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Sample records for sublethally injured cells

  1. Detection of low numbers of healthy and sub-lethally injured Salmonella enterica in chocolate.

    Science.gov (United States)

    Jasson, Vicky; Baert, Leen; Uyttendaele, Mieke

    2011-02-28

    The capacity to detect low levels of healthy and sub-lethally injured Salmonella enterica cells in chocolate by two alternative rapid detection methods iQ-Check(TM)Salmonella II real-time PCR (Bio-Rad) and VIDAS® Easy SLM (BioMérieux) was assessed and compared with ISO 6579:2005. Chocolate, a low moisture food known to support the survival of Salmonella, was challenged as food matrix. Buffered peptone water (BPW) did not support the recovery of low levels of sub-lethally injured S. enterica independent of the detection method, while BPW supplemented with milk powder enabled detection by the three examined methods. However, inhibition of real-time PCR was observed since for one out of three repetitions of chocolate inoculated with a low number of sub-lethally injured S. enterica cells, no PCR signal was obtained. Therefore, attention should be paid to the enrichment step to avoid false negative results due to the presence of especially sub-lethally injured Salmonella cells in chocolate. An appropriate sample preparation (such as enrichment media and conditions for incubation) remains the key factor for reliable detection including sub-lethally injured cells and should be evaluated, if necessary optimized, for each detection assay. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. iTRAQ-Based Proteomic Analysis of Sublethally Injured Escherichia coli O157:H7 Cells Induced by High Pressure Carbon Dioxide

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    Xiufang Bi

    2017-12-01

    Full Text Available High pressure carbon dioxide (HPCD could cause sublethally injured cells (SICs, which may cause food poisoning and spoilage during food storage and limit its application. Therefore, the formation of SICs of Escherichia coli O157:H7 was investigated by isobaric tag for relative and absolute quantification (iTRAQ proteomic methods in this study for better controlling the SICs induced by HPCD. A total of 2,446 proteins was identified by iTRAQ, of which 93 and 29 were significantly differentially expressed in the SICs compared with live control cells (CKL and dead control cells (CKD, respectively. Among the 93 differentially expressed proteins (DEP in the SICs compared with CKL, 65 proteins showed down-regulation and 28 showed up-regulation. According to the comprehensive proteome coverage analysis, the SICs survived under HPCD by reducing carbohydrate decomposing, lipid transport and metabolism, amino acid transport and metabolism, transcription and translation, DNA replication and repair. Besides, the SICs showed stress response, DNA damage response and an increased carbohydrate transport, peptidoglycan synthesis and disulfide bond formation to HPCD. Among the 29 DEP in the SICs compared with CKD, 12 proteins showed down-regulation and 17 showed up-regulation. According to the comprehensive proteome coverage analysis, the SICs survived under HPCD by accumulation of cell protective agents like carbohydrates and amino acids, and decreasing transcription and translation activities. Results showed that the formation of the SICs with low metabolic activity and high survival ability was a survival strategy for E. coli O157:H7 against HPCD.

  3. Viability of sublethally injured coliform bacteria on fresh-cut cabbage stored in high CO2atmospheres following rinsing with electrolyzed water.

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    Izumi, Hidemi; Inoue, Ayano

    2018-02-02

    The extent of sublethally injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, was evaluated during storage in air and high CO 2 controlled atmospheres (5%, 10%, and 15%) at 5°C and 10°C using the thin agar layer (TAL) method. Sublethally injured coliform bacteria on nonrinsed shredded cabbage were either absent or they were injured at a 64-65% level when present. Rinsing of shredded cabbage with electrolyzed water containing 25ppm available chlorine reduced the coliform counts by 0.4 to 1.1 log and caused sublethal injury ranging from 42 to 77%. Pantoea ananatis was one of the species injured by chlorine stress. When shredded cabbage, nonrinsed or rinsed with electrolyzed water, was stored in air and high CO 2 atmospheres at 5°C for 7days and 10°C for 5days, coliform counts on TAL plates increased from 3.3-4.5 to 6.5-9.0 log CFU/g during storage, with the increase being greater at 10°C than at 5°C. High CO 2 of 10% and 15% reduced the bacterial growth on shredded cabbage during storage at 5°C. Although injured coliform bacteria were not found on nonrinsed shredded cabbage on the initial day, injured coliforms at a range of 49-84% were detected on samples stored in air and high CO 2 atmospheres at 5°C and 10°C. Injured cells were detected more frequently during storage at both temperatures irrespective of the CO 2 atmosphere when shredded cabbage was rinsed with electrolyzed water. These results indicated that injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, exhibited different degrees of injury during storage regardless of the CO 2 atmosphere and temperature tested. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Comparison and Improvement of Detection Media for Sub-lethal Injured Clostridium perfringens

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    清水, 茂雅; 堀口, 明日香; 山崎, 浩司; 川合, 祐史

    2009-01-01

    Recovery of heat-injured vegetative cells and spores of Clostridium perfringens was evaluated on selective media, CW Egg-yolk agar with kanamycin (ECW+A), Tryptose sulfite cycloserine agar(TSC+A)and modified Handford agar (mHFA). As a result of heat treatment at 54℃ for 15 min, viable counts of C. perfringens vegetative cells on a selective medium (ECW+A)was significantly less than those on non-selective media. This means C. perfringens vegetative cells should be in an injured state. Comparin...

  5. Relationship between Sublethal Injury and Inactivation of Yeast Cells by the Combination of Sorbic Acid and Pulsed Electric Fields▿

    Science.gov (United States)

    Somolinos, M.; García, D.; Condón, S.; Mañas, P.; Pagán, R.

    2007-01-01

    The objective of this study was to investigate the occurrence of sublethal injury after the pulsed-electric-field (PEF) treatment of two yeasts, Dekkera bruxellensis and Saccharomyces cerevisiae, as well as the relation of sublethal injury to the inactivating effect of the combination of PEF and sorbic acid. PEF caused sublethal injury in both yeasts: more than 90% of surviving D. bruxellensis cells and 99% of surviving S. cerevisiae cells were sublethally injured after 50 pulses at 12 kV/cm in buffer at pHs of both 7.0 and 4.0. The proportion of sublethally injured cells reached a maximum after 50 pulses at 12.0 kV/cm (S. cerevisiae) or 16.5 kV/cm (D. bruxellensis), and it kept constant or progressively decreased at greater electric field strengths and with longer PEF treatments. Sublethally PEF-injured cells showed sensitivity to the presence of sorbic acid at a concentration of 2,000 ppm. A synergistic inactivating effect of the combination of PEF and sorbic acid was observed. Survivors of the PEF treatment were progressively inactivated in the presence of 2,000 ppm of sorbic acid at pH 3.8, with the combined treatments achieving more than log10 5 cycles of dead cells under the conditions investigated. This study has demonstrated the occurrence of sublethal injury after exposure to PEF, so yeast inactivation by PEF is not an all-or-nothing event. The combination of PEF and sorbic acid has proven to be an effective method to achieve a higher level of yeast inactivation. This work contributes to the knowledge of the mechanism of microbial inactivation by PEF, and it may be useful for improving food preservation by PEF technology. PMID:17468278

  6. Relationship between sublethal injury and inactivation of yeast cells by the combination of sorbic acid and pulsed electric fields.

    Science.gov (United States)

    Somolinos, M; García, D; Condón, S; Mañas, P; Pagán, R

    2007-06-01

    The objective of this study was to investigate the occurrence of sublethal injury after the pulsed-electric-field (PEF) treatment of two yeasts, Dekkera bruxellensis and Saccharomyces cerevisiae, as well as the relation of sublethal injury to the inactivating effect of the combination of PEF and sorbic acid. PEF caused sublethal injury in both yeasts: more than 90% of surviving D. bruxellensis cells and 99% of surviving S. cerevisiae cells were sublethally injured after 50 pulses at 12 kV/cm in buffer at pHs of both 7.0 and 4.0. The proportion of sublethally injured cells reached a maximum after 50 pulses at 12.0 kV/cm (S. cerevisiae) or 16.5 kV/cm (D. bruxellensis), and it kept constant or progressively decreased at greater electric field strengths and with longer PEF treatments. Sublethally PEF-injured cells showed sensitivity to the presence of sorbic acid at a concentration of 2,000 ppm. A synergistic inactivating effect of the combination of PEF and sorbic acid was observed. Survivors of the PEF treatment were progressively inactivated in the presence of 2,000 ppm of sorbic acid at pH 3.8, with the combined treatments achieving more than log10 5 cycles of dead cells under the conditions investigated. This study has demonstrated the occurrence of sublethal injury after exposure to PEF, so yeast inactivation by PEF is not an all-or-nothing event. The combination of PEF and sorbic acid has proven to be an effective method to achieve a higher level of yeast inactivation. This work contributes to the knowledge of the mechanism of microbial inactivation by PEF, and it may be useful for improving food preservation by PEF technology.

  7. Decreased resistance of sublethally injured Escherichia coli O157:H7 to salt, mild heat, nisin and acids induced by high pressure carbon dioxide.

    Science.gov (United States)

    Bi, Xiufang; Wang, Yongtao; Hu, Xiaosong; Liao, Xiaojun

    2018-02-01

    Resistance of sublethally injured cells (SICs) of Escherichia coli O157:H7 induced by high pressure carbon dioxide (HPCD) to salt, low temperature, mild heat, nisin, acids and low pHs was investigated in this study. The SICs of E. coli were obtained following HPCD at 5 MPa and 25 °C for 40-60 min or 5 MPa and 45 °C for 20 min. The untreated cells could survive normally while the HPCD-treated cells showed 2.87 log 10  cycles' reduction on tryptic soy agar (TSA) with 3% NaCl. The counts of the untreated cells were not significantly changed during 5 h incubation at 4 °C or 40 min incubation at 45 °C, and the HPCD-treated cells were also not affected by 5 h incubation at 4 °C but showed 1.75 log 10  cycles' reduction at 45 °C for 40 min. The antimicrobial nisin caused an extra 0.25-1.0 log 10  cycles' reduction of the HPCD-treated cells while the untreated cells was not inactivated by nisin. Except for oxalic acid (OA), citric acid (CA), malic acid (MA), tartaric acid (TA), lactic acid (LA), acetic acid (AA) and hydrochloric acid (HCl) did not inactivate the untreated cells, but all these seven acids caused a 0.74-1.53 log 10  cycles' reduction of HPCD-treated cells. These results indicated that HPCD-induced SICs had a decreased resistance to salt, mild heat, nisin and acids. Moreover, the recovery test was used to investigate the sensitivity of the SICs to different pHs. Results showed that the SICs could not recover below or equal to pH 4.0. These promising results would open up the possibility of exploring the combination of other technologies (eg. mild heat, nisin and acids) with HPCD as hurdle approaches to inactivate target pathogens in foods. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Enumeration of sublethally injured Escherichia coli O157:H7 ATCC 43895 and Escherichia coli strain B-41560 using selective agar overlays versus commercial methods.

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    Smith, Amanda R; Ellison, Alysha L; Robinson, Amanda L; Drake, Maryanne; McDowell, Susan A; Mitchell, James K; Gerard, Patrick D; Heckler, Rachel A; McKillip, John L

    2013-04-01

    Quality control procedures during food processing may involve direct inoculation of food samples onto appropriate selective media for subsequent enumeration. However, sublethally injured bacteria often fail to grow, enabling them to evade detection and intervention measures and ultimately threaten the health of consumers. This study compares traditional selective and nonselective agar-based overlays versus two commercial systems (Petrifilm and Easygel) for recovery of injured E. coli B-41560 and O157:H7 strains. Bacteria were propagated in tryptic soy broth (TSB), ground beef slurry, and infant milk formula to a density of 10(6) to 10(8) CFU/ml and then were stressed for 6 min either in lactic acid (pH 4.5) or heat shocked for 3 min at 60°C. Samples were pour plated in basal layers of either tryptic soy agar (TSA), sorbitol MacConkey agar (SMAC), or violet red bile agar (VRB) and were resuscitated for 4 h prior to addition of agar overlays. Other stressed bacteria were plated directly onto Petrifilm and Easygel. Results indicate that selective and nonselective agar overlays recovered significantly higher numbers (greater than 1 log) of acid- and heat-injured E. coli O157:H7 from TSB, ground beef, and infant milk formula compared with direct plating onto selective media, Petrifilm, or Easygel, while no significant differences among these media combinations were observed for stressed E. coli B-41560. Nonstressed bacteria from TSB and ground beef were also recovered at densities significantly higher in nonselective TSA-TSA and in VRB-VRB and SMAC-SMAC compared with Petrifilm and Easygel. These data underscore the need to implement food safety measures that address sublethally injured pathogens such as E. coli O157:H7 in order to avoid underestimation of true densities for target pathogens.

  9. The gestational power of mast cells in the injured tissue.

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    Aller, Maria-Angeles; Arias, Natalia; Martínez, Vicente; Vergara, Patri; Arias, Jaime

    2018-02-01

    The inflammatory response expressed after wound healing would be the recapitulation of systemic extra-embryonic functions, which would focus on the interstitium of the injured tissue. In the injured tissue, mast cells, provided for a great functional heterogeneity, could play the leading role in the re-expression of extra-embryonic functions, i.e., coelomic-amniotic and trophoblastic-vitelline. Moreover, mast cells would favor the production of a gastrulation-like process, which in certain tissues and organs would induce the regeneration of the injured tissue. Therefore, the engraftment of mesenchymal stem cells and mast cells, both with an extra-embryonic regenerative phenotype, would achieve a blastema, from the repaired and regenerated injured tissue, rather than by fibrosis, which is commonly made through wound-healing.

  10. Sublethal Photothermal Stimulation with a Micropulse Laser Induces Heat Shock Protein Expression in ARPE-19 Cells

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    Keiji Inagaki

    2015-01-01

    Full Text Available Purpose/Aim of the Study. Subthreshold micropulse diode laser photocoagulation is an effective treatment for macular edema. The molecular mechanisms underlying treatment success are poorly understood. Therefore, we investigated the effects of sublethal laser energy doses on a single layer of densely cultured ARPE-19 cells as a model of the human retinal pigment epithelium (RPE. Materials and Methods. A single layer of densely cultured human ARPE-19 cells was perpendicularly irradiated with a micropulse diode laser. Nonirradiated cells served as controls. Sublethal laser energy was applied to form a photocoagulation-like area in the cultured cell layers. Hsp70 expression was evaluated using quantitative polymerase chain reaction and immunocytochemistry. Results. Photocoagulation-like areas were successfully created in cultured ARPE-19 cell layers using sublethal laser energy with our laser irradiation system. Hsp70 mRNA expression in cell layers was induced within 30 min of laser irradiation, peaking at 3 h after irradiation. This increase was dependent on the number of laser pulses. Hsp70 upregulation was not observed in untreated cell layers. Immunostaining indicated that Hsp70 expression occurred concentrically around laser irradiation sites and persisted for 24 h following irradiation. Conclusion. Sublethal photothermal stimulation with a micropulse laser may facilitate Hsp70 expression in the RPE without inducing cellular damage.

  11. Single-cell analysis of S. cerevisiae growth recovery after a sublethal heat-stress applied during an alcoholic fermentation.

    Science.gov (United States)

    Tibayrenc, Pierre; Preziosi-Belloy, Laurence; Ghommidh, Charles

    2011-06-01

    Interest in bioethanol production has experienced a resurgence in the last few years. Poor temperature control in industrial fermentation tanks exposes the yeast cells used for this production to intermittent heat stress which impairs fermentation efficiency. Therefore, there is a need for yeast strains with improved tolerance, able to recover from such temperature variations. Accordingly, this paper reports the development of methods for the characterization of Saccharomyces cerevisiae growth recovery after a sublethal heat stress. Single-cell measurements were carried out in order to detect cell-to-cell variability. Alcoholic batch fermentations were performed on a defined medium in a 2 l instrumented bioreactor. A rapid temperature shift from 33 to 43 °C was applied when ethanol concentration reached 50 g l⁻¹. Samples were collected at different times after the temperature shift. Single cell growth capability, lag-time and initial growth rate were determined by monitoring the growth of a statistically significant number of cells after agar medium plating. The rapid temperature shift resulted in an immediate arrest of growth and triggered a progressive loss of cultivability from 100 to 0.0001% within 8 h. Heat-injured cells were able to recover their growth capability on agar medium after a lag phase. Lag-time was longer and more widely distributed as the time of heat exposure increased. Thus, lag-time distribution gives an insight into strain sensitivity to heat-stress, and could be helpful for the selection of yeast strains of technological interest.

  12. Characterization of high hydrostatic pressure-injured Bacillus subtilis cells.

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    Inaoka, Takashi; Kimura, Keitarou; Morimatsu, Kazuya; Yamamoto, Kazutaka

    2017-06-01

    High hydrostatic pressure (HHP) affects various cellular processes. Using a sporulation-deficient Bacillus subtilis strain, we characterized the properties of vegetative cells subjected to HHP. When stationary-phase cells were exposed to 250 MPa of HHP for 10 min at 25 °C, approximately 50% of cells were viable, although they exhibited a prolonged growth lag. The HHP-injured cells autolyzed in the presence of NaCl or KCl (at concentrations ≥100 mM). Superoxide dismutase slightly protected the viability of HHP-treated cells, whereas vegetative catalases had no effect. Thus, unlike HHP-injured Escherichia coli, oxidative stress only slightly affected vegetative B. subtilis subjected to HHP.

  13. Mitochondrial redox signaling enables repair of injured skeletal muscle cells.

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    Horn, Adam; Van der Meulen, Jack H; Defour, Aurelia; Hogarth, Marshall; Sreetama, Sen Chandra; Reed, Aaron; Scheffer, Luana; Chandel, Navdeep S; Jaiswal, Jyoti K

    2017-09-05

    Strain and physical trauma to mechanically active cells, such as skeletal muscle myofibers, injures their plasma membranes, and mitochondrial function is required for their repair. We found that mitochondrial function was also needed for plasma membrane repair in myoblasts as well as nonmuscle cells, which depended on mitochondrial uptake of calcium through the mitochondrial calcium uniporter (MCU). Calcium uptake transiently increased the mitochondrial production of reactive oxygen species (ROS), which locally activated the guanosine triphosphatase (GTPase) RhoA, triggering F-actin accumulation at the site of injury and facilitating membrane repair. Blocking mitochondrial calcium uptake or ROS production prevented injury-triggered RhoA activation, actin polymerization, and plasma membrane repair. This repair mechanism was shared between myoblasts, nonmuscle cells, and mature skeletal myofibers. Quenching mitochondrial ROS in myofibers during eccentric exercise ex vivo caused increased damage to myofibers, resulting in a greater loss of muscle force. These results suggest a physiological role for mitochondria in plasma membrane repair in injured cells, a role that highlights a beneficial effect of ROS. Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

  14. Endovascular transplantation of stem cells to the injured rat CNS

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    Lundberg, Johan; Soederman, Mikael; Andersson, Tommy; Holmin, Staffan [Karolinska University Hospital, Department of Clinical Neuroscience, Karolinska Institutet, Department of Neuroradiology, Stockholm (Sweden); Le Blanc, Katarina [Karolinska University Hospital, Department of Stem Cell Research, Karolinska Institutet, Department of Clinical Immunology, Stockholm (Sweden)

    2009-10-15

    Transplantation procedures using intraparenchymal injection of stem cells result in tissue injury in addition to associated surgical risks. Intravenous injection of mesenchymal stem cells gives engraftment to lesions, but the method has low efficiency and specificity. In traumatic brain injuries (TBI), there is a transient breakdown of the blood-brain barrier and an inflammatory response, which increase migration of cells from blood to parenchyma. The aim of this investigation was to analyze the effect of intra-arterial administration on cellular engraftment. Experimental TBI was produced in a rat model. Endovascular technique was used to administer human mesenchymal stem cells in the ipsilateral internal carotid artery. Evaluation of engraftment and side effects were performed by immunohistochemical analysis of the brain and several other organs. The results were compared to intravenous administration of stem cells. Intra-arterial transplantion of mesenchymal stem cells resulted in central nervous system (CNS) engraftment without thromboembolic ischemia. We observed a significantly higher number of transplanted cells in the injured hemisphere after intra-arterial compared to intravenous administration both 1 day (p<0.01) and 5 days (p<0.05) after the transplantation. Some cells were also detected in the spleen but not in the other organs analyzed. Selective intra-arterial administration of mesenchymal stem cells to the injured CNS is a minimally invasive method for transplantation. The method is significantly more efficient than the intravenous route and causes no side effects in the current model. The technique can potentially be used for repeated transplantation to the CNS after TBI and in other diseases. (orig.)

  15. Molecular Genetic Analysis of Human Endometrial Mesenchymal Stem Cells That Survived Sublethal Heat Shock

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    A. E. Vinogradov

    2017-01-01

    Full Text Available High temperature is a critical environmental and personal factor. Although heat shock is a well-studied biological phenomenon, hyperthermia response of stem cells is poorly understood. Previously, we demonstrated that sublethal heat shock induced premature senescence in human endometrial mesenchymal stem cells (eMSC. This study aimed to investigate the fate of eMSC-survived sublethal heat shock (SHS with special emphasis on their genetic stability and possible malignant transformation using methods of classic and molecular karyotyping, next-generation sequencing, and transcriptome functional analysis. G-banding revealed random chromosome breakages and aneuploidy in the SHS-treated eMSC. Molecular karyotyping found no genomic imbalance in these cells. Gene module and protein interaction network analysis of mRNA sequencing data showed that compared to untreated cells, SHS-survived progeny revealed some difference in gene expression. However, no hallmarks of cancer were found. Our data identified downregulation of oncogenic signaling, upregulation of tumor-suppressing and prosenescence signaling, induction of mismatch, and excision DNA repair. The common feature of heated eMSC is the silence of MYC, AKT1/PKB oncogenes, and hTERT telomerase. Overall, our data indicate that despite genetic instability, SHS-survived eMSC do not undergo transformation. After long-term cultivation, these cells like their unheated counterparts enter replicative senescence and die.

  16. ATP released by injured neurons activates Schwann cells

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    Samuele eNegro

    2016-05-01

    Full Text Available Injured nerve terminals of neuromuscular junctions (NMJs can regenerate. This remarkable and complex response is governed by molecular signals that are exchanged among the cellular components of this synapse: motor axon nerve terminal (MAT, perisynaptic Schwann cells (PSCs, and muscle fibre. The nature of signals that govern MAT regeneration is ill-known. In the present study the spider toxin α-Latrotoxin has been used as tool to investigate the mechanisms underlying peripheral neuroregeneration. Indeed this neurotoxin induces an acute, specific, localized and fully reversible damage of the presynaptic nerve terminal, and its action mimics the cascade of events that leads to nerve terminal degeneration in injured patients and in many neurodegenerative conditions. Here we provide evidence of an early release by degenerating neurons of ATP as alarm messenger, that contributes to the activation of a series of intracellular pathways within SCs that are crucial for nerve regeneration: Ca2+, cAMP, ERK1/2, and CREB. These results contribute to define the cross-talk taking place among degenerating nerve terminals and PSCs, involved in the functional recovery of the NMJ.

  17. Enhancement of committed hematopoietic stem cell colony formation by nandrolone decanoate after sublethal whole body irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Gallicchio, V.S.; Chen, M.G.; Watts, T.D.

    1984-11-01

    The ability of an anabolic steroid, nandrolone decanoate, to increase committed topoietic stem cell (CFU-gm, CFU-e, and BFU-e) colony formation after sublethal irradiation was evaluated. Immediately after receiving whole body irradiation and on the next two days, each mouse was injected intraperitoneally with nandrolone decanoate (1.25 mg) in propylene glycol. Irradiated control mice received only propylene glycol. Compared to controls, drug-treated mice showed marked peripheral blood leukocytosis and more stable packed red cell volume. Drug-treated mice also demonstrated increased erythropoiesis, as CFU-e/BFU-e concentrations from both marrow (9% to 581%) and spleen (15% to 797%) were elevated. Granulopoiesis was increased similarly, as CFU-gm concentrations from marrow (38% to 685%) and spleen (9% to 373%) were elevated. These results demonstrate that nandrolone decanoate enhances hematopoietic stem cell recovery after sublethal whole body irradiation. This suggests that following hematopoietic suppression, nandrolone decanoate may stimulate the recovery of hematopoiesis at the stem cell level and in peripheral blood.

  18. Transmission electron microscopy study of Listeria monocytogenes serotype 1/2a cells exposed to sublethal heat stress and carvacrol

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    The objective of this study was to investigate the morphological changes that occurred in Listeria monocytogenes serotype 1/2a cells as visualized by transmission electron microscopy (TEM) after exposure to sublethal heat stress at 48°C for 60 min and in combination with lethal concentration of carv...

  19. Arsenic-induced sub-lethal stress reprograms human bronchial epithelial cells to CD61¯ cancer stem cells.

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    Chang, Qingshan; Chen, Bailing; Thakur, Chitra; Lu, Yongju; Chen, Fei

    2014-03-15

    In the present report, we demonstrate that sub-lethal stress induced by consecutive exposure to 0.25 µM arsenic (As3+) for six months can trigger reprogramming of the human bronchial epithelial cell (BEAS-2B) to form cancer stem cells (CSCs) without forced introduction of the stemness transcription factors. These CSCs formed from As3+-induced sub-lethal stress featured with an increased expression of the endogenous stemness genes, including Oct4, Sox2, Klf4, Myc, and others that are associated with the pluripotency and self-renewal of the CSCs. Flow cytometry analysis indicated that 90% of the CSC cells are CD61¯, whereas 100% of the parental cells are CD61+. These CD61¯ CSCs are highly tumorigenic and metastatic to the lung in xenotransplantation tests in NOD/SCID Il2rγ-/- mice. Additional tests also revealed that the CD61¯ CSCs showed a significant decrease in the expression of the genes important for DNA repair and oxidative phosphorylation. To determine the clinical relevance of the above findings, we stratified human lung cancers based on the level of CD61 protein and found that CD61low cancer correlates with poorer survival of the patients. Such a correlation was also observed in human breast cancer and ovarian cancer. Taken together, our findings suggest that in addition to the traditional approaches of enforced introduction of the exogenous stemness circuit transcription factors, sub-lethal stress induced by consecutive low dose As3+ is also able to convert non-stem cells to the CSCs.

  20. Differentiation of different mixed Listeria strains and also acid-injured, heat-injured, and repaired cells of Listeria monocytogenes using fourier transform infrared spectroscopy.

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    Nyarko, Esmond; Donnelly, Catherine

    2015-03-01

    Fourier transform infrared (FT-IR) spectroscopy was used to differentiate mixed strains of Listeria monocytogenes and mixed strains of L. monocytogenes and Listeria innocua. FT-IR spectroscopy was also applied to investigate the hypothesis that heat-injured and acid-injured cells would return to their original physiological integrity following repair. Thin smears of cells on infrared slides were prepared from cultures for mixed strains of L. monocytogenes, mixed strains of L. monocytogenes and L. innocua, and each individual strain. Heat-injured and acid-injured cells were prepared by exposing harvested cells of L. monocytogenes strain R2-764 to a temperature of 56 ± 0.2°C for 10 min or lactic acid at pH 3 for 60 min, respectively. Cellular repair involved incubating aliquots of acid-injured and heat-injured cells separately in Trypticase soy broth supplemented with 0.6% yeast extract for 22 to 24 h; bacterial thin smears on infrared slides were prepared for each treatment. Spectral collection was done using 250 scans at a resolution of 4 cm(-1) in the mid-infrared wavelength region. Application of multivariate discriminant analysis to the wavelength region from 1,800 to 900 cm(-1) separated the individual L. monocytogenes strains. Mixed strains of L. monocytogenes and L. monocytogenes cocultured with L. innocua were successfully differentiated from the individual strains when the discriminant analysis was applied. Different mixed strains of L. monocytogenes were also successfully separated when the discriminant analysis was applied. A data set for injury and repair analysis resulted in the separation of acid-injured, heat-injured, and intact cells; repaired cells clustered closer to intact cells when the discriminant analysis (1,800 to 600 cm(-1)) was applied. FT-IR spectroscopy can be used for the rapid source tracking of L. monocytogenes strains because it can differentiate between different mixed strains and individual strains of the pathogen.

  1. [Protective effect of six Kaixin San formulas on nerve cells injured by different materials].

