates between virulent and avirulent Listeria species as defined by the mouse 50% lethal doses (LD50S). The assay is based on trypan blue-revealed cytopathogenic effects that are produced during the infection of the human enterocytelike cell line Caco-2. These effects were elicited only by Listeria strains that had an intraperitoneal mouse LD50 less than 10(8) and were not produced by nonhemolytic, avirulent strains of Listeria monocytogenes gener...

MOCK OBSERVATIONS OF BLUE STRAGGLERS IN GLOBULAR CLUSTER MODELS

International Nuclear Information System (INIS)

Sills, Alison; Glebbeek, Evert; Chatterjee, Sourav; Rasio, Frederic A.

2013-01-01

We created artificial color-magnitude diagrams of Monte Carlo dynamical models of globular clusters and then used observational methods to determine the number of blue stragglers in those clusters. We compared these blue stragglers to various cluster properties, mimicking work that has been done for blue stragglers in Milky Way globular clusters to determine the dominant formation mechanism(s) of this unusual stellar population. We find that a mass-based prescription for selecting blue stragglers will select approximately twice as many blue stragglers than a selection criterion that was developed for observations of real clusters. However, the two numbers of blue stragglers are well-correlated, so either selection criterion can be used to characterize the blue straggler population of a cluster. We confirm previous results that the simplified prescription for the evolution of a collision or merger product in the BSE code overestimates their lifetimes. We show that our model blue stragglers follow similar trends with cluster properties (core mass, binary fraction, total mass, collision rate) as the true Milky Way blue stragglers as long as we restrict ourselves to model clusters with an initial binary fraction higher than 5%. We also show that, in contrast to earlier work, the number of blue stragglers in the cluster core does have a weak dependence on the collisional parameter Γ in both our models and in Milky Way globular clusters

Alcian blue-stained particles in a eutrophic lake

DEFF Research Database (Denmark)

Worm, J.; Søndergaard, Morten

1998-01-01

We used a neutral solution of Alcian Blue to stain transparent particles in eutrophic Lake Frederiksborg Slotss0, Denmark. Alcian Blue-stained particles (ABSP) appeared to be similar to the so-called transparent exopolymer particles (TEP) identified with an acidic solution of Alcian Blue. Our...

FIXED-BED COLUMN ADSORPTION OF METHYL BLUE USING ...

African Journals Online (AJOL)

userpc

Axle Wood Carbon (AWC) was used to study the removal of Methyl Blue (MB) from ... height, initial methyl blue (MB) concentration, .... colour from blue to dark purple- .... Environ. Earth Sci. 13; 1–13. Yagub, M. T., Sen, T. K., Afroze, S., and Ang,.

Curcumin inhibits growth potential by G1 cell cycle arrest and induces apoptosis in p53-mutated COLO 320DM human colon adenocarcinoma cells.

Science.gov (United States)

Dasiram, Jade Dhananjay; Ganesan, Ramamoorthi; Kannan, Janani; Kotteeswaran, Venkatesan; Sivalingam, Nageswaran

2017-02-01

Curcumin, a natural polyphenolic compound and it is isolated from the rhizome of Curcuma longa, have been reported to possess anticancer effect against stage I and II colon cancer. However, the effect of curcumin on colon cancer at Dukes' type C metastatic stage III remains still unclear. In the present study, we have investigated the anticancer effects of curcumin on p53 mutated COLO 320DM human colon adenocarcinoma cells derived from Dukes' type C metastatic stage. The cellular viability and proliferation were assessed by trypan blue exclusion assay and MTT assay, respectively. The cytotoxicity effect was examined by lactate dehydrogenase (LDH) cytotoxicity assay. Apoptosis was analyzed by DNA fragmentation analysis, Hoechst and propidium iodide double fluorescent staining and confocal microscopy analysis. Cell cycle distribution was performed by flow cytometry analysis. Here we have observed that curcumin treatment significantly inhibited the cellular viability and proliferation potential of p53 mutated COLO 320DM cells in a dose- and time-dependent manner. In addition, curcumin treatment showed no cytotoxic effects to the COLO 320DM cells. DNA fragmentation analysis, Hoechst and propidium iodide double fluorescent staining and confocal microscopy analysis revealed that curcumin treatment induced apoptosis in COLO 320DM cells. Furthermore, curcumin caused cell cycle arrest at the G1 phase, decreased the cell population in the S phase and induced apoptosis in COLO 320DM colon adenocarcinoma cells. Together, these data suggest that curcumin exerts anticancer effects and induces apoptosis in p53 mutated COLO 320DM human colon adenocarcinoma cells derived from Dukes' type C metastatic stage. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

HYPOLIPIDEMIC EFFECT OF ARGLABIN IN HEPATOMA TISSUE CULTURE

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A. V. Ratkin

2015-01-01

Full Text Available Objective. Investigation of hypolipidemic effect of sesquiterpene γ-lactone Arglabin in hepatoma tissue culture (HTC.Materials and methods. In this study we’ve evaluated the effect of sesquiterpene γ-lactone Arglabin and gemfibrozil (reference drug on the lipid content in the hepatoma tissue culture (HTC which were incubated with a fat emulsion “Lipofundin” by fluorescent method with vital dye Nile Red. The cell viability was investigated using the MTT-test and staining by Trypan blue.Results. Cultivation of cell cultures of rat’s hepatoma cell line HTC with Arglabin and gemfibrozil in concentrations from 10 to 50 μmol and from 0.25 to 0.5 mmol, respectively, had no cytotoxic effect. HTC cell viability did not change compared with the corresponding rate in the control culture. Experimental hyperlipidemia in hepatoma culture was induced by the addition in the incubation medium of fat emulsion “Lipofundin” in a final concentration of 0.05 %. The fluorescence intensity of Nile Red in the cells was increased 4-fold (p < 0.05, which indicates a significant accumulation of lipids in the cytosol of cells. In these steady-state Arglabin and gemfibrozil at concentrations 75–100 μM and 0.25–1.0 mM, respectively, reduced the content of lipid in cells. Conclusion. In the model of hyperlipidemia induced by lipofundin, sesquiterpene γ-lactone Arglabin prevents the accumulation of lipids in the HTC cell line, as evidenced by a decrease in Nile Red fluorescence. However hypolipidemic effect of Arglabin is associated with cytotoxic effects, which is typical for anticancer drugs.

Effects of RF-EMF Exposure from GSM Mobile Phones on Proliferation Rate of Human Adipose-derived Stem Cells: An In-vitro Study

Directory of Open Access Journals (Sweden)

Shahbazi-Gahrouei D

2016-12-01

Full Text Available Background: As the use of mobile phones is increasing, public concern about the harmful effects of radiation emitted by these devices is also growing. In addition, protection questions and biological effects are among growing concerns which have remained largely unanswered. Stem cells are useful models to assess the effects of radiofrequency electromagnetic fields (RF-EMF on other cell lines. Stem cells are undifferentiated biological cells that can differentiate into specialized cells. Adipose tissue represents an abundant and accessible source of adult stem cells. The aim of this study is to investigate the effects of GSM 900 MHz on growth and proliferation of mesenchymal stem cells derived from adipose tissue within the specific distance and intensity. Materials and Methods: ADSCs were exposed to GSM mobile phones 900 MHz with intensity of 354.6 µW/cm2 square waves (217 Hz pulse frequency, 50% duty cycle, during different exposure times ranging from 6 to 21 min/day for 5 days at 20 cm distance from the antenna. MTT assay was used to determine the growth and metabolism of cells and trypan blue test was also done for cell viability. Statistical analyses were carried out using analysis of one way ANOVA. P<0.05 was considered to be statistically significant. Results: The proliferation rates of human ADSCs in all exposure groups were significantly lower than control groups (P<0.05 except in the group of 6 minutes/day which did not show any significant difference with control groups. Conclusion: The results show that 900 MHz RF signal radiation from antenna can reduce cell viability and proliferation rates of human ADSCs regarding the duration of exposure.

L-Carnitine halts apoptosis and myelosuppression induced by carboplatin in rat bone marrow cell cultures (BMC).

Science.gov (United States)

Abd-Allah, Adel R A; Al-Majed, Abdulhakeem A; Al-Yahya, Abdulaziz A; Fouda, Soliman I; Al-Shabana, Othman A

2005-07-01

Carboplatin (CP), a second generation platinum compound, is effective against various types of cancers, producing less nephrotoxicity and ototoxicity but more myelotoxicity than cisplatinum. CP-myelosuppression is the rate-limiting step of its clinical use. Prevention of CP-myelosuppression is a major target in the field of chemotherapy. Therefore, the present study investigates the use of L-carnitine (LCR)-an antioxidant, cardioprotective, neuroprotective, and immunostimulant nontoxic natural compound-to protect against CP-induced myelosuppression. The viability of BMC was studied using a trypan blue exclusion technique following incubation with CP and/or LCR as a function of time and concentration. Apoptosis was tested for by detecting the amount of DNA fragmentation and the visualization of DNA ladders upon gel electrophoresis. Bone marrow progenitor cell function was examined by colony forming unit assay. Cellular contents of glutathione (GSH) and malondialdehyde (MDA) were also estimated. Results revealed that LC50 of CP is 4.7 mM and the highest safe concentration of LCR is 5 mM. Co-exposure of LCR+CP rescued BMC viability by 37% compared to the CP-treated cultures. The LCR halts CP-induced apoptosis and it significantly improves the function of the bone marrow progenitors by increasing the number of colony-forming units as a response to granulocyte/macrophage colony stimulating factors. Finally, LCR restores CP-induced GSH depletion and prevents MDA elevation in BMC. In summary, the results suggest that LCR is able to protect against CP-induced myelosuppression, which suggests its use as an adjuvant therapy. This finding merits further investigation into the mechanism(s) of such protection as well as its interaction with CP antitumor activity.

Manganese oxidation state mediates toxicity in PC12 cells

International Nuclear Information System (INIS)

Reaney, S.H.; Smith, D.R.

2005-01-01

The role of the manganese (Mn) oxidation state on cellular Mn uptake and toxicity is not well understood. Therefore, undifferentiated PC12 cells were exposed to 0-200 μM Mn(II)-chloride or Mn(III)-pyrophosphate for 24 h, after which cellular manganese levels were measured along with measures of cell viability, function, and cytotoxicity (trypan blue exclusion, medium lactate dehydrogenase (LDH), 8-isoprostanes, cellular ATP, dopamine, serotonin, H-ferritin, transferrin receptor (TfR), Mn-superoxide dismutase (MnSOD), and copper-zinc superoxide dismutase (CuZnSOD) protein levels). Exposures to Mn(III) >10 μM produced 2- to 5-fold higher cellular manganese levels than equimolar exposures to Mn(II). Cell viability and ATP levels both decreased at the highest Mn(II) and Mn(III) exposures (150-200 μM), while Mn(III) exposures produced increases in LDH activity at lower exposures (≥50 μM) than did Mn(II) (200 μM only). Mn(II) reduced cellular dopamine levels more than Mn(III), especially at the highest exposures (50% reduced at 200 μM Mn(II)). In contrast, Mn(III) produced a >70% reduction in cellular serotonin at all exposures compared to Mn(II). Different cellular responses to Mn(II) exposures compared to Mn(III) were also observed for H-ferritin, TfR, and MnSOD protein levels. Notably, these differential effects of Mn(II) versus Mn(III) exposures on cellular toxicity could not simply be accounted for by the different cellular levels of manganese. These results suggest that the oxidation state of manganese exposures plays an important role in mediating manganese cytotoxicity

Ascorbic acid attenuates endothelial permeability triggered by cell-free hemoglobin.

Science.gov (United States)

Kuck, Jamie L; Bastarache, Julie A; Shaver, Ciara M; Fessel, Joshua P; Dikalov, Sergey I; May, James M; Ware, Lorraine B

2018-01-01

Increased endothelial permeability is central to shock and organ dysfunction in sepsis but therapeutics targeted to known mediators of increased endothelial permeability have been unsuccessful in patient studies. We previously reported that cell-free hemoglobin (CFH) is elevated in the majority of patients with sepsis and is associated with organ dysfunction, poor clinical outcomes and elevated markers of oxidant injury. Others have shown that Vitamin C (ascorbate) may have endothelial protective effects in sepsis. In this study, we tested the hypothesis that high levels of CFH, as seen in the circulation of patients with sepsis, disrupt endothelial barrier integrity. Human umbilical vein endothelial cells (HUVEC) were grown to confluence and treated with CFH with or without ascorbate. Monolayer permeability was measured by Electric Cell-substrate Impedance Sensing (ECIS) or transfer of 14 C-inulin. Viability was measured by trypan blue exclusion. Intracellular ascorbate was measured by HPLC. CFH increased permeability in a dose- and time-dependent manner with 1 mg/ml of CFH increasing inulin transfer by 50% without affecting cell viability. CFH (1 mg/ml) also caused a dramatic reduction in intracellular ascorbate in the same time frame (1.4 mM without CFH, 0.23 mM 18 h after 1 mg/ml CFH, p < 0.05). Pre-treatment of HUVECs with ascorbate attenuated CFH induced permeability. CFH increases endothelial permeability in part through depletion of intracellular ascorbate. Supplementation of ascorbate can attenuate increases in permeability mediated by CFH suggesting a possible therapeutic approach in sepsis. Copyright © 2017 Elsevier Inc. All rights reserved.

Chinchilla laniger can be used as an experimental model for Taenia solium taeniasis.

Science.gov (United States)

Maravilla, Pablo; Garza-Rodriguez, Adriana; Gomez-Diaz, Benjamin; Jimenez-Gonzalez, Diego Emiliano; Toral-Bastida, Elizabeth; Martinez-Ocaña, Joel; West, Brett; Molina, Nadia; Garcia-Cortes, Ramon; Kawa-Karasik, Simon; Romero-Valdovinos, Mirza; Avila-Ramirez, Guillermina; Flisser, Ana

2011-12-01

Chinchilla laniger has been reported as an experimental definitive host for Taenia solium; however no information about its suitability and yield of gravid tapeworm proglottids containing viable and infective eggs has been published. In total 55 outbred female chinchillas were infected with 4 cysticerci each; hosts were immunodeppressed with 6 or 8 mg of methyl-prednisolone acetate every 14 days starting the day of infection and their discomfort was followed. Kinetics of coproantigen ELISA or expelled proglottids was used to define the infection status. Efficiency of tapeworm establishment was 21% and of parasite gravidity was 8%; chinchillas showed some degree of suffering along the infection. Viability of eggs obtained from gravid proglottids was tested comparing methods previously published, our results showed 62% viability with propidium iodide, 54% with trypan blue, 34% with neutral red, 30% by oncosphere activation and 7% with bromide 3-(4,5-dimetil-tiazol-2-il)-2,5-difenil-tetrazolio (MTT) reduction; no statistical differences were obtained between most techniques, except activation. Four piglets were infected with 50,000 eggs each, necropsy was performed 3 months later and, after counting the number of cysticerci recovered, the percentage of infection was similar to data obtained with T. solium eggs recovered from humans. Our results demonstrate that the experimental model of T. solium taeniasis in C. laniger is a good alternative for providing eggs and adult tapeworms to be used in different types of experiments; optimization of the model probably depends on the use of inbred hosts and on the reduction of infected animals' suffering. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Pollen diversity, viability and floral structure of some Musa genotypes

African Journals Online (AJOL)

Pollen diversity, viability and floral structure of some Musa genotypes. ... This experiment was designed to study the floral structure, pollen morphology and the potential pollen viability of five Musa genotypes obtained ... HOW TO USE AJOL.

Blue light-induced oxidative stress in live skin.

Science.gov (United States)

Nakashima, Yuya; Ohta, Shigeo; Wolf, Alexander M

2017-07-01

Skin damage from exposure to sunlight induces aging-like changes in appearance and is attributed to the ultraviolet (UV) component of light. Photosensitized production of reactive oxygen species (ROS) by UVA light is widely accepted to contribute to skin damage and carcinogenesis, but visible light is thought not to do so. Using mice expressing redox-sensitive GFP to detect ROS, blue light could produce oxidative stress in live skin. Blue light induced oxidative stress preferentially in mitochondria, but green, red, far red or infrared light did not. Blue light-induced oxidative stress was also detected in cultured human keratinocytes, but the per photon efficacy was only 25% of UVA in human keratinocyte mitochondria, compared to 68% of UVA in mouse skin. Skin autofluorescence was reduced by blue light, suggesting flavins are the photosensitizer. Exposing human skin to the blue light contained in sunlight depressed flavin autofluorescence, demonstrating that the visible component of sunlight has a physiologically significant effect on human skin. The ROS produced by blue light is probably superoxide, but not singlet oxygen. These results suggest that blue light contributes to skin aging similar to UVA. Copyright © 2017 Elsevier Inc. All rights reserved.

Compact and efficient blue laser sheet for measurement

Science.gov (United States)

Qi, Yan; Wang, Yu; Wu, Bin; Wang, Yanwei; Yan, Boxia

2017-10-01

Compact and efficient blue laser sheet has important applications in the field of measurement, with laser diode end pumped Nd:YAG directly and LBO intracavity frequency doubling, a compact and efficient CW 473nm blue laser sheet composed of dual path liner blue laser is realized. At an incident pump power of 12.4W, up to 1.4W output power of the compound blue laser is achieved, the optical-to-optical conversion efficiency is as high as 11.3%.

A Blue Lagoon Function

DEFF Research Database (Denmark)

Markvorsen, Steen

2007-01-01

We consider a specific function of two variables whose graph surface resembles a blue lagoon. The function has a saddle point $p$, but when the function is restricted to any given straight line through $p$ it has a {\\em{strict local minimum}} along that line at $p$.......We consider a specific function of two variables whose graph surface resembles a blue lagoon. The function has a saddle point $p$, but when the function is restricted to any given straight line through $p$ it has a {\\em{strict local minimum}} along that line at $p$....

Survey of the Definition of Fetal Viability and the Availability, Indications, and Decision Making Processes for Post-Viability Termination of Pregnancy for Fetal Abnormalities and Health Conditions in Canada.

Science.gov (United States)

Hull, Danna; Davies, Gregory; Armour, Christine M

2016-06-01

The purpose of this study was to explore the definition of fetal viability and the availability, indications, and decision making processes for post-viability termination of pregnancy for fetal abnormalities and health conditions in Canada. An online survey of members of the Canadian Association of Genetic Counsellors, the Canadian College of Medical Geneticists, and the Canadian Society for Maternal-Fetal Medicine who provide direct counselling to, or management of, prenatal patients in Canada (total sample size 815). Results of this study showed that the majority of respondents indicated that their centre will offer post-viability termination of pregnancy (98/123; 80 %). Sixty-seven percent (68/101) of respondents reported the definition of fetal viability to be 24 weeks' gestation. Most respondents reported that a collaborative decision making process was used to determine if post-viability termination of pregnancy would be offered (136/170; 80 %). For conditions presumed to be lethal/likely lethal, the majority of respondents would "sometimes" or "always" offer post-viability termination of pregnancy, whereas for conditions presumed to have a mild effect, the majority of respondents would "rarely" or "never" offer post-viability termination of pregnancy. Ninety percent (77/86) of respondents reported that perinatal hospice is offered as an alternative to termination of pregnancy. In conclusion, this study suggests that although post-viability termination is available in many provinces in Canada, variation in the definition of fetal viability and indications appear to exist. While these variations may lead to unequal access to post-viability termination of pregnancy across Canada, they might also represent the complexity of the decision making process and the importance of examining individual factors to ensure that the most appropriate decision is made in each case.

The Structure of the Blue Whirl

Science.gov (United States)

Hariharan, Sriram Bharath; Hu, Yu; Xiao, Huahua; Gollner, Michael; Oran, Elaine

2017-11-01

Recent experiments have led to the discovery of the blue whirl, a small, stable regime of the fire whirl that burns typically sooty liquid hydrocarbons without producing soot. The physical structure consists of three regions - the blue cone, the vortex rim and the purple haze. The physical nature of the flame was further investigated through digital imaging techniques, which suggest that the transition (from the fire whirl to the blue whirl) and shape of the flame may be influenced by vortex breakdown. The flame was found to develop over a variety of surfaces, which indicates that the formation of the blue whirl is strongly influenced by the flow structure over the incoming boundary layer. The thermal structure was investigated using micro-thermocouples, thin-filament pyrometry and OH* spectroscopy. These revealed a peak temperature around 2000 K, and that most of the combustion occurs in the relatively small, visibly bright vortex rim. The results of these investigations provide a platform to develop a theory on the structure of the blue whirl, a deeper understanding of which may affirm potential for applications in the energy industry. This work was supported by an NSF EAGER award and Minta Martin Endowment Funds in the Department of Aerospace Engineering at the University of Maryland.

Blue whales Balaenoptera musculus off Angola: recent sightings ...

African Journals Online (AJOL)

Further survey work is required to better clarify the status of blue whales in Angolan waters, particularly with regard to population structure and potential calving grounds. Keywords: Antarctic blue whale, calving, catch data, pygmy blue whale, South-East Atlantic, stomach contents. African Journal of Marine Science 2014, ...

Antimicrobial blue light inactivation of Methicillin-resistant Staphylococcus aureus

Science.gov (United States)

Wang, Yucheng; Dai, Tianhong; Gu, Ying

2016-10-01

Background: With the increasing emergence of multidrug-resistant (MDR) bacterial strains, there is a pressing need for the development of alternative treatment for infections. Antimicrobial blue light (aBL) has provided a simple and effective approach. Methods: We first investigated the effectiveness of aBL (415 nm) inactivation of USA300 LAClux (a communityacquired Methicillin-resistant Staphylococcus aureus strain) both in the planktonic and biofilm forms. The survival of the bacteria in suspensions was determined by serial dilution and that of the biofilm-embedded bacteria was determined by bioluminescence quantification. Using a mouse model of thermal burn infected with USA300 LAClux, we further assessed the effectiveness of aBL for treating localized infections. Bioluminescence imaging was performed to monitor in real time bacterial viability in vivo. Results: In vitro study showed that, for the planktonic counterpart of the bacteria or the 24-h-old biofilms, an irradiance of 55 mW/cm2 for 60 min resulted in a 4.61 log10 or 2.56 log10 inactivation, respectively. In vivo study using infected mouse burns demonstrated that a 2.56-log10 inactivation was achieved after 100-mW/cm2 irradiation for 62 min. Conclusions: aBL is a potential alternative approach for treating Methicillin-resistant Staphylococcus aureus infections.

Immune modulation in the blue mussel Mytilus edulis exposed to North Sea produced water

Energy Technology Data Exchange (ETDEWEB)

Hannam, M.L., E-mail: marie.hannam@plymouth.ac.u [Ecotoxicology and Stress Biology Research Centre, School of Biological Sciences, University of Plymouth, Drake Circus, Plymouth, Devon, PL4 8AA (United Kingdom); Bamber, S.D.; Sundt, R.C. [IRIS - Biomiljo, Mekjarvik 12, 4070 Randaberg (Norway); Galloway, T.S. [School of Biosciences, University of Exeter, Hatherly Laboratories, Prince of Wales Road, Exeter, EX4 4PS (United Kingdom)

2009-06-15

The discharge of oil well produced water (PW) provides a constant source of contaminants to the marine environment including polycyclic aromatic hydrocarbons, alkylated phenols, metals and production chemicals. High concentrations of PW cause adverse effects to exposed biota, including reduced survival, growth and reproduction. Here we explore the effects of PW on immune function in the blue mussel, Mytilus edulis. Mussels were exposed for 21 days to sublethal PW concentrations (0.125-0.5%) and cellular parameters were measured. Cell viability, phagocytosis and cytotoxicity were inhibited after exposure to 0.25% and 0.5% PW, whilst the 0.125% PW treatment produced significant increases in these biomarker responses. This biphasic response was only observed after 7 days exposure; longer exposure periods led to a reduction in immune parameters. Results indicate that PW concentrations close to the discharge point cause modulation to cellular immunity. The implications for longer-term disease resistance are discussed. - Exposure to produced water alters immune function in the sentinel species Mytilus edulis.

Immune modulation in the blue mussel Mytilus edulis exposed to North Sea produced water

International Nuclear Information System (INIS)

Hannam, M.L.; Bamber, S.D.; Sundt, R.C.; Galloway, T.S.

2009-01-01

The discharge of oil well produced water (PW) provides a constant source of contaminants to the marine environment including polycyclic aromatic hydrocarbons, alkylated phenols, metals and production chemicals. High concentrations of PW cause adverse effects to exposed biota, including reduced survival, growth and reproduction. Here we explore the effects of PW on immune function in the blue mussel, Mytilus edulis. Mussels were exposed for 21 days to sublethal PW concentrations (0.125-0.5%) and cellular parameters were measured. Cell viability, phagocytosis and cytotoxicity were inhibited after exposure to 0.25% and 0.5% PW, whilst the 0.125% PW treatment produced significant increases in these biomarker responses. This biphasic response was only observed after 7 days exposure; longer exposure periods led to a reduction in immune parameters. Results indicate that PW concentrations close to the discharge point cause modulation to cellular immunity. The implications for longer-term disease resistance are discussed. - Exposure to produced water alters immune function in the sentinel species Mytilus edulis.

An updated methodology to review developing-country vaccine manufacturer viability.

Science.gov (United States)

Luter, Nicholas; Kumar, Ritu; Hozumi, Dai; Lorenson, Tina; Larsen, Shannon; Gowda, Bhavya; Batson, Amie

2017-07-05

In 1997, Milstien, Batson, and Meaney published "A Systematic Method for Evaluating the Potential Viability of Local Vaccine Producers." The paper identified characteristics of successful vaccine manufacturers and developed a viability framework to evaluate their performance. This paper revisits the original study after two decades to determine the ability of the framework to predict manufacturer success. By reconstructing much of the original dataset and conducting in-depth interviews, the authors developed informed views on the continued viability of manufacturers in low- and middle-income country markets. Considering the marked changes in the market and technology landscape since 1997, the authors find the viability framework to be predictive and a useful lens through which to evaluate manufacturer success or failure. Of particular interest is how incumbent and potentially new developing-country vaccine manufacturers enter and sustain production in competitive international markets and how they integrate (or fail to integrate) new technology into the production process. Ultimately, most manufacturers will need to meet global quality standards to be viable. As governments and donors consider investments in vaccine producers, the updated viability factors will be a useful tool in evaluating the prospects of manufacturers over the mid to long term. The paper emphasizes that while up-front investments are important, other critical factors-including investments in a national regulatory authority, manufacturer independence, and ability to adapt and adopt new technology-are necessary to ensure viability. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Geothermal Technologies Program Blue Ribbon Panel Recommendations

Energy Technology Data Exchange (ETDEWEB)

none,

2011-06-17

The Geothermal Technologies Program assembled a geothermal Blue Ribbon Panel on March 22-23, 2011 in Albuquerque, New Mexico for a guided discussion on the future of geothermal energy in the United States and the role of the DOE Program. The Geothermal Blue Ribbon Panel Report captures the discussions and recommendations of the experts. An addendum is available here: http://www.eere.energy.gov/geothermal/pdfs/gtp_blue_ribbon_panel_report_addendum10-2011.pdf

PBMC and MDAMB-231 cellular viability after telecobalt irradiation

International Nuclear Information System (INIS)

Andrade, Lidia M.; Campos, Tarcisio P.R.

2002-01-01

Radiotherapy by gamma rays are used for cancer treatment. Ionizing radiation effects on cells has been investigated. Studies in vitro were developed using tumor cell lines and ionizing radiation. The aim of this research was to test the cellular viability response of two cell types through MTT assay: human peripheral blood mononuclear cell (PBMC) and human breast carcinoma cell line MDAMB-231. These cells were irradiated with 60 Co source Theratron 80 radiotherapy machine from Atomic Energy Canada Limited with 20 x 20 cm field at 136.4 cGy/min, surface source distance 70 cm. Culture flasks contained 10 4 , 10 5 and 10 6 cells were irradiated with 100 Gy, 25 Gy, and 50 Gy using non fractionated doses. Cellular viability were evaluated 1h, 24h, 48h and 72h after irradiation and samples were measured by optical density at 595nm. Our results show that PBMC cells present lower cellular viability post irradiation. On the other hand, MDAMB-231 cells maintain viability after 50 Gy irradiation at 72h indicating cellular radioresistance. (author)

Viability of seeds of two representatives from Apocynaceae family

Directory of Open Access Journals (Sweden)

Y.A. Aviekin

2015-05-01

Full Text Available The viability of some Apocynaceae seeds depending on the duration of storage under conditions of low temperature was studied. Extracted embryos from the seeds of Pachypodium lamerei Drake and Adenium obesum (Forssk. Roem. et Schult with different storage history were analyzed. Embryos were stained by acidic fuchsine what allows identification of viable and destructed cells. Destructed cells were stained much more intensively, while viable cells remained unstained. Observed results showed dependence of viability of P. lamerei and A. obesum seeds from term of storage. It was obtained that the seeds of investigated succulent species, just like in many other tropical plants, rapidly lost their viability and should be described as microbiotic.

Spectra and ages of blue stragglers

International Nuclear Information System (INIS)

Abt, H.A.

1985-01-01

A mechanism similar to Wheeler's quasi-homogeneous evolution and Finzi and Wolf's proposal for blue stragglers is proposed as the origin of the blue stragglers in intermediate-age clusters. Blue stragglers are stars whose positions in color-magnitude diagrams of open and globular clusters are significantly above the turn-off points and in the region of the (former) main sequence; they seem to represent a conflict with the general conclusion that all stars in a cluster originated at about the same time. It is concluded that there are at least two kinds of blue stragglers: (1) those stars of types about B3-A2 are primarily Ap stars and slow rotators, occur in the intermediate age clusters and remain in the main sequence region probably through magnetic mixing; and (2) the stars of type O6-B2 frequently have emission lines, are rapid rotators, occur in the young cluster, and remain in the main sequence region probably by rotational mixing. 30 references

Characteristics of Alcian-blue Dye Adsorption of Natural Biofilm Matrix

Science.gov (United States)

Kurniawan, A.; Yamamoto, T.; Sukandar; Guntur

2018-01-01

In this study, natural biofilm matrices formed on stones have been used for the adsorption of Alcian blue dye. Alcian blue is a member of polyvalent basic dyes that largely used from laboratory until industrial dying purposes. The adsorption of the dye onto the biofilm matrix has been carried out at different experimental conditions such as adsorption isotherm and kinetic of adsorption. The electric charge properties of biofilm matrix and its changes related to the adsorption of Alcian blue have been also investigated. Moreover, the results of Alcian blue adsorption to the biofilm were compared to those onto the acidic and neutral resin. The kinetics of adsorption result showed that the adsorption of the Alcian blue dye reached to a maximum adsorption amount within 60 minutes. The adsorption amount of Alcian blue to biofilm increased monotonously, and the maximum adsorption amount was greater compared to the resins. On the contrary, Alcian blue did not attach to the neutral resin having no electric charge. It seems that Alcian blue attached to the acidic resins due to electrostatic attractive force, and the same seems to be the case for adsorption of Alcian blue to biofilm. The adsorption of Alcian blue to the biofilm and acidic resins fitted to Langmuir type indicates that the binding of Alcian blue to the biofilm and acidic resins occurred in a monolayer like form. The maximum adsorption amount of Alcian blue on the biofilm (0.24 mmol/dry-g) was greater than those of acidic resin (0.025 mmol/dry-g). This indicates that the biofilm has many more sites for Alcian blue attachment than acidic resins. According to the result of this study, the biofilm matrix can be a good adsorbent for dye such as Alcian blue or other dyes that causing hazards in nature.

Effects of blue light and caffeine on mood.

Science.gov (United States)

Ekström, Johan G; Beaven, C Martyn

2014-09-01

Both short wavelength (blue) light and caffeine have been studied for their mood enhancing effects on humans. The ability of blue light to increase alertness, mood and cognitive function via non-image forming neuropathways has been suggested as a non-pharmacological countermeasure for depression across a range of occupational settings. This experimental study compared blue light and caffeine and aimed to test the effects of blue light/placebo (BLU), white light/240-mg caffeine (CAF), blue light/240-mg caffeine (BCAF) and white light/placebo (PLA), on mood. A randomised, controlled, crossover design study was used, in a convenience population of 20 healthy volunteers. The participants rated their mood on the Swedish Core Affect Scales (SCAS) prior to and after each experimental condition to assess the dimensions of valence and activation. There was a significant main effect of light (p = 0.009), and the combination of blue light and caffeine had clear positive effects on core effects (ES, ranging from 0.41 to 1.20) and global mood (ES, 0.61 ± 0.53). The benefits of the combination of blue light and caffeine should be further investigated across a range of applications due to the observed effects on the dimensions of arousal, valence and pleasant activation.

Occupational contact dermatitis in blue-collar workers

DEFF Research Database (Denmark)

Schwensen, Jakob F; Menné, Torkil; Veien, Niels K

2014-01-01

BACKGROUND: Blue-collar workers have a high risk of occupational contact dermatitis, but epidemiological studies are scarce. OBJECTIVES: To investigate allergic contact dermatitis in blue-collar workers with dermatitis registered by the Danish Contact Dermatitis Group. METHODS: A retrospective...... analysis of patch test data from 1471 blue-collar workers and 1471 matched controls tested between 2003 and 2012 was performed. A logistic regression was used to test for associations. RESULTS: The blue-collar workers often had occupational hand dermatitis (p dermatitis was less commonly......, and methylchloroisothiazolinone (MCI)/methylisothiazolinone (MI). The following occupations were additionally identified as risk factors for contact sensitization to MCI/MI and MI, epoxy resins, and potassium dichromate, respectively: painting, construction work, and tile setting/terrazzo work. CONCLUSION: Contact allergy...

Degradation of phosphorescent blue organic light-emitting diodes

Energy Technology Data Exchange (ETDEWEB)

Chiu, Chien-Shu [Department of Electrical Engineering and Information Technology, Technical University of Braunschweig (Germany); Siemens AG, CT MM 1, Erlangen (Germany); Steinbacher, Frank [Department of Materials Science VI, University of Erlangen-Nuernberg (Germany); Siemens AG, CT MM 1, Erlangen (Germany); Krause, Ralf; Hunze, Arvid [Siemens AG, CT MM 1, Erlangen (Germany); Kowalsky, Wolfgang [Department of Electrical Engineering and Information Technology, Technical University of Braunschweig (Germany)

2009-07-01

Development of phosphorescent materials has significantly improved the efficiency of organic light-emitting diodes (OLEDs). By using efficient red, green and blue phosphorescent emitter materials high efficient white OLEDs can be achieved. However, due to low stability of blue phosphorescent materials the lifetime of phosphorescent white OLEDs remains an issue. As a result, degradation of blue phosphorescent materials needs to be further investigated and improved. In this work, blue OLED devices based on the phosphorescent emitter FIrpic were investigated. Single-carrier hole-only as well as electron-only devices were fabricated. For investigation of degradation process the devices were stressed with electrical current and UV-light to study the impact of charge carriers as well as excitons and exciton-polaron quenching on the stability of the blue dye.

Viability Test Device for anisakid nematodes

Directory of Open Access Journals (Sweden)

Michael Kroeger

2018-03-01

Full Text Available Up to now the visual inspection of mobility of isolated anisakid larvae serves as a measure of viability and possible risk of infection. This paper presents a new method to rule out unreliability – caused by the temporary immobility of the larvae and by the human uncertainty factor of visual observation. By means of a Near infrared (NIR imaging method, elastic curvature energies and geometric shape parameters were determined from contours, and used as a measure of viability. It was based on the modelling of larvae as a cylindrical membrane system. The interaction between curvatures, contraction of the longitudinal muscles, and inner pressure enabled the derivation of viability from stationary form data. From series of spectrally signed images within a narrow wavelength range, curvature data of the larvae were determined. Possible mobility of larvae was taken into account in statistical error variables. Experiments on individual living larvae, long-term observations of Anisakis larvae, and comparative studies of the staining method and the VTD measurements of larvae from the tissue of products confirmed the effectiveness of this method. The VTD differentiated clearly between live and dead nematode larvae isolated from marinated, deep-frozen and salted products. The VTD has been proven as excellent method to detect living anisakid nematode larvae in fishery products and is seen as useful tool for fish processing industry and control authorities. Keywords: Biophysics

Crystalline liquids: the blue phases

Science.gov (United States)

Wright, David C.; Mermin, N. David

1989-04-01

The blue phases of cholesteric liquid crystals are liquids that exhibit orientational order characterized by crystallographic space-group symmetries. We present here a pedagogical introduction to the current understanding of the equilibrium structure of these phases accompanied by a general overview of major experimental results. Using the Ginzburg-Landau free energy appropriate to the system, we first discuss in detail the character and stability of the usual helical phase of cholesterics, showing that for certain parameter ranges the helical phase is unstable to the appearance of one or more blue phases. The two principal models for the blue phases are two limiting cases of the Ginzburg-Landau theory. We explore each limit and conclude with some general considerations of defects in both models and an exact minimization of the free energy in a curved three-dimensional space.

Intraoral blue (Jadassohn-Tieche) nevus.

Science.gov (United States)

Hasse, C D; Zoutendam, G L; Gombas, O F

1978-05-01

Blue nevus of the oral mucosa is a distinctly uncommon clincial entity. Careful review of the literature yielded thirty-one previously reported cases. The present article reports the occurrence of a blue nevus of the hard palate in a 58-year-old man. It is of interest since it is the smallest (1 by 1 mm.) intraoral blue nevus to be reported. A clinicopathologic study of the previous thirty-one cases and of our case suggests that this lesion has no age or sex predilection. The most common site of occurrence was the hard palate. There appears to be no tendency toward recurrence. A brief review of the historical background, clinical features, theories of possible origin, and differential diagnosis is presented. Excisional biopsy of localized areas of oral pibmentation, together with histopathologic study, is indicated to rule out melanoma.

New York Blue

Data.gov (United States)

Federal Laboratory Consortium — New York Blue is used cooperatively by the Laboratory and Stony Brook University as part of the New York Center for Computation Sciences. Ranked as the 28th fastest...

The secret of the blue fog

Science.gov (United States)

Henrich, Oliver; Marenduzzo, Davide

2017-04-01

Why certain liquids turn blue when cooled was a mystery that stumped scientists for more than a century. As Oliver Henrich and Davide Marenduzzo explain, solving the secret of the “blue fog” proved to be an intellectual tour de force - and one that could lead to new types of display devices

Local anti-fertility effect of inhibin-enriched preparation (IEP) in female hamsters.

Science.gov (United States)

Bapat, B V; Nandedkar, T D; Sheth, A R

1984-04-01

An inhibin-enriched preparation (IEP) involved in the regulation of follicle stimulating hormone (FSH) is known to play an important role in the normal ovarian cycle. In utero administration of 10 micrograms of IEP on day 3 of pregnancy completely prevented implantation in hamsters. No toxic effect of IEP was observed on the blastocysts as indicated by the dye exclusion test performed with Trypan blue. Thus, the results of the present study indicate an extra-pituitary site of action for the anti-implantation effect of IEP.

Tapered photonic crystal fibers for blue-enhanced supercontinuum generation

DEFF Research Database (Denmark)

Møller, Uffe; Sørensen, Simon Toft; Larsen, Casper

2012-01-01

Tapering of photonic crystal fibers is an effective way of shifting the blue edge of a supercontinuum spectrum down in the deep-blue. We discuss the optimum taper profile for enhancing the power in the blue edge....

A Rare Case of Multifocal Prostatic Blue Nevus

Directory of Open Access Journals (Sweden)

Elias J. Farran

2018-01-01

Full Text Available Prostatic blue nevus is a rare benign pathologic diagnosis most commonly diagnosed incidentally on many different types of prostate specimens. Blue nevus is the deposition of stromal melanin characterized by spindle cells within the fibromuscular stroma which stains positive for melanin-specific stains Fontana-Masson and S100 and stains negative for CD68, HMB45, and iron stains. We report the case of a multifocal and bilateral blue nevus in a 52-year-old Hispanic male who presented with an elevated prostate-specific antigen of 4.3 and mild obstructive lower urinary tract symptoms, found by transrectal ultrasound-guided prostate needle biopsy. The biopsy also revealed benign prostatic tissue with postatrophic hyperplasia and chronic inflammation. This is the 35th reported case of prostatic blue nevus and the third to show multifocal blue nevus.

Detecting viability transitions of umbilical cord mesenchymal stem cells by Raman micro-spectroscopy

International Nuclear Information System (INIS)

Bai, H; Chen, P; Fang, H; Lin, L; Tang, G Q; Mu, G G; Gong, W; Liu, Z P; Wu, H; Zhao, H; Han, Z C

2011-01-01

Recent research suggests that human umbilical cord derived mesenchymal stem cells (hUC-MSCs) can be promising candidates for cell-based therapy. Since large population and high viability are generally required, detecting viability transitions of these cells is crucial for their population expansion and quality control. Here, as a non-invasive method, Raman micro-spectroscopy is applied to examine hUC-MSCs with different viability. Using peak fitting and statistic t-test, the Raman peaks with obvious differences between the cells with high viability (> 90%) and low viability ( -1 , symmetric stretching of C–C in lipids at 877 cm -1 and CH deformation in proteins at 1342 cm -1 show the most significant changes (p < 0.001). When the cell viability decreases, the intensities of the former two peaks are both about doubled while that of the latter peak reduces by about 30%. Based on these results, we propose that the viability of hUC-MSCs can be characterized by these three peaks. And their intensity changes can be understood from the model of excessive reactive oxygen species interacting with the bio-macromolecules

Blue Light Protects Against Temporal Frequency Sensitive Refractive Changes.

Science.gov (United States)

Rucker, Frances; Britton, Stephanie; Spatcher, Molly; Hanowsky, Stephan

2015-09-01

Time spent outdoors is protective against myopia. The outdoors allows exposure to short-wavelength (blue light) rich sunlight, while indoor illuminants can be deficient at short-wavelengths. In the current experiment, we investigate the role of blue light, and temporal sensitivity, in the emmetropization response. Five-day-old chicks were exposed to sinusoidal luminance modulation of white light (with blue; N = 82) or yellow light (without blue; N = 83) at 80% contrast, at one of six temporal frequencies: 0, 0.2, 1, 2, 5, 10 Hz daily for 3 days. Mean illumination was 680 lux. Changes in ocular components and corneal curvature were measured. Refraction, eye length, and choroidal changes were dependent on the presence of blue light (P light, refraction did not change across frequencies (mean change -0.24 [diopters] D), while in the absence of blue light, we observed a hyperopic shift (>1 D) at high frequencies, and a myopic shift (>-0.6 D) at low frequencies. With blue light there was little difference in eye growth across frequencies (77 μm), while in the absence of blue light, eyes grew more at low temporal frequencies and less at high temporal frequencies (10 vs. 0.2 Hz: 145 μm; P light. Illuminants rich in blue light can protect against myopic eye growth when the eye is exposed to slow changes in luminance contrast as might occur with near work.

Morphology and viability of castor bean genotypes pollen grains

Directory of Open Access Journals (Sweden)

Maria Selma Alves Silva Diamantino

2016-01-01

Full Text Available The objective of this work was to characterize the morphology and viability of the pollen of 15 genotypes of castor bean (Ricinus communis L. and to generate information that can assist in the selection of highly promising male parents for future use in genetic improvement programs aimed at producing seeds for oil extraction. Acetolysis and scanning electron microscopy was used to characterize the morphology of the pollen. The viability of the pollen grains was estimated by in vitro germination and colorimetric analysis (acetocarmine 2% and 2, 3, 5-triphenyltetrazolium chloride 1%. For the in vitro germination, pollen grains were grown in 10 types of solidified culture medium consisting of different concentrations of sucrose, boric acid, calcium nitrate, magnesium sulfate and potassium nitrate. The pollen grains had the following characteristics: medium size, isopolar and subspheroidal shape, radial symmetry, circular ambit, 3-colporate, elongated endoapertures, tectate exine and granulated sexine. The acetocarmine dye overestimated pollen viability. The media M5 and M8 were the most efficient at promoting the germination of pollen grains. The studied genotypes had high levels of viability and can therefore be used as male parents in genetic improvement programs.

Preservation of seed viability during 25 years of storage under standard genebank conditions

NARCIS (Netherlands)

Treuren, van R.; Groot, de E.C.; Hintum, van T.J.L.

2013-01-01

Maintaining sufficient viability is critical to the sustainability of ex situ conserved seed collections. For this reason, accessions are regenerated when viability falls below a predefined threshold. Viability is monitored by determining the germination ability of accessions at predefined time

Blue whales respond to anthropogenic noise.

Directory of Open Access Journals (Sweden)

Mariana L Melcón

Full Text Available Anthropogenic noise may significantly impact exposed marine mammals. This work studied the vocalization response of endangered blue whales to anthropogenic noise sources in the mid-frequency range using passive acoustic monitoring in the Southern California Bight. Blue whales were less likely to produce calls when mid-frequency active sonar was present. This reduction was more pronounced when the sonar source was closer to the animal, at higher sound levels. The animals were equally likely to stop calling at any time of day, showing no diel pattern in their sensitivity to sonar. Conversely, the likelihood of whales emitting calls increased when ship sounds were nearby. Whales did not show a differential response to ship noise as a function of the time of the day either. These results demonstrate that anthropogenic noise, even at frequencies well above the blue whales' sound production range, has a strong probability of eliciting changes in vocal behavior. The long-term implications of disruption in call production to blue whale foraging and other behaviors are currently not well understood.

Blue-light emitting triazolopyridinium and triazoloquinolinium salts

KAUST Repository

Carboni, Valentina; Su, Xin; Qian, Hai; Aprahamian, Ivan; Credi, Alberto

2017-01-01

Compounds that emit blue light are of interest for applications that include optoelectronic devices and chemo/biosensing and imaging. The design and synthesis of small organic molecules that can act as high-efficiency deep-blue-light emitters

Metabolic Abnormalities Detected in Phase II Evaluation of Doxycycline in Dogs with Multicentric B-Cell Lymphoma

Directory of Open Access Journals (Sweden)

Kelly R. Hume

2018-02-01

Full Text Available Doxycycline has antiproliferative effects in human lymphoma cells and in murine xenografts. We hypothesized that doxycycline would decrease canine lymphoma cell viability and prospectively evaluated its clinical tolerability in client-owned dogs with spontaneous, nodal, multicentric, substage a, B-cell lymphoma, not previously treated with chemotherapy. Treatment duration ranged from 1 to 8 weeks (median and mean, 3 weeks. Dogs were treated with either 10 (n = 6 or 7.5 (n = 7 mg/kg by mouth twice daily. One dog had a stable disease for 6 weeks. No complete or partial tumor responses were observed. Five dogs developed grade 3 and/or 4 metabolic abnormalities suggestive of hepatopathy with elevations in bilirubin, ALT, ALP, and/or AST. To evaluate the absorption of oral doxycycline in our study population, serum concentrations in 10 treated dogs were determined using liquid chromatography tandem mass spectrometry. Serum levels were variable and ranged from 3.6 to 16.6 µg/ml (median, 7.6 µg/ml; mean, 8.8 µg/ml. To evaluate the effect of doxycycline on canine lymphoma cell viability in vitro, trypan blue exclusion assay was performed on canine B-cell lymphoma cell lines (17-71 and CLBL and primary B-cell lymphoma cells from the nodal tissue of four dogs. A doxycycline concentration of 6 µg/ml decreased canine lymphoma cell viability by 80%, compared to matched, untreated, control cells (mixed model analysis, p < 0.0001; Wilcoxon signed rank test, p = 0.0313. Although the short-term administration of oral doxycycline is not associated with the remission of canine lymphoma, combination therapy may be worthwhile if future research determines that doxycycline can alter cell survival pathways in canine lymphoma cells. Due to the potential for metabolic abnormalities, close monitoring is recommended with the use of this drug in tumor-bearing dogs. Additional research is needed to assess the tolerability of chronic

Determinants of Colour Constancy and the Blue Bias

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Gegenfurtner, Karl

2017-01-01

We investigated several sensory and cognitive determinants of colour constancy across 40 illumination hues. In the first experiment, we measured colour naming for the illumination and for the colour induced by the illumination on the colorimetric grey. Results confirmed that the induced colours are approximately complementary to the colour of the illumination. In the second experiment, we measured colour constancy using achromatic adjustments. Average colour constancy was perfect under the blue daylight illumination and decreased in colour directions away from the blue daylight illumination due to undershooting and a strong blue bias. Apart from this blue bias, colour constancy was not related to illumination discrimination and to chromatic detection measured previously with the same setup and stimuli. We also observed a strong negative relationship between the degree of colour constancy and the consensus of naming the illumination colour. Constancy coincided with a low naming consensus, in particular because bluish illumination colours were sometimes seen as achromatic. Blue bias and category consensus alone explained >68%, and all determinants together explained >94% of the variance of achromatic adjustments. These findings suggest that colour constancy is optimised for blue daylight. PMID:29348910

Hypnosis-associated blue-tinted vision: a case report

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Savedoff Aaron D

2005-12-01

Full Text Available Abstract Background Self-hypnosis has been taught routinely at the SUNY Upstate Medical University for treatment of pulmonary symptoms thought to be amenable to psychological therapy. While using hypnosis for relaxation, four individuals, including a patient with cystic fibrosis, reported development of blue-tinted vision. Based on a search of the literature, we believe this is the first published report of hypnosis-associated blue-tinted vision. Case presentation The patient reported blue-tinted vision when he used hypnosis on an almost daily basis for seven years. The visual change typically occurred when he was relaxed. Moreover, a concurrent erection in the absence of sexual thoughts usually was present. The other three individuals reported blue-tinted vision after learning how to use hypnosis for relaxation as part of a group hypnosis instruction. Conclusion The blue-tinted vision experienced by the individuals in this report may be the result of an hypnosis-induced primary change in cognitive processing. Additionally, as the relaxing effect of hypnosis can be associated with a reduction in blood pressure and increased blood flow, hypnosis-associated blue-tinted vision also may be related to retinal vasodilation.

Effect of Chlorine on Giardia lamblia Cyst Viability

OpenAIRE

Jarroll, Edward L.; Bingham, Alan K.; Meyer, Ernest A.

1981-01-01

The effect of chlorine concentration on Giardia lamblia cyst viability was tested under a variety of conditions. The ability of Giardia cysts to undergo excystation was used as the criterion of viability. The experimental variables employed included temperature (25, 15, and 5°C), pH (6, 7, and 8), chlorine-cyst contact time (10, 30, and 60 min), and chlorine concentration (1 to 8 mg/liter). In the pH range studied, cyst survival generally was observed to increase as buffer pH increased. Water...

How does the blue-ringed octopus (Hapalochlaena lunulata) flash its blue rings?

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Mäthger, Lydia M; Bell, George R R; Kuzirian, Alan M; Allen, Justine J; Hanlon, Roger T

2012-11-01

The blue-ringed octopus (Hapalochlaena lunulata), one of the world's most venomous animals, has long captivated and endangered a large audience: children playing at the beach, divers turning over rocks, and biologists researching neurotoxins. These small animals spend much of their time in hiding, showing effective camouflage patterns. When disturbed, the octopus will flash around 60 iridescent blue rings and, when strongly harassed, bite and deliver a neurotoxin that can kill a human. Here, we describe the flashing mechanism and optical properties of these rings. The rings contain physiologically inert multilayer reflectors, arranged to reflect blue-green light in a broad viewing direction. Dark pigmented chromatophores are found beneath and around each ring to enhance contrast. No chromatophores are above the ring; this is unusual for cephalopods, which typically use chromatophores to cover or spectrally modify iridescence. The fast flashes are achieved using muscles under direct neural control. The ring is hidden by contraction of muscles above the iridophores; relaxation of these muscles and contraction of muscles outside the ring expose the iridescence. This mechanism of producing iridescent signals has not previously been reported in cephalopods and we suggest that it is an exceptionally effective way to create a fast and conspicuous warning display.

High-content analysis of single cells directly assembled on CMOS sensor based on color imaging.

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Tanaka, Tsuyoshi; Saeki, Tatsuya; Sunaga, Yoshihiko; Matsunaga, Tadashi

2010-12-15

A complementary metal oxide semiconductor (CMOS) image sensor was applied to high-content analysis of single cells which were assembled closely or directly onto the CMOS sensor surface. The direct assembling of cell groups on CMOS sensor surface allows large-field (6.66 mm×5.32 mm in entire active area of CMOS sensor) imaging within a second. Trypan blue-stained and non-stained cells in the same field area on the CMOS sensor were successfully distinguished as white- and blue-colored images under white LED light irradiation. Furthermore, the chemiluminescent signals of each cell were successfully visualized as blue-colored images on CMOS sensor only when HeLa cells were placed directly on the micro-lens array of the CMOS sensor. Our proposed approach will be a promising technique for real-time and high-content analysis of single cells in a large-field area based on color imaging. Copyright © 2010 Elsevier B.V. All rights reserved.

Noninvasive Real-Time Assessment of Cell Viability in a Three-Dimensional Tissue.

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Mahfouzi, Seyed Hossein; Amoabediny, Ghassem; Doryab, Ali; Safiabadi-Tali, Seyed Hamid; Ghanei, Mostafa

2018-04-01

Maintaining cell viability within 3D tissue engineering scaffolds is an essential step toward a functional tissue or organ. Assessment of cell viability in 3D scaffolds is necessary to control and optimize tissue culture process. Monitoring systems based on respiration activity of cells (e.g., oxygen consumption) have been used in various cell cultures. In this research, an online monitoring system based on respiration activity was developed to monitor cell viability within acellular lung scaffolds. First, acellular lung scaffolds were recellularized with human umbilical cord vein endothelial cells, and then, cell viability was monitored during a 5-day period. The real-time monitoring system generated a cell growth profile representing invaluable information on cell viability and proliferative states during the culture period. The cell growth profile obtained by the monitoring system was consistent with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis and glucose consumption measurement. This system provided a means for noninvasive, real-time, and repetitive investigation of cell viability. Also, we showed the applicability of this monitoring system by introducing shaking as an operating parameter in a long-term culture.