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    Zhao, Hai-Xia; Zhou, Xiao-Jiang; Hu, Yuan; Dong, Xian-Zhe; Cao, Yin; Liu, Ping

    2012-11-01

    To investigate the protective effect of six Kaixin San formulas on simulated cells model of depression, Alzheimer's disease and Parkinson's disease. The in vitro simulated cells model of depression, Alzheimer's disease and Parkinson's disease was established by injuring SH-SY5Y cells with corticosterone (0.4 mmol x L(-1)) , injuring PC12 cells with neurotoxic amyloid peptide (Abeta25-35) (20 micromol x L(-1)) and injuring SH-SY5Y cells with 1-methyl-4-phenylpyridinium ion (MPP+) (250 micromol x L(-1)). The cell survival rate was assayed with MTT method and the degree of cell injury was detected with LDH. 100, 500 mg x L(-1) Dingzhixiao Wan prepared as mentioned in Beiji Qianjin Yaofang could significantly increase the survival ratio of SH-SY5Y cells injured by corticosterone and reduce LDH concentration released. All of the Kaixin San formulas could significantly increase the survival ratio of PC12 cells injured by Abeta25-35 and reduce LDH concentration released. Particularly, Kaixin San (10, 100, 500 mg L(-1)) prepared as mentioned in Beiji Qianjin Yaofang shown the best effect. And 500 mg x L(-1) Fushen Wan prepared as mentioned in Gujin Luyan could significantly increase survival ratio of SH-SY5Y cell injured by MPP and reduce LDH concentration released. Dingzhixiao Wan prepared as mentioned in Beiji Qianjin Yaofang could protect corticosterone-induced SH-SY5Ycells injury, showing a potential antidepressant effect. All of the six Kaixin San formulas could protect Abeta25-35-induced PC12 cells injury, but Kaixin San prepared as mentioned in Beiji Qianjin Yaofang shown the best potential effect for Alzheimer's disease. Fushen Wan prepared as mentioned in Gujin Luyan could protect MPP(+)-induced SH-SY5Y cells injury to some extent.

  2. Protective effect of a laser-induced sub-lethal temperature rise on RPE cells from oxidative stress.

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    Iwami, Hisashi; Pruessner, Joachim; Shiraki, Kunihiko; Brinkmann, Ralf; Miura, Yoko

    2014-07-01

    Recently introduced new technologies that enable temperature-controlled laser irradiation on the RPE allowed us to investigate temperature-resolved RPE cell responses. In this study we aimed primarily to establish an experimental setup that can realize laser irradiation on RPE cell culture with the similar temperature distribution as in the clinical application, with a precise time/temperature history. With this setup, we conducted investigations to elucidate the temperature-dependent RPE cell biochemical responses and the effect of transient hyperthermia on the responses of RPE cells to the secondary-exposed oxidative stress. Porcine RPE cells cultivated in a culture dish (inner diameter = 30 mm) with culture medium were used, on which laser radiation (λ = 1940 nm, spot diameter = 30 mm) over 10 s was applied as a heat source. The irradiation provides a radially decreasing temperature profile which is close to a Gaussian shape with the highest temperature in the center. Power setting for irradiation was determined such that the peak temperature (Tmax) in the center of the laser spot at the cells reaches from 40 °C to 58 °C (40, 43, 46, 50, 58 °C). Cell viability was investigated with ethidium homodimer III staining at the time points of 3 and 24 h following laser irradiation. Twenty four hours after laser irradiation the cells were exposed to hydrogen peroxide (H2O2) for 5 h, followed by the measurement of intracellular glutathione, intracellular 4-hydroxynonenal (HNE) protein adducts, and secreted vascular endothelial growth factor (VEGF). The mean temperature threshold for RPE cell death after 3 h was found to be around 52 °C, and for 24 h around 50 °C with the current irradiation setting. A sub-lethal preconditioning on Tmax = 43 °C significantly induced the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio, and decreased H2O2-induced increase of intracellular 4-HNE protein adducts. Although sub-lethal hyperthermia (Tmax

  3. Radiosensitivity and capacity for radiation-induced sublethal damage repair of canine transitional cell carcinoma (TCC) cell lines.

    Science.gov (United States)

    Parfitt, S L; Milner, R J; Salute, M E; Hintenlang, D E; Farese, J P; Bacon, N J; Bova, F J; Rajon, D A; Lurie, D M

    2011-09-01

    Understanding the inherent radiosensitivity and repair capacity of canine transitional cell carcinoma (TCC) can aid in optimizing radiation protocols to treat this disease. The objective of this study was to evaluate the parameters surviving fraction at 2 Gy (SF(2) ), α/β ratio and capacity for sublethal damage repair (SLDR) in response to radiation. Dose-response and split-dose studies were performed using the clonogenic assay. The mean SF(2) for three established TCC cell lines was high at 0.61. All the three cell lines exhibited a low to moderate α/β ratio, with the mean being 3.27. Two cell lines exhibited statistically increased survival at 4 and 24 h in the dose-response assay. Overall, our results indicate that the cell lines are moderately radioresistant, have a high repair capacity and behave similarly to a late-responding normal tissue. These findings indicate that the radiation protocols utilizing higher doses with less fractionation may be more effective for treating TCC. © 2011 Blackwell Publishing Ltd.

  4. Real-time monitoring of cell viability and cell density on the basis of a three dimensional optical reflectance method (3D-ORM): investigation of the effect of sub-lethal and lethal injuries.

    Science.gov (United States)

    Brognaux, Alison; Bugge, Jörg; Schwartz, Friedel H; Thonart, Philippe; Telek, Samuel; Delvigne, Frank

    2013-07-01

    Cell density and cell viability have been followed on-line by using a three-dimensional optical reflectance method (3D-ORM) probe. This method has allowed to highlight the differences between a well-mixed and a scale-down bioreactor configured in order to reproduce mixing deficiencies during a fed-batch culture of Escherichia coli. These differences have been observed both for the obscuration factor (OBF) and the coincidence probability delivered by the probe. These parameters are correlated to flow cytometry measurement based on the PI-uptake test and cell density based on optical density measurement. This first set of results has pointed out the fact that the 3D-ORM probe is sensitive to sub-lethal injuries encountered by microbial cells in process-related conditions. The effect of lethal injuries has been further investigated on the basis of additional experiments involving heat stress and a sharp increase of the OBF has been observed indicating that cells are effectively injured by the increase of temperature. However, further improvement of the probe are needed in order to give access to single-cell measurements.

  5. DETECTION OF THERMAL SUBLETHAL INJURY IN ESCHERICHIA COLI VIA THE SELECTIVE MEDIUM PLATING TECHNIQUE: MECHANISMS AND IMPROVEMENTS

    Directory of Open Access Journals (Sweden)

    Laura Espina

    2016-08-01

    Full Text Available In food preservation, the selective medium plating technique (SMPT is commonly used in order to detect and quantify the amount of sublethally injured cells in their bacterial cytoplasmic membranes after inimical treatments. From an applicative point of view, this information is of use in the synergistic combination of different preservation technologies, so that cells that are sublethally injured after one or more processes can end up being entirely inactivated by other hurdle(s. However, little work has been done to explain the reasons for the inability of sublethally injured cells to outgrow in selective agar media (containing the osmolyte NaCl as a selective agent, whereas they are able to grow in non-selective agar media. This research could contribute to explain this technique’s limits. In the present paper, the performance of SMPT on Escherichia coli cells after heat treatments is explored by applying different selective agents in the recovery media, using several mutants lacking factors involved in osmoregulation, and also by examining the integrity of the cytoplasmic membrane. In view of the results, the possibility of a specific toxic effect of Na+ as the main mechanism under SMPT is discarded, and the same level of sublethal injury is detected using KCl instead of NaCl. The synthesis of the osmoprotectant trehalose determined the maximum osmotolerance of intact cells to the selective agents, but was not crucial in the quantification of sublethal injury. Moreover, the extent of sublethal injury detected via SMPT was directly correlated with the physical loss of integrity of the cell membrane as measured with the propidium iodide-exclusion technique when that dye was added before thermal treatments. The present work confirms the adequacy of SMPT as a tool for detecting the occurrence and quantity of sublethally injured cells and thus, for efficiently designing combined preservation treatments. Additionally, we propose the combination

  6. Plasticity and regeneration in the injured spinal cord after cell transplantation therapy.

    Science.gov (United States)

    Nori, Satoshi; Nakamura, Masaya; Okano, Hideyuki

    2017-01-01

    Spinal cord injury (SCI) typically damages the long axonal tracts of the spinal cord which results in permanent disability. However, regeneration of the injured spinal cord is approaching reality according to the advances in stem cell biology. Cell transplantation therapy holds potential to lead to recovery following SCI through some positive mechanisms. Grafted cells induce plasticity and regeneration in the injured spinal cord by promoting remyelination of damaged axons, reconstruction of neural circuits by synapse formation between host neurons and graft-derived neurons, and secreting neurotrophic factors to promote axonal elongation as well as reduce retrograde axonal degeneration. In this review, we will delineate (1) the microenvironment of the injured spinal cord that influence the plasticity and regeneration capacity after SCI, (2) a number of different kinds of cell transplantation therapies for SCI that has been extensively studied by researchers, and (3) potential mechanisms of grafted cell-induced regeneration and plasticity in the injured spinal cord. © 2017 Elsevier B.V. All rights reserved.

  7. Functional Electrical Stimulation Helps Replenish Progenitor Cells in the Injured Spinal Cord of Adult Rats

    Science.gov (United States)

    Becker, Daniel; Gary, Devin S.; Rosenzweig, Ephron S.; Grill, Warren M.; McDonald, John W.

    2010-01-01

    Functional electrical stimulation (FES) can restore control and offset atrophy to muscles after neurological injury. However, FES has not been considered as a method for enhancing CNS regeneration. This paper demonstrates that FES dramatically enhanced progenitor cell birth in the spinal cord of rats with a chronic spinal cord injury (SCI). A complete SCI at thoracic level 8/9 was performed on 12 rats. Three weeks later, a FES device to stimulate hindlimb movement was implanted into these rats. Twelve identically-injured rats received inactive FES implants. An additional control group of uninjured rats were also examined. Ten days after FES implantation, dividing cells were marked with bromodeoxyuridine (BrdU). The ‘cell birth’ subgroup (half the animals in each group) was sacrificed immediately after completion of BrdU administration, and the ‘cell survival’ subgroup was sacrificed 7 days later. In the injuredcell birth’ subgroup, FES induced an 82-86 % increase in cell birth in the lumbar spinal cord. In the injuredcell survival’ subgroup, the increased lumbar newborn cell counts persisted. FES doubled the proportion of the newly-born cells which expressed nestin and other markers suggestive of tripotential progenitors. In uninjured rats, FES had no effect on cell birth/survival. This report suggests that controlled electrical activation of the CNS may enhance spontaneous regeneration after neurological injuries. PMID:20059998

  8. Cell death in the injured brain: roles of metallothioneins

    DEFF Research Database (Denmark)

    Pedersen, Mie Ø; Larsen, Agnete; Stoltenberg, Meredin

    2009-01-01

    oxygen species (ROS). ROS promote oxidative stress, which leads to neurodegeneration and ultimately results in programmed cell death (secondary injury). Since this delayed, secondary tissue loss occurs days to months following the primary injury it provides a therapeutic window where potential...... neuroprotective treatment could alleviate ongoing neurodegeneration, cell death and neurological impairment following TBI. Various neuroprotective drug candidates have been described, tested and proven effective in pre-clinical studies, including glutamate receptor antagonists, calcium-channel blockers......, and caspase inhibitors. However, most of the scientific efforts have failed in translating the experimental results into clinical trials. Despite intensive research, effective neuroprotective therapies are lacking in the clinic, and TBI continues to be a major cause of morbidity and mortality. This paper...

  9. Tracking of CFSE-labeled endothelial progenitor cells in laser-injured mouse retina.

    Science.gov (United States)

    Shi, Hui; Yang, Wei; Cui, Zhi-Hua; Lu, Cheng-Wei; Li, Xiao-Hong; Liang, Ling-Ling; Song, E

    2011-03-01

    Endothelial progenitor cells (EPCs) transplantation is a promising therapeutic strategy for ischemic retinopathy. The current study aimed to establish a simple, reliable and fluorescent labeling method for tracking EPCs with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) in laser-injured mouse retina. EPCs were isolated from human umbilical cord blood mononuclear cells, cultivated, and labeled with various concentrations of CFSE. Based on fluorescence intensity and cell morphology, a 15 minutes incubation with 5 µmol/L CFSE at 37°C was selected as the optimal labeling condition. The survival capability and the apoptosis rate of CFSE-labeled EPCs were measured by Trypan blue staining and Annexin V/PI staining assay respectively. Fluorescence microscopy was used to observe the label stability during the extended culture period. Labeled EPCs were transplanted into the vitreous cavity of pigmented mice injured by retinal laser photocoagulation. Evans Blue angiography and flat mounted retinas were examined to track the labeled cells. EPCs labeled with 5 µmol/L CFSE presented an intense green fluorescence and maintained normal morphology, with no significant changes in the survival capability or apoptosis rate after being labeled for 2 days, 1 and 4 weeks. The fluorescence intensity gradually decreased in the cells at the end of 4 weeks. Evans Blue angiography of the retina displayed the retinal capillarity network clearly and fluorescence leakage was observed around photocoagulated spots in the laser-injured mouse model. One week after transplantation of labeled EPCs, the fluorescent cells were identified around the photocoagulated lesions. Four weeks after transplantation, fluorescent tube-like structures were observed in the retinal vascular networks. EPCs could be labeled by CFSE in vitro and monitored in vivo for at least 4 weeks, and participate in the repair of injured retinal vessels.

  10. Differential protection of injured retinal ganglion cell dendrites by brimonidine.

    Science.gov (United States)

    Lindsey, James D; Duong-Polk, Karen X; Hammond, Dustin; Chindasub, Panida; Leung, Christopher Kai-Shun; Weinreb, Robert N

    2015-01-29

    To determine whether brimonidine protects against the retraction and loss of retinal ganglion cell (RGC) dendrites after optic nerve crush (ONC). Fluorescent RGCs of mice expressing yellow fluorescent protein (YFP) under the control of the Thy-1 promoter (Thy1-YFP mice) were imaged in vivo and assigned to one of six groups according to dendrite structure. The mice then received brimonidine every other day starting 2 days before, or 2 or 6 days after, unilateral ONC. Control animals received vehicle every other day starting 2 days before ONC. Control animals received vehicle every other day starting 2 days before ONC. Total dendrite length, dendrite branching complexity, and the time until complete loss of dendrites were assessed weekly for 4 weeks. Overall, brimonidine treatment significantly slowed the complete loss of RGC dendrites and significantly slowed the reduction of total dendrite length and branching complexity. Separate analysis of each RGC group showed brimonidine significantly delayed the time until complete loss of dendrites in four of the RGC groups. These delays generally were similar when treatment started either 2 days before or 2 days after ONC, but were smaller or absent when treatment started 6 days after ONC Protection against loss of total dendrite length and loss of branching complexity was observed in three of the RGC groups. In two of these RGC groups, protective effects persisted until the end of the study. Brimonidine protects many RGC types against dendrite retraction, loss of branching complexity, and complete loss of dendrites following ONC. However, the pattern and magnitude of this protection differs substantially among different RGC types. These results indicate that requirements for RGC-protective therapies following optic nerve injury may differ among RGC types. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.

  11. Evaluation of a multiplex selective enrichment broth SEL for simultaneous detection of injured Salmonella, Escherichia coli O157:H7 and Listeria monocytogenes.

    Science.gov (United States)

    Suo, Biao; Wang, Yuexia

    2013-01-01

    Although many rapid and high throughput molecular methods have been developed in the recent years for the multiplex detection of foodborne pathogens, the simultaneous recovery and enrichment of sublethally injured cells is still a problem that needs to be considered. Combined with previous established multiplex real-time PCR assay, the capability of simultaneous recovery and enrichment of sublethally injured Salmonella, E. coli O157:H7 and L. monocytogenes cells was evaluated in a multiplex selective enrichment broth SEL. The injured cells were obtained by heat shock. After evaluation of different procedures, 1 h of recovery period prior to 20 h of enrichment was proved to be necessary for the detection of less than 10 CFU/5 mL broth of injured L. monocytogenes. When the detection method was applied to artificially contaminated ground beef, all the three injured pathogens could be simultaneously detected without discrimination by real-time PCR combined with SEL broth, the detection limit was SEL broth herein appears to be a promising tool for high-throughput screening of a large number of processed food samples, which require either single or multiple pathogen detection. More important, the sublethally injured foodborne pathogen cells were also detectable.

  12. Engraftment of enteric neural progenitor cells into the injured adult brain.

    Science.gov (United States)

    Belkind-Gerson, Jaime; Hotta, Ryo; Whalen, Michael; Nayyar, Naema; Nagy, Nandor; Cheng, Lily; Zuckerman, Aaron; Goldstein, Allan M; Dietrich, Jorg

    2016-01-25

    A major area of unmet need is the development of strategies to restore neuronal network systems and to recover brain function in patients with neurological disease. The use of cell-based therapies remains an attractive approach, but its application has been challenging due to the lack of suitable cell sources, ethical concerns, and immune-mediated tissue rejection. We propose an innovative approach that utilizes gut-derived neural tissue for cell-based therapies following focal or diffuse central nervous system injury. Enteric neuronal stem and progenitor cells, able to differentiate into neuronal and glial lineages, were isolated from the postnatal enteric nervous system and propagated in vitro. Gut-derived neural progenitors, genetically engineered to express fluorescent proteins, were transplanted into the injured brain of adult mice. Using different models of brain injury in combination with either local or systemic cell delivery, we show that transplanted enteric neuronal progenitor cells survive, proliferate, and differentiate into neuronal and glial lineages in vivo. Moreover, transplanted cells migrate extensively along neuronal pathways and appear to modulate the local microenvironment to stimulate endogenous neurogenesis. Our findings suggest that enteric nervous system derived cells represent a potential source for tissue regeneration in the central nervous system. Further studies are needed to validate these findings and to explore whether autologous gut-derived cell transplantation into the injured brain can result in functional neurologic recovery.

  13. Recruitment of stem cells into the injured retina after laser injury.

    Science.gov (United States)

    Singh, Tajinder; Prabhakar, Sudesh; Gupta, Amod; Anand, Akshay

    2012-02-10

    Retinal degeneration is a devastating complication of diabetes and other disorders. Stem cell therapy for retinal degeneration has shown encouraging results but functional regeneration has not been yet achieved. Our study was undertaken to evaluate the localization of stem cells delivered to the retina by intravenous versus intravitreal infusion, because stem cell localization is a key factor in ultimate in vivo function. We used lineage-negative bone marrow-derived stem cells in a model wherein retina of mice was induced by precise and reproducible laser injury. Lin(-ve) bone marrow cells (BMCs) were labeled with a tracking dye and their homing capacity was analyzed at time points after infusion. We found that Lin(-ve) BMCs get incorporated into laser-injured retina when transplanted through either the intravitreal or intravenous route. The intravenous route resulted in optimal localization of donor cells at the site of injury. These cells incorporated into injured retina in a dose-dependent manner. The data presented in this study reflect the importance of dose and route for stem cell-based treatment designed to result in retinal regeneration.

  14. The effects of the stem cell on ciliary regeneration of injured rabbit sinonasal epithelium.

    Science.gov (United States)

    Kavuzlu, Ali; Tatar, Emel Çadallı; Karagöz, Tuğba; Pınarlı, Ferda Alpaslan; Tatar, İlkan; Bayır, Ömer; Korkmaz, Mehmet Hakan

    2017-08-01

    Defects in mucosal healing after sinonasal surgery cause infection, scar formation causing obstruction, relapse of the disease within a shorter period and revision surgery. The present study aimed to create a functional ciliated epithelium using a stem cell and stem cell sheet of adipose tissue origin and to show such regeneration ultra-structurally on experimentally injured rabbit nasal epithelium. This was an experimental animal study and basic research. A total of 18 white New Zealand rabbits were divided into three groups. The medial wall of the maxillary sinus of the subjects was peeled off bilaterally. No additional procedure was applied to the subjects in Group 1. In Group 2, adipose tissue-derived mesenchymal stem cell was implanted on the wound edges of the subjects. In Group 3, a stem cell sheet of three layers was laid onto the defect area. All subjects were killed after 3 weeks. The presence of the stem cell stained with bromo-deoxyuridine was assessed with a light microscope, whereas cilia density, ciliated orientation and cilia structure were evaluated with a scanning electron microscope. Ciliary densities in Group 2 and Group 3 were statistically superior compared to the control group (p stem cell increased the healing of the injured maxillary sinus mucosa of the rabbits in terms of cilia presence, density and morphology regardless of the implementation technique. Level of evidence NA.

  15. Tendon progenitor cells in injured tendons have strong chondrogenic potential: the CD105-negative subpopulation induces chondrogenic degeneration.

    Science.gov (United States)

    Asai, Shuji; Otsuru, Satoru; Candela, Maria Elena; Cantley, Leslie; Uchibe, Kenta; Hofmann, Ted J; Zhang, Kairui; Wapner, Keith L; Soslowsky, Louis J; Horwitz, Edwin M; Enomoto-Iwamoto, Motomi

    2014-12-01

    To study the cellular mechanism of the tendon repair process, we used a mouse Achilles tendon injury model to focus on the cells recruited to the injured site. The cells isolated from injured tendon 1 week after the surgery and uninjured tendons contained the connective tissue progenitor populations as determined by colony-forming capacity, cell surface markers, and multipotency. When the injured tendon-derived progenitor cells (inTPCs) were transplanted into injured Achilles tendons, they were not only integrated in the regenerating area expressing tenogenic phenotype but also trans-differentiated into chondrogenic cells in the degenerative lesion that underwent ectopic endochondral ossification. Surprisingly, the micromass culture of the inTPCs rapidly underwent chondrogenic differentiation even in the absence of exogenous bone morphogenetic proteins or TGFβs. The cells isolated from human ruptured tendon tissues also showed connective tissue progenitor properties and exhibited stronger chondrogenic ability than bone marrow stromal cells. The mouse inTPCs contained two subpopulations one positive and one negative for CD105, a coreceptor of the TGFβ superfamily. The CD105-negative cells showed superior chondrogenic potential in vitro and induced larger chondroid degenerative lesions in mice as compared to the CD105-positive cells. These findings indicate that tendon progenitor cells are recruited to the injured site of tendons and have a strong chondrogenic potential and that the CD105-negative population of these cells would be the cause for chondroid degeneration in injured tendons. The newly identified cells recruited to the injured tendon may provide novel targets to develop therapeutic strategies to facilitate tendon repair. © 2014 AlphaMed Press.

  16. Suspension Matrices for Improved Schwann-Cell Survival after Implantation into the Injured Rat Spinal Cord

    Science.gov (United States)

    Patel, Vivek; Joseph, Gravil; Patel, Amit; Patel, Samik; Bustin, Devin; Mawson, David; Tuesta, Luis M.; Puentes, Rocio; Ghosh, Mousumi

    2010-01-01

    Abstract Trauma to the spinal cord produces endogenously irreversible tissue and functional loss, requiring the application of therapeutic approaches to achieve meaningful restoration. Cellular strategies, in particular Schwann-cell implantation, have shown promise in overcoming many of the obstacles facing successful repair of the injured spinal cord. Here, we show that the implantation of Schwann cells as cell suspensions with in-situ gelling laminin:collagen matrices after spinal-cord contusion significantly enhances long-term cell survival but not proliferation, as well as improves graft vascularization and the degree of axonal in-growth over the standard implantation vehicle, minimal media. The use of a matrix to suspend cells prior to implantation should be an important consideration for achieving improved survival and effectiveness of cellular therapies for future clinical application. PMID:20144012

  17. The radiobiology of laser-driven particle beams: focus on sub-lethal responses of normal human cells

    Science.gov (United States)

    Manti, L.; Perozziello, F. M.; Borghesi, M.; Candiano, G.; Chaudhary, P.; Cirrone, G. A. P.; Doria, D.; Gwynne, D.; Leanza, R.; Prise, K. M.; Romagnani, L.; Romano, F.; Scuderi, V.; Tramontana, A.

    2017-03-01

    Accelerated proton beams have become increasingly common for treating cancer. The need for cost and size reduction of particle accelerating machines has led to the pioneering investigation of optical ion acceleration techniques based on laser-plasma interactions as a possible alternative. Laser-matter interaction can produce extremely pulsed particle bursts of ultra-high dose rates (>= 109 Gy/s), largely exceeding those currently used in conventional proton therapy. Since biological effects of ionizing radiation are strongly affected by the spatio-temporal distribution of DNA-damaging events, the unprecedented physical features of such beams may modify cellular and tissue radiosensitivity to unexplored extents. Hence, clinical applications of laser-generated particles need thorough assessment of their radiobiological effectiveness. To date, the majority of studies have either used rodent cell lines or have focussed on cancer cell killing being local tumour control the main objective of radiotherapy. Conversely, very little data exist on sub-lethal cellular effects, of relevance to normal tissue integrity and secondary cancers, such as premature cellular senescence. Here, we discuss ultra-high dose rate radiobiology and present preliminary data obtained in normal human cells following irradiation by laser-accelerated protons at the LULI PICO2000 facility at Laser Lab Europe, France.

  18. Thermal inactivation and sublethal injury kinetics of Salmonella enterica and Listeria monocytogenes in broth versus agar surface.

    Science.gov (United States)

    Wang, Xiang; Devlieghere, Frank; Geeraerd, Annemie; Uyttendaele, Mieke

    2017-02-21

    The objective of the present study was to compare the thermal inactivation and sublethal injury kinetics of Salmonella enterica and Listeria monocytogenes in broth (suspended cells) and on solid surface (agar-seeded cells). A 3-strain cocktail of S. enterica or L. monocytogenes inoculated in broth or on agar was subjected to heating in a water bath at various set temperatures (55.0, 57.5 and 60.0°C for S. enterica and 60.0, 62.5 and 65°C for L. monocytogenes). The occurrence of sublethally injured cells was determined by comparing enumerations on nonselective (TSAYE) and selective (XLD or ALOA) media. Results showed that the inactivation curves obtained from selective media were log-linear, and significant shoulders (pagar surface exhibited higher heat resistance than those in broth. For S. enterica, cell injury increased with the exposure time, no difference was observed when treated at temperatures from 55.0 to 60.0°C, while for L. monocytogenes, cell injury increased significantly with heating time and treatment temperature (from 60.0 to 65°C). Moreover, the degree of sublethal injury affected by thermal treatment in broth or on agar surface depended upon the target microorganism. Higher proportions of injured S. enterica cells were observed for treatment in broth than on agar surface, while the opposite was found for L. monocytogenes. The provided information may be used to assess the efficacy of thermal treatment processes on surfaces for inactivation of S. enterica and L. monocytogenes, and it provides insight into the sublethally injured survival state of S. enterica and L. monocytogenes treated in liquid or on solid food. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Autoserum: An Optimal Supplement for Bone Marrow Mesenchymal Stem Cells of Liver-Injured Rats

    Directory of Open Access Journals (Sweden)

    Qinglin Zhang

    2015-01-01

    Full Text Available Mesenchymal stem cells (MSCs are an attractive source for the clinical cell therapy of liver injury. Although the use of adult serum, platelet lysate, or cord blood serum solves some of the problems caused by fetal bovine serum (FBS, the allogeneic immune response, contamination, and donor-to-donor and donor-to-receptor differences still obstruct the application of MSCs. In this study, the influences of autoserum from liver-injured rats (LIRs and allogeneic serum from healthy rats on the isolation and culture of bone marrow MSCs (BMSCs were examined and compared to FBS. The results showed that BMSCs cultured with autoserum or allogeneic serum exhibited better MSC-specific morphology, lower rate of cell senescent, and higher proliferation kinetics than those with FBS. In addition, autoserum promoted the osteogenic differentiation potential of BMSCs as allogeneic serum did. Although there were no significant differences in proliferation activity, immunophenotypic characterization, and differentiation potential between BMSCs cultured with autoserum and those with allogeneic serum, the potential adverse immunological reactions in patients with allogeneic material transplantation must be considered. We therefore believe that the autoserum from liver-injured patients may be a better choice for MSC expansion to meet the needs of liver injury therapy.

  20. Crosstalk Between Activated Myofibroblasts and β Cells in Injured Mouse Pancreas.

    Science.gov (United States)

    Bayan, Jennifer-Ann; Peng, Zhechu; Zeng, Ni; He, Lina; Chen, Jingyu; Stiles, Bangyan L

    2015-10-01

    In injury conditions, myofibroblasts are induced to lay down matrix proteins and support the repair process. In this study, we investigated the role of myofibroblasts, particularly stellate cells, in the growth and regeneration of pancreatic β cells. We used both in vitro and in vivo approaches to address whether stellate cells may promote the growth of β cells. Our experiments demonstrated that activated stellate cells support the proliferation of β cells in vitro. In vivo, mesenchymals surrounding the pancreatic islets are activated (induced to proliferate) in the islet regeneration model of Pten null mice. These mesenchymals display markers of pancreatic stellate cells, such as desmin and to a lesser extent, smooth muscle actin α. We have shown previously that targeted β-cell deletion of Pten lead to a significant increase in total islet mass. This phenotype was accompanied by an increase in peri-islet mitotic activity, particularly in islets injured by streptozotocin, a β cell-specific toxin. Together with the in vitro observations, our data, here, suggest that that these mesenchymal cells may support the regeneration of the islets. Identifying how the communication occurs may provide clinically relevant mechanism for inducing β-cell regeneration.