Pretreatment with apoaequorin protects hippocampal CA1 neurons from oxygen-glucose deprivation.

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Detert, Julia A; Adams, Erin L; Lescher, Jacob D; Lyons, Jeri-Anne; Moyer, James R

2013-01-01

Ischemic stroke affects ∼795,000 people each year in the U.S., which results in an estimated annual cost of $73.7 billion. Calcium is pivotal in a variety of neuronal signaling cascades, however, during ischemia, excess calcium influx can trigger excitotoxic cell death. Calcium binding proteins help neurons regulate/buffer intracellular calcium levels during ischemia. Aequorin is a calcium binding protein isolated from the jellyfish Aequorea victoria, and has been used for years as a calcium indicator, but little is known about its neuroprotective properties. The present study used an in vitro rat brain slice preparation to test the hypothesis that an intra-hippocampal infusion of apoaequorin (the calcium binding component of aequorin) protects neurons from ischemic cell death. Bilaterally cannulated rats received an apoaequorin infusion in one hemisphere and vehicle control in the other. Hippocampal slices were then prepared and subjected to 5 minutes of oxygen-glucose deprivation (OGD), and cell death was assayed by trypan blue exclusion. Apoaequorin dose-dependently protected neurons from OGD--doses of 1% and 4% (but not 0.4%) significantly decreased the number of trypan blue-labeled neurons. This effect was also time dependent, lasting up to 48 hours. This time dependent effect was paralleled by changes in cytokine and chemokine expression, indicating that apoaequorin may protect neurons via a neuroimmunomodulatory mechanism. These data support the hypothesis that pretreatment with apoaequorin protects neurons against ischemic cell death, and may be an effective neurotherapeutic.

Kaempferol nanoparticles achieve strong and selective inhibition of ovarian cancer cell viability

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Luo, Haitao; Jiang, Bingbing; Li, Bingyun; Li, Zhaoliang; Jiang, Bing-Hua; Chen, Yi Charlie

2012-01-01

Ovarian cancer is one of the leading causes of cancer death for women throughout the Western world. Kaempferol, a natural flavonoid, has shown promise in the chemoprevention of ovarian cancer. A common concern about using dietary supplements for chemoprevention is their bioavailability. Nanoparticles have shown promise in increasing the bioavailability of some chemicals. Here we developed five different types of nanoparticles incorporating kaempferol and tested their efficacy in the inhibition of viability of cancerous and normal ovarian cells. We found that positively charged nanoparticle formulations did not lead to a significant reduction in cancer cell viability, whereas nonionic polymeric nanoparticles resulted in enhanced reduction of cancer cell viability. Among the nonionic polymeric nanoparticles, poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) nanoparticles incorporating kaempferol led to significant reduction in cell viability of both cancerous and normal cells. Poly(DL-lactic acid-co-glycolic acid) (PLGA) nanoparticles incorporating kaempferol resulted in enhanced reduction of cancer cell viability together with no significant reduction in cell viability of normal cells compared with kaempferol alone. Therefore, both PEO-PPO-PEO and PLGA nanoparticle formulations were effective in reducing cancer cell viability, while PLGA nanoparticles incorporating kaempferol had selective toxicity against cancer cells and normal cells. A PLGA nanoparticle formulation could be advantageous in the prevention and treatment of ovarian cancers. On the other hand, PEO-PPO-PEO nanoparticles incorporating kaempferol were more effective inhibitors of cancer cells, but they also significantly reduced the viability of normal cells. PEO-PPO-PEO nanoparticles incorporating kaempferol may be suitable as a cancer-targeting strategy, which could limit the effects of the nanoparticles on normal cells while retaining their potency against cancer cells. We

Space mutagenic effects on cistanche deserticola seed viability and parasitic condition

International Nuclear Information System (INIS)

Xu Rong; Zhou Feng; Yu Jing; Chen Jun; Sun Suqin; Liu Yougang; Liu Tongning

2009-01-01

The seeds of Cistanche deserticola which from a single plant with fine properties were carried to the space by the recoverable experiment satellite 'Shijian No.8'. After space loading, the seed viability and characters of infrared spectroscopy were analyzed and then planted to observe and investigate the variation and heredity. The results showed that compared to the control group, the seed viability and germination rate increased observably after space loading. As well, the plants grew much healthier at the seedling stage. The results indicated that space loading has obvious promote effects on the seed viability, germination rate and disease resistance of Cistanche deserticola. The analysis results of infrared spectroscopy showed the contents of protein and carbohydrate were increased distinctly and the contents of oil or fat reduced somewhat in the seeds after space loading. The intensity ratio between characteristic absorption peak of protein and characteristic absorption peak of fat (I 1625 /I 1745 ) were increased from 1.07 to 1.16, which could be related to the enhancement of seed viability and the reduction of germination restraint substances. It could be concluded that microgravity and intense radiation in the space caused seed viability and material metabolism change. (authors)

Challenges for the next generation of BlueTEC emission technology; Anforderungen und Weiterentwicklungen zur naechsten Generation der BlueTEC-Antriebstechnologie

Energy Technology Data Exchange (ETDEWEB)

Enderle, C.; Binz, R.; Paule, M.; Mackensen, A.; Lindemann, B. [Daimler AG, Stuttgart (Germany)

2010-07-01

Mercedes-Benz BlueTEC vehicles have been on the cutting edge of clean diesel technology since 2006. BlueTEC vehicles furthermore passed millions of kilometres in the hands of customers without any problems. Of course SCR systems already meet the most stringent exhaust emissions standards in international markets such as the USA, Europe and Japan. Diesel engines with BlueTEC technology also reduce CO{sub 2} emissions and provide the high torque and performance associated with the diesel engine in addition to keeping exhaust emissions at the lowest possible level. The following challenges are the focus of efforts to further advance the BlueTEC drives: - Reduce development and calibration outlay. - Standardise and reduce the costs of SCR components. - Improve performance by further reducing exhaust emissions (e.g. SULEV) and meeting the special requirements associated with the ever lower exhaust temperatures of vehicles designed to minimise CO{sub 2} emissions. - Expand on BlueTEC technology by integrating additional emissions components and combining these with other CO{sub 2} technology modules such as hybrid systems. The BlueTEC diesel engines from Mercedes-Benz represents ultra-clean drive technology, with low CO{sub 2} emissions, that can be adapted for specific markets and vehicles in a modular fashion, ideally combined with other CO{sub 2} technologies, and refined to meet future requirements. (orig.)

Relationship between humidity and influenza A viability in droplets and implications for influenza's seasonality.

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Wan Yang

Full Text Available Humidity has been associated with influenza's seasonality, but the mechanisms underlying the relationship remain unclear. There is no consistent explanation for influenza's transmission patterns that applies to both temperate and tropical regions. This study aimed to determine the relationship between ambient humidity and viability of the influenza A virus (IAV during transmission between hosts and to explain the mechanisms underlying it. We measured the viability of IAV in droplets consisting of various model media, chosen to isolate effects of salts and proteins found in respiratory fluid, and in human mucus, at relative humidities (RH ranging from 17% to 100%. In all media and mucus, viability was highest when RH was either close to 100% or below ∼50%. When RH decreased from 84% to 50%, the relationship between viability and RH depended on droplet composition: viability decreased in saline solutions, did not change significantly in solutions supplemented with proteins, and increased dramatically in mucus. Additionally, viral decay increased linearly with salt concentration in saline solutions but not when they were supplemented with proteins. There appear to be three regimes of IAV viability in droplets, defined by humidity: physiological conditions (∼100% RH with high viability, concentrated conditions (50% to near 100% RH with lower viability depending on the composition of media, and dry conditions (<50% RH with high viability. This paradigm could help resolve conflicting findings in the literature on the relationship between IAV viability in aerosols and humidity, and results in human mucus could help explain influenza's seasonality in different regions.

Lithium attenuates lead induced toxicity on mouse non-adherent bone marrow cells.

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Banijamali, Mahsan; Rabbani-Chadegani, Azra; Shahhoseini, Maryam

2016-07-01

Lead is a poisonous heavy metal that occurs in all parts of environment and causes serious health problems in humans. The aim of the present study was to investigate the possible protective effect of lithium against lead nitrate induced toxicity in non-adherent bone marrow stem cells. Trypan blue and MTT assays represented that exposure of the cells to different concentrations of lead nitrate decreased viability in a dose dependent manner, whereas, pretreatment of the cells with lithium protected the cells against lead toxicity. Lead reduced the number and differentiation status of bone marrow-derived precursors when cultured in the presence of colony stimulating factor (CSF), while the effect was attenuated by lithium. The cells treated with lead nitrate exhibited cell shrinkage, DNA fragmentation, anion superoxide production, but lithium prevented lead action. Moreover, apoptotic indexes such as PARP cleavage and release of HMGB1 induced by lead, were protected by lithium, suggesting anti-apoptotic effect of lithium. Immunoblot analysis of histone H3K9 acetylation indicated that lithium overcame lead effect on acetylation. In conclusion, lithium efficiently reduces lead toxicity suggesting new insight into lithium action which may contribute to increased cell survival. It also provides a potentially new therapeutic strategy for lithium and a cost-effective approach to minimize destructive effects of lead on bone marrow stem cells. Copyright © 2016 Elsevier GmbH. All rights reserved.

Assessment of anti-mutagenic effects of stobadine dihydro-chloride on MNNG-induced mutations in Chinese hamster cells V79

International Nuclear Information System (INIS)

Horvathova, E.; Slamenova, D.; Chorvatovicova, D.; Wsolova, L.

1995-01-01

Mutagenicity of N-methyl-n'-nitro-N-nitrosoguanidine (MNNG) and anti-mutagenic effect of antioxidant stobadine (STB) were investigated by so called HGPRT/V79 system. Cells were treated by STB before, during and after MNNG-treatment. Our results showed that the highest anti-mutagenic effect of STB was observed if the drug was given as a pretreatment before exposure of cells to MNNG. This effect was not concentration dependent within the framework of 1.5 - 9 mmol. All other combinations of MNNG- and STB-treatment led to the weaker but statistically significant decrease of 6-TG r mutations. Inhibition of proteosynthesis induced by methylxanthine pentoxifylline in the time of pre-MNNG-treatment removed completely anti-mutagenic effects of STB. In addition of mutagenicity assays, cytotoxicity of STB and combined effects of MNNG and STB were studied. Trypan blue exclusion and growth activity of influenced cells showed that application of STB (1.5 mmol) before or after MNNG (0.5 μg/cm 3 ) treatment had a similar toxic effects as MNNG alone. Application of STB during MNNG-treatment or pretreatment of cells with STB followed by combined treatment of cells by STB + MNNG statistically significantly decreased viability of cells. There are probably no relationship between the anti-mutagenic and the toxic effects of combined influence of STB and MNNG on V79 cells. (author)

Detection and characterisation of multi-drug resistance protein 1 (MRP-1) in human mitochondria.

Science.gov (United States)

Roundhill, E A; Burchill, S A

2012-03-13

Overexpression of plasma membrane multi-drug resistance protein 1 (MRP-1) can lead to multidrug resistance. In this study, we describe for the first time the expression of mitochondrial MRP-1 in untreated human normal and cancer cells and tissues. MRP-1 expression and subcellular localisation in normal and cancer cells and tissues was examined by differential centrifugation and western blotting, and immunofluorescence microscopy. Viable mitochondria were isolated and MRP-1 efflux activity measured using the calcein-AM functional assay. MRP-1 expression was increased using retroviral infection and specific overexpression confirmed by RNA array. Cell viability was determined by trypan blue exclusion and annexin V-propidium iodide labelling of cells. MRP-1 was detected in the mitochondria of cancer and normal cells and tissues. The efflux activity of mitochondrial MRP-1 was more efficient (55-64%) than that of plasma membrane MRP-1 (11-22%; PMRP-1 expression resulted in a preferential increase in mitochondrial MRP-1, suggesting selective targeting to this organelle. Treatment with a non-lethal concentration of doxorubicin (0.85 nM, 8 h) increased mitochondrial and plasma membrane MRP-1, increasing resistance to MRP-1 substrates. For the first time, we have identified MRP-1 with efflux activity in human mitochondria. Mitochondrial MRP-1 may be an exciting new therapeutic target where historically MRP-1 inhibitor strategies have limited clinical success.

Effects of aflibercept on primary RPE cells: toxicity, wound healing, uptake and phagocytosis.

Science.gov (United States)

Klettner, Alexa; Tahmaz, Nihat; Dithmer, Michaela; Richert, Elisabeth; Roider, Johann

2014-10-01

Anti-VEGF treatment is the therapy of choice in age-related macular degeneration, and is also applied in diabetic macular oedema or retinal vein occlusion. Recently, the fusion protein, aflibercept, has been approved for therapeutic use. In this study, we investigate the effects of aflibercept on primary RPE cells. Primary RPE cells were prepared from freshly slaughtered pigs' eyes. The impact of aflibercept on cell viability was investigated with MTT and trypan blue exclusion assay. The influence of aflibercept on wound healing was assessed with a scratch assay. Intracellular uptake of aflibercept was investigated in immunohistochemistry and its influence on phagocytosis with a phagocytosis assay using opsonised latex beads. Aflibercept displays no cytotoxicity on RPE cells but impairs its wound healing ability. It is taken up into RPE cells and can be intracellularly detected for at least 7 days. Intracellular aflibercept impairs the phagocytic capacity of RPE cells. Aflibercept interferes with the physiology of RPE cells, as it is taken up into RPE cells, which is accompanied by a reduction of the phagocytic ability. Additionally, it impairs the wound healing capacity of RPE cells. These effects on the physiology of RPE cells may indicate possible side effects. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Involvement of direct inhibition of NMDA receptors in the effects of sigma-receptor ligands on glutamate neurotoxicity in vitro.

Science.gov (United States)

Nishikawa, H; Hashino, A; Kume, T; Katsuki, H; Kaneko, S; Akaike, A

2000-09-15

This study was performed to examine the roles of the N-methyl-D-aspartate (NMDA) receptor/phencyclidine (PCP) channel complex in the protective effects of sigma-receptor ligands against glutamate neurotoxicity in cultured cortical neurons derived from fetal rats. A 1-h exposure of cultures to glutamate caused a marked loss of viability, as determined by Trypan blue exclusion. This acute neurotoxicity of glutamate was prevented by NMDA receptor antagonists. Expression of sigma(1) receptor mRNA in cortical cultures was confirmed by reverse transcription polymerase chain reaction (RT-PCR). sigma Receptor ligands with affinity for NMDA receptor channels including the PCP site, such as (+)-N-allylnormetazocine ((+)-SKF10,047), haloperidol, and R(-)-N-(3-phenyl-1-propyl)-1-phenyl-2-aminopropane ((-)-PPAP), prevented glutamate neurotoxicity in a concentration-dependent manner. In contrast, other sigma-receptor ligands without affinity for NMDA receptors, such as carbetapentane and R(+)-3-(3-hydroxyphenyl)-N-propylpiperidine ((+)-3-PPP), did not show neuroprotective effects. Putative endogenous sigma receptor ligands such as pregnenolone, progesterone, and dehydroepiandrosterone did not affect glutamate neurotoxicity. The protective effects of (+)-SKF10,047, haloperidol, and (-)-PPAP were not affected by the sigma(1) receptor antagonist rimcazole. These results suggested that a direct interaction with NMDA receptors but not with sigma receptors plays a crucial role in the neuroprotective effects of sigma receptor ligands with affinity for NMDA receptors.

Probing the Oncolytic and Chemosensitizing Effects of Dihydrotanshinone in an In Vitro Glioblastoma Model.

Science.gov (United States)

Kumar, Varun; Radin, Daniel; Leonardi, Donna

2017-11-01

Temozolomide is the primary chemotherapeutic agent used to treat glioblastoma. However, many tumors are initially resistant to or develop resistance to temozolomide, mainly due to high levels of O 6 -methylguanine DNA transferase (MGMT) which repairs DNA damage traditionally caused by temozolomide. Dihydrotanshinone (DHT) is extracted from Salvia miltiorrhiza, a Chinese medicinal plant, and has also been shown to have antiproliferative effects on various cancer cell lines. DHT has been to shown to induce apoptosis via induction endoplasmic reticulum stress, that can reportedly sensitize cells to temozolomide. MTS cellular proliferation assays or trypan blue viability assays were used to determine the effects of DHT/temozolomide combinatorial treatment. Enzyme-linked immunosorbent assay (ELISA) was used to determine effects on MGMT and P-glycoprotein levels after singular and combinatorial treatment. DHT had a synergistic oncolytic effect in a MGMT-deficient cell line and a sensitizing effect in a MGMT-expressing cell line. Cytotoxicity due to DHT was shown to be reactive oxygen species-dependent, while the combinatorial effect of DHT and temozolomide synergistically reduced MGMT and P-glycoprotein levels. DHT was shown to augment temozolomide efficacy, indicating that, since DHT can penetrate the blood-brain barrier, temozolomide in combination with DHT may represent a promising therapeutic option for glioblastoma. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Evaluation of Pistacia lentiscus seed oil and phenolic compounds for in vitro antiproliferative effects against BHK21 cells.

Science.gov (United States)

Mezni, Faten; Shili, Sarra; Ben Ali, Nejia; Larbi Khouja, Mohamed; Khaldi, Abdelhamid; Maaroufi, Abderrazak

2016-01-01

Within the global context of increasing cancer diseases, natural products are important in devising new drugs and providing unique ideas in cancer therapy. In Tunisian folk medicine, Pistacia lentiscus L. (Anacardiaceae) fixed oil is used for cancer treatment. This investigation studied, for the first time, the antiproliferative effect of Pistacia lentiscus fixed oil and its phenolic extract on BHK21 cancer cells. Oil was extracted from fruits harvested in northwest Tunisia and the phenolic fraction was obtained by mixing with methanol. The anti-proliferative activity of the two tested substances on BHK 21 cells were investigated in vitro using trypan blue assays. Cells were treated with different concentrations of P. lentiscus oil (0.009, 0.018, 0.036, and 0.09 g/mL) and the phenolic extract (0.007, 0.014, 0.03, and 0.07 g/mL) for 24, 48, and 72 h. The inhibitory effect of Pistacia lentiscus fixed oil increases with the increase in dose. The IC50 value was estimated at 0.029 g/mL. The percentage of cell viability was 42.46 ± 3.4% at a dose of 0.09 g/mL and was significantly lower than that of the untreated control (96.24 ± 2.5%, pPistacia lentiscus fixed oil in treating cancer, as it is used in traditional medicine.

Induction of Mitochondrial Dependent Apoptosis in Human Leukemia K562 Cells by Meconopsis integrifolia: A Species from Traditional Tibetan Medicine

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Jianping Fan

2015-06-01

Full Text Available Objectives: Meconopsis integrifolia (M. integrifolia is one of the most popular members in Traditional Tibetan Medicine. This study aimed to investigate the anticancer effect of M. integrifolia and to detect the underlying mechanisms of these effects. Methods: 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT assay and trypan blue assay were used to evaluate the cytotoxicity of M. integrifolia. Changes in cell nuclear morphology and reactive oxygen species (ROS level were observed by fluorescent microscopy. Apoptosis ratio, DNA damage and mitochondrial membrane potential (MMP loss were analyzed by flow cytometry. Western blotting assay was adopted to detect the proteins related to apoptosis. Immunofluorescence was used to observe the release of cytochrome C. Results: The obtained data revealed that M. integrifolia could significantly inhibit K562 cell viability, mainly by targeting apoptosis induction and cell cycle arrest in G2/M phase. Collapse in cell morphology, chromatin condensation, DNA damage and ROS accumulation were observed. Further mechanism detection revealed that mitochondrion might be a key factor in M. integrifolia-induced apoptosis. Conclusions: M. integrifolia could induce mitochondria mediated apoptosis and cell cycle arrest in G2/M phase with little damage to normal cells, suggesting that M. integrifolia might be a potential and efficient anticancer agent that deserves further investigation.

Study of the antioxidant effects of Eremostachys laciniata rhizome extracts in isolated rat hepatocytes

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Haleh Vaez

2015-09-01

Full Text Available Eremostachys laciniata, having rich flavonoid content, is expected to have a considerable antioxidant effect. In this study We used ACMS (Accelerated cytotoxic or protective mechanism screening technique to evaluate the possible antioxidant effect of E. laciniata rhizome against oxidative cell damages induced by different types of oxidative stress such as iron-8-hydroxyquinolin (IQ complex and copper in freshly isolated liver cells. The extracts were prepared with n-hexane, dichloromethane and methanol. Hepatocytes were isolated from male Sprague-Dawley rats by a two-step collagenase perfusion. Cell viability was measured by trypan blue exclusion method. DPPH (2, 2-diphenyl-1-picrylhydrazyl assay was used to evaluate the antioxidant activity. ROS formation was measured by using DCFDA (2, 7-dichlorofluorescin diacetate probe, mitochondrial membrane potential (MMP was assessed by rhodamine 123 fluorescence and lipid peroxidation was determined by thiobarbituric acid reactive substances (TBARS assay. The MET extract was demonstrated to possess a significant radical scavenging activity (RC50%=0.212. Unlike MET extract, the n-hexane and dichloromethane extracts showed toxic effects in cell suspensions. The MET extract significantly decreased cell death and ROS formation induced by IQ complex and copper and demonstrated protective effects against copper-induced mitochondrial membrane potential collapse and lipid peroxidation. The protection induced by MET extract can be attributed to antioxidant characteristics of the phenylethanoids content.

Functional loss of GABA transaminase (GABA-T expressed early leaf senescence under various stress conditions in Arabidopsis thaliana

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Syed Uzma Jalil

2017-06-01

Full Text Available GABA-transaminase (GABA-T involved in carbon and nitrogen metabolism during the plant development process via GABA shunt and GABA-T mutant, which is defective in GABA catabolism, is ideal model to examine the role of GABA-T in plant development and leaf senescence of plant. We have characterized GABA transaminase knock out mutant pop2-1 that is transition and pop2-3 which is T-DNA insertion mutant of Arabidopsis thaliana during various stress conditions.The GABA-T knockout mutant plants displayed precocious leaf senescence, which was accompanied by the assays of physiological parameters of leaf senescence during various stress conditions. Furthermore, our physiological evidence indicates that pop2-1 and pop2-3 mutations rapidly decreased the efficiency of leaf photosynthesis, chlorophyll content, GABA content, GABA-T, and glutamate decarboxylase (GAD activity and on the other hand increases membrane ion leakage, malondialdehyde (MDA level in stress induced leaves. However, cell viability assay by trypan blue and insitu Hydrogen peroxidation assay by 3,3-diaminobenzidine (DAB in stress induced leaves also display that pop2-1 and pop2-3 mutant leaves show oversensitivity in response to different stress conditions as compared to wild type. These results strongly indicate that the loss-of-function of GABA transaminase gene induces early leaf senescence in Arabidopsis thaliana during various stress conditions.

Down-regulation of Akt by methanol extracts of Impatiens balsamina L. promotes apoptosis in human oral squamous cell carcinoma cell lines.

Science.gov (United States)

Shin, Ji-Ae; Ryu, Mi Heon; Kwon, Ki-Han; Choi, BuYoung; Cho, Sung-Dae

2015-07-01

The apoptotic activity of methanol extracts of Impatiens balsamina L. (MEIB) and related mechanisms in human oral squamous cell carcinoma (OSCC) cells have been systematically investigated. The effects of MEIB on human OSCC cell lines were investigated using trypan blue exclusion assay, MTS assay, Western blot, 4'-6-diamidino-2-phenylindole (DAPI) staining, Live/Dead assay, Immunohistochemistry, reverse transcription-polymerase chain reaction, and promoter assay. MEIB decreased cell viability and induced apoptosis in HSC-4 cells. Higher levels of p-Akt expression were observed in OSCC than in normal oral mucosa (NOM), and it correlated with poor survival of the patients. MEIB dephosphorylated p-Akt and decreased Akt expression through proteasome-dependent degradation. LY294002 (PI3K inhibitor) decreased p-Akt and Akt, resulting in enhancing MEIB-induced apoptosis. MEIB down-regulated the expression level of survivin protein at the transcriptional level and YM155 (survivin inhibitor) decreased survivin, which facilitated MEIB-induced apoptosis. MEIB and LY294002 significantly increased Bax, thereby inducing the conformational change, mitochondrial translocation, and oligomerization. In addition, MEIB-induced growth inhibition and apoptosis in OSC-20, another human OSCC cells were mediated by regulating Akt and it downstream targets, survivin and Bax. These results suggest that MEIB may serve as a potential drug candidate for the treatment of human OSCC. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Epigallocatechin-3-Gallate Suppresses Human Herpesvirus 8 Replication and Induces ROS Leading to Apoptosis and Autophagy in Primary Effusion Lymphoma Cells

Directory of Open Access Journals (Sweden)

Ching-Yi Tsai

2017-12-01

Full Text Available Epigallocatechin-3-gallate (EGCG, the major constituent of green tea, has been shown to induce cell death in cancer cells. Primary effusion lymphoma (PEL is an aggressive neoplasm caused by human herpesvirus 8 (HHV8. In this study, we examined the role of EGCG on PEL cells in cell death and HHV8 replication. We performed trypan blue exclusion assay to assess the cell viability of PEL cells, flow cytometry analysis to examine the cell cycle distribution and reactive oxygen species (ROS generation, caspase-3 activity to assay apoptosis, acridine orange staining to determine autophagy, and immunoblotting to detect the protein levels involved in apoptosis and autophagy as well as mitogen activated protein kinases (MAPKs activation upon EGCG treatment. The expression of the HHV8 lytic gene was determined by luciferase reporter assay and reverse transcription-PCR, and viral progeny production was determined by PCR. Results revealed that EGCG induced cell death and ROS generation in PEL cells in a dose-dependent manner. N-acetylcysteine (NAC inhibited the EGCG-induced ROS and rescued the cell from EGCG-induced cell death. Even though EGCG induced ROS generation in PEL cells, it reduced the production of progeny virus from PEL cells without causing HHV8 reactivation. These results suggest that EGCG may represent a novel strategy for the treatment of HHV8 infection and HHV8-associated lymphomas.