  1. Effect of prolonged exposure to sublethal concentrations of DDT and DDE on protein expression in human pancreatic beta cells.

    Science.gov (United States)

    Pavlikova, Nela; Smetana, Pavel; Halada, Petr; Kovar, Jan

    2015-10-01

    Pollution of the environment represents one of less explored potential reasons for the worldwide epidemic of type 2 diabetes. One of the most prevalent organochlorine pollutants remains the pesticide DDT and its degradation product DDE. Despite some epidemiologic correlations between levels of DDT and DDE in human organism and the prevalence of diabetes, there is almost no information about the exact targets of these compounds inside pancreatic beta cells. To detect functional areas of pancreatic beta cells that could be affected by exposure to DDT and DDE, we analyzed changes in protein expression in the NES2Y human pancreatic beta cell line exposed to three sublethal concentrations (0.1 μM, 1 μM, 10 μM) of DDT and DDE for 1 month. Protein separation and identification was achieved using high-resolution 2D-electrophoresis, computer analysis and mass spectrometry. With these techniques, four proteins were found downregulated after exposure to 10 μM DDT: three cytoskeletal proteins (cytokeratin 8, cytokeratin 18 and actin) and one protein involved in glycolysis (alpha-enolase). Two proteins were downregulated after exposure to 10 μM DDE: cytokeratin 18 and heterogenous nuclear ribonucleoprotein H1 (HNRH1). These changes correlate with previously described effects of other stress conditions (e.g. exposure to palmitate, hyperglycemia, imidazoline derivative, and cytokines) on protein expression in pancreatic beta cells. We conclude that cytoskeletal proteins and their processing, glucose metabolism, and mRNA processing may represent targets affected by exposure to conditions hostile to pancreatic beta cells, including exposure to DDT and DDE. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Muscle side population cells from dystrophic or injured muscle adopt a fibro-adipogenic fate.

    Science.gov (United States)

    Penton, Christopher M; Thomas-Ahner, Jennifer M; Johnson, Eric K; McAllister, Cynthia; Montanaro, Federica

    2013-01-01

    Muscle side population (SP) cells are rare multipotent stem cells that can participate in myogenesis and muscle regeneration upon transplantation. While they have been primarily studied for the development of cell-based therapies for Duchenne muscular dystrophy, little is known regarding their non-muscle lineage choices or whether the dystrophic muscle environment affects their ability to repair muscle. Unfortunately, the study of muscle SP cells has been challenged by their low abundance and the absence of specific SP cell markers. To address these issues, we developed culture conditions for the propagation and spontaneous multi-lineage differentiation of muscle SP cells. Using this approach, we show that SP cells from wild type muscle robustly differentiate into satellite cells and form myotubes without requiring co-culture with myogenic cells. Furthermore, this myogenic activity is associated with SP cells negative for immune (CD45) and vascular (CD31) markers but positive for Pax7, Sca1, and the mesenchymal progenitor marker PDGFRα. Additionally, our studies revealed that SP cells isolated from dystrophic or cardiotoxin-injured muscle fail to undergo myogenesis. Instead, these SP cells rapidly expand giving rise to fibroblast and adipocyte progenitors (FAPs) and to their differentiated progeny, fibroblasts and adipocytes. Our findings indicate that muscle damage affects the lineage choices of muscle SP cells, promoting their differentiation along fibro-adipogenic lineages while inhibiting myogenesis. These results have implications for a possible role of muscle SP cells in fibrosis and fat deposition in muscular dystrophy. In addition, our studies provide a useful in vitro system to analyze SP cell biology in both normal and pathological conditions.

  3. Gene expression changes in the injured spinal cord following transplantation of mesenchymal stem cells or olfactory ensheathing cells.

    Directory of Open Access Journals (Sweden)

    Abel Torres-Espín

    Full Text Available Transplantation of bone marrow derived mesenchymal stromal cells (MSC or olfactory ensheathing cells (OEC have demonstrated beneficial effects after spinal cord injury (SCI, providing tissue protection and improving the functional recovery. However, the changes induced by these cells after their transplantation into the injured spinal cord remain largely unknown. We analyzed the changes in the spinal cord transcriptome after a contusion injury and MSC or OEC transplantation. The cells were injected immediately or 7 days after the injury. The mRNA of the spinal cord injured segment was extracted and analyzed by microarray at 2 and 7 days after cell grafting. The gene profiles were analyzed by clustering and functional enrichment analysis based on the Gene Ontology database. We found that both MSC and OEC transplanted acutely after injury induce an early up-regulation of genes related to tissue protection and regeneration. In contrast, cells transplanted at 7 days after injury down-regulate genes related to tissue regeneration. The most important change after MSC or OEC transplant was a marked increase in expression of genes associated with foreign body response and adaptive immune response. These data suggest a regulatory effect of MSC and OEC transplantation after SCI regarding tissue repair processes, but a fast rejection response to the grafted cells. Our results provide an initial step to determine the mechanisms of action and to optimize cell therapy for SCI.

  4. MRI of Auto-Transplantation of Bone Marrow-Derived Stem-Progenitor Cells for potential Repair of Injured Arteries

    NARCIS (Netherlands)

    Meng, Y.; Zhang, F.; Blair, T.; Gu, H.; Feng, H.; Wang, J.; Yuan, C.; Zhang, Z.; Qiu, B.; Yang, X.

    2012-01-01

    Backgroud: This study was to validate the feasibility of using clinical 3.0T MRI to monitor the migration of autotransplanted bone marrow (BM)-derived stem-progenitor cells (SPC) to the injured arteries of near-human sized swine for potential cell-based arterial repair.Methodology: The study was

  5. Activated platelet-rich plasma improves adipose-derived stem cell transplantation efficiency in injured articular cartilage

    Science.gov (United States)

    2013-01-01

    Introduction Adipose-derived stem cells (ADSCs) have been isolated, expanded, and applied in the treatment of many diseases. ADSCs have also been used to treat injured articular cartilage. However, there is controversy regarding the treatment efficiency. We considered that ADSC transplantation with activated platelet-rich plasma (PRP) may improve injured articular cartilage compared with that of ADSC transplantation alone. In this study, we determined the role of PRP in ADSC transplantation to improve the treatment efficiency. Methods ADSCs were isolated and expanded from human adipose tissue. PRP was collected and activated from human peripheral blood. The effects of PRP were evaluated in vitro and in ADSC transplantation in vivo. In vitro, the effects of PRP on ADSC proliferation, differentiation into chondrogenic cells, and inhibition of angiogenic factors were investigated at three concentrations of PRP (10%, 15% and 20%). In vivo, ADSCs pretreated with or without PRP were transplanted into murine models of injured articular cartilage. Results PRP promoted ADSC proliferation and differentiation into chondrogenic cells that strongly expressed collagen II, Sox9 and aggrecan. Moreover, PRP inhibited expression of the angiogenic factor vascular endothelial growth factor. As a result, PRP-pretreated ADSCs improved healing of injured articular cartilage in murine models compared with that of untreated ADSCs. Conclusion Pretreatment of ADSCs with PRP is a simple method to efficiently apply ADSCs in cartilage regeneration. This study provides an important step toward the use of autologous ADSCs in the treatment of injured articular cartilage. PMID:23915433

  6. Paracrine Effects of the Pluripotent Stem Cell-Derived Cardiac Myocytes Salvage the Injured Myocardium.

    Science.gov (United States)

    Tachibana, Atsushi; Santoso, Michelle R; Mahmoudi, Morteza; Shukla, Praveen; Wang, Lei; Bennett, Mihoko; Goldstone, Andrew B; Wang, Mouer; Fukushi, Masahiro; Ebert, Antje D; Woo, Y Joseph; Rulifson, Eric; Yang, Phillip C

    2017-09-01

    Cardiac myocytes derived from pluripotent stem cells have demonstrated the potential to mitigate damage of the infarcted myocardium and improve left ventricular ejection fraction. However, the mechanism underlying the functional benefit is unclear. To evaluate whether the transplantation of cardiac-lineage differentiated derivatives enhance myocardial viability and restore left ventricular ejection fraction more effectively than undifferentiated pluripotent stem cells after a myocardial injury. Herein, we utilize novel multimodality evaluation of human embryonic stem cells (hESCs), hESC-derived cardiac myocytes (hCMs), human induced pluripotent stem cells (iPSCs), and iPSC-derived cardiac myocytes (iCMs) in a murine myocardial injury model. Permanent ligation of the left anterior descending coronary artery was induced in immunosuppressed mice. Intramyocardial injection was performed with (1) hESCs (n=9), (2) iPSCs (n=8), (3) hCMs (n=9), (4) iCMs (n=14), and (5) PBS control (n=10). Left ventricular ejection fraction and myocardial viability, measured by cardiac magnetic resonance imaging and manganese-enhanced magnetic resonance imaging, respectively, was significantly improved in hCM- and iCM-treated mice compared with pluripotent stem cell- or control-treated mice. Bioluminescence imaging revealed limited cell engraftment in all treated groups, suggesting that the cell secretions may underlie the repair mechanism. To determine the paracrine effects of the transplanted cells, cytokines from supernatants from all groups were assessed in vitro. Gene expression and immunohistochemistry analyses of the murine myocardium demonstrated significant upregulation of the promigratory, proangiogenic, and antiapoptotic targets in groups treated with cardiac lineage cells compared with pluripotent stem cell and control groups. This study demonstrates that the cardiac phenotype of hCMs and iCMs salvages the injured myocardium effectively than undifferentiated stem cells through

  7. Transplantation of erythropoietin gene-modified neural stem cells improves the repair of injured spinal cord

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    Min-fei Wu

    2015-01-01

    Full Text Available The protective effects of erythropoietin on spinal cord injury have not been well described. Here, the eukaryotic expression plasmid pcDNA3.1 human erythropoietin was transfected into rat neural stem cells cultured in vitro. A rat model of spinal cord injury was established using a free falling object. In the human erythropoietin-neural stem cells group, transfected neural stem cells were injected into the rat subarachnoid cavity, while the neural stem cells group was injected with non-transfected neural stem cells. Dulbecco′s modified Eagle′s medium/F12 medium was injected into the rats in the spinal cord injury group as a control. At 1-4 weeks post injury, the motor function in the rat lower limbs was best in the human erythropoietin-neural stem cells group, followed by the neural stem cells group, and lastly the spinal cord injury group. At 72 hours, compared with the spinal cord injury group, the apoptotic index and Caspase-3 gene and protein expressions were apparently decreased, and the bcl-2 gene and protein expressions were noticeably increased, in the tissues surrounding the injured region in the human erythropoietin-neural stem cells group. At 4 weeks, the cavities were clearly smaller and the motor and somatosensory evoked potential latencies were remarkably shorter in the human erythropoietin-neural stem cells group and neural stem cells group than those in the spinal cord injury group. These differences were particularly obvious in the human erythropoietin-neural stem cells group. More CM-Dil-positive cells and horseradish peroxidase-positive nerve fibers and larger amplitude motor and somatosensory evoked potentials were found in the human erythropoietin-neural stem cells group and neural stem cells group than in the spinal cord injury group. Again, these differences were particularly obvious in the human erythropoietin-neural stem cells group. These data indicate that transplantation of erythropoietin gene-modified neural stem

  8. Transplantation of erythropoietin gene-modified neural stem cells improves the repair of injured spinal cord.

    Science.gov (United States)

    Wu, Min-Fei; Zhang, Shu-Quan; Gu, Rui; Liu, Jia-Bei; Li, Ye; Zhu, Qing-San

    2015-09-01

    The protective effects of erythropoietin on spinal cord injury have not been well described. Here, the eukaryotic expression plasmid pcDNA3.1 human erythropoietin was transfected into rat neural stem cells cultured in vitro. A rat model of spinal cord injury was established using a free falling object. In the human erythropoietin-neural stem cells group, transfected neural stem cells were injected into the rat subarachnoid cavity, while the neural stem cells group was injected with non-transfected neural stem cells. Dulbecco's modified Eagle's medium/F12 medium was injected into the rats in the spinal cord injury group as a control. At 1-4 weeks post injury, the motor function in the rat lower limbs was best in the human erythropoietin-neural stem cells group, followed by the neural stem cells group, and lastly the spinal cord injury group. At 72 hours, compared with the spinal cord injury group, the apoptotic index and Caspase-3 gene and protein expressions were apparently decreased, and the bcl-2 gene and protein expressions were noticeably increased, in the tissues surrounding the injured region in the human erythropoietin-neural stem cells group. At 4 weeks, the cavities were clearly smaller and the motor and somatosensory evoked potential latencies were remarkably shorter in the human erythropoietin-neural stem cells group and neural stem cells group than those in the spinal cord injury group. These differences were particularly obvious in the human erythropoietin-neural stem cells group. More CM-Dil-positive cells and horseradish peroxidase-positive nerve fibers and larger amplitude motor and somatosensory evoked potentials were found in the human erythropoietin-neural stem cells group and neural stem cells group than in the spinal cord injury group. Again, these differences were particularly obvious in the human erythropoietin-neural stem cells group. These data indicate that transplantation of erythropoietin gene-modified neural stem cells into the

  9. Effects of sub-lethal neurite outgrowth inhibitory concentrations of chlorpyrifos oxon on cytoskeletal proteins and acetylcholinesterase in differentiating N2a cells

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    Flaskos, J., E-mail: flaskos@vet.auth.gr [Laboratory of Biochemistry and Toxicology, School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Nikolaidis, E. [Laboratory of Biochemistry and Toxicology, School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Harris, W. [School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS (United Kingdom); Sachana, M. [Laboratory of Biochemistry and Toxicology, School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Hargreaves, A.J., E-mail: alan.hargreaves@ntu.ac.uk [School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS (United Kingdom)

    2011-11-15

    Previous work in our laboratory has shown that sub-lethal concentrations (1-10 {mu}M) of chlorpyrifos (CPF), diazinon (DZ) and diazinon oxon (DZO) inhibit the outgrowth of axon-like neurites in differentiating mouse N2a neuroblastoma cells concomitant with altered levels and/or phosphorylation state of axonal cytoskeleton and growth-associated proteins. The aim of the present work was to determine whether chlorpyrifos oxon (CPO) was capable of inhibiting N2a cell differentiation in a similar manner. Using experimental conditions similar to our previous work, sub-lethal concentrations (1-10 {mu}M) of CPO were found to inhibit N2a cell differentiation. However, unlike previous studies with DZ and DZO, there was a high level of sustained inhibition of acetylcholinesterase (AChE) in CPO treated cells. Impairment of neurite outgrowth was also associated with reduced levels of growth associated protein-43 and neurofilament heavy chain (NFH), and the distribution of NFH in cells stained by indirect immunofluorescence was disrupted. However, in contrast to previous findings for DZO, the absolute level of phosphorylated NFH was unaffected by CPO exposure. Taken together, the findings suggest that sub-lethal concentrations of CPO inhibit axon outgrowth in differentiating N2a cells and that this effect involves reduced levels of two proteins that play key roles in axon outgrowth and maintenance. Although the inhibition of neurite outgrowth is unlikely to involve AChE inhibition directly, further work will help to determine whether the persistent inhibition of AChE by CPO can account for the different effects induced by CPO and DZO on the levels of total and phosphorylated NFH. -- Highlights: Black-Right-Pointing-Pointer Sub-lethal levels of chlorpyrifos oxon inhibit neurite outgrowth in N2a cells Black-Right-Pointing-Pointer Acetylcholinesterase exhibits sustained inhibition throughout exposure Black-Right-Pointing-Pointer The levels of neurofilament heavy chain and GAP-43

  10. MRI of auto-transplantation of bone marrow-derived stem-progenitor cells for potential repair of injured arteries.

    Directory of Open Access Journals (Sweden)

    Yanfeng Meng

    Full Text Available This study was to validate the feasibility of using clinical 3.0T MRI to monitor the migration of autotransplanted bone marrow (BM-derived stem-progenitor cells (SPC to the injured arteries of near-human sized swine for potential cell-based arterial repair.The study was divided into two phases. For in vitro evaluation, BM cells were extracted from the iliac crests of 13 domestic pigs and then labeled with a T2 contrast agent, Feridex, and/or a fluorescent tissue marker, PKH26. The viability, the proliferation efficiency and the efficacies of Feridex and/or PKH26 labeling were determined. For in vivo validation, the 13 pigs underwent endovascular balloon-mediated intimal damages of the iliofemoral arteries. The labeled or un-labeled BM cells were autotransplanted back to the same pig from which the BM cells were extracted. Approximately three weeks post-cell transplantation, 3.0T T2-weighted MRI was performed to detect Feridex-created signal voids of the transplanted BM cells in the injured iliofemoral arteries, which was confirmed by subsequent histologic correlation.Of the in vitro study, the viability of dual-labeled BM cells was 95-98%. The proliferation efficiencies of dual-labeled BM cells were not significantly different compared to those of non-labeled cells. The efficacies of Feridex- and PKH26 labeling were 90% and 100%, respectively. Of the in vivo study, 3.0T MRI detected the auto-transplanted BM cells migrated to the injured arteries, which was confirmed by histologic examinations.This study demonstrates the capability of using clinical 3.0T MRI to monitor the auto-transplantation of BM cells that migrate to the injured arteries of large animals, which may provide a useful MRI technique to monitor cell-based arterial repair.

  11. Combination of edaravone and neural stem cell transplantation repairs injured spinal cord in rats.

    Science.gov (United States)

    Song, Y Y; Peng, C G; Ye, X B

    2015-12-29

    This study sought to observe the effect of the combination of edaravone and neural stem cell (NSC) transplantation on the repair of complete spinal cord transection in rats. Eighty adult female Sprague-Dawley (SD) rats were used to establish the injury model of complete spinal cord transection at T9. Animals were divided randomly into four groups (N = 20 each): control, edaravone, transplantation, and edaravone + transplantation. The recovery of spinal function was evaluated with the Basso, Beattie, Bresnahan (BBB) rating scale on days 1, 3, and 7 each week after the surgery. After 8 weeks, the BBB scores of the control, edaravone, transplantation, and combination groups were 4.21 ± 0.11, 8.46 ± 0.1, 8.54 ± 0.13, and 11.21 ± 0.14, respectively. At 8 weeks after surgery, the spinal cord was collected; the survival and transportation of transplanted cells were observed with PKH-26 labeling, and the regeneration and distribution of spinal nerve fibers with fluorescent-gold (FG) retrograde tracing. Five rats died due to the injury. PKH-26-labeled NSCs had migrated into the spinal cord. A few intact nerve fibers and pyramidal neurons passed the injured area in the transplantation and combination groups. The numbers of PKH-26-labeled cells and FG-labeled nerve fibers were in the order: combination group > edaravone group and transplantation group > control group (P transplantation can improve the effectiveness of spinal cord injury repair in rats.

  12. Direct evaluation of myocardial viability and stem cell engraftment demonstrates salvage of the injured myocardium.

    Science.gov (United States)

    Kim, Paul J; Mahmoudi, Morteza; Ge, Xiaohu; Matsuura, Yuka; Toma, Ildiko; Metzler, Scott; Kooreman, Nigel G; Ramunas, John; Holbrook, Colin; McConnell, Michael V; Blau, Helen; Harnish, Phillip; Rulifson, Eric; Yang, Phillip C

    2015-03-27

    The mechanism of functional restoration by stem cell therapy remains poorly understood. Novel manganese-enhanced MRI and bioluminescence reporter gene imaging were applied to follow myocardial viability and cell engraftment, respectively. Human-placenta-derived amniotic mesenchymal stem cells (AMCs) demonstrate unique immunoregulatory and precardiac properties. In this study, the restorative effects of 3 AMC-derived subpopulations were examined in a murine myocardial injury model: (1) unselected AMCs, (2) ckit(+)AMCs, and (3) AMC-derived induced pluripotent stem cells (MiPSCs). To determine the differential restorative effects of the AMC-derived subpopulations in the murine myocardial injury model using multimodality imaging. SCID (severe combined immunodeficiency) mice underwent left anterior descending artery ligation and were divided into 4 treatment arms: (1) normal saline control (n=14), (2) unselected AMCs (n=10), (3) ckit(+)AMCs (n=13), and (4) MiPSCs (n=11). Cardiac MRI assessed myocardial viability and left ventricular function, whereas bioluminescence imaging assessed stem cell engraftment during a 4-week period. Immunohistological labeling and reverse transcriptase polymerase chain reaction of the explanted myocardium were performed. The unselected AMC and ckit(+)AMC-treated mice demonstrated transient left ventricular functional improvement. However, the MiPSCs exhibited a significantly greater increase in left ventricular function compared with all the other groups during the entire 4-week period. Left ventricular functional improvement correlated with increased myocardial viability and sustained stem cell engraftment. The MiPSC-treated animals lacked any evidence of de novo cardiac differentiation. The functional restoration seen in MiPSCs was characterized by increased myocardial viability and sustained engraftment without de novo cardiac differentiation, indicating salvage of the injured myocardium. © 2015 American Heart Association, Inc.

  13. Stem cell recruitment of newly formed host cells via a successful seduction? Filling the gap between neurogenic niche and injured brain site.

    Science.gov (United States)

    Tajiri, Naoki; Kaneko, Yuji; Shinozuka, Kazutaka; Ishikawa, Hiroto; Yankee, Ernest; McGrogan, Michael; Case, Casey; Borlongan, Cesar V

    2013-01-01

    Here, we report that a unique mechanism of action exerted by stem cells in the repair of the traumatically injured brain involves their ability to harness a biobridge between neurogenic niche and injured brain site. This biobridge, visualized immunohistochemically and laser captured, corresponded to an area between the neurogenic subventricular zone and the injured cortex. That the biobridge expressed high levels of extracellular matrix metalloproteinases characterized initially by a stream of transplanted stem cells, but subsequently contained only few to non-detectable grafts and overgrown by newly formed host cells, implicates a novel property of stem cells. The transplanted stem cells manifest themselves as pathways for trafficking the migration of host neurogenic cells, but once this biobridge is formed between the neurogenic site and the injured brain site, the grafted cells disappear and relinquish their task to the host neurogenic cells. Our findings reveal that long-distance migration of host cells from the neurogenic niche to the injured brain site can be achieved through transplanted stem cells serving as biobridges for initiation of endogenous repair mechanisms. This is the first report of a stem cell-paved "biobridge". Indeed, to date the two major schools of discipline in stem cell repair mechanism primarily support the concept of "cell replacement" and bystander effects of "trophic factor secretion". The present novel observations of a stem cell seducing a host cell to engage in brain repair advances basic science concepts on stem cell biology and extracellular matrix, as well as provokes translational research on propagating this stem cell-paved biobridge beyond cell replacement and trophic factor secretion for the treatment of traumatic brain injury and other neurological disorders.

  14. Inactivation and sublethal injury of Escherichia coli and Listeria innocua by high hydrostatic pressure in model suspensions and beetroot juice

    Science.gov (United States)

    Sokołowska, Barbara; Skąpska, Sylwia; Niezgoda, Jolanta; Rutkowska, Małgorzata; Dekowska, Agnieszka; Rzoska, Sylwester J.

    2014-01-01

    Cells exposed to different physical and chemical treatments, including high hydrostatic pressure (HHP), suffer from injuries that could be reversible in food materials when stored. Escherichia coli and Listeria innocua cells suspended in phosphate-buffered saline (PBS) (model suspensions), and acidified beetroot juice were subjected to a pressure of 400 MPa at a temperature of 20°C for up to 10 min. The difference between the viable and non-injured cells was used to estimate the number of injured survivors. The reduction in E. coli cell number was 3.4-4.1 log after 10 min pressurization in model suspensions and 6.2 log in beetroot juice. Sublethally injured cells in PBS accounted for up to 2.7 log after 10 min HHP treatment and 0.8 log in beetroot juice. The reduction in L. innocua cell number after 10 min pressure treatment reached from 3.8 to 4.8 log, depending on the initial concentration in model suspensions. Among the surviving L. innocua cells, even up to 100% were injured. L. innocua cells were completely inactivated after 1 min HHP treatment in beetroot juice.

  15. Subchronic exposure to sublethal dose of imidacloprid changes electrophysiological properties and expression pattern of nicotinic acetylcholine receptor subtypes in insect neurosecretory cells.

    Science.gov (United States)

    Benzidane, Yassine; Goven, Delphine; Abd-Ella, Aly Ahmed; Deshayes, Caroline; Lapied, Bruno; Raymond, Valérie

    2017-09-01

    Neonicotinoids are the most important class of insecticides used in agriculture over the last decade. They act as selective agonists of insect nicotinic acetylcholine receptors (nAChRs). The emergence of insect resistance to these insecticides is one of the major problems, which limit the use of neonicotinoids. The aim of our study is to better understand physiological changes appearing after subchronic exposure to sublethal doses of insecticide using complementary approaches that include toxicology, electrophysiology, molecular biology and calcium imaging. We used cockroach neurosecretory cells identified as dorsal unpaired median (DUM) neurons, known to express two α-bungarotoxin-insensitive (α-bgt-insensitive) nAChR subtypes, nAChR1 and nAChR2, which differ in their sensitivity to imidacloprid. Although nAChR1 is sensitive to imidacloprid, nAChR2 is insensitive to this insecticide. In this study, we demonstrate that subchronic exposure to sublethal dose of imidacloprid differentially changes physiological and molecular properties of nAChR1 and nAChR2. Our findings reported that this treatment decreased the sensitivity of nAChR1 to imidacloprid, reduced current density flowing through this nAChR subtype but did not affect its subunit composition (α3, α8 and β1). Subchronic exposure to sublethal dose of imidacloprid also affected nAChR2 functions. However, these effects were different from those reported on nAChR1. We observed changes in nAChR2 conformational state, which could be related to modification of the subunit composition (α1, α2 and β1). Finally, the subchronic exposure affecting both nAChR1 and nAChR2 seemed to be linked to the elevation of the steady-state resting intracellular calcium level. In conclusion, under subchronic exposure to sublethal dose of imidacloprid, cockroaches are capable of triggering adaptive mechanisms by reducing the participation of imidacloprid-sensitive nAChR1 and by optimizing functional properties of nAChR2, which is

  16. Stromal Derived Factor-1/CXCR4 Axis Involved in Bone Marrow Mesenchymal Stem Cells Recruitment to Injured Liver

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    Kuai Xiao Ling

    2016-01-01

    Full Text Available The molecular mechanism of bone marrow mesenchymal stromal stem cells (BMSCs mobilization and migration to the liver was poorly understood. Stromal cell-derived factor-1 (SDF-1 participates in BMSCs homing and migration into injury organs. We try to investigate the role of SDF-1 signaling in BMSCs migration towards injured liver. The expression of CXCR4 in BMSCs at mRNA level and protein level was confirmed by RT-PCR, flow cytometry, and immunocytochemistry. The SDF-1 or liver lysates induced BMSCs migration was detected by transwell inserts. CXCR4 antagonist, AMD3100, and anti-CXCR4 antibody were used to inhibit the migration. The Sprague-Dawley rat liver injury model was established by intraperitoneal injection of thioacetamide. The concentration of SDF-1 increased as modeling time extended, which was determined by ELISA method. The Dir-labeled BMSCs were injected into the liver of the rats through portal vein. The cell migration in the liver was tracked by in vivo imaging system and the fluorescent intensity was measured. In vivo, BMSCs migrated into injured liver which was partially blocked by AMD3100 or anti-CXCR4 antibody. Taken together, the results demonstrated that the migration of BMSCs was regulated by SDF-1/CXCR4 signaling which involved in BMSCs recruitment to injured liver.

  17. Akhirin regulates the proliferation and differentiation of neural stem cells in intact and injured mouse spinal cord.