Population viability analysis on domestic horse breeds (Equus caballus)

DEFF Research Database (Denmark)

Thirstrup, Janne Pia; Bach, Lars; Loeschcke, Volker

2009-01-01

simulation package was used for the population viability analysis. First, we investigated the future viability of these breeds based on present demographic and environmental parameters. Second, a sensitivity analysis revealed the most important variables for the viability of these breeds. Third, we examined...... concerning reproduction of the mares had the greatest impact, with the number of mares actively breeding being the most influential on the population forecasts. The results suggest that closing the Knabstrupper studbooks can be done only if increasing the number of mares actively breeding counteracts...... the loss of genetic variation attributable to such a management strategy. It is recommended, based on these results, that the number of Frederiksborg and Knabstrupper mares actively breeding must be increased to approximately 30% in the 2 breeds that are presently using only 13%, while leaving the third...

The inhibitory effect of convulsant agents on the enzyme in brain which inactivates nerveside.

Science.gov (United States)

Toh, C C

1969-07-01

1. An enzyme which can be extracted from brain inactivates nerveside in the optimum pH range 5.8-7.0.2. The polybasic acids trypan blue and its analogue trypan red, bromphenol blue and its analogue bromthymol blue at concentrations of 0.22 mM and ethylenediaminetetra-acetic acid (EDTA) at a concentration of 1 mM are strong inhibitors of the enzyme.3. Penicillin which is a monobasic carboxylic acid also inhibits the enzyme but only if concentrations as high as 3.6 mM are used. The antibiotic streptomycin which is a basic substance does not inhibit the enzyme.4. Caffeine at a concentration of 7.2 mM only weakly inhibits the enzyme.5. Chymotrypsin and wheat germ acid phosphatase also inactivate nerveside at pH 5.9 and are inhibited by the acidic dyes and penicillin. EDTA inhibits wheat germ phosphatase but activates chymotrypsin.6. Inactivation of nerveside by the brain enzyme and by wheat germ phosphatase is different from the action of chymotrypsin. Nerveside solutions incubated with chymotrypsin completely lose all biological activity whereas if incubation is carried out with either the brain enzyme or wheat germ acid phosphatase a residual biological activity remains even when the concentration of these two enzymes is increased. This residual biological activity is due to a peptide as it is destroyed by chymotrypsin.7. The manner in which nerveside is inactivated by the brain enzyme is uncertain as the preparation of the latter contained phosphodiesterase and protease activities which were similarly inhibited by the acid dyes, penicillin and EDTA.8. Pentylenetetrazole, picrotoxin, strychnine and tetanus toxin do not inhibit the brain enzyme.9. The nerveside-inactivating enzyme is not identical with the Substance P-inactivating enzyme in brain as the former is inhibited by EDTA while the latter is not.

QCD on the BlueGene/L Supercomputer

International Nuclear Information System (INIS)

Bhanot, G.; Chen, D.; Gara, A.; Sexton, J.; Vranas, P.

2005-01-01

In June 2004 QCD was simulated for the first time at sustained speed exceeding 1 TeraFlops in the BlueGene/L supercomputer at the IBM T.J. Watson Research Lab. The implementation and performance of QCD in the BlueGene/L is presented

QCD on the BlueGene/L Supercomputer

Science.gov (United States)

Bhanot, G.; Chen, D.; Gara, A.; Sexton, J.; Vranas, P.

2005-03-01

In June 2004 QCD was simulated for the first time at sustained speed exceeding 1 TeraFlops in the BlueGene/L supercomputer at the IBM T.J. Watson Research Lab. The implementation and performance of QCD in the BlueGene/L is presented.

Blue-light emitting triazolopyridinium and triazoloquinolinium salts

KAUST Repository

Carboni, Valentina

2017-01-27

Compounds that emit blue light are of interest for applications that include optoelectronic devices and chemo/biosensing and imaging. The design and synthesis of small organic molecules that can act as high-efficiency deep-blue-light emitters in the solid state and can be easily processed from solutions represents a significant challenge. Herein we present the preparation and photophysical, photochemical and electrochemical properties of a series of triazolopyridinium and triazoloquinolinium compounds. The compounds are soluble in water or polar organic solvents and exhibit photoluminescence in the blue region of the spectrum in fluid solution, in the solid state and in a frozen matrix.

Effect of high hydrostatic pressure on mycelial development, spore viability and enzyme activity of Penicillium Roqueforti.

Science.gov (United States)

Martínez-Rodríguez, Yamile; Acosta-Muñiz, Carlos; Olivas, Guadalupe I; Guerrero-Beltrán, José; Rodrigo-Aliaga, Dolores; Mujica-Paz, Hugo; Welti-Chanes, Jorge; Sepulveda, David R

2014-01-03

This study investigated the effect of high hydrostatic pressure treatments on mycelial development, spore viability, and total proteolytic and lipolytic activity of Penicillium roqueforti PV-LYO 10 D. Fungus growing in liquid medium was pressure-treated at 300, 400, and 500 MPa for 10 min at 20°C following seven days of incubation at 25°C and analyzed periodically up to day 9 after treatments to evaluate the effect on fungal growth. Mycelial mass of P. roqueforti was significantly affected at all pressure treatments evaluated, being 15.48%, 22.28%, 30.03%, and 12.53% lower than controls on day 1, 3, 6, and 9 after 300 MPa treatment, respectively. In a similar way, at 400 and 500 MPa, mycelial mass was 31.08% and 60.34% lower than controls one day after treatments and 49.74% and 80.85% lower on day 9, respectively. The viability of P. roqueforti spores decreased by 36.53% at 300 MPa, and complete inactivation took place at ≥400 MPa from an initial count of 7 log cfu/mL. Total proteolytic activity was not significantly affected at 300 MPa but was reduced by 18.22% at 400 MPa and by 43.18% at 500 MPa. Total lipolytic activity also decreased as the intensity of the pressure treatments increased. 21.69%, 39.12%, and 56.26% activity reductions were observed when treatments of 300, 400 and 500 MPa were applied, respectively. The results from this study show that pressure treatments are able to control growth, inactivate spores, and alter enzyme activity of P. roqueforti, which could be of interest in extending the shelf-life of blue-veined cheeses and other food products. © 2013.

Turnover of Glycerolipid Metabolite Pool and Seed Viability

Directory of Open Access Journals (Sweden)

Xiao-Long Hu

2018-05-01

Full Text Available Hydration–dehydration cycles can frequently cause stress to seeds, but can also be used to improve germination. However, the molecular basis of the stress caused is poorly understood. Herein, we examine the effects of hydration–dehydration cycles on seed viability and profile the membrane glycerolipid molecular species. We find that seed viability was not affected during the first two cycles, but significantly decreased as further cycles were applied, until all viability was lost. The abundances of seven glycerolipid classes increased and decreased through hydration and dehydration, respectively, but the phosphatidic acid and diacylglycerol abundances changed in the opposite sense, while total glycerolipid contents remained constant. This suggests that during hydration–dehydration cycles, turnover of glycerolipid metabolite pools take place, while no significant lipid synthesis or degradation is involved. As further hydration–dehydration cycles occurred, lipid unsaturation increased, plastidic lipids decreased, and phosphatidylserine acyl chains lengthened. The latter two could be lethal for seeds. Our findings reveal a novel model of membrane lipid changes, and provide new insights into the responses of seeds to hydration–dehydration cycles.

Ca-Lignosulphonate and sclerotial viability of Sclerotinia sclerotiorum

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MATTEO MONTANARI

2012-01-01

Full Text Available Lignosulphonates, low cost by-products of the pulping process, have shown suppressive effects against some diseases caused by soil-borne pathogens. In this study, the effect of 1.5% v/v calcium lignosulphonate (Ca-Ls amendment to two commercial potting mixes (peat + coconut fibres; PC; and municipal compost + peat + pumice; MCPP on the viability of Sclerotinia sclerotiorum sclerotia was investigated. Sclerotia were buried in the Ca-Ls amended substrates for 30 days. Non-amended PC and MCPP, sterile sand and sterile PC with and without Ca-Ls were used as controls. The viability of sclerotia recovered from PC and MCPP amended with Ca-Ls was reduced by 50 and 42% respectively compared to control treatments. Ca-Ls amendment decreased sclerotial viability by enhancing the activity of the indigenous mycoparasitic fungi, Fusarium oxysporum, Mucor spp. and Trichoderma spp. The biocontrol ability of Ca-Ls against sclerotia was due to the stimulation of microbial activity and is, therefore, strictly dependent on the microbial composition of the substrate.

Molecular assays for determining Mycobacterium leprae viability in tissues of experimentally infected mice.

Science.gov (United States)

Davis, Grace L; Ray, Nashone A; Lahiri, Ramanuj; Gillis, Thomas P; Krahenbuhl, James L; Williams, Diana L; Adams, Linda B

2013-01-01

The inability of Mycobacterium leprae to grow on axenic media has necessitated specialized techniques in order to determine viability of this organism. The purpose of this study was to develop a simple and sensitive molecular assay for determining M. leprae viability directly from infected tissues. Two M. leprae-specific quantitative reverse transcription PCR (qRT-PCR) assays based on the expression levels of esxA, encoding the ESAT-6 protein, and hsp18, encoding the heat shock 18 kDa protein, were developed and tested using infected footpad (FP) tissues of both immunocompetent and immunocompromised (athymic nu/nu) mice. In addition, the ability of these assays to detect the effects of anti-leprosy drug treatment on M. leprae viability was determined using rifampin and rifapentine, each at 10 mg/kg for 1, 5, or 20 daily doses, in the athymic nu/nu FP model. Molecular enumeration (RLEP PCR) and viability determinations (qRT-PCR) were performed via Taqman methodology on DNA and RNA, respectively, purified from ethanol-fixed FP tissue and compared with conventional enumeration (microscopic counting of acid fast bacilli) and viability assays (radiorespirometry, viability staining) which utilized bacilli freshly harvested from the contralateral FP. Both molecular and conventional assays demonstrated growth and high viability of M. leprae in nu/nu FPs over a 4 month infection period. In contrast, viability was markedly decreased by 8 weeks in immunocompetent mice. Rifapentine significantly reduced bacterial viability after 5 treatments, whereas rifampin required up to 20 treatments for the same efficacy. Neither drug was effective after a single treatment. In addition, host gene expression was monitored with the same RNA preparations. hsp18 and esxA qRT-PCR are sensitive molecular indicators, reliably detecting viability of M. leprae in tissues without the need for bacterial isolation or immediate processing, making these assays applicable for in vivo drug screening and

The cybernetics of viability: an overview

Science.gov (United States)

Nechansky, Helmut

2011-10-01

A three-level approach to viability is developed, considering (1) living systems, (2) a niche, understood as the area within the reach of their actions, and (3) an environment. A systematic analysis of the interrelations between these levels shows that living systems emerge with matter/energy processing systems. These can add controller structures when producing excess energy. A three-sensor controller structure enables a living system to deal with unfavourable and scarce environments. Further evolution of these controller structures offers improved ways to act on niches. Maintaining niches in scarce environments can require technology or economy. So social systems emerge, which are understood as aggregates of living systems. Basic patterns of interactions within social systems are analysed. So the introduction of the notion of the niche into the discussion of viability allows us to explain phenomena ranging from properties of single living systems to societal organization.

A Classification Method for Seed Viability Assessment with Infrared Thermography

Directory of Open Access Journals (Sweden)

Sen Men

2017-04-01

Full Text Available This paper presents a viability assessment method for Pisum sativum L. seeds based on the infrared thermography technique. In this work, different artificial treatments were conducted to prepare seeds samples with different viability. Thermal images and visible images were recorded every five minutes during the standard five day germination test. After the test, the root length of each sample was measured, which can be used as the viability index of that seed. Each individual seed area in the visible images was segmented with an edge detection method, and the average temperature of the corresponding area in the infrared images was calculated as the representative temperature for this seed at that time. The temperature curve of each seed during germination was plotted. Thirteen characteristic parameters extracted from the temperature curve were analyzed to show the difference of the temperature fluctuations between the seeds samples with different viability. With above parameters, support vector machine (SVM was used to classify the seed samples into three categories: viable, aged and dead according to the root length, the classification accuracy rate was 95%. On this basis, with the temperature data of only the first three hours during the germination, another SVM model was proposed to classify the seed samples, and the accuracy rate was about 91.67%. From these experimental results, it can be seen that infrared thermography can be applied for the prediction of seed viability, based on the SVM algorithm.

Efficient fluorescent red, green, and blue organic light-emitting devices with a blue host of spirobifluorene derivative

Energy Technology Data Exchange (ETDEWEB)

Lee, R.-H. [Department of Chemical and Material Engineering, National Yunlin University of Science and Technology, Yunlin 640, Taiwan (China)], E-mail: lerongho@yuntech.edu.tw; Huang, Y.-W.; Wang, Y.-Y. [Department of Chemical and Material Engineering, National Yunlin University of Science and Technology, Yunlin 640, Taiwan (China); Chang, H.-Y. [EChem Hightech CO., LTD, Hsin-Chu Industrial Park, Hu-Kou, Hsin-Chu, Taiwan (China)

2008-06-02

Efficient fluorescent blue, green, and red (RGB) organic light-emitting devices (OLEDs) were fabricated using a blue host material of pyrimidine-containing spirobifluorene derivative 2,7-bis[2-(4-tert-butylphenyl)pyrimidine-5-yl]-9,9'-spirobifluorene (TBPSF) doped with blue dye perylene, green dye 10-(2-benzothiazolyl)-1,1,7,7-tetramethyl-2,3,6,7-tetrahydro-1H,5H, 11H-benzo[l] pyrano[6,7,8-ij] quinolizin-11-one (C545T), and red dye 4-(dicyanomethylene)-2-t-butyl-6-(1,1,7,7-tetramethyljulolidyl-9-enyl) -4H-pyran (DCJTB), respectively. The brightness and current efficiency of the perylene doped blue device reached 10117 cd/m{sup 2} and 2.97 cd/A. Green emission of the C545T doped device reached 8500 cd/m{sup 2} and 13.0 cd/A. Red emission of the DCJTB doped device can be as high as 9000 cd/m{sup 2} and 2.0 cd/A, respectively. High color purity of the blue (Commission Internationale de L'Eclairage (CIE{sub x,y}) coordinates (CIE, x = 0.27, y = 0.24)), green (CIE, x = 0.19, y = 0.63) and red (CIE, x = 0.62, y = 0.37) emissions were achieved for RGB dyes doped TBPSF OLEDs. High brightness, large current efficiency, and good color purity of TBPSF-based RGB OLEDs were obtained by the configuration optimization device, such as inserting the hole and electron-injection materials, and suitable dopant content and light emitting layer thickness.

Blue Pseudoazulene-Skeleton Pigments of Natural Origin

OpenAIRE

井上, 謙一郎; イノウエ, ケンイチロウ; KENICHIRO, INOUE

1993-01-01

Genipin, an iridoid constituent of Genipa americana, readily reacts with amino acids in the presence of oxygen to give a mixture of polymeric blue pigments whose structures are not determined. In the basic studies to elucidate the structure and formation mechnism of blue pigments, the reaction of genipin with methylamine in the absence of oxygen yielded 9 red compounds leading to blue pigments. In this article, the structures and spectroscopic properties of these red compounds were described....

Analytical Research to Determine the effects of the Components of ONGABO on the Viability of HepG2 Cancer Cells by Using the Sovereign, Minister, Assistant and Courier Principle (君臣佐使論

Directory of Open Access Journals (Sweden)

Shin Jeong-Hun

2012-12-01

Full Text Available Objectives: This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle (君臣佐使論 to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng, Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Methods: Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa, Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Results: Although Ginseng Radix (Red Ginseng and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. Conclusions: In the sovereign, minister, assistant and courier principle (君臣佐使論, Ginseng Radix (Red Ginseng corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in

Economic Viability and Marketing Strategies of Periwinkle ...

African Journals Online (AJOL)

Economic Viability and Marketing Strategies of Periwinkle Tympanotonus Fuscatus in Rivers State, Nigeria. ... The results indicated that marketing strategies are enroute, through harvesters (collectors), ... EMAIL FULL TEXT EMAIL FULL TEXT

Challenge testing of gametes to enhance their viability

DEFF Research Database (Denmark)

Callesen, Henrik

2010-01-01

of survival mechanism that enables them to come through the process. The details of the mechanism remain unknown but, if identified, it could have immense potential as a new way to improve the viability of embryos produced by ART. However, few publications describe systematic ways to challenge test gametes...... and then to use the results as a basis for improving gamete viability. Furthermore, new methods to monitor the reactions of gametes to such challenge tests are needed. In the present review, these two issues are discussed, as are some of the conditions necessary before a challenge test protocol can be part...

Viability and Biomechanics of Diced Cartilage Blended With Platelet-Rich Plasma and Wrapped With Poly (Lactic-Co-Glycolic) Acid Membrane.

Science.gov (United States)

Liao, Jun-Lin; Chen, Jia; He, Bin; Chen, Yong; Xu, Jia-Qun; Xie, Hong-Ju; Hu, Feng; Wang, Ai-Jun; Luo, ChengQun; Li, Qing-Feng; Zhou, Jian-Da

2017-09-01

The objective of this study was to investigate the viability and biomechanics of diced cartilage blended with platelet-rich plasma (PRP) and wrapped with poly (lactic-co-glycolic) acid (PLGA) membrane in a rabbit model. A total of 10 New Zealand rabbits were used for the study. Cartilage grafts were harvested from 1 side ear. The grafts were divided into 3 groups for comparison: bare diced cartilage, diced cartilage wrapped with PLGA membrane, and diced cartilage blended with PRP and wrapped with PLGA membrane. Platelet-rich plasma was prepared using 8 mL of auricular blood. Three subcutaneous pockets were made in the backs of the rabbits, and the grafts were placed in these pockets. The subcutaneous implant tests were conducted for safety assessment of the PLGA membrane in vivo. All of the rabbits were sacrificed at the end of 3 months, and the specimens were collected. The sections were stained with hematoxylin and eosin, toluidin blue, and collagen II immunohistochemical. Simultaneously, biomechanical properties of grafts were assessed. This sample of PLGA membrane was conformed to the current standard of biological evaluation of medical devices. Moderate resorption was seen at the end of 3 months in the gross assessment in diced cartilage wrapped with PLGA membrane, while diced cartilage blended with PRP had no apparent resorption macroscopically and favorable viability in vivo after 3 months, and the histological parameters supported this. Stress-strain curves for the compression test indicated that the modulus of elasticity of bare diced cartilage was 7.65 ± 0.59 MPa; diced cartilage wrapped with PLGA membrane was 5.98 ± 0.45 MPa; and diced cartilage blended with PRP and wrapped with PLGA membrane was 7.48 ± 0.55 MPa, respectively. Diced cartilage wrapped with PLGA membrane had moderate resorption macroscopically after 3 months. However, blending with PRP has beneficial effects in improving the viability of diced cartilages. Additionally, the

Assessment of the technical viability of reactor options for plutonium disposition

International Nuclear Information System (INIS)

Primm, R.T. III.

1996-01-01

Various reactor concepts for the disposition of surplus Pu have been proposed by reactor vendors; not all have attained the same level of technical viability. Studies were performed to differentiate between reactor concepts by devising a quantitative index for technical viability. For a quantitative assessment, three issues required resolution: the definition of a technical maturity scale, the treatment of ''subjective'' factors which cannot be easily represented in a quantitative format, and the protocol for producing a single technical viability figure-of-merit for each alternative. Alternatives involving the use of foreign facilities were found to be the most technically viable

Quirks of dye nomenclature. 1. Evans blue.

Science.gov (United States)

Cooksey, C J

2014-02-01

The history, origin, identity, chemistry and use of Evans blue dye are described along with the first application to staining by Herbert McLean Evans in 1914. In the 1930s, the dye was marketed under the name, Evans blue dye, which was profoundly more acceptable than the ponderous chemical name.

A comparison of assays measuring the viability of Legionella ...

Science.gov (United States)

Background: The relatively high prevalence of Legionella pneumophila in premise plumbing systems has been widely reported. Published reports indicate Legionella has a comparatively high resistance to chlorine and moreover has the ability to grow in phagocytic amoeba which could provide additional protection in chlorinated drinking water distribution systems. Copper-Silver (Cu-Ag) ionization treatment systems are commercially available for use in large building water systems to help control the risks from Legionella bacteria. The objectives of this study were to develop and optimize Legionella viability assays and use them to investigate the viability of Legionella bacteria after exposure to water treated with coppper and silver ions. Methods: Log phase L. pneumophila cells were used in all experiments and were generated by incubation at 35C for 48 hours in buffered yeast extract broth. Viability assays used included plating on buffered charcoal yeast extract agar to determine the number of culturable cells and treating cells with propidium monoazide (PMA) or ethidium monoazide (EMA) followed by quantitative PCR targeting mip gene of L. pneumophila. The qPCR viability assays were optimized using L. pneumophila inactivated by heat treatment at 65C for 60 min. The effectiveness of Cu-Ag ionization treatment was studied by inoculating L. pneumonia at 105 CFU/mL in water collected directly from a building water system that employed this technology and incubat

Effect of salt hyperosmotic stress on yeast cell viability

Directory of Open Access Journals (Sweden)

Logothetis Stelios

2007-01-01

Full Text Available During fermentation for ethanol production, yeasts are subjected to different kinds of physico-chemical stresses such as: initially high sugar concentration and low temperature; and later, increased ethanol concentrations. Such conditions trigger a series of biological responses in an effort to maintain cell cycle progress and yeast cell viability. Regarding osmostress, many studies have been focused on transcriptional activation and gene expression in laboratory strains of Saccharomyces cerevisiae. The overall aim of this present work was to further our understanding of wine yeast performance during fermentations under osmotic stress conditions. Specifically, the research work focused on the evaluation of NaCl-induced stress responses of an industrial wine yeast strain S. cerevisiae (VIN 13, particularly with regard to yeast cell growth and viability. The hypothesis was that osmostress conditions energized specific genes to enable yeast cells to survive under stressful conditions. Experiments were designed by pretreating cells with different sodium chloride concentrations (NaCl: 4%, 6% and 10% w/v growing in defined media containing D-glucose and evaluating the impact of this on yeast growth and viability. Subsequent fermentation cycles took place with increasing concentrations of D-glucose (20%, 30%, 40% w/v using salt-adapted cells as inocula. We present evidence that osmostress induced by mild salt pre-treatments resulted in beneficial influences on both cell viability and fermentation performance of an industrial wine yeast strain.

How bees distinguish patterns by green and blue modulation.

Science.gov (United States)

Horridge, Adrian

2015-01-01

In the 1920s, Mathilde Hertz found that trained bees discriminated between shapes or patterns of similar size by something related to total length of contrasting contours. This input is now interpreted as modulation in green and blue receptor channels as flying bees scan in the horizontal plane. Modulation is defined as total contrast irrespective of sign multiplied by length of edge displaying that contrast, projected to vertical, therefore, combining structure and contrast in a single input. Contrast is outside the eye; modulation is a phasic response in receptor pathways inside. In recent experiments, bees trained to distinguish color detected, located, and measured three independent inputs and the angles between them. They are the tonic response of the blue receptor pathway and modulation of small-field green or (less preferred) blue receptor pathways. Green and blue channels interacted intimately at a peripheral level. This study explores in more detail how various patterns are discriminated by these cues. The direction of contrast at a boundary was not detected. Instead, bees located and measured total modulation generated by horizontal scanning of contrasts, irrespective of pattern. They also located the positions of isolated vertical edges relative to other landmarks and distinguished the angular widths between vertical edges by green or blue modulation alone. The preferred inputs were the strongest green modulation signal and angular width between outside edges, irrespective of color. In the absence of green modulation, the remaining cue was a measure and location of blue modulation at edges. In the presence of green modulation, blue modulation was inhibited. Black/white patterns were distinguished by the same inputs in blue and green receptor channels. Left-right polarity and mirror images could be discriminated by retinotopic green modulation alone. Colors in areas bounded by strong green contrast were distinguished as more or less blue than the

Global monthly water scarcity: Blue water footprints versus blue water availability

NARCIS (Netherlands)

Hoekstra, Arjen Ysbert; Mekonnen, Mesfin; Chapagain, Ashok; Mathews, R.E.; Richter, B.D.

2012-01-01

Freshwater scarcity is a growing concern, placing considerable importance on the accuracy of indicators used to characterize and map water scarcity worldwide. We improve upon past efforts by using estimates of blue water footprints (consumptive use of ground- and surface water flows) rather than

G2 arrest and apoptosis of cultured Raji cells by continuous low dose rate beta irradiation therapy with 188Re-perrhenate

International Nuclear Information System (INIS)

Yim, S. J.; Kim, E. H.; Lee, T. S.; Woo, K. S.; Jeong, W. S.; Choi, C. W.; Yim, S. M.

2001-01-01

Beta emitting radionuclide therapy gives exponentially decreasing radiation dose rate and results in cell death presumably by apoptosis. We observed changes in DNA content and apoptosis in relatively low dose rate beta irradiation. Raji cells were cultured and incubated with 188Re-perrhenate (3.7MBq, or 370MBq/ml) for 4 hours to give irradiation dose of 0.4, 4, or 40 Gy. After changing the culture media, cells were cultured for 2,4,8,16, and 24 hours. The cells were stained with Trypan blue, Annexin-V and Propidium Iodide (PI) to observe cell viability, cell membrane alternation by apoptosis and changes in DNA content respectively. Flowcytometry was done for Annexin-V and PI to quantitate apoptosis and necrosis in the irradiated cells. DAPI(4,6-diamidino-2-phenylindole) stain was also done to observe the damage in the nucleus. Cell viability decreased with an increasing radiation dose. Cells irradiated in 40 Gy showed early uptake of both Annexin-V and PI suggesting cell death by necrosis. Cells irradiated in 0.4 Gy showed delayed uptake of Annexin-V only, and later on PI uptake suggesting cell death mainly by apoptosis. The cells irradiated in 0.4 Gy showed G2 arrest in 16 hours after irradiation, but the cells irradiated in 40 Gy showed early DNA fragmentation within 2 hours after irradiation. In DAPI stain, early nucleus damage was observed in the cells irradiated in 40 Gy. On the other hand, slowly increasing apoptotic bodies were observed in the cells irradiated in 0.4 Gy. These results suggest that continuous low-dose irradiation induces G2 arrest and progressive apoptosis in cells while continuous high-dose irradiation induces rapid necrosis. Therefore, we expect therapeutic effect by continuous low-dose rate irradiation with beta emitting radiopharmaceuticals

Elevation of cAMP Levels Inhibits Doxorubicin-Induced Apoptosis in Pre- B ALL NALM- 6 Cells Through Induction of BAD Phosphorylation and Inhibition of P53 Accumulation.