    Science.gov (United States)

    Abdulhaleem, Felemban Athary M; Song, Xiaohong; Kawano, Rie; Uezono, Naohiro; Ito, Ayako; Ahmed, Giasuddin; Hossain, Mahmud; Nakashima, Kinichi; Tanaka, Hideaki; Ohta, Kunimasa

    2015-05-01

    Although the central nervous system is considered a comparatively static tissue with limited cell turnover, cells with stem cell properties have been isolated from most neural tissues. The spinal cord ependymal cells show neural stem cell potential in vitro and in vivo in injured spinal cord. However, very little is known regarding the ependymal niche in the mouse spinal cord. We previously reported that a secreted factor, chick Akhirin, is expressed in the ciliary marginal zone of the eye, where it works as a heterophilic cell-adhesion molecule. Here, we describe a new crucial function for mouse Akhirin (M-AKH) in regulating the proliferation and differentiation of progenitors in the mouse spinal cord. During embryonic spinal cord development, M-AKH is transiently expressed in the central canal ependymal cells, which possess latent neural stem cell properties. Targeted inactivation of the AKH gene in mice causes a reduction in the size of the spinal cord and decreases BrdU incorporation in the spinal cord. Remarkably, the expression patterns of ependymal niche molecules in AKH knockout (AKH-/-) mice are different from those of AKH+/+, both in vitro and in vivo. Furthermore, we provide evidence that AKH expression in the central canal is rapidly upregulated in the injured spinal cord. Taken together, these results indicate that M-AKH plays a crucial role in mouse spinal cord formation by regulating the ependymal niche in the central canal. © 2014 Wiley Periodicals, Inc.

  18. In vivo serial MR imaging of magnetically labeled endothelial progenitor cells homing to the endothelium injured artery in mice.

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    Jun Chen

    Full Text Available BACKGROUND: Emerging evidence of histopathological analyses suggests that endothelial progenitor cells (EPCs play an important role in vascular diseases. Neointimal hyperplasia can be reduced by intravenous transfusion of EPCs after vascular injury in mice. Therefore, it would be advantageous to develop an in vivo technique that can explore the temporal and spatial migration of EPCs homing to the damaged endothelium noninvasively. METHODOLOGY/PRINCIPAL FINDINGS: The left carotid common artery (LCCA was injured by removal of endothelium with a flexible wire in Kunming mice. EPCs were collected by in vitro culture of spleen-derived mouse mononuclear cells (MNCs. EPCs labeling was carried out in vitro using Fe₂O₃-poly-L-lysine (Fe₂O₃-PLL. In vivo serial MR imaging was performed to follow-up the injured artery at different time points after intravenous transfusion of EPCs. Vessel wall areas of injured artery were computed on T₂WI. Larger MR signal voids of vessel wall on T₂WI was revealed in all 6 mice of the labeled EPC transfusion group 15 days after LCCA injury, and it was found only in 1 mouse in the unlabeled EPC transfusion group (p = 0.015. Quantitative analyses of vessel wall areas on T₂WI showed that the vessel wall areas of labeled EPC transfusion group were less than those of unlabeled EPC transfusion group and control group fifteen days after artery injury (p<0.05. Histopathological analyses confirmed accumulation and distribution of transfused EPCs at the injury site of LCCA. CONCLUSIONS/SIGNIFICANCE: These data indicate that MR imaging might be used as an in vivo method for the tracking of EPCs homing to the endothelium injured artery.

  19. Smooth Muscle Progenitor Cells Derived From Human Pluripotent Stem Cells Induce Histologic Changes in Injured Urethral Sphincter.

    Science.gov (United States)

    Li, Yanhui; Wen, Yan; Wang, Zhe; Wei, Yi; Wani, Prachi; Green, Morgaine; Swaminathan, Ganesh; Ramamurthi, Anand; Pera, Renee Reijo; Chen, Bertha

    2016-12-01

    : Data suggest that myoblasts from various sources, including bone marrow, skeletal muscle, and adipose tissue, can restore muscle function in patients with urinary incontinence. Animal data have indicated that these progenitor cells exert mostly a paracrine effect on the native tissues rather than cell regeneration. Limited knowledge is available on the in vivo effect of human stem cells or muscle progenitors on injured muscles. We examined in vivo integration of smooth muscle progenitor cells (pSMCs) derived from human pluripotent stem cells (hPSCs). pSMCs were derived from a human embryonic stem cell line (H9-ESCs) and two induced pluripotent stem cell (iPSC) lines. pSMCs were injected periurethrally into urethral injury rat models (2 × 10(6) cells per rat) or intramuscularly into severe combined immunodeficiency mice. Histologic and quantitative image analysis revealed that the urethras in pSMC-treated rats contained abundant elastic fibers and thicker muscle layers compared with the control rats. Western blot confirmed increased elastin/collagen III content in the urethra and bladder of the H9-pSMC-treated rats compared with controls. iPSC-pSMC treatment also showed similar trends in elastin and collagen III. Human elastin gene expression was not detectable in rodent tissues, suggesting that the extracellular matrix synthesis resulted from the native rodent tissues rather than from the implanted human cells. Immunofluorescence staining and in vivo bioluminescence imaging confirmed long-term engraftment of pSMCs into the host urethra and the persistence of the smooth muscle phenotype. Taken together, the data suggest that hPSC-derived pSMCs facilitate restoration of urethral sphincter function by direct smooth muscle cell regeneration and by inducing native tissue elastin/collagen III remodeling. The present study provides evidence that a pure population of human smooth muscle progenitor cells (pSMCs) derived from human pluripotent stem cells (hPSCs) (human

  20. Enumeration and Identification of Coliform Bacteria Injured by Chlorine or Fungicide Mixed with Agricultural Water.

    Science.gov (United States)

    Izumi, Hidemi; Nakata, Yuji; Inoue, Ayano

    2016-10-01

    Chemical sanitizers may induce no injury (bacteria survive), sublethal injury (bacteria are injured), or lethal injury (bacteria die). The proportion of coliform bacteria that were injured sublethally by chlorine and fungicide mixed with agricultural water (pond water), which was used to dilute the pesticide solution, was evaluated using the thin agar layer (TAL) method. In pure cultures of Enterobacter cloacae , Escherichia coli , and E. coli O157:H7 (representing a human pathogen), the percentage of chlorine-injured cells was 69 to 77% for dilute electrolyzed water containing an available chlorine level of 2 ppm. When agricultural water was mixed with electrolyzed water, the percentage of injured coliforms in agricultural water was 75%. The isolation and identification of bacteria on TAL and selective media suggested that the chlorine stress caused injury to Enterobacter kobei . Of the four fungicide products tested, diluted to their recommended concentrations, Topsin-M, Sumilex, and Oxirane caused injury to coliform bacteria in pure cultures and in agricultural water following their mixture with each pesticide, whereas Streptomycin did not induce any injury to the bacteria. The percentage of injury was 45 to 97% for Topsin-M, 80 to 87% for Sumilex, and 50 to 97% for Oxirane. A comparison of the coliforms isolated from the pesticide solutions and then grown on either TAL or selective media indicated the possibility of fungicide-injured Rahnella aquatilis , Yersinia mollaretii , and E. coli . These results suggest the importance of selecting a suitable sanitizer and the necessity of adjusting the sanitizer concentration to a level that will kill the coliforms rather than cause sanitizer-induced cell injury that can result in the recovery of the coliforms.

  1. Protective effect of milk constituents and sublethal injuries limiting process effectiveness during PEF inactivation of Lb. rhamnosus.

    Science.gov (United States)

    Jaeger, H; Schulz, A; Karapetkov, N; Knorr, D

    2009-08-31

    The inactivation of Lb. rhamnosus by pulsed electric field treatment (PEF) was studied in different fractions of raw milk and Ringer solution in order to evaluate the protective effect of nutrient rich media in comparison to aqueous buffer solutions. Apart from monitoring of culturability, analysis of the physiological fitness of Lb. rhamnosus was conducted aiming to identify sublethally damaged cells. Therefore, flow cytometry and a selective medium plating technique were used and compared to each other. The goal of the study was to apply three different parameters describing the physiological fitness of the model organism Lb. rhamnosus after PEF treatment such as culturability, membrane permeability and metabolic activity depending on treatment media and parameters. A concentration dependent protective effect of the milk protein fraction could be shown and allocated to micellar casein as the major milk protein. Increasing the concentration of whey proteins up to 2% showed a similar impact on limiting the PEF inactivation of Lb. rhamnosus. The evaluation of physiological fitness of cells was based on a determination of structural and functional characteristics by rapid cellular staining using carboxyfluorescein diacetate and propidium iodide. This approach showed good accordance to the conventional selective medium plating technique for the enumeration of sublethally-injured bacteria but flow cytometry provided additional information for the characterisation of this fraction. The extent of occurrence of dead, sublethal and vital fractions of cells was found dependent on the PEF treatment parameters such as electrical field strength and energy input as well as the different milk fractions used as treatment media.

  2. Cell-seeded alginate hydrogel scaffolds promote directed linear axonal regeneration in the injured rat spinal cord.

    Science.gov (United States)

    Günther, Manuel Ingo; Weidner, Norbert; Müller, Rainer; Blesch, Armin

    2015-11-01

    Despite recent progress in enhancing axonal growth in the injured spinal cord, the guidance of regenerating axons across an extended lesion site remains a major challenge. To determine whether regenerating axons can be guided in rostrocaudal direction, we implanted 2mm long alginate-based anisotropic capillary hydrogels seeded with bone marrow stromal cells (BMSCs) expressing brain-derived neurotrophic factor (BDNF) or green fluorescent protein (GFP) as control into a C5 hemisection lesion of the rat spinal cord. Four weeks post-lesion, numerous BMSCs survived inside the scaffold channels, accompanied by macrophages, Schwann cells and blood vessels. Quantification of axons growing into channels demonstrated 3-4 times more axons in hydrogels seeded with BMSCs expressing BDNF (BMSC-BDNF) compared to control cells. The number of anterogradely traced axons extending through the entire length of the scaffold was also significantly higher in scaffolds with BMSC-BDNF. Increasing the channel diameters from 41μm to 64μm did not lead to significant differences in the number of regenerating axons. Lesions filled with BMSC-BDNF without hydrogels exhibited a random axon orientation, whereas axons were oriented parallel to the hydrogel channel walls. Thus, alginate-based scaffolds with an anisotropic capillary structure are able to physically guide regenerating axons. After injury, regenerating axons have to extend across the lesion site in the injured spinal cord to reestablish lost neuronal connections. While cell grafting and growth factor delivery can promote growth of injured axons, without proper guidance, axons rarely extend across the lesion site. Here, we show that alginate biomaterials with linear channels that are filled with cells expressing the growth-promoting neurotrophin BDNF promote linear axon extension throughout the channels after transplantation to the injured rat spinal cord. Animals that received the same cells but no alginate guidance structure did not

  3. A Single-Cell Resolution Imaging Protocol of Mitochondrial DNA Dynamics in Physiopathology, mTRIP, Which Also Evaluates Sublethal Cytotoxicity.

    Science.gov (United States)

    Chatre, Laurent; Montagne, Benjamin; Ricchetti, Miria

    2016-01-01

    Mitochondria autonomously replicate and transcribe their own genome, which is present in multiple copies in the organelle. Transcription and replication of the mitochondrial DNA (mtDNA), which are defined here as mtDNA processing, are essential for mitochondrial function. The extent, efficiency, and coordination of mtDNA processing are key parameters of the mitochondrial state in living cells. Recently, single-cell analysis of mtDNA processing revealed a large and dynamic heterogeneity of mitochondrial populations in single cells, which is linked to mitochondrial function and is altered during disease. This was achieved using mitochondrial Transcription and Replication Imaging Protocol (mTRIP), a modified fluorescence in situ hybridization (FISH) approach that simultaneously reveals the mitochondrial RNA content and mtDNA engaged in initiation of replication at the single-cell level. mTRIP can also be coupled to immunofluorescence or MitoTracker, resulting in the additional labeling of proteins or active mitochondria, respectively. Therefore, mTRIP detects quantitative and qualitative alterations of the dynamics of mtDNA processing in human cells that respond to physiological changes or result from diseases. In addition, we show here that mTRIP is a rather sensitive tool for detecting mitochondrial alterations that may lead to loss of cell viability, and is thereby a useful tool for monitoring sublethal cytotoxicity for instance during chronic drug treatment.

  4. Relationship between Sublethal Injury and Microbial Inactivation by the Combination of High Hydrostatic Pressure and Citral or tert-Butyl Hydroquinone ▿

    Science.gov (United States)

    Somolinos, Maria; García, Diego; Pagán, Rafael; Mackey, Bernard

    2008-01-01

    The aim was to investigate (i) the occurrence of sublethal injury in Listeria monocytogenes, Escherichia coli, and Saccharomyces cerevisiae after high hydrostatic pressure (HHP) treatment as a function of the treatment medium pH and composition and (ii) the relationship between the occurrence of sublethal injury and the inactivating effect of a combination of HHP and two antimicrobial compounds, tert-butyl hydroquinone (TBHQ) and citral. The three microorganisms showed a high proportion of sublethally injured cells (up to 99.99% of the surviving population) after HHP. In E. coli and L. monocytogenes, the extent of inactivation and sublethal injury depended on the pH and the composition of the treatment medium, whereas in S. cerevisiae, inactivation and sublethal injury were independent of medium pH or composition under the conditions tested. TBHQ alone was not lethal to E. coli or L. monocytogenes but acted synergistically with HHP and 24-h refrigeration, resulting in a viability decrease of >5 log10 cycles of both organisms. The antimicrobial effect of citral depended on the microorganism and the treatment medium pH. Acting alone for 24 h under refrigeration, 1,000 ppm of citral caused a reduction of 5 log10 cycles of E. coli at pH 7.0 and almost 3 log10 cycles of L. monocytogenes at pH 4.0. The combination of citral and HHP also showed a synergistic effect. Our results have confirmed that the detection of sublethal injury after HHP may contribute to the identification of those treatment conditions under which HHP may act synergistically with other preserving processes. PMID:18952869

  5. Critical analysis of the maximum non inhibitory concentration (MNIC) method in quantifying sub-lethal injury in Saccharomyces cerevisiae cells exposed to either thermal or pulsed electric field treatments.

    Science.gov (United States)

    Kethireddy, V; Oey, I; Jowett, Tim; Bremer, P

    2016-09-16

    Sub-lethal injury within a microbial population, due to processing treatments or environmental stress, is often assessed as the difference in the number of cells recovered on non-selective media compared to numbers recovered on a "selective media" containing a predetermined maximum non-inhibitory concentration (MNIC) of a selective agent. However, as knowledge of cell metabolic response to injury, population diversity and dynamics increased, the rationale behind the conventional approach of quantifying sub-lethal injury must be scrutinized further. This study reassessed the methodology used to quantify sub-lethal injury for Saccharomyces cerevisiae cells (≈ 4.75 Log CFU/mL) exposed to either a mild thermal (45°C for 0, 10 and 20min) or a mild pulsed electric field treatment (field strengths of 8.0-9.0kV/cm and energy levels of 8, 14 and 21kJ/kg). Treated cells were plated onto either Yeast Malt agar (YM) or YM containing NaCl, as a selective agent at 5-15% in 1% increments. The impact of sub-lethal stress due to initial processing, the stress due to selective agents in the plating media, and the subsequent variation of inhibition following the treatments was assessed based on the CFU count (cell numbers). ANOVA and a generalised least squares model indicated significant effects of media, treatments, and their interaction effects (P<0.05) on cell numbers. It was shown that the concentration of the selective agent used dictated the extent of sub-lethal injury recorded owing to the interaction effects of the selective component (NaCl) in the recovery media. Our findings highlight a potential common misunderstanding on how culture conditions impact on sub-lethal injury. Interestingly for S. cerevisiae cells the number of cells recovered at different NaCl concentrations in the media appears to provide valuable information about the mode of injury, the comparative efficacy of different processing regimes and the inherent degree of resistance within a population. This

  6. Adult skin-derived precursor Schwann cell grafts form growths in the injured spinal cord of Fischer rats.

    Science.gov (United States)

    May, Zacnicte; Kumar, Ranjan; Führmann, Tobias; Tam, Roger; Vulic, Katarina; Forero, Juan; Lucas-Osma, Ana M; Fenrich, Keith; Assinck, Peggy; Lee, Michael J; Moulson, Aaron; Shoichet, Molly S; Tetzlaff, Wolfram; Biernaskie, Jeff; Fouad, Karim

    2017-10-25

    In this study, GFP+ skin-derived precursor Schwann cells (SKP-SCs) from adult rats were grafted into the injured spinal cord of immunosuppressed rats. Our goal was to improve grafted cell survival in the injured spinal cord, which is typically low. Cells were grafted in hyaluronan-methylcellulose hydrogel (HAMC) or hyaluronan-methylcellulose modified with laminin- and fibronectin-derived peptide sequences (eHAMC). The criteria for selection of hyaluronan was for its shear-thinning properties, making the hydrogel easy to inject, methylcellulose for its inverse thermal gelation, helping to keep grafted cells in situ, and fibronectin and laminin to improve cell attachment and, thus, prevent cell death due to dissociation from substrate molecules (i.e., anoikis). Post-mortem examination revealed large masses of GFP+ SKP-SCs in the spinal cords of rats that received cells in HAMC (5 out of n = 8) and eHAMC (6 out of n = 8). Cell transplantation in eHAMC caused significantly greater spinal lesions compared to lesion and eHAMC only control groups. A parallel study showed similar masses in the contused spinal cord of rats after transplantation of adult GFP+ SKP-SCs without a hydrogel or immunosuppression. These findings suggest that adult GFP+ SKP-SCs, cultured/transplanted under the conditions described here, have a capacity for uncontrolled proliferation. Growth-formation in pre-clinical research has also been documented after transplantation of: human induced pluripotent stem cell-derived neural stem cells (Itakura et al., 2015), embryonic stem cells and embryonic stem cell-derived neurons (Brederlau et al., 2006; Dressel et al., 2008), bone marrow derived mesenchymal stem cells (Jeong et al., 2011) and rat nerve-derived SCs following in vitro expansion for ˃11 passages (Funk et al., 2007; Langford et al., 1988; Morrissey et al., 1991). It is of upmost importance to define the precise culture/transplantation parameters for maintenance of normal cell function and safe

  7. Role of protein synthesis in the repair of sublethal x-ray damage in a mutant Chinese hamster ovary cell line

    Energy Technology Data Exchange (ETDEWEB)

    Yezzi, M.J.

    1985-04-01

    A temperature-sensitive mutant for protein synthesis, CHO-TSH1, has been compared to the wild-type cell, CHO-sC1, in single- and split-radiation-dose schemes. When the exponentially growing TS mutant and the wild-type cells were treated at 40/sub 0/C for up to 2 hrs prior to graded doses of x rays, the survival curves were identical and were the same as those obtained without heat treatment. If the cultures were incubated at 40/sup 0/C for 2 hrs before a first dose and maintained at 40/sup 0/C during a 2 hr dose fractionation interval, repair of radiation damage was reduced in the mutant compared to the wild type. These observations implied that a pool of proteins was involved in the repair of sublethal x-ray damage. However, if repair was measured by the alkaline-unwinding technique under the same time and temperature schemes, no difference in the kientics of DNA strand rejoining was observed. Misrepair processes may permit restoration of DNA strand integrity but not allow functional repair. The effect of diminished repair under conditions of inhibition of protein synthesis was found to be cell-cycle dependent in survival studies with synchronized mutant cell populations. Repair was found to be almost completely eliminated if the temperature sequence described above was applied in the middle of the DNA synthetic phase. Treatment of cell populations in the middle of G/sub 1/-phase yielded repair inhibition comparable to that observed with the asynchronous cells. Splitdose experiments were done using pre-incubation with cycloheximide to chemically inhibit protein synthesis. WT cells and TS cells were treated with cycloheximide at 35/sup 0/C for 2 hrs before a first dose and during a 2 hr dose fractionation interval. 23 figs., 7 tabs.

  8. Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties

    Science.gov (United States)

    Jin, Hua; Yang, Qi; Ji, Feng; Zhang, Ya-jie; Zhao, Yan; Luo, Min

    2015-01-01

    The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as embryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C6 root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C6 brachial plexus injury site (1 × 106 cells/mL, 3 μL/injection, 25 injections) immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also significantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effectively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals. PMID:25883625

  9. Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties

    Directory of Open Access Journals (Sweden)

    Hua Jin

    2015-01-01

    Full Text Available The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as embryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C 6 root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C 6 brachial plexus injury site (1 × 10 6 cells/mL, 3 μL/injection, 25 injections immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C 6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also significantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effectively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals.

  10. Reinjury risk of nano-calcium oxalate monohydrate and calcium oxalate dihydrate crystals on injured renal epithelial cells: aggravation of crystal adhesion and aggregation.

    Science.gov (United States)

    Gan, Qiong-Zhi; Sun, Xin-Yuan; Bhadja, Poonam; Yao, Xiu-Qiong; Ouyang, Jian-Ming

    2016-01-01

    Renal epithelial cell injury facilitates crystal adhesion to cell surface and serves as a key step in renal stone formation. However, the effects of cell injury on the adhesion of nano-calcium oxalate crystals and the nano-crystal-induced reinjury risk of injured cells remain unclear. African green monkey renal epithelial (Vero) cells were injured with H2O2 to establish a cell injury model. Cell viability, superoxide dismutase (SOD) activity, malonaldehyde (MDA) content, propidium iodide staining, hematoxylin-eosin staining, reactive oxygen species production, and mitochondrial membrane potential (Δψm) were determined to examine cell injury during adhesion. Changes in the surface structure of H2O2-injured cells were assessed through atomic force microscopy. The altered expression of hyaluronan during adhesion was examined through laser scanning confocal microscopy. The adhesion of nano-calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) crystals to Vero cells was observed through scanning electron microscopy. Nano-COM and COD binding was quantitatively determined through inductively coupled plasma emission spectrometry. The expression of hyaluronan on the cell surface was increased during wound healing because of Vero cell injury. The structure and function of the cell membrane were also altered by cell injury; thus, nano-crystal adhesion occurred. The ability of nano-COM to adhere to the injured Vero cells was higher than that of nano-COD crystals. The cell viability, SOD activity, and Δψm decreased when nano-crystals attached to the cell surface. By contrast, the MDA content, reactive oxygen species production, and cell death rate increased. Cell injury contributes to crystal adhesion to Vero cell surface. The attached nano-COM and COD crystals can aggravate Vero cell injury. As a consequence, crystal adhesion and aggregation are enhanced. These findings provide further insights into kidney stone formation.

  11. From transplanting Schwann cells in experimental rat spinal cord injury to their transplantation into human injured spinal cord in clinical trials.

    Science.gov (United States)

    Bunge, Mary B; Monje, Paula V; Khan, Aisha; Wood, Patrick M

    2017-01-01

    Among the potential therapies designed to repair the injured spinal cord is cell transplantation, notably the use of autologous adult human Schwann cells (SCs). Here, we detail some of the critical research accomplished over the last four decades to establish a foundation that enables these cells to be tested in clinical trials. New culture systems allowed novel information to be gained about SCs, including discovering ways to stimulate their proliferation to acquire adequately large numbers for transplantation into the injured human spinal cord. Transplantation of rat SCs into rat models of spinal cord injury has demonstrated that SCs promote repair of injured spinal cord. Additional work required to gain approval from the Food and Drug Administration for the first SC trial in the Miami Project is disclosed. This trial and a second one now underway are described. © 2017 Elsevier B.V. All rights reserved.

  12. [Repair mechanism of frozen sublethally damaged Staphylococcus aureus].

    Science.gov (United States)

    Huang, Zhongmin; Lv, Haipeng; Ai, Zhilu; Wang, Na; Xie, Xinhua; Fan, Huiping; Pan, Zhili; Suo, Biao

    2015-11-04

    To study the repair mechanisms of frozen sublethally damaged Staphylococcus aurous cells. We resuscitated frozen sublethally damaged S. aureus at 37 degrees C for different time within 3 h. Meanwhile, we compared the morphological changes of the frozen sublethally damaged cells after 1 h of resuscitation using transmission electron microscopy assay (TEM). The expressions of the transcriptional attenuator MsrR (msrR), iron (Fe3+) ABC transporter ATP-binding protein (fhuC), and cytochrome b (cytB) genes were quantitatively analyzed by real-time fluorescence quantitative PCR (Real-time PCR) method. The content of cells outside leakage, active oxygen (ROS), and superoxide dismutase (SOD) activity were also determined by ultraviolet spectrophotometry. More than 99% of the frozen sublethally damaged S. aureus repaired after 3 h. The resuscitated cells expressed an equal resistance to high concentration of NaCl. Real-time PCR results showed that the msrR and fhuC genes expressions were down-regulated, whereas the cytB gene expression was up-regulated significantly. The frozen sublethally damaged S. aureus cellar surface ultrastructure significant changed during resuscitation. The cell surface became compact and sturdy from smooth and transparent. The cell leakage rate of ultraviolet absorption material gradually decreased. Meanwhile, the intracellular ROS level declined along with the decrease of SOD activity. Frozen sublethally damaged cells may regain the capability of resistance to high salt stress by repairing cell membrane integrity, reducing the content of ROS through gene regulation, inhibiting the toxicity of active oxygen to the cells. Meanwhile, the regulation of metabolism related genes (cytB) provides the energy for the requirement of cells, therefore, the frozen sublethally damaged cells were repaired finally.

  13. Synaptic inputs from stroke-injured brain to grafted human stem cell-derived neurons activated by sensory stimuli.

    Science.gov (United States)

    Tornero, Daniel; Tsupykov, Oleg; Granmo, Marcus; Rodriguez, Cristina; Grønning-Hansen, Marita; Thelin, Jonas; Smozhanik, Ekaterina; Laterza, Cecilia; Wattananit, Somsak; Ge, Ruimin; Tatarishvili, Jemal; Grealish, Shane; Brüstle, Oliver; Skibo, Galina; Parmar, Malin; Schouenborg, Jens; Lindvall, Olle; Kokaia, Zaal

    2017-03-01

    Transplanted neurons derived from stem cells have been proposed to improve function in animal models of human disease by various mechanisms such as neuronal replacement. However, whether the grafted neurons receive functional synaptic inputs from the recipient's brain and integrate into host neural circuitry is unknown. Here we studied the synaptic inputs from the host brain to grafted cortical neurons derived from human induced pluripotent stem cells after transplantation into stroke-injured rat cerebral cortex. Using the rabies virus-based trans-synaptic tracing method and immunoelectron microscopy, we demonstrate that the grafted neurons receive direct synaptic inputs from neurons in different host brain areas located in a pattern similar to that of neurons projecting to the corresponding endogenous cortical neurons in the intact brain. Electrophysiological in vivo recordings from the cortical implants show that physiological sensory stimuli, i.e. cutaneous stimulation of nose and paw, can activate or inhibit spontaneous activity in grafted neurons, indicating that at least some of the afferent inputs are functional. In agreement, we find using patch-clamp recordings that a portion of grafted neurons respond to photostimulation of virally transfected, channelrhodopsin-2-expressing thalamo-cortical axons in acute brain slices. The present study demonstrates, for the first time, that the host brain regulates the activity of grafted neurons, providing strong evidence that transplanted human induced pluripotent stem cell-derived cortical neurons can become incorporated into injured cortical circuitry. Our findings support the idea that these neurons could contribute to functional recovery in stroke and other conditions causing neuronal loss in cerebral cortex. © The Author (2017). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. Human mesenchymal cells from adipose tissue deposit laminin and promote regeneration of injured spinal cord in rats.

    Science.gov (United States)

    Menezes, Karla; Nascimento, Marcos Assis; Gonçalves, Juliana Pena; Cruz, Aline Silva; Lopes, Daiana Vieira; Curzio, Bianca; Bonamino, Martin; de Menezes, João Ricardo Lacerda; Borojevic, Radovan; Rossi, Maria Isabel Doria; Coelho-Sampaio, Tatiana

    2014-01-01

    Cell therapy is a promising strategy to pursue the unmet need for treatment of spinal cord injury (SCI). Although several studies have shown that adult mesenchymal cells contribute to improve the outcomes of SCI, a description of the pro-regenerative events triggered by these cells is still lacking. Here we investigated the regenerative properties of human adipose tissue derived stromal cells (hADSCs) in a rat model of spinal cord compression. Cells were delivered directly into the spinal parenchyma immediately after injury. Human ADSCs promoted functional recovery, tissue preservation, and axonal regeneration. Analysis of the cord tissue showed an abundant deposition of laminin of human origin at the lesion site and spinal midline; the appearance of cell clusters composed of neural precursors in the areas of laminin deposition, and the appearance of blood vessels with separated basement membranes along the spinal axis. These effects were also observed after injection of hADSCs into non-injured spinal cord. Considering that laminin is a well-known inducer of axonal growth, as well a component of the extracellular matrix associated to neural progenitors, we propose that it can be the paracrine factor mediating the pro-regenerative effects of hADSCs in spinal cord injury.

  15. Mesenchymal Stem Cell Therapy for Protection and Repair of Injured Vital Organs

    NARCIS (Netherlands)

    van Poll, D.; Parekkadan, B.; Rinkes, I. H. M. Borel; Tilles, A. W.; Yarmush, M. L.

    Recently there has been a paradigm shift in what is considered to be the therapeutic promise of mesenchymal stem cells (MSCs) in diseases of vital organs. Originally, research focused on MSCs as a source of regenerative cells by differentiation of transplanted cells into lost cell types. It is now

  16. Characterization of a novel fibroblast-like cell line from rainbow trout and responses to sublethal anoxia

    DEFF Research Database (Denmark)

    Ossum, Carlo Gunnar; Hoffmann, Else Kay; Vijayan, M.M.

    2004-01-01

    A novel fibroblast-like cell line RTHDF was established from hypodermal connective tissue of rainbow trout Oncorhynchus mykiss and telomerase activity was demonstrated early and late in cell line development. When RTHDF cells were exposed to bioenergetic stress, i.e. anoxia, activation of the str......A novel fibroblast-like cell line RTHDF was established from hypodermal connective tissue of rainbow trout Oncorhynchus mykiss and telomerase activity was demonstrated early and late in cell line development. When RTHDF cells were exposed to bioenergetic stress, i.e. anoxia, activation...... rapidly, with maximal activity after 10 min of anoxia. Hsp70 was induced after 30 min of anoxia, followed by overnight recovery in growth medium at 21° C. Using the p38MAPK-specific inhibitor SB203580, the enhanced expression of Hsp70 occurred independently of p38MAPK activation in RTHDF. These data...

  17. Pax7 is necessary and sufficient for the myogenic specification of CD45+:Sca1+ stem cells from injured muscle.

    Directory of Open Access Journals (Sweden)

    Patrick Seale

    2004-05-01

    Full Text Available CD45(+:Sca1(+ adult stem cells isolated from uninjured muscle do not display any myogenic potential, whereas those isolated from regenerating muscle give rise to myoblasts expressing the paired-box transcription factor Pax7 and the bHLH factors Myf5 and MyoD. By contrast, CD45(+:Sca1(+ isolated from injured Pax7( -/- muscle were incapable of forming myoblasts. Infection of CD45(+:Sca1(+ cells from uninjured muscle with retrovirus expressing Pax7 efficiently activated the myogenic program. The resulting myoblasts expressed Myf5 and MyoD and differentiated into myotubes that expressed myogenin and myosin heavy chain. Infection of CD45(-:Sca1(- cells from Pax7( -/- muscle similarly gave rise to myoblasts. Notably, infection of Pax7-deficient muscle with adenoviral Pax7 resulted in the de novo formation of regenerated myofibers. Taken together, these results indicate that Pax7 is necessary and sufficient to induce the myogenic specification of CD45(+ stem cells resident in adult skeletal muscle. Moreover, these experiments suggest that viral transduction of Pax7 is a potential therapeutic approach for the treatment of neuromuscular degenerative diseases.

  18. Monitoring Notch Signaling-Associated Activation of Stem Cell Niches within Injured Dental Pulp.

    Science.gov (United States)

    Mitsiadis, Thimios A; Catón, Javier; Pagella, Pierfrancesco; Orsini, Giovanna; Jimenez-Rojo, Lucia

    2017-01-01

    Dental pulp stem/progenitor cells guarantee tooth homeostasis, repair and regeneration throughout life. The decision between renewal and differentiation of these cells is influenced by physical and molecular interactions with stromal cells and extracellular matrix molecules forming the specialized microenvironment of dental pulp stem cell niches. Here we study the activation of putative pulp niches after tooth injury through the upregulation of Notch signaling pathway. Notch1, Notch2, and Notch3 molecules were used as markers of dental pulp stem/progenitor cells. Upon dental injury, Notch1 and Notch3 are detected in cells related to vascular structures suggesting a role of these proteins in the activation of specific pulpal perivascular niches. In contrast, a population of Notch2-positive cells that are actively proliferative is observed in the apical part of the pulp. Kinetics of these cells is followed up with a lipophilic DiI labeling, showing that apical pulp cells migrate toward the injury site where dynamic regenerative/repair events occur. The knowledge of the activation and regulation of dental pulp stem/progenitor cells within their niches in pathologic conditions may be helpful for the realization of innovative dental treatments in the near future.

  19. Behavior of pulsed electric field injured Escherichia coli O157:H7 cells in apple juice amended with pyruvate and catalase

    Science.gov (United States)

    Pulse Electric Field (PEF) treatment has been used to inactivate bacteria in liquid foods. However, information on the behavior of PEF injured Escherichia coli bacteria in media during storage at 5 and 23C are limited. In this study, we investigated the fate of E. coli O157:H7 cells at 6.8 log CFU/m...

  20. Possible Local Stem Cells Activation by Microcurrent Application in Experimentally Injured Soleus Muscle

    OpenAIRE

    Zickri, Maha Baligh

    2014-01-01

    Background: Severe injuries in skeletal muscle result in muscle weakness that delays recovery and contribute to progressive decline in muscle function. Microcurrent therapy (MCT) is a novel treatment method used in soft tissue injury and tissue regeneration therapy. The regenerative capacity of skeletal muscle tissue resides in satellite cells, the quiescent adult stem cells. Aim: The present work aimed at investigating the relation between microcurrent therapy and local stem cells in regener...

  1. A single exposure to a sublethal pediocin concentration initiates a resistance-associated temporal cell envelope and general stress response in Listeria monocytogenes

    DEFF Research Database (Denmark)

    Laursen, Martin Frederik; Bahl, Martin Iain; Licht, Tine Rask

    2015-01-01

    was to determine if exposure to sublethal concentrations of pediocin-containing Lactobacillus plantarum WHE 92 supernatant could prime L. monocytogenes for resistance. By transcriptomic analysis, we found two, 55 and 539 genes differentially expressed after 10, 60 and 180 min of exposure to L. plantarum WHE 92...... resistant than wild types to L. plantarum WHE 92 supernatant. LisRK, SigB and SigL regulation and genes associated with resistance are involved in the temporal adaptive response to pediocin and all three regulatory systems affect pediocin resistance. Thus, a single exposure to a sublethal pediocin...

  2. Possible local stem cells activation by microcurrent application in experimentally injured soleus muscle.

    Science.gov (United States)

    Zickri, Maha Baligh

    2014-11-01

    Severe injuries in skeletal muscle result in muscle weakness that delays recovery and contribute to progressive decline in muscle function. Microcurrent therapy (MCT) is a novel treatment method used in soft tissue injury and tissue regeneration therapy. The regenerative capacity of skeletal muscle tissue resides in satellite cells, the quiescent adult stem cells. The present work aimed at investigating the relation between microcurrent therapy and local stem cells in regeneration of induced skeletal muscle injury in albino rat. Twenty six adult male albino rats were divided into Sham group, Injury group (I): subjected to soleus muscle injury and subdivided into subgroups I1 & I2 sacrificed 2 and 4 weeks after injury respectively. Microcurrent group (M): subjected to muscle injury and micro-current was applied. The animals were subdivided into subgroups M1 and M2 sacrificed 2 and 4 weeks after injury. Histological, immunohistochemical and morphometric studies were performed. Atypical fibers widely separated by infiltrating cells and strong acidophilic sarcoplasm with focal vacuolations were found in injury group. In M1 subgroup few atypical fibers were found. In M2 subgroup multiple typical fibers were detected. A significant decrease in the mean area of atypical fibers, a significant increase in the mean area% of alpha SMA+ve cells and that of CD34+ve cells were found in microcurrent group compared to injury group. A definite therapeutic effect of the microcurrent was found on induced skeletal muscle injury. This effect was proved to be related to satellite cell activation.

  3. Cell counts and survival to vitrification of bovine in vitro produced blastocysts subjected to sublethal high hydrostatic pressure.

    Science.gov (United States)

    Trigal, B; Muñoz, M; Gómez, E; Caamaño, J N; Martin, D; Carrocera, S; Casais, R; Diez, C

    2013-04-01

    This work analyses the effects of a high hydrostatic pressure (HHP) treatment on in vitro survival of in vitro produced (IVP) bovine embryos vitrified with the Cryologic Vitrification Method (CVM). Consequences on embryo quality in terms of cell proliferation and differentiation, and levels of embryonic Heat Shock Protein 70 (Hsp-70) were also examined. Day 7 and 8 bovine in vitro-produced blastocysts were submitted to an HHP treatment (60 MPa, at 32 °C for 1 h) and allowed to recover for 1 or 2 h in culture medium. The HHP treatment did not improve blastocyst survival rates after vitrification/warming. Survival (24 h post-warming) and hatching (48 h post-warming) rates were 79.3 ± 4.9 and 51.8 ± 4.2 vs 73.9 ± 4.2 and 44.7 ± 4.1 for untreated controls and HHP-treated embryos, respectively. Total cell numbers measured in fresh embryos were reduced after 1 h at 32 °C, with or without HHP treatment, indicating that cell proliferation was stopped as a result of stress. Vitrified HHP-treated embryos that hatched at 48 h after warming showed increased cell numbers in their ICM compared with untreated controls (50.2 ± 3.1 vs 38.8 ± 2.7), indicating higher embryo quality. Treatment of blastocysts with HHP did not alter the level of the Hsp-70 protein. In our conditions, HHP treatment did not affect the cryoresistance of these embryos. However, combination of HHP treatment and vitrification in fibreplugs resulted in an increase in the ICM cell number of hatched embryos 48 h post-warming. © 2012 Blackwell Verlag GmbH.

  4. Single dose of inducible nitric oxide synthase inhibitor induces prolonged inflammatory cell accumulation and fibrosis around injured tendon and synovium

    Directory of Open Access Journals (Sweden)

    Homa Darmani

    2004-01-01

    Full Text Available THE aim of the current study was to investigate the effect of inhibition of nitric oxide (NO production after injury on inflammatory cell accumulation and fibrosis around digital flexor tendon and synovium. A standard crush injury was applied to the flexor tendons of the middle digit of the hindpaw and the overlying muscle and synovium of female Wistar rats. Thirty animals received an intraperitoneal injection of either isotonic saline or N(G-nitro-l-arginine methyl ester (L-NAME; 5 mg/kg immediately following the crush injury, and five animals were then sacrificed at various intervals and the paws processed for histology. Another group of five animals was sacrificed after 3 days for nitrite determinations. The results showed that nitrite production and hence NO synthase activity is doubled at the acute phase of tendon wound healing, and we can prevent this by administering a single dose of L-NAME immediately after injury. The incidence and severity of fibrocellular adhesions between tendon and synovium was much more marked in animals treated with L-NAME. Treatment with L-NAME elicited a chronic inflammatory response characterised by a persistent and extraordinarily severe accumulation of large numbers of inflammatory cells in the subcutaneous tissues, in muscle and in tendon. These findings indicate that in the case of injured tendon and synovium, NO could act to protect the healing tissue from an uncontrolled inflammatory response.

  5. Carvacrol and 1,8-cineole alone or in combination at sublethal concentrations induce changes in the cell morphology and membrane permeability of Pseudomonas fluorescens in a vegetable-based broth.

    Science.gov (United States)

    de Sousa, Jossana Pereira; Torres, Rayanne de Araújo; de Azerêdo, Geíza Alves; Figueiredo, Regina Célia Bressan Queiroz; Vasconcelos, Margarida Angélica da Silva; de Souza, Evandro Leite

    2012-08-01

    This study aimed to investigate the effects of sublethal concentrations of carvacrol (CAR) and 1,8-cineole (CIN) alone and in combination on the morphology, cell viability and membrane permeability of Pseudomonas fluorescens ATCC 11253 cultivated in a vegetable-based broth. Transmission and scanning electron microscopy images of bacterial cells exposed to CAR and CIN alone or in combination showed marked ultrastructural changes after 1h of exposure. These changes included shrunken protoplasm, discontinuity of the outer and cytoplasmic membranes and leakage of the intracellular material. Confocal scanning laser microscopy images corroborated the electron microscopy data, showing a decrease in the number of SYTO-9 cells (intact cells) with a concomitant increase in the number of PI-positive cells (dead cells). All of these morphological changes are indicative of increased membrane permeability and the loss of bacterial envelope integrity, which ultimately lead to cell death. The combination of sublethal concentrations of CAR and CIN could be applied to inhibit the growth of P. fluorescens on vegetables. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Modelling the effect of sub(lethal) heat treatment of Bacillus subtilis spores on germination rate and outgrowth to exponentially growing vegetative cells.

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    Smelt, J P P M; Bos, A P; Kort, R; Brul, S

    2008-11-30

    Spores of Bacillus subtilis were subjected to relatively mild heat treatments in distilled water and properties of these spores were studied. These spores had lost all or part of their dipicolinic acid (DPA) depending on the severity of the heat treatment. Even after relatively mild heat treatments these spore lost already a small but significant amount of DPA. When these spores were inoculated in nutrient medium-tryptone soy broth (TSA)-the non-lethally heated spores started to germinate. Results of classical optical density measurements showed that both phase darkening and subsequent outgrowth could be affected by sub-lethal heat. A study of single cells in TSB showed that lag times originating from exponentially growing cells followed a normal distribution, whereas lag times originating from spores followed a Weibull distribution. Besides classical optical density measurements were made to study the effect of previous heating on the kinetics of the first stages of germination. The germination kinetics could be described by the model as was proposed by Geeraerd et al. [Geeraerd, A.H., Herremans, C.H. and Van Impe, J.F., 2000. Structural model requirements to describe microbial inactivation during a mild heat treatment. International Journal of Food Microbiology 59, 185-209]. Two of the 4 parameters of the sigmoid model of Geeraerd were dependent on heating time and heating temperature, whereas the two other parameters were considered as independent of the heating conditions. Based on these observations, a secondary model could be developed that describes the combined effect of heating temperature and heating time on the kinetics of germination. To have more detailed information of the kinetics of germination samples incubated in TSB were tested at regular time intervals by flow cytometry. To that end the cells were stained with syto 9 to distinguish between the various germination stages. There was a qualitative agreement between the results of flow cytometry and

  7. Knockout of CD8 delays reendothelialization and accelerates neointima formation in injured arteries of mouse via TNF-α inhibiting the endothelial cells migration.

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    Jun-Meng Zhang

    Full Text Available OBJECTIVE: Delayed or impaired reendothelialization is a major cause of stent thrombosis in the interventional treatment of coronary heart disease. T cells are involved in neointima formation of injured arteries. However, the regulated mechanism of reendothelialization and the role of CD8 T cell in reendothelialization are unclear. METHODS AND RESULTS: Immunofluorescence staining showed that CD8 positive cells were increased in wire injured femoral artery of mice. On day 21 after injury, elastin staining showed that knockout of CD8 (CD8(-/- significantly increased intimal thickness and a ratio of intima to media by 1.8 folds and 1.9 folds respectively in injured arteries. Evans blue staining showed that knockout of CD8 delayed the reendothelialization area on day 7 after injury (18.8±0.5% versus 42.1±5.6%, p<0.05. In vitro, a migration assay revealed that CD8(-/- T cells co-cultured with WT macrophages significantly inhibited the migration of the endothelial cells (ECs; compared to CD4(+ T cells, and CD8(+ T cells could promote the ECs migration. Furthermore, real-time PCR analysis showed that knockout of CD8 increased the level of tumor necrosis factor α (TNF-α in injured arteries and cytometric bead cytokine array showed that TNF-α was elevated in cultured CD8(-/- T cells. Finally, a wound-healing assay showed that recombinant TNF-α significantly inhibited the migration of ECs. CONCLUSION: Our study suggested that CD8(+ T cells could promote the reendothelialization and inhibit the neointima formation after the artery wire injury, and this effect is at least partly dependent on decreasing TNF-α production promoting ECs migration.

  8. Rejuvenation of the aged muscle stem cell population restores strength to injured aged muscles

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    Cosgrove, Benjamin D.; Gilbert, Penney M.; Porpiglia, Ermelinda; Mourkioti, Foteini; Lee, Steven P.; Corbel, Stephane Y.; Llewellyn, Michael E.; Delp, Scott L.; Blau, Helen M.

    2014-01-01

    The aged suffer from progressive muscle weakness and regenerative failure. We demonstrate that muscle regeneration is impaired with aging due in part to a cell-autonomous functional decline in skeletal muscle stem cells (MuSCs). Two-thirds of aged MuSCs are intrinsically defective relative to young MuSCs, with reduced capacity to repair myofibers and repopulate the stem cell reservoir in vivo following transplantation due to a higher incidence of cells that express senescence markers and that have elevated p38α/β MAPK activity. We show that these limitations cannot be overcome by transplantation into the microenvironment of young recipient muscles. In contrast, subjecting the aged MuSC population to transient inhibition of p38α/β in conjunction with culture on soft hydrogel substrates rapidly expands the residual functional aged MuSC population, rejuvenating its potential for regeneration, serial transplantation, and strengthening damaged muscles of aged mice. These findings reveal a synergy between biophysical and biochemical cues that provides a paradigm for a localized autologous muscle stem cell therapy in aged individuals. PMID:24531378

  9. Rejuvenation of the muscle stem cell population restores strength to injured aged muscles.

    Science.gov (United States)

    Cosgrove, Benjamin D; Gilbert, Penney M; Porpiglia, Ermelinda; Mourkioti, Foteini; Lee, Steven P; Corbel, Stephane Y; Llewellyn, Michael E; Delp, Scott L; Blau, Helen M

    2014-03-01

    The elderly often suffer from progressive muscle weakness and regenerative failure. We demonstrate that muscle regeneration is impaired with aging owing in part to a cell-autonomous functional decline in skeletal muscle stem cells (MuSCs). Two-thirds of MuSCs from aged mice are intrinsically defective relative to MuSCs from young mice, with reduced capacity to repair myofibers and repopulate the stem cell reservoir in vivo following transplantation. This deficiency is correlated with a higher incidence of cells that express senescence markers and is due to elevated activity of the p38α and p38β mitogen-activated kinase pathway. We show that these limitations cannot be overcome by transplantation into the microenvironment of young recipient muscles. In contrast, subjecting the MuSC population from aged mice to transient inhibition of p38α and p38β in conjunction with culture on soft hydrogel substrates rapidly expands the residual functional MuSC population from aged mice, rejuvenating its potential for regeneration and serial transplantation as well as strengthening of damaged muscles of aged mice. These findings reveal a synergy between biophysical and biochemical cues that provides a paradigm for a localized autologous muscle stem cell therapy for the elderly.

  10. Neural Stem Cell Plasticity: Advantages in Therapy for the Injured Central Nervous System.

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    Ottoboni, Linda; Merlini, Arianna; Martino, Gianvito

    2017-01-01

    The physiological and pathological properties of the neural germinal stem cell niche have been well-studied in the past 30 years, mainly in animals and within given limits in humans, and knowledge is available for the cyto-architectonic structure, the cellular components, the timing of development and the energetic maintenance of the niche, as well as for the therapeutic potential and the cross talk between neural and immune cells. In recent years we have gained detailed understanding of the potentiality of neural stem cells (NSCs), although we are only beginning to understand their molecular, metabolic, and epigenetic profile in physiopathology and, further, more can be invested to measure quantitatively the activity of those cells, to model in vitro their therapeutic responses or to predict interactions in silico . Information in this direction has been put forward for other organs but is still limited in the complex and very less accessible context of the brain. A comprehensive understanding of the behavior of endogenous NSCs will help to tune or model them toward a desired response in order to treat complex neurodegenerative diseases. NSCs have the ability to modulate multiple cellular functions and exploiting their plasticity might make them into potent and versatile cellular drugs.

  11. CCND2 Overexpression Enhances the Regenerative Potency of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Remuscularization of Injured Ventricle.

    Science.gov (United States)

    Zhu, Wuqiang; Zhao, Meng; Mattapally, Saidulu; Chen, Sifeng; Zhang, Jianyi

    2017-10-10

    Rationale: The effectiveness of transplanted, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) for treatment of ischemic myocardial injury is limited by the exceptionally low engraftment rate. Objective: To determine whether overexpression of the cell-cycle activator CCND2 in hiPSC-CMs can increase the graft size and improve myocardial recovery in a mouse model of myocardial infarction (MI) by increasing the proliferation of grafted cells. Methods and Results: Human CCND2 was delivered to hiPSCs via lenti-viral mediated gene transfection. In cultured cells, markers for cell-cycle activation and proliferation were ~3-7 folds higher in CCND2-overexpressing hiPSC-CMs (hiPSC-CCND2(OE)CMs) than in hiPSC-CMs with normal levels of CCND2 (hiPSC-CCND2(WT)CMs) (p<0.01). The pluripotent genes (Oct 4, Sox2, Nanog) decrease to minimal- and undetectable -levels at day 1 and 10 after differentiate to CMs. In the mouse MI model, cardiac function, infarct size, and the number of engrafted cells were similar at week 1 after treatment with hiPSC-CCND2(OE)CMs or hiPSC-CCND2(WT)CMs but was about tripled in hiPSC-CCND2(OE)CM-treated than in hiPSC-CCND2(WT)CM-treated animals at week 4 (p<0.01). The cardiac function and infarct size were significantly better in both cell-treatment groups hearts than in control hearts, which was most prominent in hiPSC-CCND2(OE)CM-treated animals (p<0.05, each). No tumor formation was observed in any hearts. Conclusions: CCND2 overexpression activates cell-cycle progression in hiPSC-CMs that results in a significant enhanced potency for myocardial repair as evidenced by remuscularization of injured myocardium. This LV muscle regeneration and increased angiogenesis in border zone are accompanied by a significant improvement of LV chamber function.

  12. Intraovarian Transplantation of Female Germline Stem Cells Rescue Ovarian Function in Chemotherapy-Injured Ovaries.

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    Jiaqiang Xiong

    Full Text Available Early menopause and infertility often occur in female cancer patients after chemotherapy (CTx. For these patients, oocyte/embryo cryopreservation or ovarian tissue cryopreservation is the current modality for fertility preservation. However, the above methods are limited in the long-term protection of ovarian function, especially for fertility preservation (very few females with cancer have achieved pregnancy with cryopreserved ovarian tissue or eggs until now. In addition, the above methods are subject to their scope (females with no husband or prepubertal females with no mature oocytes. Thus, many females who suffer from cancers would not adopt the above methods pre- and post-CTx due to their uncertainty, safety and cost-effectiveness. Therefore, millions of women have achieved long-term survival after thorough CTx treatment and have desired to rescue their ovarian function and fertility with economic, durable and reliable methods. Recently, some studies showed that mice with infertility caused by CTx can produce normal offspring through intraovarian injection of exogenous female germline stem cells (FGSCs. Though exogenous FGSC can be derived from mice without immune rejection in the same strain, it is difficult to obtain human female germline stem cells (hFGSCs, and immune rejection could occur between different individuals. In this study, infertility in mice was caused by CTx, and the ability of FGSCs to restore ovarian function or even produce offspring was assessed. We had successfully isolated and purified the FGSCs from adult female mice two weeks after CTx. After infection with GFP-carrying virus, the FGSCs were transplanted into ovaries of mice with infertility caused by CTx. Finally, ovarian function was restored and the recipients produced offspring long-term. These findings showed that mice with CTx possessed FGSCs, restoring ovarian function and avoiding immune rejection from exogenous germline stem cells.

  13. Dental Pulp Stem Cell Recruitment Signals within Injured Dental Pulp Tissue

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    Charlotte Rombouts

    2016-03-01

    Full Text Available The recruitment of dental pulp stem cells (DPSC is a prerequisite for the regeneration of dentin damaged by severe caries and/or mechanical injury. Understanding the complex process of DPSC recruitment will benefit future in situ tissue engineering applications based on the stimulation of endogenous DPSC for dentin pulp regeneration. The current known mobilization signals and subsequent migration of DPSC towards the lesion site, which is influenced by the pulp inflammatory state and the application of pulp capping materials, are reviewed. The research outcome of migration studies may be affected by the applied methodology, which should thus be chosen with care. Both the advantages and disadvantages of commonly used assays for investigating DPSC migration are discussed. This review highlights the fact that DPSC recruitment is dependent not only on the soluble chemotactic signals, but also on their interaction with neighboring cells and the extracellular matrix, which can be modified under pathological conditions. These are discussed to explain how these modifications lead to the stimulation of DPSC recruitment.

  14. 28Si total body irradiation injures bone marrow hematopoietic stem cells via induction of cellular apoptosis

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    Chang, Jianhui; Feng, Wei; Wang, Yingying; Allen, Antiño R.; Turner, Jennifer; Stewart, Blair; Raber, Jacob; Hauer-Jensen, Martin; Zhou, Daohong; Shao, Lijian

    2017-05-01

    Long-term space mission exposes astronauts to a radiation environment with potential health hazards. High-energy charged particles (HZE), including 28Si nuclei in space, have deleterious effects on cells due to their characteristics with high linear energy transfer and dense ionization. The influence of 28Si ions contributes more than 10% to the radiation dose equivalent in the space environment. Understanding the biological effects of 28Si irradiation is important to assess the potential health hazards of long-term space missions. The hematopoietic system is highly sensitive to radiation injury and bone marrow (BM) suppression is the primary life-threatening injuries after exposure to a moderate dose of radiation. Therefore, in the present study we investigated the acute effects of low doses of 28Si irradiation on the hematopoietic system in a mouse model. Specifically, 6-month-old C57BL/6 J mice were exposed to 0.3, 0.6 and 0.9 Gy 28Si (600 MeV) total body irradiation (TBI). The effects of 28Si TBI on BM hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs) were examined four weeks after the exposure. The results showed that exposure to 28Si TBI dramatically reduced the frequencies and numbers of HSCs in irradiated mice, compared to non-irradiated controls, in a radiation dose-dependent manner. In contrast, no significant changes were observed in BM HPCs regardless of radiation doses. Furthermore, irradiated HSCs exhibited a significant impairment in clonogenic ability. These acute effects of 28Si irradiation on HSCs may be attributable to radiation-induced apoptosis of HSCs, because HSCs, but not HPCs, from irradiated mice exhibited a significant increase in apoptosis in a radiation dose-dependent manner. However, exposure to low doses of 28Si did not result in an increased production of reactive oxygen species and DNA damage in HSCs and HPCs. These findings indicate that exposure to 28Si irradiation leads to acute HSC damage.

  15. Importance of considering injured microorganisms in sterilization validation.

    Science.gov (United States)

    Shintani, Hideharu

    2006-09-01

    Disinfection or sterilization treatment by heating, irradiation, or chemicals can cause injury to microorganisms at sublethal levels. Microbial injury is the inability to grow under conditions suitable for the uninjured microorganisms. This inability of injured microorganisms to grow is explained in terms of more complex or different nutritional requirements or in terms of increased sensitivity to environmental conditions such as incubation conditions (time or temperature) or to chemical agents such as halogen compounds. Injured microorganisms can be distinguished from those that are dead or mutated by their ability to regain normal physiological activity when placed in appropriate conditions for cultivation. The return to normal physiological function has been termed repair. The extent and severity of sublethal injury, the mechanisms of injury, and the mechanisms and degree of recovery vary with the sterilization procedures, the species, the strains, the condition of the microorganism, and the methods of repair. Injury to spore formers has been detected at different stages of the spore cycle. The sites of injury include damage to enzymes, membrane disruption, and/or damage to DNA or RNA. Information on the sublethal injury and recovery of microorganisms is very important in evaluating sterilization/disinfection procedures. This paper supplies academic as well as practical information dealing with the repair, and detection of injured microorganisms for performing reproducible sterilization validation.

  16. Septin4 as a novel binding partner of PARP1 contributes to oxidative stress induced human umbilical vein endothelial cells injure.

    Science.gov (United States)

    Zhang, Naijin; Zhang, Ying; Zhao, Sichao; Sun, Yingxian

    2018-02-05

    Oxidative stress induced vascular endothelial cell injure is one of the key and initial event in the development of atherosclerosis. Septin4, as a member of GTP binding protein family, is widely expressed in the eukaryotic cells and considered to be an essential component of the cytoskeleton which is involved in many important physiological processes. However, whether Septin4 is involved in cardiovascular diseases, such as oxidative stress inducted endothelial cell injury still unclear. PARP1 as a DNA repair enzyme can be activated by identifying DNA damaged fragments, which consumes high levels of energy and leads to vascular endothelial cell apoptosis. Here, our results first found that Septin4 is involved in oxidative stress induced endothelial cell ROS production and apoptosis through knock-down and over-expression Septin4 approaches. Furthermore, to explore how Septin4 is involved in oxidative stress induced endothelial cells injure, we first identified that Septin4 is a novel PARP1 interacting protein and the interaction is enhanced under oxidative stress. In conclusions, our founding indicates that Septin4 is a novel essential factor involved in oxidative stress induced vascular endothelial cell injury by interacting with apoptosis-related protein PARP1. Copyright © 2018 Elsevier Inc. All rights reserved.