Science.gov (United States)

Fatemi, Ahmad; Kazemi, Ahmad; Kashiri, Meysam; Safa, Majid

2015-01-01

Recognition of the molecular mechanisms of cAMP action against DNA damage-induced apoptosis can be useful to improve the efficacy of DNA damaging therapeutic agents. Considering the critical role of bcl-2-associated death promoter (BAD) and p53 proteins in DNA damage -induced apoptosis, the aim of this study was to assess the effect of cAMP-elevating agents on these proteins in doxorubicin-treated pre-B acute lymphoblastic leukemia (pre-B ALL) NALM-6 cells.The pre-B ALL cell line NALM-6 was cultured and treated with doxorubicin in combination with or without cAMP-elevating agents forskolin and 3-isobutyl-1-methylxanthine (IBMX). Cell viability was measured by trypan blue staining and MTT assay. For evaluation of apoptosis, annexin-V staining by flow cytometry and caspase-3 activity assay were used. Protein expression of p53, BAD and phoshorylated BAD was detected by western blotting analysis.cAMP-increasing agents diminished the doxorubicin-mediated cytotoxicity in NALM-6 cells as indicated by the viability assays. Annexin-V apoptosis assay showed that the cAMP-elevating agents decreased doxorubicin-induced apoptosis. Moreover, doxorubicin-induced caspase-3 activity was attenuated in the presence of cAMP-increasing agents. Western blot results revealed the reduced expression of p53 protein in cells treated with combination of cAMP-elevating agents and doxorubicin in contrast to cells treated with doxorubicin alone. Expression of total BAD protein was not affected by doxorubicin and cAMP-elevating agents. However, phosphorylation of BAD protein was induced in the presence of cAMP-elevating agents. Our study suggests that elevated cAMP levels inhibit doxorubicin-induced apoptosis in pre-B ALL cells through induction of BAD phosphorylation and abrogation of p53 accumulation.

A Modified Ficoll-Paque Gradient Method for Isolating Mononuclear Cells from the Peripheral and Umbilical Cord Blood of Humans for Biobanks and Clinical Laboratories.

Science.gov (United States)

Jia, Yanjuan; Xu, Hui; Li, Yonghong; Wei, Chaojun; Guo, Rui; Wang, Fang; Wu, Yu; Liu, Jing; Jia, Jing; Yan, Junwen; Qi, Xiaoming; Li, Yuanting; Gao, Xiaoling

2018-04-01

Although the Ficoll-Paque method is classically used to isolate peripheral blood mononuclear cells (PBMCs), modifications in this method are required for a more rapid and economic output for biobanks and clinical laboratories, particularly in developing countries. In this study, we addressed this issue by modifying the Ficoll-Paque method for the isolation of PBMCs or mononuclear cells from the peripheral and the umbilical cord blood of healthy and diseased (infected, anemic, and chronic obstructive pulmonary disease) adult individuals. In the modified method, we initiated the cell isolation process from the buffy coat layer, which appears in the interface between the plasma and sediments after centrifugation, instead of using the whole blood as described in the classic method. Although the PBMC yield by the modified method was about 12% less than in the classic method, the number of PBMCs isolated by the modified method was more than one million, which is enough for different research/diagnostic purposes, such as multi-omics detection. Assessment of cell viability and purity by hematology analyzer and trypan blue showed no significant difference between the viability and purity of the PBMCs isolated by these two methods in almost all groups, except samples from the infected and cord blood groups, where lower PBMC purity with higher granulocyte contamination were observed. In addition, at delayed processing time points, all parameters for the two methods were decreased in a time-dependent manner, especially at 8, 12, or 24 hours after the sample collection. In summary, the performance of PBMC isolation by the classic and modified methods mainly relies on the PBMC ratio in original samples. The modified method could be preferred for PBMC isolation because of its time and cost savings, especially for the biobanks and clinical laboratories in developing countries.

Vitamins C and K3: A Powerful Redox System for Sensitizing Leukemia Lymphocytes to Everolimus and Barasertib.

Science.gov (United States)

Ivanova, Donika; Zhelev, Zhivko; Lazarova, Dessislava; Getsov, Plamen; Bakalova, Rumiana; Aoki, Ichio

2018-03-01

Recent studies provided convincing evidence for the anticancer activity of combined application of vitamin C and pro-vitamin K3 (menadione). The molecular pathways underlying this process are still not well established. The present study aimed to investigate the effect of the combination of vitamin C plus pro-vitamin K3 on the redox status of leukemia and normal lymphocytes, as well as their sensitizing effect for a variety of anticancer drugs. Cytotoxicity of the substances was analyzed by trypan blue staining and automated counting of live and dead cells. Apoptosis was analyzed by fluorescein isothiocyanate-annexin V test. Oxidative stress was evaluated by the intracellular levels of reactive oxygen and nitrogen species and protein-carbonyl products. Combined administration of 300 μM vitamin C plus 3 μM pro-vitamin K3 reduced the viability of leukemia lymphocytes by ~20%, but did not influence the viability of normal lymphocytes. All combinations of anticancer drug plus vitamins C and K3 were characterized by synergistic cytotoxicity towards Jurkat cells, compared to cells treated with drug alone for 24 h. In the case of barasertib and everolimus, this synergistic cytotoxicity increased within 72 hours. It was accompanied by strong induction of apoptosis, but a reduction of level of hydroperoxides and moderately increased protein-carbonyl products in leukemia cells. Leukemia lymphocytes were more sensitive to combined administration of anticancer drug (everolimus or barasertib) plus vitamins C and K3, compared to normal lymphocytes. The combination of vitamin C plus K3 seems to be a powerful redox system that could specifically influence redox homeostasis of leukemia cells and sensitize them to conventional chemotherapy. Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Propofol ameliorates doxorubicin-induced oxidative stress and cellular apoptosis in rat cardiomyocytes

Energy Technology Data Exchange (ETDEWEB)

Lai, H.C. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Medicine and Cardiovascular Research Center, National Yang-Ming University School of Medicine, Taipei, Taiwan (China); Yeh, Y.C. [Graduate Institute of Natural Healing Sciences, Nanhua University, Chiayi, Taiwan (China); Wang, L.C. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Ting, C.T.; Lee, W.L. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Medicine and Cardiovascular Research Center, National Yang-Ming University School of Medicine, Taipei, Taiwan (China); Lee, H.W. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Wang, K.Y. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Medicine, Chung-Shan Medical University, Taichung, Taiwan (China); Wu, A. [College of Biological Science, University of California, Davis (United States); Su, C.S. [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Medicine and Cardiovascular Research Center, National Yang-Ming University School of Medicine, Taipei, Taiwan (China); Liu, T.J., E-mail: trliu@vghtc.gov.tw [Cardiovascular Center and Department of Anesthesiology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Medicine and Cardiovascular Research Center, National Yang-Ming University School of Medicine, Taipei, Taiwan (China)

2011-12-15

Background: Propofol is an anesthetic with pluripotent cytoprotective properties against various extrinsic insults. This study was designed to examine whether this agent could also ameliorate the infamous toxicity of doxorubicin, a widely-used chemotherapeutic agent against a variety of cancer diseases, on myocardial cells. Methods: Cultured neonatal rat cardiomyocytes were administrated with vehicle, doxorubicin (1 {mu}M), propofol (1 {mu}M), or propofol plus doxorubicin (given 1 h post propofol). After 24 h, cells were harvested and specific analyses regarding oxidative/nitrative stress and cellular apoptosis were conducted. Results: Trypan blue exclusion and MTT assays disclosed that viability of cardiomyocytes was significantly reduced by doxorubicin. Contents of reactive oxygen and nitrogen species were increased and antioxidant enzymes SOD1, SOD2, and GPx were decreased in these doxorubicin-treated cells. Mitochondrial dehydrogenase activity and membrane potential were also depressed, along with activation of key effectors downstream of mitochondrion-dependent apoptotic signaling. Besides, abundance of p53 was elevated and cleavage of PKC-{delta} was induced in these myocardial cells. In contrast, all of the above oxidative, nitrative and pro-apoptotic events could be suppressed by propofol pretreatment. Conclusions: Propofol could extensively counteract oxidative/nitrative and multiple apoptotic effects of doxorubicin in the heart; hence, this anesthetic may serve as an adjuvant agent to assuage the untoward cardiac effects of doxorubicin in clinical application. -- Highlights: Black-Right-Pointing-Pointer We evaluate how propofol prevents doxorubicin-induced toxicity in cardiomyocytes. Black-Right-Pointing-Pointer Propofol reduces doxorubicin-imposed nitrative and oxidative stress. Black-Right-Pointing-Pointer Propofol suppresses mitochondrion-, p53- and PKC-related apoptotic signaling. Black-Right-Pointing-Pointer Propofol ameliorates apoptosis and

In vitro and in vivo evaluation of a new nanocomposite, containing high density polyethylene, tricalcium phosphate, hydroxyapatite, and magnesium oxide nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Pourdanesh, Fereydoun [Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran 8916733754 (Iran, Islamic Republic of); Jebali, Ali, E-mail: alijebal2011@gmail.com [Department of Medical Physics and Biomedical Engineering, Shahid Beheshti University of Medical Sciences, Tehran (Iran, Islamic Republic of); Hekmatimoghaddam, Seyedhossein [Department of Laboratory Sciences, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd (Iran, Islamic Republic of); Allaveisie, Azra [Department of Genetics, Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd (Iran, Islamic Republic of)

2014-07-01

In this study, a new nanocomposite, which contained high density polyethylene (HDPE), tricalcium phosphate (Ca{sub 3}(PO{sub 4}){sub 2}) nanoparticles (TCP NPs), hydroxyapatite nanoparticles (HA NPs), and magnesium oxide nanoparticles (MgO NPs) was prepared. As in vitro experiment, human osteoblasts (HOB) cells were exposed to pristine HDPE and its nanocomposite for a period of 1, 4, and 7 days at 37 °C, and then different assays were carried out, including osteoblast cell proliferation, Trypan blue staining, cell viability, alkaline phosphatase (ALP), and cell adhesion. Antibacterial property of pristine HDPE and its nanocomposite was evaluated, and also their mechanical properties were measured after 2 and 4 months. As in vivo experiment, pristine HDPE and its nanocomposite were separately implanted on calvarium bone of rabbits, and tissue inflammation and osteogenesis were investigated after 2, 4, and 6 months. In case of HOB cells treated with HDPE or nanocomposite, as incubation time was increased, cell proliferation, live/dead ratio, and cell viability were decreased. But, the ALP activity and cell adhesion of HOB cells which treated with nanocomposite were raised after increase of incubation time. This study demonstrated that although the mechanical properties of nanocomposite were similar to HDPE sheet, but their antibacterial property was not similar. The in vivo experiment showed that both pristine HDPE and its nanocomposite had same inflammation responses. Interestingly, osteogenesis was observed after 2 months at bone/nanocomposite interface, and was highly increased after 4 and 6 months. It must be noted that such pattern was not seen at bone/HDPE interface. - Highlights: • The effect of various nanoparticles like as Ca{sub 3}(PO{sub 4}){sub 2}, hydroxyapatite, and MgO was studied. • HDPE/TCP/HA/MgO nanocomposite was biocompatible. • The effect of nanoparticles showed high antibacterial property.

In vitro and in vivo evaluation of a new nanocomposite, containing high density polyethylene, tricalcium phosphate, hydroxyapatite, and magnesium oxide nanoparticles

International Nuclear Information System (INIS)

Pourdanesh, Fereydoun; Jebali, Ali; Hekmatimoghaddam, Seyedhossein; Allaveisie, Azra

2014-01-01

In this study, a new nanocomposite, which contained high density polyethylene (HDPE), tricalcium phosphate (Ca 3 (PO 4 ) 2 ) nanoparticles (TCP NPs), hydroxyapatite nanoparticles (HA NPs), and magnesium oxide nanoparticles (MgO NPs) was prepared. As in vitro experiment, human osteoblasts (HOB) cells were exposed to pristine HDPE and its nanocomposite for a period of 1, 4, and 7 days at 37 °C, and then different assays were carried out, including osteoblast cell proliferation, Trypan blue staining, cell viability, alkaline phosphatase (ALP), and cell adhesion. Antibacterial property of pristine HDPE and its nanocomposite was evaluated, and also their mechanical properties were measured after 2 and 4 months. As in vivo experiment, pristine HDPE and its nanocomposite were separately implanted on calvarium bone of rabbits, and tissue inflammation and osteogenesis were investigated after 2, 4, and 6 months. In case of HOB cells treated with HDPE or nanocomposite, as incubation time was increased, cell proliferation, live/dead ratio, and cell viability were decreased. But, the ALP activity and cell adhesion of HOB cells which treated with nanocomposite were raised after increase of incubation time. This study demonstrated that although the mechanical properties of nanocomposite were similar to HDPE sheet, but their antibacterial property was not similar. The in vivo experiment showed that both pristine HDPE and its nanocomposite had same inflammation responses. Interestingly, osteogenesis was observed after 2 months at bone/nanocomposite interface, and was highly increased after 4 and 6 months. It must be noted that such pattern was not seen at bone/HDPE interface. - Highlights: • The effect of various nanoparticles like as Ca 3 (PO 4 ) 2 , hydroxyapatite, and MgO was studied. • HDPE/TCP/HA/MgO nanocomposite was biocompatible. • The effect of nanoparticles showed high antibacterial property

Influence of Cryopreservation Solution on the In Vitro Culture of Skin Tissues Derived from Collared Peccary (Pecari tajacu Linnaeus, 1758).

Science.gov (United States)

Borges, Alana A; Lira, Gabriela P O; Nascimento, Lucas E; Queiroz Neta, Luiza B; Santos, Maria V O; Oliveira, Moacir F; Silva, Alexandre R; Pereira, Alexsandra F

2018-04-01

Skin vitrification is a promising and alternative tool for the conservation of biodiversity, especially for wild mammals, such as collared peccaries. Several factors can affect the success of this procedure, such as the cryoprotectant solution used. Therefore, this study was carried out to compare the efficiency of various vitrification solutions for recovery of viable cells after in vitro culture of cryopreserved skin tissues derived from the collared peccary, aiming to study the application in biobanking, where cellular use is not immediately required. Then, Dulbecco's modified Eagle's medium (DMEM) composed of 2.2 g/L sodium bicarbonate and 10% fetal bovine serum (FBS) was supplemented with 3.0 M ethylene glycol (EG) or 3.0 M dimethyl sulfoxide (DMSO) or 1.5 M EG plus 1.5 M DMSO with or without sucrose (SUC; 0.25 M) to produce six solutions for solid-surface vitrification. After warming, skin tissues were cultured in vitro and recovered cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity for developing the growth curve and determining the population doubling time (PDT), and viability by Trypan Blue. The vitrification did not alter the ability of the tissues to adhere to the culture dish, as well as the day of all explants with cell growth, subconfluence samples, subconfluence total time, and PDT (p > 0.05). Moreover, independent of the cryoprotectant solution used, the vitrification altered the day of all attached explants (p  0.05). Additionally, for viability after the third passage, only the EG-SUC group maintained the cell quality (88.3%), when compared with the nonvitrified (97.8%, p > 0.05). In conclusion, DMEM with 10% FBS, 3.0 M EG, and 0.25 M sucrose was the most efficient solution for vitrifying collared peccary skin tissues, leading to the in vitro culture of viable cells.

TiO{sub 2} nanoparticle-induced ROS correlates with modulated immune cell function

Energy Technology Data Exchange (ETDEWEB)

Maurer-Jones, Melissa A.; Christenson, Jenna R.; Haynes, Christy L., E-mail: chaynes@umn.edu [University of Minnesota, Department of Chemistry (United States)

2012-12-15

Design of non-toxic nanoparticles will be greatly facilitated by understanding the nanoparticle-cell interaction mechanism on a cell function level. Mast cells are important cells for the immune system's first line of defense, and we can utilize their exocytotic behavior as a model cellular function as it is a conserved process across cell types and species. Perturbations in exocytosis can also have implications for whole organism health. One proposed mode of toxicity is nanoparticle-induced reactive oxygen species (ROS), particularly for titanium dioxide (TiO{sub 2}) nanoparticles. Herein, we have correlated changes in ROS with the perturbation of the critical cell function of exocytosis, using UV light to induce greater levels of ROS in TiO{sub 2} exposed cells. The primary culture mouse peritoneal mast cells (MPMCs) were exposed to varying concentrations of TiO{sub 2} nanoparticles for 24 h. ROS content was determined using 2,7-dihydrodichlorofluorescein diacetate (DCFDA). Cellular viability was determined with the MTT and Trypan blue assays, and exocytosis was measured by the analytical electrochemistry technique of carbon-fiber microelectrode amperometry. MPMCs exposed to TiO{sub 2} nanoparticles experienced a dose-dependent increase in total ROS content. While there was minimal impact of ROS on cellular viability, there is a correlation between ROS amount and exocytosis perturbation. As nanoparticle-induced ROS increases, there is a significant decrease (45 %) in the number of serotonin molecules being released during exocytosis, increase (26 %) in the amount of time for each exocytotic granule to release, and decrease (28 %) in the efficiency of granule trafficking and docking. This is the first evidence that nanoparticle-induced ROS correlates with chemical messenger molecule secretion, possibly making a critical connection between functional impairment and mechanisms contributing to that impairment.

The ancient blue oak woodlands of California: longevity and hydroclimatic history

Science.gov (United States)

Stahle, D.W.; Griffin, R.D.; Meko, D.M.; Therrell, M.D.; Edmondson, J.R.; Cleaveland, M.K.; Burnette, D.J.; Abatzoglou, J.T.; Redmond, K.T.; Dettinger, M.D.; Cayan, D.R.

2013-01-01

Ancient blue oak trees are still widespread across the foothills of the Coast Ranges, Cascades, and Sierra Nevada in California. The most extensive tracts of intact old-growth blue oak woodland appear to survive on rugged and remote terrain in the south Coast Ranges and on the foothills west and southwest of Mt. Lassen. In our sampling of old-growth stands, most blue oak appear to have recruited to the canopy in the mid- to late-19th century. The oldest living blue oak tree sampled was over 459-years old and several dead blue oak logs had over 500 annual rings. Precipitation sensitive tree-ring chronologies up to 700-years long have been developed from old blue oak trees and logs. Annual ring-width chronologies of blue oak are strongly correlated with cool season precipitation totals, streamflow in the major rivers of California, and the estuarine water quality of San Francisco Bay. A new network of 36 blue oak chronologies records spatial anomalies in growth that arise from latitudinal changes in the mean storm track and location of landfalling atmospheric rivers. These long, climate-sensitive blue oak chronologies have been used to reconstruct hydroclimatic history in California and will help to better understand and manage water resources. The environmental history embedded in blue oak growth chronologies may help justify efforts to conserve these authentic old-growth native woodlands.

Suppressed speckle contrast of blue light emission out of white lamp with phosphors excited by blue laser diodes for high-brightness lighting applications

Science.gov (United States)

Kinoshita, Junichi; Ikeda, Yoshihisa; Takeda, Yuji; Ueno, Misaki; Kawasaki, Yoji; Matsuba, Yoshiaki; Heike, Atsushi

2012-11-01

The speckle contrast of blue light emission out of high-brightness white lamps using phosphors excited by InGaN/GaN blue laser diodes is evaluated as a measure of coherence. As a result, speckle contrast of as low as 1.7%, the same level as a blue light emitting diode, is obtained. This implies that the original blue laser light can be converted into incoherent light through lamp structures without any dynamic mechanisms. This unique speckle-free performance is considered to be realized by multiple scattering inside the lamp structure, the multi-longitudinal mode operation of the blue laser diodes, and the use of multiple laser diodes. Such almost-incoherent white lamps can be applied for general lighting without any nuisance of speckle noise and should be categorized as lamps rather than lasers in terms of laser safety regulation.

Viability of infrared FEL facilities

International Nuclear Information System (INIS)

Schwettman, H.A.

2004-01-01

Infrared FELs have broken important ground in optical science in the past decade. The rapid development of optical parametric amplifiers and oscillators, and THz sources, however, has changed the competitive landscape and compelled FEL facilities to identify and exploit their unique advantages. The viability of infrared FEL facilities depends on targeting unique world-class science and providing adequate experimental beam time at competitive costs

Heterozygous effects of irradiated chromosomes on viability in Drosophila melanogaster

International Nuclear Information System (INIS)

Simmons, M.J.

1976-01-01

Two large experiments were conducted in order to evaluate the heterozygous effects of irradiated chromosomes on viability. Mutations were accumulated on several hundred second chromosomes by delivering doses of 2,500R over either two or four generations for total x-ray exposures of 5,000R or 10,000R. Chromosomes treated with 5,000R were screened for lethals after the first treatment, and surviving nonlethals were used to generate families of fully treated chromosomes. The members of these families shared the effects of the first irradiation, but differed with respect to those of the second. The chromosomes treated with 10,000R were not grouped into families since mutations were accumulated independently on each chromosome in that experiment. Heterozygous effects on viability of the irradiated chromosomes were tested in both isogenic (homozygous) and nonisogenic (heterozygous) genetic backgrounds. In conjunction with these tests, homozygous viabilities were determined by the marked-inversion technique. This permitted a separation of the irradiated chromosomes into those which were drastic when made homozygous and those which were not. The results indicate that drastic chromosomes have deleterious effects in heterozygous condition, since viability was reduced by 2 to 4 percent in tests performed with the 10,000R chromosomes, and by 1 percent in those involving the 5,000R material. Within a series of tests, the effects were more pronounced when the genetic background was homozygous. These results suggest that the mutants induced by high doses of x-rays are principally drastic ones which show deleterious effects on viability in heterozygous condition

Curcumin Modulates Pancreatic Adenocarcinoma Cell-Derived Exosomal Function

Science.gov (United States)

Osterman, Carlos J. Diaz; Lynch, James C.; Leaf, Patrick; Gonda, Amber; Ferguson Bennit, Heather R.; Griffiths, Duncan; Wall, Nathan R.

2015-01-01

Pancreatic cancer has the highest mortality rates of all cancer types. One potential explanation for the aggressiveness of this disease is that cancer cells have been found to communicate with one another using membrane-bound vesicles known as exosomes. These exosomes carry pro-survival molecules and increase the proliferation, survival, and metastatic potential of recipient cells, suggesting that tumor-derived exosomes are powerful drivers of tumor progression. Thus, to successfully address and eradicate pancreatic cancer, it is imperative to develop therapeutic strategies that neutralize cancer cells and exosomes simultaneously. Curcumin, a turmeric root derivative, has been shown to have potent anti-cancer and anti-inflammatory effects in vitro and in vivo. Recent studies have suggested that exosomal curcumin exerts anti-inflammatory properties on recipient cells. However, curcumin’s effects on exosomal pro-tumor function have yet to be determined. We hypothesize that curcumin will alter the pro-survival role of exosomes from pancreatic cancer cells toward a pro-death role, resulting in reduced cell viability of recipient pancreatic cancer cells. The main objective of this study was to determine the functional alterations of exosomes released by pancreatic cancer cells exposed to curcumin compared to exosomes from untreated pancreatic cancer cells. We demonstrate, using an in vitro cell culture model involving pancreatic adenocarcinoma cell lines PANC-1 and MIA PaCa-2, that curcumin is incorporated into exosomes isolated from curcumin-treated pancreatic cancer cells as observed by spectral studies and fluorescence microscopy. Furthermore, curcumin is delivered to recipient pancreatic cancer cells via exosomes, promoting cytotoxicity as demonstrated by Hoffman modulation contrast microscopy as well as AlamarBlue and Trypan blue exclusion assays. Collectively, these data suggest that the efficacy of curcumin may be enhanced in pancreatic cancer cells through

A binary origin for 'blue stragglers' in globular clusters.

Science.gov (United States)

Knigge, Christian; Leigh, Nathan; Sills, Alison

2009-01-15

Blue stragglers in globular clusters are abnormally massive stars that should have evolved off the stellar main sequence long ago. There are two known processes that can create these objects: direct stellar collisions and binary evolution. However, the relative importance of these processes has remained unclear. In particular, the total number of blue stragglers found in a given cluster does not seem to correlate with the predicted collision rate, providing indirect support for the binary-evolution model. Yet the radial distributions of blue stragglers in many clusters are bimodal, with a dominant central peak: this has been interpreted as an indication that collisions do dominate blue straggler production, at least in the high-density cluster cores. Here we report that there is a clear, but sublinear, correlation between the number of blue stragglers found in a cluster core and the total stellar mass contained within it. From this we conclude that most blue stragglers, even those found in cluster cores, come from binary systems. The parent binaries, however, may themselves have been affected by dynamical encounters. This may be the key to reconciling all of the seemingly conflicting results found to date.

Screening of oral premalignant lesions in smokers using toluidine blue

Directory of Open Access Journals (Sweden)

Yanti Leosari

2009-06-01

Full Text Available Background: A smoker is associated with the risk of developing oral premalignant lesions due to the cacinogenic contents in cigarette. Toluidine blue is a basic chromatic dye used in screening the presence of premalignant lesions due to its ability to detect acidic components in cells and tissues. Purpose: This study was purposed to observe the outcomes of toluidine blue staining on oral mucosa of smokers and non smokers and to find out whether quantity and duration of smoking affect the final results of toluidine blue staining. Methods: Forty male subjects, aged 20-60 years old were involved in this study, consisted of 10 heavy smokers, 10 moderate smokers, 10 light smokers and 10 non smokers. Subjects were instructed to rinse their mouths with mineral water for 20 seconds followed by acetic acid 1% for another 20 seconds. Toluidine blue stain was applied in excess and left on site for 1 minute. Subjects were instructed to rinse with acetic acid 1% and sufficient water consecutively for 20 seconds each. The areas of oral mucosa that stained blue were captured with intraoral camera and transferred to the computer unit. The staining procedure was repeated after 14 days. Results: Chi-square test showed that toluidine blue positive staining dominates the smokers group. Regression and correlation test indicate that Toluidine blue staining is more obvious in subjects who consume more cigarettes. Conclusion: It was concluded that oral mucosa of smokers absorbed more toluidine blue than that of non smokers and retention of toluidine blue is affected by quantity and duration of smoking.

Comparative study of eco- and cytotoxicity during biotransformation of anthraquinone dye Alizarin Blue Black B in optimized cultures of microscopic fungi.

Science.gov (United States)

Rybczyńska-Tkaczyk, Kamila; Święciło, Agata; Szychowski, Konrad A; Korniłłowicz-Kowalska, Teresa

2018-01-01

The aim of this study was to select optimal conditions (C and N sources, initial pH and temperature) for biodecolorization of 0.03% anthraquinone dye Alizarin Blue Black B (ABBB) by microscopic fungi: Haematonectria haematococca BwIII43, K37 and Trichoderma harzianum BsIII33. The phenolic compounds, phytotoxicity (Lepidium sativum L.), biotoxicity (Microtox), cytotoxicity and yeast viability assay were performed to determine the extent of ABBB detoxification. Biodecolorization and detoxification of 0.03% ABBB in H. haematococca BwIII43 and T. harzianum BsIII33 cultures was correlated with extracellular oxidoreductases activity. In turn, secondary products, toxic to human fibroblasts and respiring sod1 Saccharomyces cerevisiae cells, were formed in H. haematococca K37 strain cultures, despite efficient decolorization. Copyright © 2017 Elsevier Inc. All rights reserved.

Evaluation of transport conditions for autologous bone marrow-derived mesenchymal stromal cells for therapeutic application in horses

Directory of Open Access Journals (Sweden)

Miguel Espina

2016-03-01

Full Text Available Background. Mesenchymal stromal cells (MSCs are increasingly used for clinical applications in equine patients. For MSC isolation and expansion, a laboratory step is mandatory, after which the cells are sent back to the attending veterinarian. Preserving the biological properties of MSCs during this transport is paramount. The goal of the study was to compare transport-related parameters (transport container, media, temperature, time, cell concentration that potentially influence characteristics of culture expanded equine MSCs. Methods. The study was arranged in three parts comparing (I five different transport containers (cryotube, two types of plastic syringes, glass syringe, CellSeal, (II seven different transport media, four temperatures (4 °C vs. room temperature; −20 °C vs. −80 °C, four time frames (24 h vs. 48 h; 48 h vs. 72 h, and (III three MSC concentrations (5 × 106, 10 × 106, 20 × 106 MSC/ml. Cell viability (Trypan Blue exclusion; percent and total number viable cell, proliferation and trilineage differentiation capacity were assessed for each test condition. Further, the recovered volume of the suspension was determined in part I. Each condition was evaluated using samples of six horses (n = 6 and differentiation protocols were performed in duplicates. Results. In part I of the study, no significant differences in any of the parameters were found when comparing transport containers at room temperature. The glass syringe was selected for all subsequent evaluations (highest recoverable volume of cell suspension and cell viability. In part II, media, temperatures, or time frames had also no significant influence on cell viability, likely due to the large number of comparisons and small sample size. Highest cell viability was observed using autologous bone marrow supernatant as transport medium, and “transport” at 4 °C for 24 h (70.6% vs. control group 75.3%; this was not significant. Contrary, viability was unacceptably

Origin of faint blue stars

International Nuclear Information System (INIS)

Tutukov, A.; Iungelson, L.

1987-01-01

The origin of field faint blue stars that are placed in the HR diagram to the left of the main sequence is discussed. These include degenerate dwarfs and O and B subdwarfs. Degenerate dwarfs belong to two main populations with helium and carbon-oxygen cores. The majority of the hot subdwarfs most possibly are helium nondegenerate stars that are produced by mass exchange close binaries of moderate mass cores (3-15 solar masses). The theoretical estimates of the numbers of faint blue stars of different types brighter than certain stellar magnitudes agree with star counts based on the Palomar Green Survey. 28 references

Blue light phototherapy for Psoriasis from a systems biology perspective

NARCIS (Netherlands)

Félix Garza, Z.C.; Liebmann, J.; Hilbers, P.A.J.; Riel, van N.A.W.

2014-01-01

This work analyses the effect of UV-free blue light (BL) irradiation of the skin using mathematical modelling. Prior research has shown that blue light reduces the proliferation of keratinocytes by inducing their differentiation, and causes apoptosis of lymphocytes. The effects of blue light on

Blue-noise remeshing with farthest point optimization

KAUST Repository

Yan, Dongming

2014-08-01

In this paper, we present a novel method for surface sampling and remeshing with good blue-noise properties. Our approach is based on the farthest point optimization (FPO), a relaxation technique that generates high quality blue-noise point sets in 2D. We propose two important generalizations of the original FPO framework: adaptive sampling and sampling on surfaces. A simple and efficient algorithm for accelerating the FPO framework is also proposed. Experimental results show that the generalized FPO generates point sets with excellent blue-noise properties for adaptive and surface sampling. Furthermore, we demonstrate that our remeshing quality is superior to the current state-of-the art approaches. © 2014 The Eurographics Association and John Wiley & Sons Ltd.

Blue-noise remeshing with farthest point optimization

KAUST Repository

Yan, Dongming; Guo, Jianwei; Jia, Xiaohong; Zhang, Xiaopeng; Wonka, Peter

2014-01-01

In this paper, we present a novel method for surface sampling and remeshing with good blue-noise properties. Our approach is based on the farthest point optimization (FPO), a relaxation technique that generates high quality blue-noise point sets in 2D. We propose two important generalizations of the original FPO framework: adaptive sampling and sampling on surfaces. A simple and efficient algorithm for accelerating the FPO framework is also proposed. Experimental results show that the generalized FPO generates point sets with excellent blue-noise properties for adaptive and surface sampling. Furthermore, we demonstrate that our remeshing quality is superior to the current state-of-the art approaches. © 2014 The Eurographics Association and John Wiley & Sons Ltd.

Red Light Combined with Blue Light Irradiation Regulates Proliferation and Apoptosis in Skin Keratinocytes in Combination with Low Concentrations of Curcumin.

Directory of Open Access Journals (Sweden)

Tianhui Niu

Full Text Available Curcumin is a widely known natural phytochemical from plant Curcuma longa. In recent years, curcumin has received increasing attention because of its capability to induce apoptosis and inhibit cell proliferation as well as its anti-inflammatory properties in different cancer cells. However, the therapeutic benefits of curcumin are severely hampered due to its particularly low absorption via trans-dermal or oral bioavailability. Phototherapy with visible light is gaining more and more support in dermatological therapy. Red light is part of the visible light spectrum, which is able to deeply penetrate the skin to about 6 mm, and directly affect the fibroblast of the skin dermis. Blue light is UV-free irradiation which is fit for treating chronic inflammation diseases. In this study, we show that curcumin at low concentrations (1.25-3.12 μM has a strong anti-proliferative effect on TNF-α-induced psoriasis-like inflammation when applied in combination with light-emitting-diode devices. The treatment was especially effective when LED blue light at 405 nm was combined with red light at 630 or 660 nm, which markedly amplified the anti-proliferative and apoptosis-inducing effects of curcumin. The experimental results demonstrated that this treatment reduced the viability of human skin keratinocytes, decreased cell proliferation, induced apoptosis, inhibited NF-κB activity and activated caspase-8 and caspase-9 while preserving the cell membrane integrity. Moreover, the combined treatment also down-regulated the phosphorylation level of Akt and ERK. Taken together, our results indicated that the combination of curcumin with LED blue light united red light irradiation can attain a higher efficiency of regulating proliferation and apoptosis in skin keratinocytes.

Red Light Combined with Blue Light Irradiation Regulates Proliferation and Apoptosis in Skin Keratinocytes in Combination with Low Concentrations of Curcumin

Science.gov (United States)

Cai, Qing; Ren, Qu; Wei, Lizhao

2015-01-01

Curcumin is a widely known natural phytochemical from plant Curcuma longa. In recent years, curcumin has received increasing attention because of its capability to induce apoptosis and inhibit cell proliferation as well as its anti-inflammatory properties in different cancer cells. However, the therapeutic benefits of curcumin are severely hampered due to its particularly low absorption via trans-dermal or oral bioavailability. Phototherapy with visible light is gaining more and more support in dermatological therapy. Red light is part of the visible light spectrum, which is able to deeply penetrate the skin to about 6 mm, and directly affect the fibroblast of the skin dermis. Blue light is UV-free irradiation which is fit for treating chronic inflammation diseases. In this study, we show that curcumin at low concentrations (1.25–3.12 μM) has a strong anti-proliferative effect on TNF-α-induced psoriasis-like inflammation when applied in combination with light-emitting-diode devices. The treatment was especially effective when LED blue light at 405 nm was combined with red light at 630 or 660 nm, which markedly amplified the anti-proliferative and apoptosis-inducing effects of curcumin. The experimental results demonstrated that this treatment reduced the viability of human skin keratinocytes, decreased cell proliferation, induced apoptosis, inhibited NF-κB activity and activated caspase-8 and caspase-9 while preserving the cell membrane integrity. Moreover, the combined treatment also down-regulated the phosphorylation level of Akt and ERK. Taken together, our results indicated that the combination of curcumin with LED blue light united red light irradiation can attain a higher efficiency of regulating proliferation and apoptosis in skin keratinocytes. PMID:26382065

THE PITFALLS OF THE BLUE OCEAN STRATEGY CANVAS

DEFF Research Database (Denmark)

Lindgren, Peter; Saghaug, Kristin Margrethe; Clemmensen, Suberia

2009-01-01

Numerous authors have developed a list of tactics and analyticaltechniques to discover new business or new business models (Markides2008)(Johnson 2008). The Blue Ocean strategy (Kim & Mauborgne 2005)have been one of these - probably one of the most important analyticaltechniques related to the area...... of innovation and new business model (BM)innovation since 2005. Today many consultancies and managersresponsible for innovation use the Blue Ocean framework as one of theirtop 5 innovation tools.Wanting to accentuate the importance of analyzing the strategy canvascarefully, when generating new business models...... - it is important tounderstand the very foundation and construction of the strategy canvas -namely value.This paper addresses the question on How is value defined, measured andfrom which viewed in the Blue Ocean theory and framework. How is valuedefined and used by companies using the Blue Ocean strategy...

Blue jay attacks and consumes cedar waxwing

Science.gov (United States)

Daniel Saenz; Joshua B. Pierce

2009-01-01

Blue Jays (Cyanocitta cristata) are known to be common predators on bird nests (Wilcove 1985, Picman and Schriml 1994). In addition to predation on eggs and nestlings, Blue Jays occasionally prey on fledgling and adult birds (Johnson and Johnson 1976, Dubowy 1985). A majority of reports involve predation on House Sparrows (Passer domesticus) and other small birds (...

Detection of viability by percent thallium uptake with conventional thallium scintigraphy

International Nuclear Information System (INIS)

Imai, Kamon; Araki, Yasushi; Horiuchi, Kou-ichi; Yumikura, Sei; Saito, Satoshi; Ozawa, Yukio; Kan-matsuse, Katsuo; Hagiwara, Kazuo.

1994-01-01

Thallium myocardial scintigraphy (TMS) is used for diagnosis of viability in infarcted myocardium before coronary revascularization. Underestimation of viability by TMS has been reported by many investigators. To evaluate viability precisely, thallium re-injection method or 24 hour delayed imaging is performed. However, these techniques are not convenient and are difficult to perform in clinical practice. Percent T1-uptake method was developed for predicting myocardial viability. To evaluate usefulness of this method, TMS was performed before and after PTCA in 23 patients with myocardial infarction. Left ventricle was divided into 3 layers, then each layer was divided into 4 segments (12 segments in total). Forth three segments showed recovery of perfusion on TMS after PTCA. Viability in infarcted myocardium is predicted by 1) redistribution (RD), 2) %T1-uptake≥45% on the image immediately after exercise (TE), and 3) %T1-uptake≥45% on delayed image (TD). Sensitivity was RD: 60%, TE: 90% and TD: 95% (p<0.001 vs. RD). Specificity was RD: 74%, TE: 68%, and TD: 60% (NS). Predictive accuracy (PA) was RD: 69%, TE: 77%, TD: 73% (NS). Compared with RD, %T1-uptake, either TE or TD, increased sensitivity with slightly improved PA, but decreased specificity slightly. Therefore %T1-uptake would be a sensitive and useful predictor to find patients who are most likely to benefit from re-vascularization. (author)

BlueHealth: a study programme protocol for mapping and quantifying the potential benefits to public health and well-being from Europe's blue spaces.

Science.gov (United States)

Grellier, James; White, Mathew P; Albin, Maria; Bell, Simon; Elliott, Lewis R; Gascón, Mireia; Gualdi, Silvio; Mancini, Laura; Nieuwenhuijsen, Mark J; Sarigiannis, Denis A; van den Bosch, Matilda; Wolf, Tanja; Wuijts, Susanne; Fleming, Lora E

2017-06-14

Proximity and access to water have long been central to human culture and accordingly deliver countless societal benefits. Over 200 million people live on Europe's coastline, and aquatic environments are the top recreational destination in the region. In terms of public health, interactions with 'blue space' (eg, coasts, rivers, lakes) are often considered solely in terms of risk (eg, drowning, microbial pollution). Exposure to blue space can, however, promote health and well-being and prevent disease, although underlying mechanisms are poorly understood. The BlueHealth project aims to understand the relationships between exposure to blue space and health and well-being, to map and quantify the public health impacts of changes to both natural blue spaces and associated urban infrastructure in Europe, and to provide evidence-based information to policymakers on how to maximise health benefits associated with interventions in and around aquatic environments. To achieve these aims, an evidence base will be created through systematic reviews, analyses of secondary data sets and analyses of new data collected through a bespoke international survey and a wide range of community-level interventions. We will also explore how to deliver the benefits associated with blue spaces to those without direct access through the use of virtual reality. Scenarios will be developed that allow the evaluation of health impacts in plausible future societal contexts and changing environments. BlueHealth will develop key inputs into policymaking and land/water-use planning towards more salutogenic and sustainable uses of blue space, particularly in urban areas. Throughout the BlueHealth project, ethics review and approval are obtained for all relevant aspects of the study by the local ethics committees prior to any work being initiated and an ethics expert has been appointed to the project advisory board. So far, ethical approval has been obtained for the BlueHealth International Survey and

Northeast Atlantic blue whiting

OpenAIRE

Heino, Mikko

2010-01-01

Heino, M. 2010. Northeast Atlantic blue whiting. In Life cycle spatial patterns of small pelagic fish in the Northeast Atlantic, pp. 59-64. Ed by P. Petitgas. ICES Cooperative Research Report 306. ICES, Copenhagen.

Manajemen Strategi Pengembangan Pariwisata dengan Pendekatan Blue Ocean Strategy

OpenAIRE

Muzha, Vianda Kushardianti

2015-01-01

Persaingan industri pariwisata di Indonesia saat ini sangatlah ketat, setiap daerah berlomba untuk menonjolkan keunikannya tersendiri. Dengan adanya persaingan yang sangat ketat tersebut, Kota Batu berusaha keluar dari persaingan (red ocean) dengan menciptkan inovasi baru melalui konsep Blue Ocean Strategy. Blue Ocean Strategy adalah istilah dalam ilmu manajemen strategi yang merujuk pada siasat untuk menciptakan pasar baru yang belum dipenuhi persaingan yang ketat. Blue Ocean Strategy pada d...

Plant responses to UV and blue light: biochemical and genetic approaches

International Nuclear Information System (INIS)

Jenkins, G.I.; Christie, J.M.; Fuglevand, G.; Long, J.C.; Jackson, J.A.

1995-01-01

UV and blue light control many aspects of plant growth and development. It is evident that several different photoreceptors mediate responses to UV and blue light, and there are reports of the functional and biochemical characterisation of a putative photoreceptor for phototropism and of the functional and molecular characterisation of the CRY1 photoreceptor, encoded by the Arabidopsis HY4 gene. The CRY1 photoreceptor mediates extension growth and gene expression responses to UV-A/blue light presumably through different or branching signal transduction pathways. Progress has been made in cell physiological and biochemical studies of UV/blue light signal transduction, but much remains to be done to relate candidate UV/blue signal transduction events to particular photoreceptors and responses. The application of a genetic approach in Arabidopsis has been responsible for many advances in understanding UV/blue responses, but further UV-B, UV-A and blue light response mutants need to be isolated. (author)

BlueHealth: a study programme protocol for mapping and quantifying the potential benefits to public health and well-being from Europe’s blue spaces

Science.gov (United States)

White, Mathew P; Albin, Maria; Bell, Simon; Elliott, Lewis R; Gascón, Mireia; Gualdi, Silvio; Mancini, Laura; Nieuwenhuijsen, Mark J; Sarigiannis, Denis A; van den Bosch, Matilda; Wolf, Tanja; Wuijts, Susanne; Fleming, Lora E

2017-01-01

Introduction Proximity and access to water have long been central to human culture and accordingly deliver countless societal benefits. Over 200 million people live on Europe’s coastline, and aquatic environments are the top recreational destination in the region. In terms of public health, interactions with ‘blue space’ (eg, coasts, rivers, lakes) are often considered solely in terms of risk (eg, drowning, microbial pollution). Exposure to blue space can, however, promote health and well-being and prevent disease, although underlying mechanisms are poorly understood. Aims and methods The BlueHealth project aims to understand the relationships between exposure to blue space and health and well-being, to map and quantify the public health impacts of changes to both natural blue spaces and associated urban infrastructure in Europe, and to provide evidence-based information to policymakers on how to maximise health benefits associated with interventions in and around aquatic environments. To achieve these aims, an evidence base will be created through systematic reviews, analyses of secondary data sets and analyses of new data collected through a bespoke international survey and a wide range of community-level interventions. We will also explore how to deliver the benefits associated with blue spaces to those without direct access through the use of virtual reality. Scenarios will be developed that allow the evaluation of health impacts in plausible future societal contexts and changing environments. BlueHealth will develop key inputs into policymaking and land/water-use planning towards more salutogenic and sustainable uses of blue space, particularly in urban areas. Ethics and dissemination Throughout the BlueHealth project, ethics review and approval are obtained for all relevant aspects of the study by the local ethics committees prior to any work being initiated and an ethics expert has been appointed to the project advisory board. So far, ethical approval

Present trends in the detection of myocardial viability using nuclear cardiology tests

International Nuclear Information System (INIS)

Peix Gonzalez, Amalia; Garcia Barreto, David

1999-01-01

The myocardial viability diagnosis is important for those who will undergo myocardial revascularization whether by surgery or coronary angioplasty. Our purpose is to present some of the present trends in the detection of myocardial viability using nuclear cardiology tests. Emphasis is made on the estimation of radiopharmaceutical uptake and the use of vasodilators in perfusion scintigraphy mainly with technetium-labeled compounds. Also, the current possibilities for a myocardial metabolism study using single-photon emission-computed tomography as well as some clinical implications of myocardial viability are set forth

Study of internalization and viability of multimodal nanoparticles for labeling of human umbilical cord mesenchymal stem cells; Estudo de internalizacao e viabilidade de nanoparticulas multimodal para marcacao de celulas-tronco mesenquimais de cordao umbilical humano

Energy Technology Data Exchange (ETDEWEB)

Miyaki, Liza Aya Mabuchi [Faculdade de Enfermagem, Hospital Israelita Albert Einstein - HIAE, Sao Paulo, SP (Brazil); Sibov, Tatiana Tais; Pavon, Lorena Favaro; Mamani, Javier Bustamante; Gamarra, Lionel Fernel, E-mail: tatianats@einstein.br [Instituto do Cerebro - InCe, Hospital Israelita Albert Einstein - HIAE, Sao Paulo, SP (Brazil)

2012-04-15

Objective: To analyze multimodal magnetic nanoparticles-Rhodamine B in culture media for cell labeling, and to establish a study of multimodal magnetic nanoparticles-Rhodamine B detection at labeled cells evaluating they viability at concentrations of 10 {mu}g Fe/mL and 100{mu}g Fe/mL. Methods: We performed the analysis of stability of multimodal magnetic nanoparticles-Rhodamine B in different culture media; the mesenchymal stem cells labeling with multimodal magnetic nanoparticles-Rhodamine B; the intracellular detection of multimodal magnetic nanoparticles-Rhodamine B in mesenchymal stem cells, and assessment of the viability of labeled cells by kinetic proliferation. Results: The stability analysis showed that multimodal magnetic nanoparticles-Rhodamine B had good stability in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium. The mesenchymal stem cell with multimodal magnetic nanoparticles-Rhodamine B described location of intracellular nanoparticles, which were shown as blue granules co-localized in fluorescent clusters, thus characterizing magnetic and fluorescent properties of multimodal magnetic nanoparticles Rhodamine B. Conclusion: The stability of multimodal magnetic nanoparticles-Rhodamine B found in cultured Dulbecco's Modified Eagle's-Low Glucose medium and RPMI 1640 medium assured intracellular mesenchymal stem cells labeling. This cell labeling did not affect viability of labeled mesenchymal stem cells since they continued to proliferate for five days. (author)

The Blue Hook Populations of Massive Globular Clusters

Science.gov (United States)

Brown, Thomas

2006-07-01

Blue hook stars are a class of hot { 35,000 K} subluminous horizontal branch stars that have been recently discovered using HST ultraviolet images of the globular clusters omega Cen and NGC 2808. These stars occupy a region of the HR diagram that is unexplained by canonical stellar evolution theory. Using new theoretical evolutionary and atmospheric models, we have shown that the blue hook stars are very likely the progeny of stars that undergo extensive internal mixing during a late helium core flash on the white dwarf cooling curve. This "flash mixing" produces an enormous enhancement of the surface helium and carbon abundances, which suppresses the flux in the far ultraviolet. Although flash mixing is more likely to occur in stars that are born with high helium abundances, a high helium abundance, by itself, does not explain the presence of a blue hook population - flash mixing of the envelope is required. We propose ACS ultraviolet {SBC/F150LP and HRC/F250W} observations of the five additional globular clusters for which the presence of blue hook stars is suspected from longer wavelength observations. Like omega Cen and NGC 2808, these five targets are also among the most massive globular clusters, because less massive clusters show no evidence for blue hook stars. Because our targets span 1.5 dex in metallicity, we will be able to test our prediction that flash-mixing should be less drastic in metal-rich blue hook stars. In addition, our observations will test the hypothesis that blue hook stars only form in globular clusters massive enough to retain the helium-enriched ejecta from the first stellar generation. If this hypothesis is correct, then our observations will yield important constraints on the chemical evolution and early formation history in globular clusters, as well as the role of helium self-enrichment in producing blue horizontal branch morphologies and multiple main sequence turnoffs. Finally, our observations will provide new insight into the

A prospective randomized trial comparing patent blue and methylene blue for the detection of the sentinel lymph node in breast cancer patients

Directory of Open Access Journals (Sweden)

Régis Resende Paulinelli

Full Text Available Summary Introduction: Methylene blue is more widely available and less expensive than patent blue, with an apparently lower risk of anaphylaxis. Objective: The two dyes were compared regarding detection of the sentinel lymph node (SLN. Method: A prospective, randomized trial involved 142 patients with invasive breast carcinoma. Sixty-nine (49.3% assigned to patent blue (group A and 71 (50.70% to methylene blue (group B. Thirty-five patients (25.0% were clinical stage III or IV; 55 (38.7% had axillary lymph nodes affected; and 69 (49.3% underwent neoadjuvant chemotherapy. Two patients were excluded because the dye type was not recorded. Results: Patients and tumor characteristics were similar in both groups. SLNs were identified in 47 women (68.1% in group A and 43 (60.6% in group B (p=0.35. SLNs were affected in 22 cases (51.2% in group A and 21 (48.8% in group B (p=0.62. The SLN was the only node affected in 12 cases (54.5% in group A and six (33.3% in group B (p=0.18. The time and degree of difficulty involved in identifying the SLN were similar in both groups. There were no complications or allergies. Conclusion: Methylene blue performed as well as patent blue in identifying the SLN in breast cancer patients.

Blue Ribbon Panel Report

Science.gov (United States)

An NCI Cancer Currents blog by the NCI acting director thanking the cancer community for contributing to the Cancer Moonshot Blue Ribbon Panel report, which was presented to the National Cancer Advisory Board on September 7.

Twisted bilayer blue phosphorene: A direct band gap semiconductor

Science.gov (United States)

Ospina, D. A.; Duque, C. A.; Correa, J. D.; Suárez Morell, Eric

2016-09-01

We report that two rotated layers of blue phosphorene behave as a direct band gap semiconductor. The optical spectrum shows absorption peaks in the visible region of the spectrum and in addition the energy of these peaks can be tuned with the rotational angle. These findings makes twisted bilayer blue phosphorene a strong candidate as a solar cell or photodetection device. Our results are based on ab initio calculations of several rotated blue phosphorene layers.

Fluctuating and Directional Asymmetry of the Blue Mussel (

NARCIS (Netherlands)

Lajus, D.; Katolikova, M.; Strelkov, P.; Hummel, H.

2015-01-01

In this work we examined morphological variation at different levels to study performance and population structuring of the blue mussel Mytilus edulis. Our objectives were: (i) to develop an integrated technique for analyzing morphological variation in blue mussels and, based on this technique; (ii)

The Blue Marble

Science.gov (United States)

2002-01-01

This spectacular Moderate Resolution Imaging Spectroradiometer (MODIS) 'blue marble' image is based on the most detailed collection of true-color imagery of the entire Earth to date. Using a collection of satellite-based observations, scientists and visualizers stitched together months of observations of the land surface, oceans, sea ice, and clouds into a seamless, true-color mosaic of every square kilometer (.386 square mile) of our planet. Most of the information contained in this image came from MODIS, illustrating MODIS' outstanding capacity to act as an integrated tool for observing a variety of terrestrial, oceanic, and atmospheric features of the Earth. The land and coastal ocean portions of this image is based on surface observations collected from June through September 2001 and combined, or composited, every eight days to compensate for clouds that might block the satellite's view on any single day. Global ocean color (or chlorophyll) data was used to simulate the ocean surface. MODIS doesn't measure 3-D features of the Earth, so the surface observations were draped over topographic data provided by the U.S. Geological Survey EROS Data Center. MODIS observations of polar sea ice were combined with observations of Antarctica made by the National Oceanic and Atmospheric Administration's AVHRR sensor-the Advanced Very High Resolution Radiometer. The cloud image is a composite of two days of MODIS imagery collected in visible light wavelengths and a third day of thermal infra-red imagery over the poles. A large collection of imagery based on the blue marble in a variety of sizes and formats, including animations and the full (1 km) resolution imagery, is available at the Blue Marble page. Image by Reto Stockli, Render by Robert Simmon. Based on data from the MODIS Science Team

Sustainable model for financial viability of decentralized biomass gasifier based power projects

NARCIS (Netherlands)

Palit, D.; Malhotra, R.; Kumar, Atul

2011-01-01

This paper made a modest attempt for designing a sustainable model for financial viability of biomass gasifier power projects for enhancing electricity access in India and other developing countries. For long term sustainability of distributed generation projects in remote rural areas, viability

Borax methylene blue: a spectroscopic and staining study.

Science.gov (United States)

Donaldson, P T; Russo, A; Reynolds, C; Lillie, R D

1978-07-01

Borax methylene blue is quite stable at room temperatures of 22-25 C. At 30 C polychroming is slow; during 50 days in a water bath at this temperature the absorption peak moves from 665 to 656 nm. At 35 C, the absorption peak reaches 660 nm in 7 days, 654 nm in 14. At 60 C polychroming is rapid, the absorption peak reaching 640-620 nm in 3 days. When the pH of the borax methylene blue solutions, normally about 9.0, is adjusted to pH 6.5, the absorption peak remains at 665 nm even when incubated at 60 C for extended periods. When used as a blood stain 0.4 ml borax methylene blue (1% methylene blue in 1% borax), 4 ml acetone, 2 ml borax-acid phosphate buffer to bring the solution to pH 6.5, and distilled water to make 40 ml, with 0.2 ml 1% eosin added just before using, an excellent Nocht-Giemsa type stain is achieved after 30 minutes staining. The material plasmodia P. falciparum, P. vivax, and P. berghei stain moderate blue with dark red chromatin and green to black pigment granules. The study confirms Malachowski's 1891 results and explains Gautier's 1896-98 failure to duplicate it.