  17. GM-CSF produced by nonhematopoietic cells is required for early epithelial cell proliferation and repair of injured colonic mucosa.

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    Egea, Laia; McAllister, Christopher S; Lakhdari, Omar; Minev, Ivelina; Shenouda, Steve; Kagnoff, Martin F

    2013-02-15

    GM-CSF is a growth factor that promotes the survival and activation of macrophages and granulocytes, as well as dendritic cell differentiation and survival in vitro. The mechanism by which exogenous GM-CSF ameliorates the severity of Crohn's disease in humans and colitis in murine models has mainly been considered to reflect its activity on myeloid cells. We used GM-CSF-deficient (GM-CSF(-/-)) mice to probe the functional role of endogenous host-produced GM-CSF in a colitis model induced after injury to the colon epithelium. Dextran sodium sulfate (DSS), at doses that resulted in little epithelial damage and mucosal ulceration in wild type mice, caused marked colon ulceration and delayed ulcer healing in GM-CSF(-/-) mice. Colon crypt epithelial cell proliferation in vivo was significantly decreased in GM-CSF(-/-) mice at early times after DSS injury. This was paralleled by decreased expression of crypt epithelial cell genes involved in cell cycle, proliferation, and wound healing. Decreased crypt cell proliferation and delayed ulcer healing in GM-CSF(-/-) mice were rescued by exogenous GM-CSF, indicating the lack of a developmental abnormality in the epithelial cell proliferative response in those mice. Nonhematopoietic cells, and not myeloid cells, produced the GM-CSF important for colon epithelial proliferation after DSS-induced injury, as revealed by bone marrow chimera and dendritic cell-depletion experiments, with colon epithelial cells being the cellular source of GM-CSF. Endogenous epithelial cell-produced GM-CSF has a novel nonredundant role in facilitating epithelial cell proliferation and ulcer healing in response to injury of the colon crypt epithelium.

  18. Acute and sublethal toxicity of seepage waters from garbage dumps to permanent cell lines and primary cultures of hepatocytes from rainbow trout (Oncorhynchus mykiss): a novel approach to environmental risk assessment for chemicals and chemical mixtures.

    Science.gov (United States)

    Zahn, T; Hauck, C; Holzschuh, J; Braunbeck, T

    1995-01-01

    In order to evaluate the suitability of cytopathological criteria in isolated fish hepatocytes as endpoints in (eco)toxicological research, liver cells isolated from rainbow trout (Oncorhynchus mykiss) by collagenase perfusion were exposed in vitro for up to 5 days to sublethal dilutions of two seepage water samples collected from garbage dumps. Hepatocytes were analysed with respect to acute (lactate dehydrogenase leakage) and sublethal toxicity (electron microscopy, stereology). In addition, acute toxicity (24 h) was tested in the piscine fibrocytic cell line R1 by means of crystal violet staining and neutral red retention. Acute toxicity in R1 cells and isolated hepatocytes could only be documented for sample I at dilutions of 1:2 and 1:4. This difference in toxicity could be corroborated by cytological alterations in isolated hepatocytes, which could be documented for dilutions of 1:100 and 1:8 in samples I and II, respectively. Ultrastructural changes were time- and dose-dependent and included reduction of hepatocellular volume, disturbance of intracellular compartmentation, modified heterochromatin distribution, transformation of rough endoplasmic reticulum into concentric membrane whorls, proliferation of lysosomes and cytoplasmic vacuoles, as well as reduction of hepatocellular glycogen. Although several hepatocellular reactions were found after exposure to either sample, the syndrome of ultrastructural alterations allowed clear differentiation between the two samples. Results illustrate that cytological effects far below macroscopically detectable damage can be discovered not only in intact fish, but also in fish cell culture systems. On the basis of the data presented, a multi-tiered test procedure for aquatic toxicity assessment exclusively based on tests with fish cell culture systems is proposed: (1) rapid screening for acute toxicity with permanent cell lines; (2) short-term tests with more complex, yet more sensitive systems such as primary

  19. Mechanisms of adhesion and subsequent actions of a haematopoietic stem cell line, HPC-7, in the injured murine intestinal microcirculation in vivo.

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    Dean P J Kavanagh

    Full Text Available Although haematopoietic stem cells (HSCs migrate to injured gut, therapeutic success clinically remains poor. This has been partially attributed to limited local HSC recruitment following systemic injection. Identifying site specific adhesive mechanisms underpinning HSC-endothelial interactions may provide important information on how to enhance their recruitment and thus potentially improve therapeutic efficacy. This study determined (i the integrins and inflammatory cyto/chemokines governing HSC adhesion to injured gut and muscle (ii whether pre-treating HSCs with these cyto/chemokines enhanced their adhesion and (iii whether the degree of HSC adhesion influenced their ability to modulate leukocyte recruitment.Adhesion of HPC-7, a murine HSC line, to ischaemia-reperfused (IR injured mouse gut or cremaster muscle was monitored intravitally. Critical adhesion molecules were identified by pre-treating HPC-7 with blocking antibodies to CD18 and CD49d. To identify cyto/chemokines capable of recruiting HPC-7, adhesion was monitored following tissue exposure to TNF-α, IL-1β or CXCL12. The effects of pre-treating HPC-7 with these cyto/chemokines on surface integrin expression/clustering, adhesion to ICAM-1/VCAM-1 and recruitment in vivo was also investigated. Endogenous leukocyte adhesion following HPC-7 injection was again determined intravitally.IR injury increased HPC-7 adhesion in vivo, with intestinal adhesion dependent upon CD18 and muscle adhesion predominantly relying on CD49d. Only CXCL12 pre-treatment enhanced HPC-7 adhesion within injured gut, likely by increasing CD18 binding to ICAM-1 and/or CD18 surface clustering on HPC-7. Leukocyte adhesion was reduced at 4 hours post-reperfusion, but only when local HPC-7 adhesion was enhanced using CXCL12.This data provides evidence that site-specific molecular mechanisms govern HPC-7 adhesion to injured tissue. Importantly, we show that HPC-7 adhesion is a modulatable event in IR injury and

  20. Derivation of injury-responsive dendritic cells for acute brain targeting and therapeutic protein delivery in the stroke-injured rat.

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    Nathan C Manley

    Full Text Available Research with experimental stroke models has identified a wide range of therapeutic proteins that can prevent the brain damage caused by this form of acute neurological injury. Despite this, we do not yet have safe and effective ways to deliver therapeutic proteins to the injured brain, and this remains a major obstacle for clinical translation. Current targeted strategies typically involve invasive neurosurgery, whereas systemic approaches produce the undesirable outcome of non-specific protein delivery to the entire brain, rather than solely to the injury site. As a potential way to address this, we developed a protein delivery system modeled after the endogenous immune cell response to brain injury. Using ex-vivo-engineered dendritic cells (DCs, we find that these cells can transiently home to brain injury in a rat model of stroke with both temporal and spatial selectivity. We present a standardized method to derive injury-responsive DCs from bone marrow and show that injury targeting is dependent on culture conditions that maintain an immature DC phenotype. Further, we find evidence that when loaded with therapeutic cargo, cultured DCs can suppress initial neuron death caused by an ischemic injury. These results demonstrate a non-invasive method to target ischemic brain injury and may ultimately provide a way to selectively deliver therapeutic compounds to the injured brain.

  1. Analysis of cell cycle regulated and regulating proteins following exposure of lung derived cells to sub-lethal doses of a-rays

    Science.gov (United States)

    Trani, D.; Claudio, P. P.; Cassone, M.; Lucchetti, C.; D'Agostino, L.; Caputi, M.; Giordano, A.

    Introduction Since the last century mankind had to face an increased exposure to man made and natural sources of radiation Radiation represents a therapeutic instrument for radiosensitive cancers as well as a cytotoxic agent for normal human tissues The effects of prolonged exposure to low doses of high energy radiation are still not well-known at the molecular and clinical level Understanding their molecular effects will aid in developing more tailored therapeutic strategies as well as implementing radio-protective measures essential prerequisite for the long-time permanence of men in space Objective of the study The general aim of this study was to evaluate the susceptibility and the response of lung epithelial cells to DNA damage induced by ionizing radiations We decided to study a panel of epithelial bronchial cell lines because of their fast-growth rate and their prominent exposure to both environmental and medical radiations The specific objective of our study was to qualitatively and semi-quantitatively assess the involvement and behaviour of selected genes in DNA damage DNA-repair mechanisms and apoptosis which follow radiation exposure with the aim to determine the involvement of the most promising targets for the early detection of radiation-mediated lung damage before chronic disease develops Methods Four epithelial cell lines one normal and three neoplastic were selected in order to detect and compare survival cell cycle and protein expression differences related to their different genetic asset

  2. Effect of Content of Sulfate Groups in Seaweed Polysaccharides on Antioxidant Activity and Repair Effect of Subcellular Organelles in Injured HK-2 Cells

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    Xiao-Tao Ma

    2017-01-01

    Full Text Available This study aims to investigate the repair effect of subcellular structure injuries of the HK-2 cells of four degraded seaweed polysaccharides (DSPs, namely, the degraded Porphyra yezoensis, Gracilaria lemaneiformis, Sargassum fusiform, and Undaria pinnatifida polysaccharides. The four DSPs have similar molecular weight, but with different content of sulfate groups (i.e., 17.9%, 13.3%, 8.2%, and 5.5%, resp.. The damaged model was established using 2.8 mmol/L oxalate to injure HK-2 cells, and 60 μg/mL of various DSPs was used to repair the damaged cells. With the increase of sulfate group content in DSPs, the scavenging activity of radicals and their reducing power were all improved. Four kinds of DSPs have repair effect on the subcellular organelles of damaged HK-2 cells. After being repaired by DSPs, the release amount of lactate dehydrogenase was decreased, the integrity of cell membrane and lysosome increased, the Δψm increased, the cell of G1 phase arrest was inhibited, the proportion of S phase increased, and cell apoptotic and necrosis rates were significantly reduced. The greater the content of sulfate group is, the stronger is the repair ability of the polysaccharide. These DSPs, particularly the polysaccharide with higher sulfate group content, may be a potential drug for the prevention and cure of kidney stones.

  3. Transplanted neurally modified bone marrow-derived mesenchymal stem cells promote tissue protection and locomotor recovery in spinal cord injured rats.

    Science.gov (United States)

    Alexanian, Arshak R; Fehlings, Michael G; Zhang, Zhiying; Maiman, Dennis J

    2011-01-01

    Stem cell-based therapy for repair and replacement of lost neural cells is a promising treatment for central nervous system (CNS) diseases. Bone marrow (BM)-derived mesenchymal stem cells (MSCs) can differentiate into neural phenotypes and be isolated and expanded for autotransplantation with no risk of rejection. The authors examined whether transplanted neurally induced human MSCs (NI hMSCs), developed by a new procedure, can survive, differentiate, and promote tissue protection and functional recovery in injured spinal cord (ISC) rats. Neural induction was achieved by exposing cells simultaneously to inhibitors of DNA methylation, histone deacetylation, and pharmacological agents that increased cAMP levels. Three groups of adult female Sprague-Dawley rats were injected immediately rostral and caudal to the midline lesion with phosphate-buffered saline, MSCs, or NI hMSCs, 1 week after a spinal cord impact injury at T-8. Functional outcome was measured using the Basso Beattie Bresnahan (BBB) locomotor rating scale and thermal sensitivity test on a weekly basis up to 12 weeks postinjury. Graft integration and anatomy of spinal cord was assessed by stereological, histochemical, and immunohistochemical techniques. The transplanted NI hMSCs survived, differentiated, and significantly improved locomotor recovery of ISC rats. Transplantation also reduced the volume of lesion cavity and white matter loss. This method of hMSC modification may provide an alternative source of autologous adult stem cells for CNS repair.

  4. Overexpression of Wnt3a facilitates the proliferation and neural differentiation of neural stem cells in vitro and after transplantation into an injured rat retina.

    Science.gov (United States)

    Yang, Xi-Tao; Bi, Yong-Yan; Chen, Er-Tao; Feng, Dong-Fu

    2014-02-01

    Neural stem cell-based therapy is a promising option for repair after injury. However, poor stem cell proliferation and insufficient differentiation of the stem cells into neurons are still difficult problems. The present study investigated whether transplantation of neural stem cells (NSCs) genetically modified to express Wnt3a is a promising approach to overcome these difficulties. We explored the possibility that Wnt3a might contribute to the therapeutic effect of NSC transplantation in retinal repair. The relative promotion of proliferation and neural differentiation by modified NSCs was investigated in a rat model of optic nerve crush. A recombinant lentivirus (Lenti-Wnt3a) was engineered to express Wnt3a. NSCs infected with control lentivirus (Lenti-GFP) or Lenti-Wnt3a were transplanted into the subretinal space immediately after the optic nerve crush. The proliferation and neural differentiation activity of the NSCs were assessed in vitro and in vivo. Overexpression of Wnt3a in NSCs induced activation of Wnt signaling, promoted proliferation, and directed the differentiation of the NSCs into neurons both in vitro and in vivo. Our study suggests that Wnt3a can potentiate the therapeutic benefits of NSC-based therapy in the injured retina. Copyright © 2013 Wiley Periodicals, Inc.

  5. Effects of CO2 on the resuscitation of Listeria monocytogenes injured by various bactericidal treatments.

    Science.gov (United States)

    Van Houteghem, Nancy; Devlieghere, Frank; Rajkovic, Andreja; Gómez, Sandra Maria Osés; Uyttendaele, Mieke; Debevere, Johan

    2008-03-31

    To assure the microbiological safety and quality of a food product, a combination of preservation hurdles is often used. Therefore, the effects of carbon dioxide at concentrations of 0, 20, 40 and 60% in modified atmospheres on the resuscitation of Listeria monocytogenes cells injured by mild bactericidal treatments during storage at 7 degrees C were examined. The bactericidal treatments were intense light pulses (ILP), chlorine dioxide (ClO(2)), lactic acid (LA) and heat. The results indicated additional bactericidal effects of CO(2) on cultures treated with LA, ClO(2) and ILP, with additional reductions in viable L. monocytogenes of 0.5-1.0 log cfu/ml. Lag phase duration was significantly different between the different treatments, with non-treated cells having the shortest lag phase, followed by that of heat, intense light pulses, lactic acid and finally ClO(2) treated cells. Maximum growth rate was also estimated and results showed a negative correlation with increasing CO(2) concentrations. A relationship was found between the amount of sub-lethally damaged cells after a mild inactivation treatment and the lag phase duration in the CO(2) environment. Current findings demonstrate the possibility that combining mild decontamination treatments and packaging in a CO(2) enriched environment could reduce the risk of L. monocytogenes infections in food due to an extension of the lag phase.

  6. Calcium oxalate crystal adherence to hyaluronan-, osteopontin-, and CD44-expressing injured/regenerating tubular epithelial cells in rat kidneys

    NARCIS (Netherlands)

    M. Asselman (Marino); A. Verhulst; M.E. de Broe; C.F. Verkoelen

    2003-01-01

    textabstractRetention of crystals in the kidney is an essential early step in renal stone formation. Studies with renal tubular cells in culture indicate that hyaluronan (HA) and osteopontin (OPN) and their mutual cell surface receptor CD44 play an important role in calcium oxalate

  7. Effects of dibutyryl cyclic-AMP on survival and neuronal differentiation of neural stem/progenitor cells transplanted into spinal cord injured rats.

    Directory of Open Access Journals (Sweden)

    Howard Kim

    Full Text Available Neural stem/progenitor cells (NSPCs have great potential as a cell replacement therapy for spinal cord injury. However, poor control over transplant cell differentiation and survival remain major obstacles. In this study, we asked whether dibutyryl cyclic-AMP (dbcAMP, which was shown to induce up to 85% in vitro differentiation of NSPCs into neurons would enhance survival of transplanted NSPCs through prolonged exposure either in vitro or in vivo through the controlled release of dbcAMP encapsulated within poly(lactic-co-glycolic acid (PLGA microspheres and embedded within chitosan guidance channels. NSPCs, seeded in fibrin scaffolds within the channels, differentiated in vitro to betaIII-tubulin positive neurons by immunostaining and mRNA expression, in response to dbcAMP released from PLGA microspheres. After transplantation in spinal cord injured rats, the survival and differentiation of NSPCs was evaluated. Untreated NSPCs, NSPCs transplanted with dbcAMP-releasing microspheres, and NSPCs pre-differentiated with dbcAMP for 4 days in vitro were transplanted after rat spinal cord transection and assessed 2 and 6 weeks later. Interestingly, NSPC survival was highest in the dbcAMP pre-treated group, having approximately 80% survival at both time points, which is remarkable given that stem cell transplantation often results in less than 1% survival at similar times. Importantly, dbcAMP pre-treatment also resulted in the greatest number of in vivo NSPCs differentiated into neurons (37±4%, followed by dbcAMP-microsphere treated NSPCs (27±14% and untreated NSPCs (15±7%. The reverse trend was observed for NSPC-derived oligodendrocytes and astrocytes, with these populations being highest in untreated NSPCs. This combination strategy of stem cell-loaded chitosan channels implanted in a fully transected spinal cord resulted in extensive axonal regeneration into the injury site, with improved functional recovery after 6 weeks in animals implanted with

  8. [Integration of Injured Hands].

    Science.gov (United States)

    Winkel, Reiner

    2017-02-01

    Any injury of a hand more or less disintegrates the injured hand. The rehabilitation of an alienated hand is prolonged. How can the alienation of the own but injured hand and the involuntary and unconscious inhibition of its perception and use be diagnosed, explained, avoided and designated? Medical observations and interviews with patients and therapists on the occasion of rehabilitation of hand injuries resulted in the development of a new concept of biopsychosocial integrating rehabilitation and a creative hand therapy with the objective of integrating injured and alienated hands. The inhibition of the gesture of thinking has been the most revealing sign for the diagnosis of a disintegration of an injured hand. Explanation: The involuntary inhibition to recognize and use an injured hand causes and implicates the alienation of the rested hand. Information by hand surgeons and hand therapists. Creative hand therapy guides the attention to complex and pleasing activities. In complex disintegrations after hand injuries the hand as well as the person need help. Designation: Hypotheses: Posttraumatic Proportionate Regional Disintegration while the tissues heal. Posttraumatic Complex Regional Disintegration, if the disintegration overruns the healing of tissues or if the person suffers. The Complex Regional Pain Syndrome is distinguished as a rare exception of a biopsychosocial disintegration. Posttraumatic regional disintegration seems to be proportionate while the tissues are healing. If the mentioned sensations of the patients and the visible signs of disintegration persist, the disorder spreads onto the biopsychosocial unit of the person. This disorder of hand and person may be designated as a complex disorder. The impact of a proportionate posttraumatic disintegration as well as the formation and exacerbation of a complex posttraumatic disintegration may be prevented by the facilitation of the integration of an injured hand. Scientific prove is needed for: the

  9. Human adipose stromal cells (ASC for the regeneration of injured cartilage display genetic stability after in vitro culture expansion.

    Directory of Open Access Journals (Sweden)

    Simona Neri

    Full Text Available Mesenchymal stromal cells are emerging as an extremely promising therapeutic agent for tissue regeneration due to their multi-potency, immune-modulation and secretome activities, but safety remains one of the main concerns, particularly when in vitro manipulation, such as cell expansion, is performed before clinical application. Indeed, it is well documented that in vitro expansion reduces replicative potential and some multi-potency and promotes cell senescence. Furthermore, during in vitro aging there is a decrease in DNA synthesis and repair efficiency thus leading to DNA damage accumulation and possibly inducing genomic instability. The European Research Project ADIPOA aims at validating an innovative cell-based therapy where autologous adipose stromal cells (ASCs are injected in the diseased articulation to activate regeneration of the cartilage. The primary objective of this paper was to assess the safety of cultured ASCs. The maintenance of genetic integrity was evaluated during in vitro culture by karyotype and microsatellite instability analysis. In addition, RT-PCR array-based evaluation of the expression of genes related to DNA damage signaling pathways was performed. Finally, the senescence and replicative potential of cultured cells was evaluated by telomere length and telomerase activity assessment, whereas anchorage-independent clone development was tested in vitro by soft agar growth. We found that cultured ASCs do not show genetic alterations and replicative senescence during the period of observation, nor anchorage-independent growth, supporting an argument for the safety of ASCs for clinical use.

  10. Sublethal injury and virulence changes in Listeria monocytogenes and Listeria innocua treated with antimicrobials carvacrol and citral.

    Science.gov (United States)

    Silva, A; Genovés, S; Martorell, P; Zanini, S F; Rodrigo, D; Martinez, A

    2015-09-01

    The aim of this study was to evaluate the effect of two antimicrobial substances, carvacrol and citral, on Listeria monocytogenes and Listeria innocua cells, as well as possible virulence changes in injured cells, using Caenorhabditis elegans as a model test. The results indicated that the percentage of sublethal damage was higher in L. monocytogenes than in L. innocua. The results of the study carried out by using C. elegans indicated that C. elegans fed in a lawn of L. monocytogenes previously treated with carvacrol showed a loss in life span (p ≤ 0.05) as compared with L. monocytogenes treated with citral, Escherichia coli OP50 as a negative control, and treated and untreated L. innocua. Egg laying was also affected: worms fed in a lawn of treated and untreated L. monocytogenes laid fewer eggs than those fed in a lawn of treated and untreated L. innocua or fed with OP50 as a negative control. Worms fed in a lawn of treated and untreated L. innocua also laid fewer eggs than those fed with OP50 as a negative control. A phenotype named bag of worms and an undescribed new one, "vulva inflammation", were also observed. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Phenotypic changes of Schwann cells on the proximal stump of injured peripheral nerve during repair using small gap conduit tube

    Directory of Open Access Journals (Sweden)

    Shi-jun Zhang

    2017-01-01

    Full Text Available Dedifferentiation of Schwann cells is an important feature of the response to peripheral nerve injury and specific negative myelination regulators are considered to have a major role in this process. However, most experiments have focused on the distal nerve stump, where the Notch signaling pathway is strongly associated with Schwann cell dedifferentiation and repair of the nerve. We observed the phenotypic changes of Schwann cells and changes of active Notch signaling on the proximal stump during peripheral nerve repair using small gap conduit tubulization. Eighty rats, with right sciatic nerve section of 4 mm, were randomly assigned to conduit bridging group and control group (epineurium suture. Glial fibrillary acidic protein expression, in myelinating Schwann cells on the proximal stump, began to up-regulate at 1 day after injury and was still evident at 5 days. Compared with the control group, Notch1 mRNA was expressed at a higher level in the conduit bridging group during the first week on the proximal stump. Hes1 mRNA levels in the conduit bridging group significantly increased compared with the control group at 3, 5, 7 and 14 days post-surgery. The change of the Notch intracellular domain shared a similar trend as Hes1 mRNA expression. Our results confirmed that phenotypic changes of Schwann cells occurred in the proximal stump. The differences in these changes between the conduit tubulization and epineurium suture groups correlate with changes in Notch signaling. This suggests that active Notch signaling might be a key mechanism during the early stage of neural regeneration in the proximal nerve stump.

  12. Intravenously delivered mesenchymal stem cell-derived exosomes target M2-type macrophages in the injured spinal cord

    OpenAIRE

    Lankford, Karen L.; Arroyo, Edgardo J.; Nazimek, Katarzyna; Bryniarski, Krzysztof; Askenase, Philip W.; Kocsis, Jeffery D.

    2018-01-01

    In a previous report we showed that intravenous infusion of bone marrow-derived mesenchymal stem cells (MSCs) improved functional recovery after contusive spinal cord injury (SCI) in the non-immunosuppressed rat, although the MSCs themselves were not detected at the spinal cord injury (SCI) site [1]. Rather, the MSCs lodged transiently in the lungs for about two days post-infusion. Preliminary studies and a recent report [2] suggest that the effects of intravenous (IV) infusion of MSCs could ...

  13. Implanted spike wave electric stimulation promotes survival of the bone marrow mesenchymal stem cells and functional recovery in the spinal cord injured rats.

    Science.gov (United States)

    Wu, Wenliang; Zhao, Hua; Xie, Bin; Liu, Haichun; Chen, Yunzhen; Jiao, Guangjun; Wang, Hongliang

    2011-03-10

    Transplantation of bone marrow-derived mesenchymal stromal cells (BMSCs) into the injured spinal cord may provide therapeutic benefit, but its application is limited by their poor survival and low differentiation rate into neurons. Electrical stimulation (ES) has been reported to promote survival and differentiation of the BMSCs. Therefore we investigated whether implanted spike wave ES could improve survival of BMSCs after transplantation and result in functional improvement in animals with spinal cord injury. Our results showed that the number and ratio of survived BMSCs near the lesion site were significantly increased in the BMSCs+ES-treated group as compared to BMSCs transplantation or ES treatment alone group. Furthermore, results from BBB scales, SSEP and DTI demonstrated a significant improved functional recovery in the BMSCs+ES group. This indicated that implanted spike wave ES could promote the bioactivity of BMSCs and their survival. This represents a new therapeutic potential of the combination of BMSCs transplantation with implanted spike wave ES to treat spinal cord injury. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  14. Functional modifications of macrophage activity after sublethal irradiation. [Toxoplasma gondii

    Energy Technology Data Exchange (ETDEWEB)

    Swartz, R.P.

    1982-01-01

    The modifications of macrophage activity following sublethal irradiation, both in vivo and in vitro, were studied using spreading and C3b-receptor-mediated ingestion assays. Nonelicited peritoneal washout cells were examined for changes in activity and selected population characteristics. The cells from irradiated mice were from a resident peritoneal population and not immigrating cells. The macrophage population showed enhanced activity early with a refractory period (24-48) when the macrophages were unresponsive to stimulation by irradiated lymphocytes. The enhanced activity was inversely dose dependent on macrophage. The lymphocytes showed a regulatory function(s) on the time post irradiation at which they were examined. Early lymphocytes exhibited the ability to enhance the activity of normal macrophages while lymphocytes removed 24 hours post irradiation could suppress the activity of already activated macrophages. The effect(s) of the various lymphocyte populations were reproduced with cell-free supernatants which was indicative of the production of lymphokines. Separation on nylon wool columns indicated that the activity resided primarily in the T-cell population of lymphocytes. In vitro irradiation indicated that stimulation of the lymphocytes is macrophage dependent. Additional work indicated that sublethally irradiated macrophages did not inhibit replication of the coccidian protozoon Toxoplasma gondii although they did show increased phagocytosis. Examination of the serum from whole body irradiated mice showed the presence of a postirradiation substance which enhanced the phagocytosis of normal macrophages. It was not present in the serum of normal mice and was not endotoxin.

  15. CXCL12α/SDF-1 from perisynaptic Schwann cells promotes regeneration of injured motor axon terminals.

    Science.gov (United States)

    Negro, Samuele; Lessi, Francesca; Duregotti, Elisa; Aretini, Paolo; La Ferla, Marco; Franceschi, Sara; Menicagli, Michele; Bergamin, Elisanna; Radice, Egle; Thelen, Marcus; Megighian, Aram; Pirazzini, Marco; Mazzanti, Chiara M; Rigoni, Michela; Montecucco, Cesare

    2017-08-01

    The neuromuscular junction has retained through evolution the capacity to regenerate after damage, but little is known on the inter-cellular signals involved in its functional recovery from trauma, autoimmune attacks, or neurotoxins. We report here that CXCL12α, also abbreviated as stromal-derived factor-1 (SDF-1), is produced specifically by perisynaptic Schwann cells following motor axon terminal degeneration induced by α-latrotoxin. CXCL12α acts via binding to the neuronal CXCR4 receptor. A CXCL12α-neutralizing antibody or a specific CXCR4 inhibitor strongly delays recovery from motor neuron degeneration in vivo Recombinant CXCL12α in vivo accelerates neurotransmission rescue upon damage and very effectively stimulates the axon growth of spinal cord motor neurons in vitro These findings indicate that the CXCL12α-CXCR4 axis plays an important role in the regeneration of the neuromuscular junction after motor axon injury. The present results have important implications in the effort to find therapeutics and protocols to improve recovery of function after different forms of motor axon terminal damage. © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

  16. Sublethal concentrations of ichthyotoxic alga Prymnesium parvum affect rainbow trout susceptibility to viral haemorrhagic septicaemia virus

    DEFF Research Database (Denmark)

    Andersen, Nikolaj Gedsted; Lorenzen, Ellen; Boutrup, Torsten Snogdal

    2016-01-01

    concentrations of the ichthyotoxic alga Prymnesium parvum affect the susceptibility of rainbow trout Oncorhynchus mykiss to viral haemorrhagic septicaemia virus (VHSV). During exposure to sublethal algal concentrations, the fish increased production of mucus on their gills. When fish were exposed to the algae......Ichthyotoxic algal blooms are normally considered a threat to maricultured fish only when blooms reach lethal cell concentrations. The degree to which sublethal algal concentrations challenge the health of the fish during blooms is practically unknown. In this study, we analysed whether sublethal...... for 12 h prior to the addition of virus, a marginal decrease in the susceptibility to VHSV was observed compared to fish exposed to VHSV without algae. If virus and algae were added simultaneously, inclusion of the algae increased mortality by 50% compared to fish exposed to virus only, depending...