The BlueSky Smoke Modeling Framework: Recent Developments

Science.gov (United States)

Sullivan, D. C.; Larkin, N.; Raffuse, S. M.; Strand, T.; ONeill, S. M.; Leung, F. T.; Qu, J. J.; Hao, X.

2012-12-01

BlueSky systems—a set of decision support tools including SmartFire and the BlueSky Framework—aid public policy decision makers and scientific researchers in evaluating the air quality impacts of fires. Smoke and fire managers use BlueSky systems in decisions about prescribed burns and wildland firefighting. Air quality agencies use BlueSky systems to support decisions related to air quality regulations. We will discuss a range of recent improvements to the BlueSky systems, as well as examples of applications and future plans. BlueSky systems have the flexibility to accept basic fire information from virtually any source and can reconcile multiple information sources so that duplication of fire records is eliminated. BlueSky systems currently apply information from (1) the National Oceanic and Atmospheric Administration's (NOAA) Hazard Mapping System (HMS), which represents remotely sensed data from the Moderate Resolution Imaging Spectroradiometer (MODIS), Advanced Very High Resolution Radiometer (AVHRR), and Geostationary Operational Environmental Satellites (GOES); (2) the Monitoring Trends in Burn Severity (MTBS) interagency project, which derives fire perimeters from Landsat 30-meter burn scars; (3) the Geospatial Multi-Agency Coordination Group (GeoMAC), which produces helicopter-flown burn perimeters; and (4) ground-based fire reports, such as the ICS-209 reports managed by the National Wildfire Coordinating Group. Efforts are currently underway to streamline the use of additional ground-based systems, such as states' prescribed burn databases. BlueSky systems were recently modified to address known uncertainties in smoke modeling associated with (1) estimates of biomass consumption derived from sparse fuel moisture data, and (2) models of plume injection heights. Additional sources of remotely sensed data are being applied to address these issues as follows: - The National Aeronautics and Space Administration's (NASA) Tropical Rainfall Measuring Mission

Effect of storage temperature on Streptococcus mutans viability

Directory of Open Access Journals (Sweden)

Ana Lídia Soares COTA

2018-04-01

Full Text Available Abstract Introduction Proper storage conditions and maintenance of viable biological material plays an important role in microbiological research, allowing for the opportunity to conduct future studies. Objective To evaluate the viability of Streptococcus mutans strains that were previously grown and stored under different temperatures for approximately eight years. Material and method In this study, we evaluated 393 bacterial isolates that were stored in a freezer at -80°C (G1 and 200 isolates stored in a freezer at -20°C (G2. Aliquots of each sample were plated on blood agar and mitis-salivarius bacitracin sucrose agar-solidified medium. After incubating under microaerophilic conditions in an incubator at 37°C for 72 hours, the presence, morphology and purity of bacterial growth was observed. The data were analyzed by means of descriptive statistics. Result Microbial viability was observed in almost all samples (99.7% in G1, whereas all isolates stored at -20°C were considered inviable. Conclusion The viability of S. mutans is influenced by the storage temperature of the samples, and the strains remain viable when stored under ideal temperature conditions (-80°C, even when stored for a long period of time.

Assessment of myocardial perfusion and metabolism for assessment of myocardial viability

International Nuclear Information System (INIS)

Beller, G.

1996-01-01

Identifying preserved myocardial viability in the presence of severe regional left ventricular dysfunction is becoming increasingly more important for clinical decision-making to better select those patients with coronary artery disease who will benefit most from revascularization. 201 Tl remains the most commonly employed radionuclide for detecting both ischemia and viability. A severe persistent defect with 201 Tl uptake compared to peak to improved perfusion and corresponding improved function after revascularisation. Detection of defect reversibility on 201 Tl imaging is enhanced by 'reinjection' of a second 201 Tl dose after acquisition of redistribution images. Initial and 4-hour rest/redistribution imaging has proven most usefull for detection of viability in the resting state in patients with ischemic cardiomyopathy. The greater the extent of preoperative viability, the greater is the improvement in regional and global function after revascularisation. 99 Tc sestamibi has also been demonstrated to be extracted by myocardial cells in proportion to regional blood flow in the presence of viable myocities. Although this agrnt does not redistribute after intravenous injection, its >50% uptake of the tracer implies viablility and predicts improved regional function after revascularisation. Finally positron emission tomography with 18 F fluorodeoxoglucose (FDG) is perhaps the most sensitive noninvasive imaging technique for detection of viability in stunned or hibernating myocardium. A mismatch pattern between regional flow and FDG uptake as approximately an 80-85% positive preicted value for predicting improved function in asynergic myocardial regions after revascualarisation

A Survey of Blue-Noise Sampling and Its Applications

KAUST Repository

Yan, Dongming; Guo, Jian-Wei; Wang, Bin; Zhang, Xiao-Peng; Wonka, Peter

2015-01-01

In this paper, we survey recent approaches to blue-noise sampling and discuss their beneficial applications. We discuss the sampling algorithms that use points as sampling primitives and classify the sampling algorithms based on various aspects, e.g., the sampling domain and the type of algorithm. We demonstrate several well-known applications that can be improved by recent blue-noise sampling techniques, as well as some new applications such as dynamic sampling and blue-noise remeshing.

A Survey of Blue-Noise Sampling and Its Applications

KAUST Repository

Yan, Dongming

2015-05-05

In this paper, we survey recent approaches to blue-noise sampling and discuss their beneficial applications. We discuss the sampling algorithms that use points as sampling primitives and classify the sampling algorithms based on various aspects, e.g., the sampling domain and the type of algorithm. We demonstrate several well-known applications that can be improved by recent blue-noise sampling techniques, as well as some new applications such as dynamic sampling and blue-noise remeshing.

Aspen biology, community classification, and management in the Blue Mountains

Science.gov (United States)

David K. Swanson; Craig L. Schmitt; Diane M. Shirley; Vicky Erickson; Kenneth J. Schuetz; Michael L. Tatum; David C. Powell

2010-01-01

Quaking aspen (Populus tremuloides Michx.) is a valuable species that is declining in the Blue Mountains of northeastern Oregon. This publication is a compilation of over 20 years of aspen management experience by USDA Forest Service workers in the Blue Mountains. It includes a summary of aspen biology and occurrence in the Blue Mountains, and a...

Implementation of EAM and FS potentials in HOOMD-blue

Science.gov (United States)

Yang, Lin; Zhang, Feng; Travesset, Alex; Wang, Caizhuang; Ho, Kaiming

HOOMD-blue is a general-purpose software to perform classical molecular dynamics simulations entirely on GPUs. We provide full support for EAM and FS type potentials in HOOMD-blue, and report accuracy and efficiency benchmarks, including comparisons with the LAMMPS GPU package. Two problems were selected to test the accuracy: the determination of the glass transition temperature of Cu64.5Zr35.5 alloy using an FS potential and the calculation of pair distribution functions of Ni3Al using an EAM potential. In both cases, the results using HOOMD-blue are indistinguishable from those obtained by the GPU package in LAMMPS within statistical uncertainties. As tests for time efficiency, we benchmark time-steps per second using LAMMPS GPU and HOOMD-blue on one NVIDIA Tesla GPU. Compared to our typical LAMMPS simulations on one CPU cluster node which has 16 CPUs, LAMMPS GPU can be 3-3.5 times faster, and HOOMD-blue can be 4-5.5 times faster. We acknowledge the support from Laboratory Directed Research and Development (LDRD) of Ames Laboratory.

Comparison of tissue viability imaging and colorimetry: skin blanching.

Science.gov (United States)

Zhai, Hongbo; Chan, Heidi P; Farahmand, Sara; Nilsson, Gert E; Maibach, Howard I

2009-02-01

Operator-independent assessment of skin blanching is important in the development and evaluation of topically applied steroids. Spectroscopic instruments based on hand-held probes, however, include elements of operator dependence such as difference in applied pressure and probe misalignment, while laser Doppler-based methods are better suited for demonstration of skin vasodilatation than for vasoconstriction. To demonstrate the potential of the emerging technology of Tissue Viability Imaging (TiVi) in the objective and operator-independent assessment of skin blanching. The WheelsBridge TiVi600 Tissue Viability Imager was used for quantification of human skin blanching with the Minolta chromameter CR 200 as an independent colorimeter reference method. Desoximetasone gel 0.05% was applied topically on the volar side of the forearm under occlusion for 6 h in four healthy adults. In a separate study, the induction of blanching in the occlusion phase was mapped using a transparent occlusion cover. The relative uncertainty in the blanching estimate produced by the Tissue Viability Imager was about 5% and similar to that of the chromameter operated by a single user and taking the a(*) parameter as a measure of blanching. Estimation of skin blanching could also be performed in the presence of a transient paradoxical erythema, using the integrated TiVi software. The successive induction of skin blanching during the occlusion phase could readily be mapped by the Tissue Viability Imager. TiVi seems to be suitable for operator-independent and remote mapping of human skin blanching, eliminating the main disadvantages of methods based on hand-held probes.

Collecting and displaying well-site data using BlueGauge technology

Energy Technology Data Exchange (ETDEWEB)

Morley, D.; Thiessen, N. [TR Labs, Calgary, AB (Canada)

2004-07-01

BlueGauge is a wireless technology solution for collecting and displaying well-site data, using off-the-shelf components and working at a distance of up to 100 meters. In addition, BlueGauge supports up to 255 sensors that monitor pressure, flow, or temperature. It was developed by TRLabs for an Edmonton-based company, for whom the prototype was built and tested. This presentation outlined the BlueGauge system architecture and hardware prototypes were demonstrated, including transceivers, embedded computers and pressure transducers. An overview of the software and its operating functions was presented, including screen shots. Operational benefits of BlueGauge technology include increased productivity through gathering process data quickly and easily, and the elimination of data entry errors. Safety was considered to be a primary benefit of BlueGauge equipment, particularly for monitoring dials and gauges in hard-to-access or hazardous locations. An additional safety benefit was the fact that BlueGauge has the ability to alert personnel when readings are outside of normal operating ranges. Details of installation and maintenance benefits include: ease of installation and relocation; support towards auto-discovery of new sensors; and the fact that no instrumentation cables or radio licence are required. Future plans were outlined and included a search for other companies interested in commercializing BlueGauge technology. tabs, figs.

Economic Viability of Brewery Spent Grain as a Biofuel

Energy Technology Data Exchange (ETDEWEB)

Morrow, Charles [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2016-01-01

This report summarizes an investigation into the technical feasibility and economic viability of use grain wastes from the beer brewing process as fuel to generate the heat needed in subsequent brewing process. The study finds that while use of spent grain as a biofuel is technically feasible, the economics are not attractive. Economic viability is limited by the underuse of capital equipment. The investment in heating equipment requires a higher utilization that the client brewer currently anticipates. It may be possible in the future that changing factors may swing the decision to a more positive one.

Viability Assessment Volume 1

International Nuclear Information System (INIS)

1998-01-01

Since May 1996, under its draft Civilian Radioactive Waste Management Program Plan (DOE 1996), DOE has been carrying out a 5-year program of work to support the decision in 2001 by the Secretary of Energy on whether or not to recommend the site to the President. Part of this program was to address major unresolved technical issues and to complete an assessment of the viability of the Yucca Mountain site by 1998. Affirming the DOE plans, Congress directed DOE in the 1997 Energy and Water Development Appropriations Act to provide a viability assessment of the Yucca Mountain site to Congress and the President. This Viability Assessment (VA) document is the DOE report to Congress and the President. They are expected to use the VA to make an informed decision about program direction and funding. Drawing on 15 years of scientific investigation and design work at Yucca Mountain, the VA summarizes a large technical basis of field investigations, laboratory tests, models, analyses, and engineering, described in cited references. The VA identifies the major uncertainties relevant to the technical defensibility of DOE analyses and designs, the DOE approach to managing these uncertainties, and the status of work toward the site recommendation and LA. The VA also identifies DOE plans for the remaining work, and the estimated costs of completing an LA and constructing and operating a repository. The attention to uncertainties is important because DOE must evaluate how the repository will perform during the next 10,000 years or longer. Uncertainties exist because of variability in the natural (geologic and hydrologic) systems at Yucca Mountain and because of imperfect scientific understanding of the natural processes that might affect the repository system. This is Volume 1 and it covers, Introduction and Site Characteristics, includes a high-level summary of the results of the VA and some additional background information. (The overview is bound separately.) Section 1 of Volume

Influence of harvesting and processing methods on organic viability of soybean seed

Directory of Open Access Journals (Sweden)

Đukanović Lana

2000-01-01

Full Text Available Organic viability of soybean seed for three soybean varieties - elite (Bosa, ZPS 015 and Nena depending on methods of manipulation with seeds during harvesting and processing phase were determined in this paper. Trial was conducted in Zemun Polje during 1999; manual and mechanized harvesting or processing methods were applied. Seed germination was tested using ISTA methods (Standard method and Cold test. Following parameters were evaluated: germination viability, germination, rate-speed of emergence, length of hypocotile and main root Rate-speed of emergence was based on number of emerged plants per day. Length of hypocotile or root and percent of germination determined vigour index. Based on obtained results it maybe concluded that methods of seed manipulation during harvesting or processing phase were influenced on soybean seed quality parameters evaluated. Ways of seed manipulation - methods evaluated were influenced organic viability of soybean seed by decreasing germination viability, total germination and length of main root.

Geology along the Blue Ridge Parkway in Virginia

Science.gov (United States)

Carter, Mark W.; Southworth, C. Scott; Tollo, Richard P.; Merschat, Arthur J.; Wagner, Sara; Lazor, Ava; Aleinikoff, John N.

2017-01-01

Detailed geologic mapping and new SHRIMP (sensitive high-resolution ion microprobe) U-Pb zircon, Ar/Ar, Lu-Hf, 14C, luminescence (optically stimulated), thermochronology (fission-track), and palynology reveal the complex Mesoproterozoic to Quaternary geology along the ~350 km length of the Blue Ridge Parkway in Virginia. Traversing the boundary of the central and southern Appalachians, rocks along the parkway showcase the transition from the para-autochthonous Blue Ridge anticlinorium of northern and central Virginia to the allochthonous eastern Blue Ridge in southern Virginia. From mile post (MP) 0 near Waynesboro, Virginia, to ~MP 124 at Roanoke, the parkway crosses the unconformable to faulted boundary between Mesoproterozoic basement in the core of the Blue Ridge anticlinorium and Neoproterozoic to Cambrian metasedimentary and metavolcanic cover rocks on the western limb of the structure. Mesoproterozoic basement rocks comprise two groups based on SHRIMP U-Pb zircon geochronology: Group I rocks (1.2-1.14 Ga) are strongly foliated orthogneisses, and Group II rocks (1.08-1.00 Ga) are granitoids that mostly lack obvious Mesoproterozoic deformational features.Neoproterozoic to Cambrian cover rocks on the west limb of the anticlinorium include the Swift Run and Catoctin Formations, and constituent formations of the Chilhowee Group. These rocks unconformably overlie basement, or abut basement along steep reverse faults. Rocks of the Chilhowee Group are juxtaposed against Cambrian rocks of the Valley and Ridge province along southeast- and northwest-dipping, high-angle reverse faults. South of the James River (MP 64), Chilhowee Group and basement rocks occupy the hanging wall of the nearly flat-lying Blue Ridge thrust fault and associated splays.South of the Red Valley high-strain zone (MP 144.5), the parkway crosses into the wholly allochthonous eastern Blue Ridge, comprising metasedimentary and meta-igneous rocks assigned to the Wills Ridge, Ashe, and Alligator

Liquid crystal blue phases: stability, field effects and alignment

OpenAIRE

Gleeson, HF; Miller, RJ; Tian, L; Görtz, V; Goodby, JW

2015-01-01

The blue phases are fascinating structures in liquid crystals, fluids that exhibit cubic structures that have true crystalline order. The blue phases were discovered in the 1970s and were the subject of extensive research in the 1980s, when a deep understanding of many of their properties was established. The discovery that the blue phases could be stabilised to exist over wide temperature ranges meant that they became more than scientific curiosities and led to a recent resurgence in researc...

Service design as an approach for recognizing blue ocean

OpenAIRE

Koskelo, Minna

2013-01-01

This thesis aims to show that service design as an approach can be utilized for discovering new market space, referred as blue ocean and new business opportunities, and thus to show that service design can be seen as an approach for bringing the new logic for value creation into strategic level. From the field of strategic management the theory of blue ocean strategy was chosen for presenting the strategic outcome and further since there are similarities be-tween blue ocean strategy approach ...

“Key to the highway”: blues records and the great migration

Directory of Open Access Journals (Sweden)

Louis Mazzari

2012-01-01

Full Text Available This paper looks at the way “race record” blues of the 1920s and 1930s reinforced the decision of poor farmers, sharecroppers, and working men and women to move to the cities of the North. The theme is the way black southerners used the blues as the soundtrack of the Great Migration. In a sense, the Delta blues was a musical travel narrative for tens of thousands of people who were leaving the rural South for an unknown, modern and industrial future. The paper will explore blues music as an expression of the fluidity of African American society and culture during the Great Depression.While avoiding direct protest, blues singers and musicians—first women, later men—crafted an art form and employed the technology of the phonograph to encourage freedom of movement and choice. At the moment the “race record” industry was being born, and black farmers and families were quietly picking up and leaving the South, the music they traveled with was the blues. The paper will look at examples of blues singers whose records dealt specifically with the Great Migration and consider their influence on listeners.Cet article considère la manière dont le « race-record » blues des années 1920 et 1930 a renforcé la décision des agriculteurs, des métayers, et des ouvriers Afro-Américains d’émigrer vers les villes du nord. L’objectif général est de montrer comment les Afro-Américains se sont servi du blues comme accompagnement musical pour cette « grande migration ». En un sens, les vinyles de blues représentaient un récit de voyage musical pour des dizaines de milliers de personnes. Les chanteurs et les musiciens de blues ont conçu leur art et ont utilisé la technologie du phonographe en ayant pour but d’encourager la liberté de mouvement et de choix. Enfin, cet article examine en détail des exemples de disques de blues qui traitaient directement de la « grande migration », ainsi que leurs influences sur les musiciens et

Blue-green photoluminescence in MCM-41 mesoporous nanotubes

CERN Document Server

Shen, J L; Lui, Y L; Cheng, P W; Cheng, C F

2003-01-01

Different photoluminescence (PL) techniques have been used to study the blue-green emission from siliceous MCM-41 nanotubes. It was found that the intensity of the blue-green PL is enhanced by rapid thermal annealing (RTA). This enhancement is explained by the generation of twofold-coordinated Si centres and non-bridging oxygen hole centres, in line with the surface properties of MCM-41. On the basis of the analysis of the PL following RTA, polarized PL, and PL excitation, we suggest that the triplet-to-singlet transition of twofold-coordinated silicon centres is responsible for the blue-green PL in MCM-41 nanotubes. (letter to the editor)

Adsorption of methylene blue onto treated activated carbon

International Nuclear Information System (INIS)

Yamin Yasin; Mohd Zobir Hussein; Faujan Ahmad

2007-01-01

The potential feasibility of treated and untreated activated carbon for removal of methylene blue from aqueous solution was investigated. The effects of various experimental parameters such as contact time, solution pH and adsorbent dosage were investigated. The extent of methylene blue removal increased with the increased in contact time, solution pH and amount of adsorbent used. Adsorption data was better fitted to the Langmuir isotherm. The results in this study indicated that the treated activated carbon was an attractive candidate for removing organic dye of methylene blue which shows great reduction of colour while reducing the time contact to achieve equilibrium. (author)

Nuclear Power Options Viability Study. Volume 4. Bibliography

Energy Technology Data Exchange (ETDEWEB)

Trauger, D B; White, J D; Sims, J W [eds.

1986-09-01

Documents in the Nuclear Power Options Viability Study (NPOVS) bibliography are classified under one of four headings or categories as follows: nuclear options; light water reactors; liquid metal reactors; and high temperature reactors. The collection and selection of these documents, beginning early in 1984 and continuing through March of 1986, was carried out in support of the study's objective: to explore the viabilities of several nuclear electric power generation options for commercial deployment in the United States between 2000 and 2010. There are approximately 550 articles, papers, reports, and books in the bibliography that have been selected from some 2000 surveyed. The citations have been made computer accessible to facilitate rapid on-line retrieval by keyword, author, corporate author, title, journal name, or document number.

Nuclear Power Options Viability Study. Volume 4. Bibliography

International Nuclear Information System (INIS)

Trauger, D.B.; White, J.D.; Sims, J.W.

1986-09-01

Documents in the Nuclear Power Options Viability Study (NPOVS) bibliography are classified under one of four headings or categories as follows: nuclear options; light water reactors; liquid metal reactors; and high temperature reactors. The collection and selection of these documents, beginning early in 1984 and continuing through March of 1986, was carried out in support of the study's objective: to explore the viabilities of several nuclear electric power generation options for commercial deployment in the United States between 2000 and 2010. There are approximately 550 articles, papers, reports, and books in the bibliography that have been selected from some 2000 surveyed. The citations have been made computer accessible to facilitate rapid on-line retrieval by keyword, author, corporate author, title, journal name, or document number

Prussian blue as an antidote for radioactive thallium and cesium poisoning

Directory of Open Access Journals (Sweden)

Altagracia-Martinez M

2012-06-01

Full Text Available Marina Altagracia-Martínez, Jaime Kravzov-Jinich, Juan Manuel Martínez-Núñez, Camilo Ríos-Castañeda, Francisco López-NaranjoDepartments of Biological Systems and Health Care, Biological and Health Sciences Division, Universidad Autónoma Metropolitana-Xochimilco, Mexico DF, MexicoBackground: Following the attacks on the US on September 11, 2001, potentially millions of people might experience contamination from radioactive metals. However, before the specter of such accidents arose, Prussian blue was known only as an investigational agent for accidental thallium and cesium poisoning. The purpose of this review is to update the state of the art concerning use of Prussian blue as an effective and safe drug against possible bioterrorism attacks and to disseminate medical information in order to contribute to the production of Prussian blue as a biodefense drug.Methods: We compiled articles from a systematic review conducted from January 1, 1960 to March 30, 2011. The electronic databases consulted were Medline, PubMed, the Cochrane Library, and Scopus.Results: Prussian blue is effective and safe for use against radioactive intoxications involving cesium-137 and thallium. The US Food and Drug Administration has approved Prussian blue as a drug, but there is only one manufacturer providing Prussian blue to the US. Based on the evidence, Prussian blue is effective for use against radioactive intoxications involving cesium-137 and thallium, but additional clinical research on and production of Prussian blue are needed.Keywords: Prussian blue, radioactive cesium, thallium, intoxication, biodefense drug

Can methylene blue dye be used as an alternative to patent blue dye to find the sentinel lymph node in breast cancer surgery?

Directory of Open Access Journals (Sweden)

Asieh Sadat Fattahi

2014-01-01

Full Text Available Background: Sentinel lymph node biopsy (SLNB is standard care to evaluate axillary involvement in early breast cancer. It has fewer complications than complete lymph node dissection; however, using blue dye in SLNB is controversial. We have evaluated the detection rate and local complications associated with methylene blue dye (MBD used in SLNB in early breast cancer patients and compared these results to patent blue dye (PBD. Materials and Methods : In a cohort prospective study, 312 patients with early breast cancer without axillary lymph node involvement were divided into two groups according to dye type. All of the patients received radiotracer and one type of blue dye. We filled out a checklist for the patients that contained demographic data, size of tumor, stage, detection of sentinel lymph node, and complications and then analyzed the data. Results: Demographic and histopathologic characteristics were not significantly different in both groups. Mean (standard deviation [SD] tumor size in all patients was 2.4 (0.8 cm. Detection rate in the MBD group was 77.5% with dye alone and 94.2% with dye and radioisotope; and in the PBD group it was 80.1% and 92.9% respectively (P > 0.05. We had blue discoloration of the skin in 23.7% in the PBD and 14.1% in the MBD group (P < 0.05 local inflammation was detected in one patient in the PBD and five in the MBD group (P < 0.05. Skin necrosis and systemic complications were not observed. Conclusion: Methylene blue has an acceptable detection rate, which may be a good alternative in SLNB. Complication such as blue discoloration of the skin was also lower with MBD.

Microwave-assisted synthesis of graphene-Prussian Blue networked nanocomposites for electrocatalysis

DEFF Research Database (Denmark)

Zhang, Minwei; Ulstrup, Jens; Chi, Qijin

There has been a great deal of interest recently in Prussian blue functional graphene. Due to they displayed advantage of both Prussian blue and graphene, we presented a one-pot and green method to synthesize interlocked graphene-Prussian Blue nanocomposites. Considering that graphene oxide (GO) ...

Radiopeptide internalisation and externalisation assays: Cell viability and radioligand integrity

International Nuclear Information System (INIS)

Raza Naqvi, Syed Ali; Sosabowski, Jane K.; Ahamad Nagra, Saeed; Ishfaq, Malik M.; Mather, Stephen J.; Matzow, Torkjel

2011-01-01

Various aspects of radiopeptide receptor-mediated cell internalisation and externalisation assays were assessed, including the integrity of externalised peptides and the effect of varying the pH and incubation time of the acid wash step (to remove surface receptor-bound ligand) on efficacy and cell viability. The observed intact proportion of externalised peptide was 5-10%, and acid wash buffers with pH 2.8 or below were found to be detrimental to cell viability and integrity, particularly following prolonged incubation times.

Is cell viability always directly related to corrosion resistance of stainless steels?

International Nuclear Information System (INIS)

Salahinejad, E.; Ghaffari, M.; Vashaee, D.; Tayebi, L.

2016-01-01

It has been frequently reported that cell viability on stainless steels is improved by increasing their corrosion resistance. The question that arises is whether human cell viability is always directly related to corrosion resistance in these biostable alloys. In this work, the microstructure and in vitro corrosion behavior of a new class of medical-grade stainless steels were correlated with adult human mesenchymal stem cell viability. The samples were produced by a powder metallurgy route, consisting of mechanical alloying and liquid-phase sintering with a sintering aid of a eutectic Mn–Si alloy at 1050 °C for 30 and 60 min, leading to nanostructures. In accordance with transmission electron microscopic studies, the additive particles for the sintering time of 30 min were not completely melted. Electrochemical impedance spectroscopic experiments suggested the higher corrosion resistance for the sample sintered for 60 min; however, a better cell viability on the surface of the less corrosion-resistant sample was unexpectedly found. This behavior is explained by considering the higher ion release rate of the Mn–Si additive material, as preferred sites to corrosion attack based on scanning electron microscopic observations, which is advantageous to the cells in vitro. In conclusion, cell viability is not always directly related to corrosion resistance in stainless steels. Typically, the introduction of biodegradable and biocompatible phases to biostable alloys, which are conventionally anticipated to be corrosion-resistant, can be advantageous to human cell responses similar to biodegradable metals. - Highlights: • Cell viability vs. corrosion resistance for medical-grade stainless steels • The stainless steel samples were prepared by powder metallurgy. • Unpenetrated additive played a critical role in the correlation.