  17. Induction and stability of oxidative stress adaptation in Listeria monocytogenes EGD (Bug600) and F1057 in sublethal concentrations of H2O2 and NaOH.

    Science.gov (United States)

    De Abrew Abeysundara, Piumi; Nannapaneni, Ramakrishna; Soni, Kamlesh A; Sharma, Chander S; Mahmoud, Barakat

    2016-12-05

    Food processing and food handling environments may contain residual levels of sanitizers or cleaners which may trigger oxidative stress adaptation in Listeria monocytogenes. The aim of this study was to determine the induction and stability of oxidative stress adaptation in L. monocytogenes EGD (Bug600) (serotype 1/2a) and F1057 (serotype 4b) at different concentrations and times of sublethal oxidative stress induced by H2O2 or sublethal alkali stress induced by NaOH at 37°C. Both L. monocytogenes Bug600 and F1057 strains showed significantly higher survival in lethal oxidative stress (1000ppm H2O2) after pre-exposure to 50ppm H2O2 for 30min compared to control cells (no pre-exposure to H2O2). When the cells were pre-exposed to sublethal alkali stress by NaOH, the oxidative stress adaptation was induced within 5min in L. monocytogenes. The survival of both L. monocytogenes strains was increased by 2 to 4.5 logs in lethal oxidative stress when the cells were pre-exposed to sublethal alkali stress at pH9 from 5 to 120min by NaOH compared to control cells (no pre-exposure to sublethal alkali pH). Two other alkali reagents tested (KOH and NH4OH) also induced oxidative stress adaptation in L. monocytogenes. For both L. monocytogenes strains, the oxidative stress adaptation induced by sublethal H2O2 was reversible in 30min and that induced by sublethal alkali stress was reversible within 60min at 37°C in the absence of such sublethal stress. These findings show that sublethal oxidative or alkali stress conditions can induce oxidative stress adaptation that may increase the risk of survival of L. monocytogenes cells in lethal oxidative stress. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. GM-CSF produced by non-hematopoietic cells is required for early epithelial cell proliferation and repair of injured colonic mucosa1,2

    Science.gov (United States)

    Egea, Laia; McAllister, Christopher S.; Lakhdari, Omar; Minev, Ivelina; Shenouda, Steve; Kagnoff, Martin F.

    2012-01-01

    GM-CSF is a growth factor that promotes the survival and activation of macrophages and granulocytes, and dendritic cell (DC) differentiation and survival in vitro. The mechanism by which exogenous GM-CSF ameliorates the severity of Crohn’s disease in humans and colitis in murine models has been considered mainly to reflect its activity on myeloid cells. We used GM-CSF deficient (GM-CSF−/−) mice to probe the functional role of endogenous host-produced GM-CSF in a colitis model induced after injury to the colon epithelium. Dextran sodium sulfate (DSS) at doses that resulted in little epithelial damage and mucosal ulceration in wild type (WT) mice resulted in marked colon ulceration and delayed ulcer healing in GM-CSF−/− mice. Colon crypt epithelial cell proliferation in vivo was significantly decreased in GM-CSF−/− mice at early times after DSS injury. This was paralleled by decreased expression of crypt epithelial cell genes involved in cell cycle, proliferation, and wound healing. Decreased crypt cell proliferation and delayed ulcer healing in GM-CSF−/− mice were rescued by exogenous GM-CSF, indicating the lack of a developmental abnormality in the epithelial cell proliferative response in those mice. Non-hematopoietic cells and not myeloid cells produced the GM-CSF important for colon epithelial proliferation after DSS-induced injury as revealed by bone marrow chimera and DC depletion experiments, with colon epithelial cells being the cellular source of GM-CSF. Endogenous epithelial cell produced GM-CSF has a novel non-redundant role in facilitating epithelial cell proliferation and ulcer healing in response to injury of the colon crypt epithelium. PMID:23325885

  19. Injured coliforms in drinking water.

    OpenAIRE

    McFeters, G A; Kippin, J S; LeChevallier, M W

    1986-01-01

    Coliforms were enumerated by using m-Endo agar LES and m-T7 agar in 102 routine samples of drinking water from three New England community water systems to investigate the occurrence and significance of injured coliforms. Samples included water collected immediately after conventional treatment, during the backwash cycle, at various points in the distribution system, and 1 week after the break and subsequent repair of a distribution main. Injured coliforms in these samples averaged greater th...

  20. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells

    DEFF Research Database (Denmark)

    Branco, Patrícia; Monteiro Lomba Viana, Tiago; Albergaria, Helena

    2015-01-01

    Hi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals...... on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability...... that these AMPs compromise the plasma membrane integrity (and possibly also the vacuole membrane) of H. guilliermondii cells, disturbing the pHi homeostasis and inducing a loss of culturability....

  1. Sialic acid accelerates the electrophoretic velocity of injured dorsal root ganglion neurons.

    Science.gov (United States)

    Li, Chen-Xu; Ma, Guo-Ying; Guo, Min-Fang; Liu, Ying

    2015-06-01

    Peripheral nerve injury has been shown to result in ectopic spontaneous discharges on soma and injured sites of sensory neurons, thereby inducing neuropathic pain. With the increase of membrane proteins on soma and injured site neurons, the negatively charged sialic acids bind to the external domains of membrane proteins, resulting in an increase of this charge. We therefore speculate that the electrophoretic velocity of injured neurons may be faster than non-injured neurons. The present study established rat models of neuropathic pain via chronic constriction injury. Results of the cell electrophoresis test revealed that the electrophoretic velocity of injured neuronal cells was faster than that of non-injured (control) cells. We then treated cells with divalent cations of Ca(2+) and organic compounds with positive charges, polylysine to counteract the negatively charged sialic acids, or neuraminidase to specifically remove sialic acids from the membrane surface of injured neurons. All three treatments significantly reduced the electrophoretic velocity of injured neuronal cells. These findings suggest that enhanced sialic acids on injured neurons may accelerate the electrophoretic velocity of injured neurons.

  2. Sialic acid accelerates the electrophoretic velocity of injured dorsal root ganglion neurons

    Directory of Open Access Journals (Sweden)

    Chen-xu Li

    2015-01-01

    Full Text Available Peripheral nerve injury has been shown to result in ectopic spontaneous discharges on soma and injured sites of sensory neurons, thereby inducing neuropathic pain. With the increase of membrane proteins on soma and injured site neurons, the negatively charged sialic acids bind to the external domains of membrane proteins, resulting in an increase of this charge. We therefore speculate that the electrophoretic velocity of injured neurons may be faster than non-injured neurons. The present study established rat models of neuropathic pain via chronic constriction injury. Results of the cell electrophoresis test revealed that the electrophoretic velocity of injured neuronal cells was faster than that of non-injured (control cells. We then treated cells with divalent cations of Ca 2+ and organic compounds with positive charges, polylysine to counteract the negatively charged sialic acids, or neuraminidase to specifically remove sialic acids from the membrane surface of injured neurons. All three treatments significantly reduced the electrophoretic velocity of injured neuronal cells. These findings suggest that enhanced sialic acids on injured neurons may accelerate the electrophoretic velocity of injured neurons.

  3. Human Wharton’s jelly-derived mesenchymal stromal cells reduce renal fibrosis through induction of native and foreign hepatocyte growth factor synthesis in injured tubular epithelial cells

    Science.gov (United States)

    2013-01-01

    Introduction Based on some well-documented reports, we attempted to clarify the antifibrotic mechanisms of human Wharton’s-jelly-derived mesenchymal stromal cells (WJ-MSCs) from the perspective of induction of hepatocyte growth factor (HGF) expression in tubular epithelial cells (TECs). Methods A rat model of acute kidney injury (AKI) was established through unilateral renal ischemia for 1 hour. Two days later, a single intravenous cell or vehicle injection, or contralateral nephrectomy, was performed. Rats were sacrificed at 1 day, 1 week, 4 weeks, or 6 weeks after the intervention. Renal fibrosis was evaluated by Masson trichrome staining and Sircol collagen assay. The upregulation of α-smooth muscle actin (α-SMA) versus E-cadherin expression was adopted as an indicator of tubular epithelial-mesenchymal transition (EMT). Gene and protein expression of HGF or transforming growth factor-beta1 (TGF-β1) was determined by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. HGF expression in TECs was detected with immunostaining. In vitro, rat TECs subjected to hypoxia injury were incubated with or without conditioned medium (CM) from WJ-MSCs for 1, 3, 24, or 48 hours. Rat or human HGF synthesis in TECs was assessed with immunostaining, RT-PCR, or ELISA. Results Cell delivery or nephrectomy led to abrogation of renal scarring. At the incipient period of AKI, through induction of HGF expression, either of them remarkably promoted the upregulation of HGF versus TGF-β1 expression in damaged kidney. Rat TECs were not only the principal cells expressing HGF but also exhibited human HGF expression after cell infusion. During fibrogenesis, the downregulation of HGF versus TGF-β1 expression was greatly prevented by WJ-MSCs or kidney removal, thereby resulting in tubular EMT delay. In vitro, after 24 or 48 hours of incubation, CM not only robustly induced the upregulation of rat HGF gene expression in TECs but substantially amplified the release

  4. Effects of anti-inflammatory compounds on sulfur mustard injured cells: Recommendations and caveats suggested by in vitro cell culture models.

    Science.gov (United States)

    Menacher, Georg; Steinritz, Dirk; Schmidt, Annette; Popp, Tanja; Worek, Franz; Gudermann, Thomas; Thiermann, Horst; Balszuweit, Frank

    2017-09-12

    Sulfur mustard (SM) is a vesicant agent who had its first military use 100 years ago, in Ypres. Since then it has been used in several conflicts like the Iran-Iraq war in the 1980s. The use of SM in Syria 2015 indicated the still existing threat. Despite decades of research no causal antidote against SM intoxication is available, so far. A SM intoxication is accompanied by necrosis, apoptosis and inflammation. To counteract the SM-induced inflammation, glucocorticoids and non-steroidal anti-inflammatory compounds (NSAIDs) are recommended. Aim of this study was to evaluate the efficacy of the anti-inflammatory compounds dexamethasone, ibuprofen and diclofenac in vitro. For that purpose, two different cell culture models were used. Firstly, a monoculture of keratinocytes (HaCaT) and secondly, an established co-culture of keratinocytes (HaCaT) and immunocompetent cells (THP-1) to identify the role of immune cells in the process and to mimic the dermal physiology more closely. Both models were challenged with different SM concentrations (100, 200 and 300μM) and treated with different anti-inflammatory compounds one hour after the SM exposure. Analytical analysis of necrosis (ToxiLight), apoptosis (CDDE) and inflammation (IL-6 and -8 ELISAs) followed 24h thereafter. Dexamethasone provided small but consistent protective effects in the monoculture. For the reduction of apoptosis, 3μM dexamethasone was sufficient. The most effective reduction regarding interleukin (IL) production was found with 6μM dexamethasone. Protective effects were less pronounced in co-culture, which implies, that the protective effects of dexamethasone are rather generic and not due to a modulation of the immune cells. Against our expectations, ibuprofen strongly amplified apoptosis and necrosis in SM exposed cells in the monoculture as well as the co-culture. Therefore, use of ibuprofen for treatment of SM intoxication should at least be considered most critically, if not even regarded as

  5. Sublethal effects of manganese on the carbohydrate metabolism of ...

    African Journals Online (AJOL)

    Carbohydrate metabolism variables of Oreochromis mossambicuswere investigated after acute and chronic sublethal manganese exposure. The sublethal concentrations were determined from the LC50 value of manganese. After the exposures, the fish were carefully netted and blood was drawn from the caudal aorta.

  6. Rescue of Injured Myocytes

    Science.gov (United States)

    1990-11-30

    of Naval Research 6c. ADDRESS (City, State, and ZIP Code) 7b. ADDRESS (City, State, and ZIP Code)Department of Cell Biology & Anatomy 800 N. Quincy...correlated linearly with LDH release. Employing this technique , two reputed protective agents, U74006F and calciphor, were evaluated during ’chemical

  7. Effect of non-homogenous thermal stress during sub-lethal photodynamic antimicrobial chemotherapy

    Science.gov (United States)

    Gadura, N.; Kokkinos, D.; Dehipawala, S.; Cheung, E.; Sullivan, R.; Subramaniam, R.; Schneider, P.; Tremberger, G., Jr.; Holden, T.; Lieberman, D.; Cheung, T.

    2012-03-01

    Pathogens could be inactivated via a light source coupled with a photosensitizing agent in photodynamic antimicrobial chemotherapy (PACT). This project studied the effect of non-homogenous substrate on cell colony. The non-homogeneity could be controlled by iron oxide nano-particles doping in porous glassy substrates such that each cell would experience tens of hot spots when illuminated with additional light source. The substrate non-homogeneity was characterized by Atomic Force Microscopy, Transmission Electron Microscopy and Extended X-Ray Absorption Fine Structure at Brookhaven Synchrotron Light Source. Microscopy images of cell motion were used to study the motility. Laboratory cell colonies on non-homogenous substrates exhibit reduced motility similar to those observed with sub-lethal PCAT treatment. Such motility reduction on non-homogenous substrate is interpreted as the presence of thermal stress. The studied pathogens included E. coli and Pseudomonas aeruginosa. Non-pathogenic microbes Bacillus subtilis was also studied for comparison. The results show that sub-lethal PACT could be effective with additional non-homogenous thermal stress. The use of non-uniform illumination on a homogeneous substrate to create thermal stress in sub-micron length scale is discussed via light correlation in propagation through random medium. Extension to sub-lethal PACT application complemented with thermal stress would be an appropriate application.

  8. Genetic regulation of allolysis in response to sub-lethal antibiotic stress in Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    MANISHA DASH

    2014-11-01

    Full Text Available Dash M, Dash HR, Das S. 2014. Genetic regulation of allolysis in response to sub-lethal antibiotic stress in Streptococcus pneumoniae. Nusantara Bioscience 6: 111-117. Allolysis is the phenomenon of cell lysis induced by other cells of the same species. Gram-positive bacterium Streptococcus pneumoniae, a major human pathogen exhibits competence induced allolysis that increases the genetic recombination and enhances the virulence. During allolysis, a group of non-competent bacterial cells are lysed by another group of competent cells in the same culture. This process is regulated by com operon as well as bacteriocin. In this study, allolysis was induced in Streptococcus pneumoniae MTCC655 by sub-lethal dose of antibiotic (chloramphenicol and the mechanism of allolysis has been deduced by amplification of lytA, lytC and cbpD genes in the bacterium. The strain was found to be resistant to a number of antibiotics including amoxicillin, cefpodoxime, erythromycin and vancomycin. The early onset of allolysis induction from 7-9 h under normal conditions to 2-3 h by sub-lethal dose of chloramphenicol was observed.

  9. Sublethal consequences of urban life for wild vertebrates

    National Research Council Canada - National Science Library

    Gallagher, Austin J; Peiman, Kathryn S; de Bruijn, Robert; Cooke, Steven J; Birnie-Gauvin, Kim

    2016-01-01

    ... — while others have not. Here we present a review of the sublethal consequences of life in the city for wild vertebrates, and demonstrate that urban animals face an almost completely different set of physiological...

  10. Busulfan administration produces sublethal effects on somatic tissues and inhibits gametogenesis in Senegalese sole juveniles.

    Science.gov (United States)

    Pacchiarini, T; Olague, E; Sarasquete, C; Cabrita, E

    2014-05-01

    Busulfan, a cytotoxic alkylating agent used for treatment of chronic myeloid leukemia has effects in mammalian germ cells. In fish species, the use of this compound is of special interest in intra and interspecies germ cell transplants. To determine the effects of busulfan in fish a previous range finding experiment was designed. Survival and growth rate of 150-days after hatching (150DAH) Senegalese sole (Solea senegalensis) juveniles was determined. In a second experiment, the effects of a sublethal busulfan dose in fish germ cell depletion and in somatic tissues were analysed. Sublethal effects of several busulfan treatments (B10-10 days after injection, B20-20 days after injection, B20÷-20 days after injection with double injection) were determined in somatic and gonadal tissues. Alterations were registered through histopathological techniques, TUNEL (cell apoptosis) and quantified at molecular level (Q-PCR analyses) using the vasa mRNAs (Ssvasa1-2 and Ssvasa3-4 mRNAs) as molecular markers for germinal cells in Senegalese sole juveniles. Several sublethal effects were observed with 40 mg kg⁻¹ busulfan, a non-lethal dose, such as: pyknosis in liver, increase of melanomacrophage centres and blood stagnation in spleen and interruption of gonadal development. Females were more affected by busulfan treatments than males in terms of germ cell disruption, since a significant decrease in the expression of both Ssvasa1-2 and Ssvasa3-4 markers was found in the gonad of treated females rather than males. At 10 days post-treatment (B10), females already presented a decrease in germ cell proliferation, as confirmed by Q-PCR. Ssvasa expression proved to be a reliable tool for the direct evaluation of the effects of busulfan on Senegalese sole gonadal development, proving that busulfan can be a suitable treatment for causing transient sterility in recipient gonads for germ cell transplantation.

  11. Promoting the Recovery of Injured Liver with Poly (3-Hydroxybutyrate-Co-3-Hydroxyvalerate-Co-3-Hydroxyhexanoate) Scaffolds Loaded with Umbilical Cord-Derived Mesenchymal Stem Cells

    OpenAIRE

    Li, Pengshan; Zhang, Jin; Liu, Jing; Ma, Huan; Liu, Jie; Lie, Puchang; Wang, Yuechun; Liu, Gexiu; Zeng, Huilan; Li, Zhizhong; Wei, Xing

    2014-01-01

    Cell-based therapies are major focus of current research for treatment of liver diseases. In this study, mesenchymal stem cells were isolated from human umbilical cord Wharton's jelly (WJ-MSCs). Results confirmed that WJ-MSCs isolated in this study could express the typical MSC-specific markers and be induced to differentiate into adipocytes, osteoblasts, and chondrocytes. They could also be induced to differentiate into hepatocyte-like cells. Poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3...

  12. Sublethal RNA Oxidation as a Mechanism for Neurodegenerative Disease

    Directory of Open Access Journals (Sweden)

    Mark A. Smith

    2008-05-01

    Full Text Available Although cellular RNA is subjected to the same oxidative insults as DNA and other cellular macromolecules, oxidative damage to RNA has not been a major focus in investigations of the biological consequences of free radical damage. In fact, because it is largely single-stranded and its bases lack the protection of hydrogen bonding and binding by specific proteins, RNA may be more susceptible to oxidative insults than is DNA. Oxidative damage to protein-coding RNA or non-coding RNA will, in turn, potentially cause errors in proteins and/or dysregulation of gene expression. While less lethal than mutations in the genome, such sublethal insults to cells might be associated with underlying mechanisms of several chronic diseases, including neurodegenerative disease. Recently, oxidative RNA damage has been described in several neurodegenerative diseases including Alzheimer disease, Parkinson disease, dementia with Lewy bodies, and prion diseases. Of particular interest, oxidative RNA damage can be demonstrated in vulnerable neurons early in disease, suggesting that RNA oxidation may actively contribute to the onset of the disease. An increasing body of evidence suggests that, mechanistically speaking, the detrimental effects of oxidative RNA damage to protein synthesis are attenuated, at least in part, by the existence of protective mechanisms that prevent the incorporation of the damaged ribonucleotides into the translational machinery. Further investigations aimed at understanding the processing mechanisms related to oxidative RNA damage and its consequences may provide significant insights into the pathogenesis of neurodegenerative and other degenerative diseases and lead to better therapeutic strategies.

  13. [CHANGES IN THE NUMBER OF REGENERATING MYELINATED FIBERS IN INJURED NERVE OF THE RAT AFTER ALLOTRANSPLANTATION OF THE DISSOCIATED CELLS OF THE EMBRYONIC CNS ANLAGES].

    Science.gov (United States)

    Petrova, Ye S; Isayeva, Ye N

    2015-01-01

    The study was conducted on 6 female and 36 male adult Wistar rats to compare the effects of dissociated cells derived from different embryonic CNS anlages, on the growth of regenerating nerve fibers in the damaged nerve of the recipient. After the sciatic nerve was damaged by ligation, part of the animals received the injection into the proximal portion of the nerve with a suspension of the cells obtained by dissociation of the fragments of spinal cord or forebrain vesicle taken from rat embryos at Day 15 of development. The analysis of transverse semithin sections of the distal part of the nerves was performed 21 and 60 days after surgery. It was found that the number of regenerating myelinated nerve fibers was increased 60 days after the injection of dissociated embryonic spinal cord cells, but not the neocortical cells into the damaged nerve of the recipient.

  14. Transplantation of Allogeneic PW1pos/Pax7neg Interstitial Cells Enhance Endogenous Repair of Injured Porcine Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    Fiona C. Lewis, BSc, PhD

    2017-12-01

    Full Text Available Skeletal muscle-derived PW1pos/Pax7neg interstitial cells (PICs express and secrete a multitude of proregenerative growth factors and cytokines. Utilizing a porcine preclinical skeletal muscle injury model, delivery of allogeneic porcine PICs (pPICs significantly improved and accelerated myofiber regeneration and neocapillarization, compared with saline vehicle control-treated muscles. Allogeneic pPICs did not contribute to new myofibers or capillaries and were eliminated by the host immune system. In conclusion, allogeneic pPIC transplantation stimulated the endogenous stem cell pool to bring about enhanced autologous skeletal muscle repair and regeneration. This allogeneic cell approach is considered a cost-effective, easy to apply, and readily available regenerative therapeutic strategy.

  15. Boundary cap neural crest stem cells homotopically implanted to the injured dorsal root transitional zone give rise to different types of neurons and glia in adult rodents

    OpenAIRE

    Trolle, Carl; Abrahamsson, Ninnie; König, Niclas; Vasylovska, Svitlana; Kozlova, Elena

    2014-01-01

    The boundary cap is a transient group of neural crest-derived cells located at the presumptive dorsal root transitional zone (DRTZ) when sensory axons enter the spinal cord during development. Later, these cells migrate to dorsal root ganglia and differentiate into subtypes of sensory neurons and glia. After birth when the DRTZ is established, sensory axons are no longer able to enter the spinal cord. Here we explored the fate of mouse bNCSCs implanted to the uninjured DRTZ after dorsal root ...

  16. Acute Putrescine Supplementation with Schwann Cell Implantation Improves Sensory and Serotonergic Axon Growth and Functional Recovery in Spinal Cord Injured Rats.

    Science.gov (United States)

    Iorgulescu, J Bryan; Patel, Samik P; Louro, Jack; Andrade, Christian M; Sanchez, Andre R; Pearse, Damien D

    2015-01-01

    Schwann cell (SC) transplantation exhibits significant potential for spinal cord injury (SCI) repair and its use as a therapeutic modality has now progressed to clinical trials for subacute and chronic human SCI. Although SC implants provide a receptive environment for axonal regrowth and support functional recovery in a number of experimental SCI models, axonal regeneration is largely limited to local systems and the behavioral improvements are modest without additional combinatory approaches. In the current study we investigated whether the concurrent delivery of the polyamine putrescine, started either 30 min or 1 week after SCI, could enhance the efficacy of SCs when implanted subacutely (1 week after injury) into the contused rat spinal cord. Polyamines are ubiquitous organic cations that play an important role in the regulation of the cell cycle, cell division, cytoskeletal organization, and cell differentiation. We show that the combination of putrescine with SCs provides a significant increase in implant size, an enhancement in axonal (sensory and serotonergic) sparing and/or growth, and improved open field locomotion after SCI, as compared to SC implantation alone. These findings demonstrate that polyamine supplementation can augment the effectiveness of SCs when used as a therapeutic approach for subacute SCI repair.

  17. Endotoxin-activated microglia injure brain derived endothelial cells via NF-κB, JAK-STAT and JNK stress kinase pathways

    Directory of Open Access Journals (Sweden)

    Yenari Midori A

    2011-03-01

    Full Text Available Abstract Background We previously showed that microglia damage blood brain barrier (BBB components following ischemic brain insults, but the underlying mechanism(s is/are not well known. Recent work has established the contribution of toll-like receptor 4 (TLR4 activation to several brain pathologies including ischemia, neurodegeneration and sepsis. The present study established the requirement of microglia for lipopolysaccharide (LPS mediated endothelial cell death, and explored pathways involved in this toxicity. LPS is a classic TLR4 agonist, and is used here to model aspects of brain conditions where TLR4 stimulation occurs. Methods/Results In monocultures, LPS induced death in microglia, but not brain derived endothelial cells (EC. However, LPS increased EC death when cocultured with microglia. LPS led to nitric oxide (NO and inducible NO synthase (iNOS induction in microglia, but not in EC. Inhibiting microglial activation by blocking iNOS and other generators of NO or blocking reactive oxygen species (ROS also prevented injury in these cocultures. To assess the signaling pathway(s involved, inhibitors of several downstream TLR-4 activated pathways were studied. Inhibitors of NF-κB, JAK-STAT and JNK/SAPK decreased microglial activation and prevented cell death, although the effect of blocking JNK/SAPK was rather modest. Inhibitors of PI3K, ERK, and p38 MAPK had no effect. Conclusions We show that LPS-activated microglia promote BBB disruption through injury to endothelial cells, and the specific blockade of JAK-STAT, NF-κB may prove to be especially useful anti-inflammatory strategies to confer cerebrovascular protection.

  18. Endotoxin-activated microglia injure brain derived endothelial cells via NF-κB, JAK-STAT and JNK stress kinase pathways.

    Science.gov (United States)

    Kacimi, Rachid; Giffard, Rona G; Yenari, Midori A

    2011-03-07

    We previously showed that microglia damage blood brain barrier (BBB) components following ischemic brain insults, but the underlying mechanism(s) is/are not well known. Recent work has established the contribution of toll-like receptor 4 (TLR4) activation to several brain pathologies including ischemia, neurodegeneration and sepsis. The present study established the requirement of microglia for lipopolysaccharide (LPS) mediated endothelial cell death, and explored pathways involved in this toxicity. LPS is a classic TLR4 agonist, and is used here to model aspects of brain conditions where TLR4 stimulation occurs. In monocultures, LPS induced death in microglia, but not brain derived endothelial cells (EC). However, LPS increased EC death when cocultured with microglia. LPS led to nitric oxide (NO) and inducible NO synthase (iNOS) induction in microglia, but not in EC. Inhibiting microglial activation by blocking iNOS and other generators of NO or blocking reactive oxygen species (ROS) also prevented injury in these cocultures. To assess the signaling pathway(s) involved, inhibitors of several downstream TLR-4 activated pathways were studied. Inhibitors of NF-κB, JAK-STAT and JNK/SAPK decreased microglial activation and prevented cell death, although the effect of blocking JNK/SAPK was rather modest. Inhibitors of PI3K, ERK, and p38 MAPK had no effect. We show that LPS-activated microglia promote BBB disruption through injury to endothelial cells, and the specific blockade of JAK-STAT, NF-κB may prove to be especially useful anti-inflammatory strategies to confer cerebrovascular protection.

  19. Stem Cell Mesenchymal Injection Increases Platelet-Derived Growth Factors Level and Percentage of Collagen in Third-Degree Burn injured Mice

    OpenAIRE

    Erna Ridawati; Taufiqurrachman Nasihun; Wiratno Wiratno

    2017-01-01

    Introduction: Mesenchymal stem cell (MSC) has been demonstrated to accelerate wound healing. Platelet-derived growth factor (PDGF) and collagen have an important role in wound healing. However, the effect of MSC on levels of PDGF and number of collagen has not been established. Objectives: To determine the effect of MSC on the levels of PDGF and the percentage of collagen in the burns healing. Methods: in this experimental study, 20 BALB /c mice were divided into 4 groups: a control group...

  20. Acellular spinal cord scaffold seeded with mesenchymal stem cells promotes long-distance axon regeneration and functional recovery in spinal cord injured rats.