Is cell viability always directly related to corrosion resistance of stainless steels?

Energy Technology Data Exchange (ETDEWEB)

Salahinejad, E., E-mail: salahinejad@kntu.ac.ir [Faculty of Materials Science and Engineering, K.N. Toosi University of Technology, Tehran (Iran, Islamic Republic of); Ghaffari, M. [Bruker AXS Inc., 5465 East Cheryl Parkway, Madison, WI 53711 (United States); Vashaee, D. [Electrical and Computer Engineering Department, North Carolina State University, Raleigh, NC 27606 (United States); Tayebi, L. [Department of Developmental Sciences, Marquette University School of Dentistry, Milwaukee, WI 53201 (United States); Department of Engineering Science, University of Oxford, Oxford OX1 3PJ (United Kingdom)

2016-05-01

It has been frequently reported that cell viability on stainless steels is improved by increasing their corrosion resistance. The question that arises is whether human cell viability is always directly related to corrosion resistance in these biostable alloys. In this work, the microstructure and in vitro corrosion behavior of a new class of medical-grade stainless steels were correlated with adult human mesenchymal stem cell viability. The samples were produced by a powder metallurgy route, consisting of mechanical alloying and liquid-phase sintering with a sintering aid of a eutectic Mn–Si alloy at 1050 °C for 30 and 60 min, leading to nanostructures. In accordance with transmission electron microscopic studies, the additive particles for the sintering time of 30 min were not completely melted. Electrochemical impedance spectroscopic experiments suggested the higher corrosion resistance for the sample sintered for 60 min; however, a better cell viability on the surface of the less corrosion-resistant sample was unexpectedly found. This behavior is explained by considering the higher ion release rate of the Mn–Si additive material, as preferred sites to corrosion attack based on scanning electron microscopic observations, which is advantageous to the cells in vitro. In conclusion, cell viability is not always directly related to corrosion resistance in stainless steels. Typically, the introduction of biodegradable and biocompatible phases to biostable alloys, which are conventionally anticipated to be corrosion-resistant, can be advantageous to human cell responses similar to biodegradable metals. - Highlights: • Cell viability vs. corrosion resistance for medical-grade stainless steels • The stainless steel samples were prepared by powder metallurgy. • Unpenetrated additive played a critical role in the correlation.

BLUE OCEAN STRATEGY (BOS SEBAGAI PARADIGMA BARU MANAJEMEN PENDIDIKAN ISLAM

Directory of Open Access Journals (Sweden)

Ikhwan Fuad

2016-08-01

Full Text Available The focus of this article is to describe the Blue Ocean Strategy as a new paradigm of Islamic education management. the author seeks to elaborate on how the adoption of the Blue Ocean strategy of the business world to the world of education and answered the principles of what is accepted and rejected away from the theory. Adoption of blue ocean strategy is done by applying the universal principles are : among others reconstruct market boundaries, focus on the big picture rather than numbers, reach beyond existing demand, perform a series of strategies with appropriate, efforts to overcome organizational constraints and integrate execution into strategy. The principles of Blue Ocean Strategy Indicators that can be absorbed is an indicator that focuses gave excellent service and were not absorbed is a strong indicator of economic motivation. Generally, Blue Ocean Strategy quite well applied as a management paradigm of Islamic education.

Impact of age and diagnosis on viability during centrifugation and cryopreservation of peripheral blood stem cell products.

Science.gov (United States)

Civriz Bozdag, S; Bay, M; Ayyıldız, E; Topcuoglu, P; Ilhan, O

2012-08-01

The viability of the hematopoietic stem cells infused to the patient is important for transplant outcome. We evaluated 31 peripheral blood stem cell product collected from 15 patients. We aimed to check the viabilities of the cells from patients with different age and diagnosis, in different stages of the cryopreservation procedure. We showed a markedly decreased viability rate after centrifugation and addition of DMSO. Percentages of viabilities were similar between young and old patients in each step. Type of hematological malignancy did not make a significant influence on the viability. High speed centrifugation has a negative impact on the viability. Copyright © 2012 Elsevier Ltd. All rights reserved.

Blue LED irradiation to hydration of skin

Science.gov (United States)

Menezes, Priscila F. C.; Requena, Michelle B.; Lizarelli, Rosane F., Z.; Bagnato, Vanderlei S.

2015-06-01

Blue LED system irradiation shows many important properties on skin as: bacterial decontamination, degradation of endogenous skin chromophores and biostimulation. In this clinical study we prove that the blue light improves the skin hydration. In the literature none authors reports this biological property on skin. Then this study aims to discuss the role of blue light in the skin hydration. Twenty patients were selected to this study with age between 25-35 years old and phototype I, II and III. A defined area from forearm was pre determined (A = 4.0 cm2). The study was randomized in two treatment groups using one blue light device (power of 5.3mW and irradiance of 10.8mW/cm2). The first treatment group was irradiated with 3J/cm2 (277seconds) and the second with 6J/cm2 (555 seconds). The skin hydration evaluations were done using a corneometer. The measurements were collected in 7, 14, 21 and 30 days, during the treatment. Statistical test of ANOVA, Tukey and T-Student were applied considering 5% of significance. In conclusion, both doses were able to improve the skin hydration; however, 6J/cm2 has kept this hydration for 30 days.

Linking population viability, habitat suitability, and landscape simulation models for conservation planning

Science.gov (United States)

Michael A. Larson; Frank R., III Thompson; Joshua J. Millspaugh; William D. Dijak; Stephen R. Shifley

2004-01-01

Methods for habitat modeling based on landscape simulations and population viability modeling based on habitat quality are well developed, but no published study of which we are aware has effectively joined them in a single, comprehensive analysis. We demonstrate the application of a population viability model for ovenbirds (Seiurus aurocapillus)...

LIVE/DEAD YEAST VIABILITY STAINING AS A TOOL FOR IMPROVING ARTISANAL PILSNER BEER PRODUCTION

Directory of Open Access Journals (Sweden)

Benedetta Bottari

2014-10-01

Full Text Available The production of an artisanal beer, made by brewers using traditional practices on a small scale, is founded on the empirical adjustment of parameters, including yeasts handling and serial repitching. The aim of this study was to monitor yeast viability during different stages of artisanal beer productions through the Live/Dead Yeast viability staining and to correlate it with fermentation dynamics in order to increase process standardization and to maintain the quality of final products. Yeast viability and fermentation activities were evaluated during seven fermentation cycles of an artisanal pilsner beer. Yeast inoculated with higher viability performed generally better in fermentation, resulting in faster sugar consumption, faster ethanol production and stability. Handling yeast and serial repitching based on Live/Dead viability measurements, could be the key way to ensure reliable manufacture of high quality beer and to improve process standardization particularly for microbreweries, where variability of production can be a challenging point.

Blue and grey water footprint of textile industry in China.

Science.gov (United States)

Wang, Laili; Ding, Xuemei; Wu, Xiongying

2013-01-01

Water footprint (WF) is a newly developed idea that indicates impacts of freshwater appropriation and wastewater discharge. The textile industry is one of the oldest, longest and most complicated industrial chains in the world's manufacturing industries. However, the textile industry is also water intensive. In this paper, we applied a bottom-up approach to estimate the direct blue water footprint (WFdir,blue) and direct grey water footprint (WFdir,grey) of China's textile industry at sector level based on WF methodology. The results showed that WFdir,blue of China's textile industry had an increasing trend from 2001 to 2010. The annual WFdir,blue surpassed 0.92 Gm(3)/yr (giga cubic meter a year) since 2004 and rose to peak value of 1.09 Gm(3)/yr in 2007. The original and residuary WFdir,grey (both were calculated based on the concentration of chemical oxygen demand (CODCr)) of China's textile industry had a similar variation trend with that of WFdir,blue. Among the three sub-sectors of China's textile industry, the manufacture of textiles sector's annual WFdir,blue and WFdir,grey were much larger than those of the manufacture of textile wearing apparel, footware and caps sector and the manufacture of chemical fibers sector. The intensities of WFdir,blue and WF(res)dir,grey of China's textile industry were year by year decreasing through the efforts of issuing restriction policies on freshwater use and wastewater generation and discharge, and popularization of water saving and wastewater treatment technologies.

Biological behaviour of buccal cells exposed to blue light

International Nuclear Information System (INIS)

Gritsch, Kerstin; Ponsonnet, Laurence; Schembri, Catherine; Farge, Pierre; Pourreyron, Laurence; Grosgogeat, Brigitte

2008-01-01

Blue light is used in dental practise to cure resin-based materials, but the path of the light often includes oral tissues such as gingival tissues. While adverse effects of blue light exposure on cells - such as retina cells - are well known, few studies have investigated the impact of blue light exposure on oral cells. The aim of the present in vitro study was to assess the biological effects of blue light emitted by two dental curing devices (a plasma-arc and a light-emitting diode curing unit) on human gingival fibroblasts. Light intensities and light-induced temperature rise were respectively measured with a radiometer and a thermocouple. Cellular response to blue light exposure was assessed by the observation of cell morphology (scanning electron microscopy) and the estimation of cell mitochondrial activity (MTT assay). Light intensities measured at the clinical distance were 488 ± 42 mW/cm 2 for the plasma-arc unit and ranged from 61 ± 5 to 140 ± 16 mW/cm 2 for the light-emitting diodes unit, according to the curing program used. The highest temperature rise was 0.5 and 3.5 deg. C for exposure to the plasma-arc light and to the light-emitting diodes light, respectively. Results showed no differences between exposed- and non-exposed cells in regards to cell morphology. However, cells exposed to blue light presented an increased mitochondrial activity compared to control cells (non-exposed), and mostly those exposed to plasma-arc light

Drying process strongly affects probiotics viability and functionalities.

Science.gov (United States)

Iaconelli, Cyril; Lemetais, Guillaume; Kechaou, Noura; Chain, Florian; Bermúdez-Humarán, Luis G; Langella, Philippe; Gervais, Patrick; Beney, Laurent

2015-11-20

Probiotic formulations are widely used and are proposed to have a variety of beneficial effects, depending on the probiotic strains present in the product. The impact of drying processes on the viability of probiotics is well documented. However, the impact of these processes on probiotics functionality remains unclear. In this work, we investigated variations in seven different bacterial markers after various desiccation processes. Markers were composed of four different viability evaluation (combining two growth abilities and two cytometric measurements) and in three in vitro functionalities: stimulation of IL-10 and IL-12 production by PBMCs (immunomodulation) and bacterial adhesion to hexadecane. We measured the impact of three drying processes (air-drying, freeze-drying and spray-drying), without the use of protective agents, on three types of probiotic bacteria: Bifidobacterium bifidum, Lactobacillus plantarum and Lactobacillus zeae. Our results show that the bacteria respond differently to the three different drying processes, in terms of viability and functionality. Drying methods produce important variations in bacterial immunomodulation and hydrophobicity, which are correlated. We also show that adherence can be stimulated (air-drying) or inhibited (spray-drying) by drying processes. Results of a multivariate analysis show no direct correlation between bacterial survival and functionality, but do show a correlation between probiotic responses to desiccation-rewetting and the process used to dry the bacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

Compete or Leapfrog: Creating Blue Ocean through Entrepreneurial Orientation

Directory of Open Access Journals (Sweden)

Arslan Ayub

2013-10-01

Full Text Available The study analyzes the role of entrepreneurial orientation with mediating effect of knowledge creation process to creating Blue Ocean in corporate sector in Pakistan There is an increasing competition among companies due to globalization and technological advancements. Thus, it requires a study to measure the multifaceted influence of entrepreneurial orientation on knowledge creation process and Blue Ocean besides the actual paradigm of this terminology. This concept has been well discussed in this research arena since its inception in 2005. Numerous such initiatives have already been taken, however this concept invites a lot more addition, related companies are still in pursuit to materialize the research concepts. We highlight the contingencies in the shift from a red ocean to Blue Ocean. The study uses exploratory approach; primary data is collected from 391 professionals working in different sectors of Pakistan. The study uses structural equation model (SEM technique to test the hypotheses. The study found a positive relationship between entrepreneurial orientation and Blue Ocean, entrepreneurial orientation, knowledge creation process, and Blue Ocean. The study throws light on the importance of entrepreneurial orientation and knowledge creation process to head on this fast-paced competition.

A complex approach to the blue-loop problem

Science.gov (United States)

Ostrowski, Jakub; Daszynska-Daszkiewicz, Jadwiga

2015-08-01

The problem of the blue loops during the core helium burning, outstanding for almost fifty years, is one of the most difficult and poorly understood problems in stellar astrophysics. Most of the work focused on the blue loops done so far has been performed with old stellar evolution codes and with limited computational resources. In the end the obtained conclusions were based on a small sample of models and could not have taken into account more advanced effects and interactions between them.The emergence of the blue loops depends on many details of the evolution calculations, in particular on chemical composition, opacity, mixing processes etc. The non-linear interactions between these factors contribute to the statement that in most cases it is hard to predict without a precise stellar modeling whether a loop will emerge or not. The high sensitivity of the blue loops to even small changes of the internal structure of a star yields one more issue: a sensitivity to numerical problems, which are common in calculations of stellar models on advanced stages of the evolution.To tackle this problem we used a modern stellar evolution code MESA. We calculated a large grid of evolutionary tracks (about 8000 models) with masses in the range of 3.0 - 25.0 solar masses from the zero age main sequence to the depletion of helium in the core. In order to make a comparative analysis, we varied metallicity, helium abundance and different mixing parameters resulting from convective overshooting, rotation etc.The better understanding of the properties of the blue loops is crucial for our knowledge of the population of blue supergiants or pulsating variables such as Cepheids, α-Cygni or Slowly Pulsating B-type supergiants. In case of more massive models it is also of great importance for studies of the progenitors of supernovae.

Evaluation of pollen viability, stigma receptivity and fertilization ...

African Journals Online (AJOL)

AJL

2013-11-13

Nov 13, 2013 ... by field artificial pollination were analyzed in this study. The maximum pollen viability .... the day before anthesis to avoid self-pollination. Subsequently, between ..... The Lagerstroemia handbook/checklist. Ameri. Association ...

Posthuman blues

CERN Document Server

Tonnies, Mac

2013-01-01

Posthuman Blues, Vol. I is first volume of the edited version of the popular weblog maintained by author Mac Tonnies from 2003 until his tragic death in 2009. Tonnies' blog was a pastiche of his original fiction, reflections on his day-to-day life, trenchant observations of current events, and thoughts on an eclectic range of material he culled from the Internet. What resulted was a remarkably broad portrait of a thoughtful man and the complex times in which he lived, rendered with intellige...

The monoamine oxidase inhibition properties of selected structural analogues of methylene blue

International Nuclear Information System (INIS)

Delport, Anzelle; Harvey, Brian H.; Petzer, Anél; Petzer, Jacobus P.

2017-01-01

The thionine dye, methylene blue (MB), is a potent inhibitor of monoamine oxidase (MAO) A, a property that may, at least in part, mediate its antidepressant effects in humans and animals. The central inhibition of MAO-A by MB has also been linked to serotonin toxicity (ST) which may arise when MB is used in combination with serotonergic drugs. Structural analogues and the principal metabolite of MB, azure B, have also been reported to inhibit the MAO enzymes, with all compounds exhibiting specificity for the MAO-A isoform. To expand on the structure-activity relationships (SARs) of MAO inhibition by MB analogues, the present study investigates the human MAO inhibition properties of five MB analogues: neutral red, Nile blue, new methylene blue, cresyl violet and 1,9-dimethyl methylene blue. Similar to MB, these analogues also are specific MAO-A inhibitors with cresyl violet (IC 50 = 0.0037 μM), Nile blue (IC 50 = 0.0077 μM) and 1,9-dimethyl methylene blue (IC 50 = 0.018 μM) exhibiting higher potency inhibition compared to MB (IC 50 = 0.07 μM). Nile blue also represents a potent MAO-B inhibitor with an IC 50 value of 0.012 μM. From the results it may be concluded that non-thionine MB analogues (e.g. cresyl violet and Nile blue) also may exhibit potent MAO inhibition, a property which should be considered when using these compounds in pharmacological studies. Benzophenoxazines such as cresyl violet and Nile blue are, similar to phenothiazines (e.g. MB), representative of high potency MAO-A inhibitors with a potential risk of ST. - Highlights: • MB analogues, cresyl violet and Nile blue, are high potency MAO-A inhibitors. • Nile blue also represents a potent MAO-B inhibitor. • Potent MAO-A inhibition should alert to potential serotonin toxicity.

The monoamine oxidase inhibition properties of selected structural analogues of methylene blue

Energy Technology Data Exchange (ETDEWEB)

Delport, Anzelle [Pharmaceutical Chemistry, School of Pharmacy, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Centre of Excellence for Pharmaceutical Sciences, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Harvey, Brian H. [Centre of Excellence for Pharmaceutical Sciences, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Pharmacology, School of Pharmacy, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Petzer, Anél [Pharmaceutical Chemistry, School of Pharmacy, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Centre of Excellence for Pharmaceutical Sciences, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Petzer, Jacobus P., E-mail: jacques.petzer@nwu.ac.za [Pharmaceutical Chemistry, School of Pharmacy, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa); Centre of Excellence for Pharmaceutical Sciences, North-West University, Private Bag X6001, Potchefstroom 2520 (South Africa)

2017-06-15

The thionine dye, methylene blue (MB), is a potent inhibitor of monoamine oxidase (MAO) A, a property that may, at least in part, mediate its antidepressant effects in humans and animals. The central inhibition of MAO-A by MB has also been linked to serotonin toxicity (ST) which may arise when MB is used in combination with serotonergic drugs. Structural analogues and the principal metabolite of MB, azure B, have also been reported to inhibit the MAO enzymes, with all compounds exhibiting specificity for the MAO-A isoform. To expand on the structure-activity relationships (SARs) of MAO inhibition by MB analogues, the present study investigates the human MAO inhibition properties of five MB analogues: neutral red, Nile blue, new methylene blue, cresyl violet and 1,9-dimethyl methylene blue. Similar to MB, these analogues also are specific MAO-A inhibitors with cresyl violet (IC{sub 50} = 0.0037 μM), Nile blue (IC{sub 50} = 0.0077 μM) and 1,9-dimethyl methylene blue (IC{sub 50} = 0.018 μM) exhibiting higher potency inhibition compared to MB (IC{sub 50} = 0.07 μM). Nile blue also represents a potent MAO-B inhibitor with an IC{sub 50} value of 0.012 μM. From the results it may be concluded that non-thionine MB analogues (e.g. cresyl violet and Nile blue) also may exhibit potent MAO inhibition, a property which should be considered when using these compounds in pharmacological studies. Benzophenoxazines such as cresyl violet and Nile blue are, similar to phenothiazines (e.g. MB), representative of high potency MAO-A inhibitors with a potential risk of ST. - Highlights: • MB analogues, cresyl violet and Nile blue, are high potency MAO-A inhibitors. • Nile blue also represents a potent MAO-B inhibitor. • Potent MAO-A inhibition should alert to potential serotonin toxicity.

Viability of Event Management Business in Batangas City, Philippine: Basis for Business Operation Initiatives

OpenAIRE

Jeninah Christia D. Borbon

2016-01-01

The research study on Viability of Event Management Business in Batangas City: Basis for Business Operation Initiatives aimed to assess the viability of this type of business using Thompson’s (2005) Dimension of Business Viability as its tool in order to create business operation initiatives. It provided a good framework for defining success factors in entrepreneurial operation initiatives in a specific business type – event management. This study utilized event organizers based i...

Gamma irradiation of yellow and blue colorants in polystyrene packaging materials

International Nuclear Information System (INIS)

Komolprasert, V.; Diel, Todd; Sadler, G.

2006-01-01

The effect of 10- and 20-kGy gamma irradiation was studied on chromophtal yellow 2RLTS (Yellow 110-2, 3, 4, 5-tetrachloro-6-cyanobenzoic acid) and Irgalite Blue GBP (copper (II) phthalocyanine blue) colorants, which were added to polystyrene (PS) material used to package food prior to irradiation. Analytical results obtained suggest that irradiation did not generate any new chemicals in the PS polymer containing either yellow or blue colorant at a concentration of up to 1% (w/w). Both yellow and blue colorants are relatively stable to gamma irradiation

China Mobile: Expanding "Blue Ocean"

Institute of Scientific and Technical Information of China (English)

无

2006-01-01

Driving force is crucial for realizing high-speed growth. The strong driving force from "Blue Ocean Strategy" is an important advantage for China Mobile to realize harmonious and leap-forward development.

Fe K-edge XANES of Maya blue pigment

Science.gov (United States)

Río, M. Sánchez del; Sodo, A.; Eeckhout, S. G.; Neisius, T.; Martinetto, P.; Dooryhée, E.; Reyes-Valerio, C.

2005-08-01

The utilization of techniques used in Materials Science for the characterization of artefacts of interest for cultural heritage is getting more and more attention nowadays. One of the products of the ancient Maya chemistry is the "Maya blue" pigment, made with natural indigo and palygorskite. This pigment is different from any other pigment used in other parts of the world. It is durable and acid-resistant, and still keeps many secrets to scientists even though it has been studied for more than 50 years. Although the pigment is basically made of palygorskite Si8(Mg2Al2)O20(OH)2(OH2)4.4H2O and an organic colourant (indigo: C16H10N2O2), a number of other compounds have been found in previous studies on archaeological samples, like other clays and minerals, iron nanoparticles, iron oxides, impurities of transition metals (Cr, Mn, Ti, V), etc. We measured at the ESRF ID26 beamline the Fe K-edge XANES spectra of the blue pigment in ancient samples. They are compared to XANES spectra of Maya blue samples synthesized under controlled conditions, and iron oxides usually employed as pigments (hematite and goethite). Our results show that the iron found in ancient Maya blue pigment is related to the Fe exchanged in the palygorskite clay. We did not find iron in metallic form or goethite in archaeological Maya blue.

Fe K-edge XANES of Maya blue pigment

International Nuclear Information System (INIS)

Rio, M. Sanchez del; Sodo, A.; Eeckhout, S.G.; Neisius, T.; Martinetto, P.; Dooryhee, E.; Reyes-Valerio, C.

2005-01-01

The utilization of techniques used in Materials Science for the characterization of artefacts of interest for cultural heritage is getting more and more attention nowadays. One of the products of the ancient Maya chemistry is the 'Maya blue' pigment, made with natural indigo and palygorskite. This pigment is different from any other pigment used in other parts of the world. It is durable and acid-resistant, and still keeps many secrets to scientists even though it has been studied for more than 50 years. Although the pigment is basically made of palygorskite Si 8 (Mg 2 Al 2 )O 20 (OH) 2 (OH 2 ) 4 .4H 2 O and an organic colourant (indigo: C 16 H 10 N 2 O 2 ), a number of other compounds have been found in previous studies on archaeological samples, like other clays and minerals, iron nanoparticles, iron oxides, impurities of transition metals (Cr, Mn, Ti, V), etc. We measured at the ESRF ID26 beamline the Fe K-edge XANES spectra of the blue pigment in ancient samples. They are compared to XANES spectra of Maya blue samples synthesized under controlled conditions, and iron oxides usually employed as pigments (hematite and goethite). Our results show that the iron found in ancient Maya blue pigment is related to the Fe exchanged in the palygorskite clay. We did not find iron in metallic form or goethite in archaeological Maya blue

Physicochemical, Thermal, and Sensory Properties of Blue Corn (Zea Mays L.).

Science.gov (United States)

Mutlu, Ceren; Arslan-Tontul, Sultan; Candal, Cihadiye; Kilic, Ozlem; Erbas, Mustafa

2018-01-01

The aim of this study was to investigate some physicochemical and sensory properties of blue corn cultivated in Turkey. The length and width of the cob with kernels, hectoliter, and 1000-kernel weight of blue corn were measured as 7.66, 2.02 mm, 84.40 kg/100 L, and 44.27 g, respectively. The gelatinization onset, peak, and end temperatures were measured as 61.12 °C, 64.35 °C, and 75.65 °C, respectively. The water activity, moisture content, total protein, lipid, and crude fiber contents of the blue corn sample were detected as 0.44, 9.39%, 13.13%, 4.30%, and 2.68%, respectively. Total starch and resistant starch contents of blue corn were determined as 63.94% and 8.89%, respectively. Also, total monomeric anthocyanin content and antioxidant capacity of blue corn were detected as 915.43 mg CGE/kg and 7.99 μmol TE/g, respectively. Additionally, the major fatty acids detected in blue corn samples were palmitic, stearic, oleic, and linoleic acids. Blue corn can be utilized in the production of enjoyable and healthier snacks, such as popcorn and chips, because of its color and high phenolic, anthocyanin, and fiber contents. © 2017 Institute of Food Technologists®.

The relationship between myocardial blood flow and myocardial viability after reperfusion. Myocardial viability assessed by 15O-water-PET

International Nuclear Information System (INIS)

Tsukagoshi, Joichi

1994-01-01

The purpose of this study was to examine the relationship between myocardial blood flow and myocardial viability in the ischemic canine myocardium after reperfusion. Transient ischemia was induced by 60-, 90-, and 180-minute occlusion of the left anterior descending coronary artery. Myocardial blood flow (MBF) was measured in the areas in which regional contractility was severely impaired (ehocardiographically akinetic or dyskinetic) in the early reperfusion period by 15 O-water positron emission tomography (PET) 12 hours and 4 weeks after reperfusion. An MBF ratio of ischemic to nonischemic regions 12 hours after reperfusion was inversely correlated with the amount of histologically determined tissue necrosis (r=-0.74). The regional contractility recovered 4 weeks later in the areas where an MBF ratio was 0.48 or greater, but did not recover in the areas with a lower MBF ratio. Thus, myocardial viability can be appropriately predicted in the early phase of myocardial perfusion by PET with 15 O-water even in the absence of metabolic imaging. (author)

THE TRANSATLANTIC BLUE DIPLOMACY

Directory of Open Access Journals (Sweden)

Ioana GUTU

2016-12-01

Full Text Available The international diplomatic environment has reached to an unprecedented development, involving one of the newly specialized diplomatic types, namely the economic diplomacy. At the core of the fast movements in the diplomatic spheres across the Globe are the international agreements like the Transatlantic Trade and Investment Partnership (TTIP that determined diplomacy to dissolve into new subtypes, evolving from ground to the ocean and implementing new ways of achieving economic and climate sustainability. One of the newly created diplomatic spheres, is the blue ocean diplomacy that acts mainly in accordance with the rules and regulations that are being applied to the transatlantic economy. Even though TTIP encourages the increase of trade flows across the Atlantic, it will also ease the foreign investment procedures that, under the approach of keeping a sustainable environment, will represent one of the most important initiatives in implementing the blue economy concept within the framework of the transatlantic diplomacy.

Echinococcus metacestode: in search of viability markers.