    Science.gov (United States)

    Liu, Jia; Chen, Jian; Liu, Bin; Yang, Cuilan; Xie, Denghui; Zheng, Xiaochen; Xu, Song; Chen, Tianyu; Wang, Liang; Zhang, Zhongmin; Bai, Xiaochun; Jin, Dadi

    2013-02-15

    The stem cell-based experimental therapies are partially successful for the recovery of spinal cord injury (SCI). Recently, acellular spinal cord (ASC) scaffolds which mimic native extracellular matrix (ECM) have been successfully prepared. This study aimed at investigating whether the spinal cord lesion gap could be bridged by implantation of bionic-designed ASC scaffold alone and seeded with human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) respectively, and their effects on functional improvement. A laterally hemisected SCI lesion was performed in adult Sprague-Dawley (SD) rats (n=36) and ASC scaffolds seeded with or without hUCB-MSCs were implanted into the lesion immediately. All rats were behaviorally tested using the Basso-Beattie-Bresnahan (BBB) test once a week for 8weeks. Behavioral analysis showed that there was significant locomotor recovery improvement in combined treatment group (ASC scaffold and ASC scaffold+hUCB-MSCs) as compared with the SCI only group (pregeneration at lesion sites. This study provides evidence for the first time that ASC scaffold seeded with hUCB-MSCs is able to bridge a spinal cord cavity and promote long-distance axon regeneration and functional recovery in SCI rats. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Sublethal effects of manganese on the haematology and ...

    African Journals Online (AJOL)

    Information concerning the sublethal effects of pollutants, such as metals, forms an integral part of ecosystem health assessment programmes and of procedures followed to develop water quality guidelines for environmetal protection. The data from this study were incorporated into a water quality index (RAUWaterz) ...

  2. Sublethal haematological effects of zinc on the freshwater fish ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... industrial and domestic wastes water discharges and animals where it ... that zinc could cause sub-acute effects that change fish behaviours. ... These include the sublethal effects of concentrations of water extracts of akee apple on C. gariepinus (Onusiriuka and Ufodike, 1998). Toxicity of cas- sava leaf ...

  3. Effects of sublethal doses of chlorfluazuron on the ovarian ...

    African Journals Online (AJOL)

    Therefore, it is concluded that sublethal doses of chlorfluazuron reduced the amounts of ovarian constituents during ovarian development and oogenesis in S. litura. These reductions increased with an increase in dose from LD10 to LD30. The effects of chlorfluazuron on the amounts of ovarian constituents are presumed to ...

  4. Histopathological effects of lethal and sub-lethal concentrations of ...

    African Journals Online (AJOL)

    The histopathological effects of lethal and sub-lethal concentrations of glyphosate on African catfish Clarias gariepinus were investigated. C. gariepinus juveniles were assessed in a static renewal bioassay for 96 hours (acute toxicity) and 28 days (chronic toxicity) using varying concentrations (0.0 mg/l 20.0 mg/l, 30.0 mg/l, ...

  5. Review: Sublethal effects of temperature on freshwater organisms ...

    African Journals Online (AJOL)

    Review: Sublethal effects of temperature on freshwater organisms, with special reference to aquatic insects. HF Dallas, V Ross-Gillespie. Abstract. Water temperature is a key variable affecting aquatic organisms. Understanding their response to elevated water temperatures is important for estimating upper thermal limits, ...

  6. Impact of sublethal concentration of triazophos on regulation of ...

    African Journals Online (AJOL)

    Exposure to sublethal doses of triazophos extract caused significant (p < 0.05) time and dose dependent reduction in the levels of total protein, acetylcholinesterase (AchE) and significant enhancement in the levels of total free amino acids, glutamine, adenosine monophosphate (AMP) deaminases, adenosine deaminases, ...

  7. Sublethal effects of industrial chemicals on fish fingerlings ( Tilapia ...

    African Journals Online (AJOL)

    Tilapia guineensis commonly found in the Niger Delta ecological zone of Nigeria was exposed to sublethal concentrations (1.56, 3.13 mg/l) of neatex (industrial detergent) and norust CR 486 (corrosion inhibitor) using the Organisation for Economic Cooperation and Development (OECD) # 203 protocol. At test termination ...

  8. Sublethal effects of carbaryl on embryonic and gonadal ...

    African Journals Online (AJOL)

    Sex reversal was delayed in the experimental groups, with a sex ratio of 13 females to 0 males, but the control group recorded 6 females to 8 males. These results suggest that sublethal doses of carbaryl in the environment, similar to those used in the current study, may have an adverse effect on the reproductive success of ...

  9. Effects Of Exposure To Sublethal Concentrations Of Azadirachta ...

    African Journals Online (AJOL)

    The physiological impairment on the fingerlings of Clarias gariepinus when exposed to sublethal concentrations of Azadirachta Indica was investigated. The fish were exposed to concentrations of 1.25, 2.50, 5.0, 10.0, 20.0 ML -1 for the period of 12 weeks. The crude protein content decreased with increased concentration ...

  10. Stem Cell Mesenchymal Injection Increases Platelet-Derived Growth Factors Level and Percentage of Collagen in Third-Degree Burn injured Mice

    Directory of Open Access Journals (Sweden)

    Erna Ridawati

    2017-03-01

    Full Text Available Introduction: Mesenchymal stem cell (MSC has been demonstrated to accelerate wound healing. Platelet-derived growth factor (PDGF and collagen have an important role in wound healing. However, the effect of MSC on levels of PDGF and number of collagen has not been established. Objectives: To determine the effect of MSC on the levels of PDGF and the percentage of collagen in the burns healing. Methods: in this experimental study, 20 BALB /c mice were divided into 4 groups: a control group (G-0 injected with 2 ml of distilled water and group 1 (G-1, Group 2 (G-2, and group 3 (G- 3 injected with MSC at the dose of 1 x 104, 2 x 104, and 4 x 104 respectively. The burn wound was made by attaching a metal nail that had been heated for 20 seconds at the feet of mice. Levels of PDGF on day 2 (PDGF-2 and 7 (PDGF-7 was assessed by ELISA, while on day 10 excision of skin was done to calculate the percentage of collagen. Results: There was a significant difference in the levels of PDGF-2, PDGF-7, and the percentage of collagen ( P <0.05. Posthoc analysis showed that the levels of PDGF-2, PDGF-7, and percentage of collagen in the G-2 and G-3 was significantly higher than that of G-0, p <0.05. While the levels of PDGF-2, PDGF-7, and the percentage of collagen in the G-3 was significantly higher than that of G-2 and G-1 (p <0.05. Conclusion: Injection of MSC at a dose of 2 x 1044 and 4 x 1044 increases the levels of PDGF and the percentage of collagen.

  11. Discrimination of intact and injured Listeria monocytogenes by Fourier transform infrared spectroscopy and principal component analysis.

    Science.gov (United States)

    Lin, Mengshi; Al-Holy, Murad; Al-Qadiri, Hamzah; Kang, Dong-Hyun; Cavinato, Anna G; Huang, Yiqun; Rasco, Barbara A

    2004-09-22

    Fourier transform infrared spectroscopy (FT-IR, 4000-600 cm(-)(1)) was used to discriminate between intact and sonication-injured Listeria monocytogenes ATCC 19114 and to distinguish this strain from other selected Listeria strains (L. innocua ATCC 51742, L. innocua ATCC 33090, and L. monocytogenes ATCC 7644). FT-IR vibrational overtone and combination bands from mid-IR active components of intact and injured bacterial cells produced distinctive "fingerprints" at wavenumbers between 1500 and 800 cm(-)(1). Spectral data were analyzed by principal component analysis. Clear segregations of different intact and injured strains of Listeria were observed, suggesting that FT-IR can detect biochemical differences between intact and injured bacterial cells. This technique may provide a tool for the rapid assessment of cell viability and thereby the control of foodborne pathogens.

  12. Can PRP effectively treat injured tendons?

    Science.gov (United States)

    Wang, James H-C

    2014-01-01

    PRP is widely used to treat tendon and other tissue injuries in orthopaedics and sports medicine; however, the efficacy of PRP treatment on injured tendons is highly controversial. In this commentary, I reason that there are many PRP- and patient-related factors that influence the outcomes of PRP treatment on injured tendons. Therefore, more basic science studies are needed to understand the mechanism of PRP on injured tendons. Finally, I suggest that better understanding of the PRP action mechanism will lead to better use of PRP for the effective treatment of tendon injuries in clinics.

  13. Transplantation of Human Dental Pulp-Derived Stem Cells or Differentiated Neuronal Cells from Human Dental Pulp-Derived Stem Cells Identically Enhances Regeneration of the Injured Peripheral Nerve.

    Science.gov (United States)

    Ullah, Imran; Park, Ju-Mi; Kang, Young-Hoon; Byun, June-Ho; Kim, Dae-Geon; Kim, Joo-Heon; Kang, Dong-Ho; Rho, Gyu-Jin; Park, Bong-Wook

    2017-09-01

    Human dental mesenchymal stem cells isolated from the dental follicle, pulp, and root apical papilla of extracted wisdom teeth have been known to exhibit successful and potent neurogenic differentiation capacity. In particular, human dental pulp-derived stem cells (hDPSCs) stand out as the most prominent source for in vitro neuronal differentiation. In this study, to evaluate the in vivo peripheral nerve regeneration potential of hDPSCs and differentiated neuronal cells from DPSCs (DF-DPSCs), a total of 1 × 106 hDPSCs or DF-hDPSCs labeled with PKH26 tracking dye and supplemented with fibrin glue scaffold and collagen tubulization were transplanted into the sciatic nerve resection (5-mm gap) of rat models. At 12 weeks after cell transplantation, both hDPSC and DF-hDPSC groups showed notably increased behavioral activities and higher muscle contraction forces compared with those in the non-cell transplanted control group. In immunohistochemical analysis of regenerated nerve specimens, specific markers for angiogenesis, axonal fiber, and myelin sheath increased in both the cell transplantation groups. Pretransplanted labeled PKH26 were also distinctly detected in the regenerated nerve tissues, indicating that transplanted cells were well-preserved and differentiated into nerve cells. Furthermore, no difference was observed in the nerve regeneration potential between the hDPSC and DF-hDPSC transplanted groups. These results demonstrate that dental pulp tissue is an excellent stem cell source for nerve regeneration, and in vivo transplantation of the undifferentiated hDPSCs could exhibit sufficient and excellent peripheral nerve regeneration potential.

  14. Pesticides and Arthropods: Sublethal Effects and Demographic Toxicology

    Directory of Open Access Journals (Sweden)

    Dejan Marčić

    2007-01-01

    Full Text Available Insecticides and acaricides designed to control primary harmful insects and mites may also variously affect some other arthopods present in an (agroecosystem (e.g. secondary pests, predators, parasitoids, saprophytes, bioindicators, pollinators. Apart from insecticides and acaricides, arthropods may also be affected by the activity of other pesticides (fungicides, herbicides, etc.. Regardless of whether they are deemed desirable or not, the effects that pesticides have on arthopods need to be quantified as closely as possible through appropriate experimental procedures. Data acquired in tests designed to determined LD50/LC50 values are inadequate for evaluation of pesticide effectiveness in the field as pesticidesalso cause various sublethal effects, generally disregarded in such investigations. The sublethal effects of pesticides refer to any altered behaviour and/or physiology of individuals that have survived exposure to pesticides at doses/concentrations that can be lethal(within range causing mortality in an experimental population that exceeds mortality in an untreated population or sublethal (below that range. Pesticides affect locomotion and mobility, stimulate dispersion of arthropods from treated areas, complicate or prevent their navigation, orientation and ability to locate hosts, and cause changes in their feeding, mating and egg-laying patterns. Sublethal pesticide effects on arthropod physiology reflect on the life span, rate of development, fecundity and/or fertility, sex ratio and immunity of surviving individuals. Different parameters are being used in arthropod bioassays to determine sublethal effects (ED50/EC50, LOEC, NOEC, total effect index. Compared to acute toxicity tests, these parameters improve the quality of evaluation and create a more accurate view of the effects of a pesticide. However, such approach covers mainly fecundity/fertility alone, while all other sublethal effects remain unaccounted for. Besides, it

  15. Mitochondria Localize to Injured Axons to Support Regeneration.

    Science.gov (United States)

    Han, Sung Min; Baig, Huma S; Hammarlund, Marc

    2016-12-21

    Axon regeneration is essential to restore the nervous system after axon injury. However, the neuronal cell biology that underlies axon regeneration is incompletely understood. Here we use in vivo, single-neuron analysis to investigate the relationship between nerve injury, mitochondrial localization, and axon regeneration. Mitochondria translocate into injured axons so that average mitochondria density increases after injury. Moreover, single-neuron analysis reveals that axons that fail to increase mitochondria have poor regeneration. Experimental alterations to axonal mitochondrial distribution or mitochondrial respiratory chain function result in corresponding changes to regeneration outcomes. Axonal mitochondria are specifically required for growth-cone migration, identifying a key energy challenge for injured neurons. Finally, mitochondrial localization to the axon after injury is regulated in part by dual-leucine zipper kinase 1 (DLK-1), a conserved regulator of axon regeneration. These data identify regulation of axonal mitochondria as a new cell-biological mechanism that helps determine the regenerative response of injured neurons. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Kinetic analysis of the swimming behavior of the goldfish, Carassius auratus, exposed to nickel: Hypoactivity induced by sublethal concentrations

    Energy Technology Data Exchange (ETDEWEB)

    Ellgaard, E.G.; Ashley, S.E.; Langford, A.E. [Tulane Univ., New Orleans, LA (United States)] [and others

    1995-12-01

    The discharge of nickel into aquatic environments from numerous industries poses a threat to fish populations because of its toxcity. Although little is known, however, about the precise mechanism of its toxicity in freshwater fish, it produces some of the symptoms associated with heavy-metal poisoning in general; it accumulates in fish tissues and results in alterations in gill structure, including hypertrophy of respiratory and mucous cells, separation of the epithelial layer from the pillar cell system, cauterization and sloughing, and necrosis of the epithelium. The destruction of the gill lamellae decreases the ventilation rate and if severe, as after acute exposure, may cause blood hypoxia and death. The effects of short-term exposure of fish to sublethal concentrations of nickel and not as well defined. The kinetic method of Ellgaard et al., which uses locomotor activity to assess the general health of fish, is ideally suited to examine whether sublethal concentrations of nickel adversely affect fish. In previous studies, the measured changes in locomotor activity observed when fish are exposed to pollutants correlate with more specific changes, e.g., physiological, biochemical, histological or neurosensory changes, which occur under the same conditions. Thus, the kinetic method also meets the criterial for pollution early warning systems as discussed by Cairns and van der Schale. This method has previously been used to demonstrate that short-term exposure to sublethal concentrations of the heavy metals cadmium, chromium, and zinc and copper are detrimental to the health of bluegills. The present study examines the effects of short-term exposures of sublethal concentrations of nickel on the locomotor activity of the goldfish, Carassius auratus. 11 refs., 1 fig., 2 tabs.

  17. Sublethal Heavy Metal Stress Stimulates Innate Immunity in Tomato

    Directory of Open Access Journals (Sweden)

    Nilanjan Chakraborty

    2015-01-01

    Full Text Available Effect of sublethal heavy metal stress as plant biotic elicitor for triggering innate immunity in tomato plant was investigated. Copper in in vivo condition induced accumulation of defense enzymes like peroxidase (PO, polyphenol oxidase (PPO, phenylalanine ammonia-lyase (PAL, and β-1,3 glucanase along with higher accumulation of total phenol, antioxidative enzymes (catalase and ascorbate peroxidase, and total chlorophyll content. Furthermore, the treatment also induced nitric oxide (NO production which was confirmed by realtime visualization of NO burst using a fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2DA and spectrophotometric analysis. The result suggested that the sublethal dose of heavy metal can induce an array of plant defense responses that lead to the improvement of innate immunity in plants.

  18. Sublethal dietary effects of Microcystis on Sacramento splittail, Pogonichthys macrolepidotus.

    Science.gov (United States)

    Acuña, Shawn; Deng, Dong-Fang; Lehman, Peggy; Teh, Swee

    2012-04-01

    The presence of the toxic cyanobacterium Microcystis in the upper San Francisco Estuary (SFE) since 1999 is a potential but to date an unquantified threat to the health and survival of aquatic organisms, such as fish and zooplankton. The microcystins (MCs) predominantly in the LR-form (MC-LR) contained in Microcystis is hepatotoxic and a potential threat to the fishery. This study was conducted to determine the effects of dietary exposure of the endemic Sacramento splittail, Pogonichthys macrolepidotus in SFE to Microcystis and its toxin, MC-LR. Juvenile splittail (12.59 ± 0.7 g fish(-1)) were exposed to five diets for 28 d with MC-LR obtained from: (1) Microcystis harvested from the SFE and (2) a synthetic purified form of MC-LR. Three of the test diets contained 3.55 (D5), 9.14 (D10) and 17.13 (D20)mg MC-LR kg(-1) from Microcystis. The other two diets contained either purified MC-LR at 3.89 mg MC-LR kg(-1) (D5R) or no MC-LR (D0). The RNA/DNA ratio of fish muscle was significantly lower for all treatments fed test diets containing MC-LR compared to the control diet D0, suggesting Microcystis adversely affected nutritional status. Protein phosphatase 2A expression in the fish from the D5, D10 and D20 treatments were inversely affected by increasing concentrations of MC-LR. Cytoplasmic inclusion bodies and single cell necrosis were more prevalent and greater in severity in the fish exposed to the diets D10 and D20 compared to fish from the D0 treatment and indicate severe liver toxicity in splittail exposed to MC-LR. The sublethal effects on splittail characterized by this study suggest cyanobacterial blooms have the potential to affect splittail nutritional status and health in SFE. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Sublethal amounts of Origanum vulgare L. essential oil and carvacrol cause injury and changes in membrane fatty acid of Salmonella Typhimurium cultivated in a meat broth.

    Science.gov (United States)

    Luz, Isabelle da Silva; de Melo, Adma Nadja Ferreira; Bezerra, Taliana Kênia Alves; Madruga, Marta Suely; Magnani, Marciane; de Souza, Evandro Leite

    2014-05-01

    This study aimed to evaluate whether sublethal concentrations of the essential oil of Origanum vulgare L. (OVEO) and its major compound carvacrol (CAR) cause injury to the cell membrane and outer membrane of Salmonella enterica serovar Typhimurium ATCC 14028 grown in a meat broth and to assess the effect of these substances on membrane fatty acid (FA) composition. Exposure of Salmonella Typhimurium ATCC 14028 to sublethal concentrations of OVEO or CAR caused damage to the cytoplasmic membrane and outer membrane. OVEO- and CAR-treated cells showed lower amounts of saturated FA than nontreated cells. Changes in membrane FA composition were mainly related to an increase of C16:1ω7c, C16:1ω7t, and C18:2ω6c, and to a decrease of C16:0, C17:0 cyclo, and C19:0 cyclo. These results indicate that exposure to sublethal concentrations of OVEO or CAR caused sublethal injury Salmonella Typhimurium ATCC 14028 and suggest that an adaptive response to these stresses is related to increased synthesis of unsaturated FA and cis-trans isomerization.

  20. Gene expression in Listeria monocytogenes exposed to sublethal concentration of benzalkonium chloride.

    Science.gov (United States)

    Tamburro, Manuela; Ripabelli, Giancarlo; Vitullo, Monia; Dallman, Timothy James; Pontello, Mirella; Amar, Corinne Francoise Laurence; Sammarco, Michela Lucia

    2015-06-01

    In this study, tolerance at sublethal concentration of benzalkonium chloride and transcription levels of mdrL, ladR, lde, sigB and bcrABC genes in Listeria monocytogenes strains were evaluated. Viable cells reduction occurred in 45% of strains and clinical isolates showed lower sensitivity than isolates from foods. An increased transcription of an efflux system encoding gene was found in 60% of strains, and simultaneous mdrL overexpression and ladR underexpression occurred in 30% of isolates. A significant association between reduced benzalkonium chloride activity and both mdrL and sigB overexpression was observed; sigB expression also correlated with both mdrL and ladR genes. The bcrABC gene was only found in six strains, all isolated from foods and sensitive to benzalkonium chloride, and in four strains an underexpression was observed. Disinfection at sublethal concentration was less effective in clinical isolates, and mdrL and sigB expression was significantly affected by disinfection. Further insights are needed to understand the adaptation to benzalkonium chloride and to evaluate whether changes in gene expression could affect the L. monocytogenes virulence traits and persistence in the environment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Sublethal Dose of Diazinon Induces Pulmonary Toxicity in Rat: Histopathological Evidence

    Directory of Open Access Journals (Sweden)

    Gholamreza Najafi

    2014-02-01

    Full Text Available Objective: Diazinon (DZ is a widely used contact organophosphorous pesticide with broad spectrum insecticide activity. The extensive use of DZ has caused great concern due to the hazardous side effects on human beings as well as wild and domestic animals. The aim of this study was to investigate the effect of sublethal dose of DZ on rat lung. Material and methods: Seven groups of male Wistar rats were used comprising control and test groups. The control group received corn oil (0.3 ml/day for 60 days by oral gavages. The test groups received DZ at a dose of 60 mg/kg body weight orally for 2, 10, 24, 30, 54 and 60 days, respectively. Results: The histopathological analysis of the lungs in DZ-treated groups revealed congestion on day 2, pulmonary edema and emphysema on day 10, congestion and atelectasia on day 24, infiltration of mononuclear cells on day 30 and pulmonary hemorrhage along with bronchial glands hyperplasia on days 54 and 60. DZ administration also caused a significant decrease in serum cholinesterase activity in a time-dependent manner. Conclusion: These findings indicate that sublethal dose of DZ can induce severe lesions in the lung of rat. [J Interdiscipl Histopathol 2014; 2(1.000: 26-31

  2. Sub-Lethal Dose of Shiga toxin 2 from Enterohemorrhagic Escherichia coli Affects Balance and Cerebellar Cythoarquitecture.

    Directory of Open Access Journals (Sweden)

    Luciana eD’Alessio

    2016-02-01

    Full Text Available Shiga toxin producing Escherichia coli may damage the central nervous system before or concomitantly to manifested hemolytic uremic syndrome symptoms. The cerebellum is frequently damaged during this syndrome, however the deleterious effects of Shiga toxin 2 has never been integrally reported by ultrastructural, physiological and behavioral means. The aim of this study was to determine the cerebellar compromise after intravenous administration of a sub-lethal dose of Shiga toxin 2 by measuring the cerebellar blood brain barrier permeability, behavioral task of cerebellar functionality (inclined plane test, and ultrastructural analysis (transmission electron microscope. Intravenous administration of vehicle (control group, sub-lethal dose of 0.5 ηg and 1 ηg of Stx2 per mouse were tested for behavioral and ultrastructural studies. A set of three independent experiments were performed for each study (n=6. Blood–Brain Barrier resulted damaged and consequently its permeability was significantly increased. Lower scores obtained in the inclined plane task denoted poor cerebellar functionality in comparison to their controls. The most significant lower score was obtained after 5 days of 1ηg of toxin administration. Transmission electron microscope micrographs from the Stx2-treated groups showed neurons with a progressive neurodegenerative condition in a dose dependent manner. As sub-lethal intravenous Shiga toxin 2 altered the blood brain barrier permeability in the cerebellum the toxin penetrated the cerebellar parenchyma and produced cell damaged with significant functional implications in the test balance.

  3. Sub-Lethal Dose of Shiga Toxin 2 from Enterohemorrhagic Escherichia coli Affects Balance and Cerebellar Cytoarchitecture

    Science.gov (United States)

    Pinto, Alipio; Cangelosi, Adriana; Geoghegan, Patricia A.; Tironi-Farinati, Carla; Brener, Gabriela J.; Goldstein, Jorge

    2016-01-01

    Shiga toxin producing Escherichia coli may damage the central nervous system before or concomitantly to manifested hemolytic–uremic syndrome symptoms. The cerebellum is frequently damaged during this syndrome, however, the deleterious effects of Shiga toxin 2 has never been integrally reported by ultrastructural, physiological and behavioral means. The aim of this study was to determine the cerebellar compromise after intravenous administration of a sub-lethal dose of Shiga toxin 2 by measuring the cerebellar blood–brain barrier permeability, behavioral task of cerebellar functionality (inclined plane test), and ultrastructural analysis (transmission electron microscope). Intravenous administration of vehicle (control group), sub-lethal dose of 0.5 and 1 ηg of Stx2 per mouse were tested for behavioral and ultrastructural studies. A set of three independent experiments were performed for each study (n = 6). Blood–brain barrier resulted damaged and consequently its permeability was significantly increased. Lower scores obtained in the inclined plane task denoted poor cerebellar functionality in comparison to their controls. The most significant lower score was obtained after 5 days of 1 ηg of toxin administration. Transmission electron microscope micrographs from the Stx2-treated groups showed neurons with a progressive neurodegenerative condition in a dose dependent manner. As sub-lethal intravenous Shiga toxin 2 altered the blood brain barrier permeability in the cerebellum the toxin penetrated the cerebellar parenchyma and produced cell damaged with significant functional implications in the test balance. PMID:26904009

  4. Impacts of Sublethal Mercury Exposure on Birds: A Detailed Review.

    Science.gov (United States)

    Whitney, Margaret C; Cristol, Daniel A

    Mercury is a ubiquitous environmental contaminant known to accumulate in, and negatively affect, fish-eating and oceanic bird species, and recently demonstrated to impact some terrestrial songbirds to a comparable extent. It can bioaccumulate to concentrations of >1 μg/g in tissues of prey organisms such as fish and insects. At high enough concentrations, exposure to mercury is lethal to birds. However, environmental exposures are usually far below the lethal concentrations established by dosing studies.The objective of this review is to better understand the effects of sublethal exposure to mercury in birds. We restricted our survey of the literature to studies with at least some exposures >5 μg/g. The majority of sublethal effects were subtle and some studies of similar endpoints reached different conclusions. Strong support exists in the literature for the conclusion that mercury exposure reduces reproductive output, compromises immune function, and causes avoidance of high-energy behaviors. For some endpoints, notably certain measures of reproductive success, endocrine and neurological function, and body condition, there is weak or contradictory evidence of adverse effects and further study is required. There was no evidence that environmentally relevant mercury exposure affects longevity, but several of the sublethal effects identified likely do result in fitness reductions that could adversely impact populations. Overall, 72% of field studies and 91% of laboratory studies found evidence of deleterious effects of mercury on some endpoint, and thus we can conclude that mercury is harmful to birds, and the many effects on reproduction indicate that bird population declines may already be resulting from environmental mercury pollution.

  5. Sublethal effect of neem extract on mediterranean fruit fly adults

    Directory of Open Access Journals (Sweden)

    Márcio Alves Silva

    2013-03-01

    Full Text Available The sublethal effect of extracts of Azadirachta indica on Ceratitis capitata was evaluated. Two pairs of flies were treated in plastic tubes with cotton placed in plastic cages. An artificial diet (hydrolyzed protein + sugar was provided ad libitum. The extracts affected significantly the longevity of C. capitata. The pre-oviposition period were not significantly affected by the extracts. The A. indica branches extracted with dichloromethane (888 ppm affected significantly the fecundity and fertility, reducing the number of eggs laid to approximately 80 % and the egg hatching by 30 % at the 8th day. Therefore, the neem branches extracted with dichloromethane affected the reproduction of C. capitata.

  6. Stability of sublethal acid stress adaptation and induced cross protection against lauric arginate in Listeria monocytogenes.

    Science.gov (United States)

    Shen, Qian; Soni, Kamlesh A; Nannapaneni, Ramakrishna

    2015-06-16

    The stability of acid stress adaptation in Listeria monocytogenes and its induced cross protection effect against GRAS (generally recognized as safe) antimicrobial compounds has never been investigated before. In the present study, the acid stress adaptation in L. monocytogenes was initially induced in pH 5.0 tryptic soy broth supplemented with 0.6% yeast extract (TSB-YE) at 37 °C. Subsequently, the stability of acid stress adaptation, which was defined as the capacity to maintain its acquired acid adaptation after induction in the absence of sublethal acid stress, was determined at 37 °C, 22 °C or 4 °C in broth and in different food substrates. Then, the acid stress adaptation induced cross protection against lauric arginate (LAE) and its stability was investigated in TSB-YE, milk and carrot juice. Our findings show that the acid stress adaptation was stable at 4 °C up to 24h but was reversed at 37 °C or 22 °C within 2h. In the cross protection assay with LAE, the acid stress adapted cells had approximately 2 log CFU/ml greater survival than non-adapted cells in broth at 22 °C or in milk and carrot juice at 4 °C. The acid adaptation induced cross protection against LAE in L. monocytogenes was reversible within 1h at 4 °C in the absence of sublethal acid stress. Our findings suggest that the stability of acid adaptation in L. monocytogenes under cold conditions should be taken into account when the risk analysis is performed during food processing. Copyright © 2015. Published by Elsevier B.V.

  7. Sublethal effect of imidacloprid on Solenopsis invicta (Hymenoptera: Formicidae) feeding, digging, and foraging behavior

    Science.gov (United States)

    There is increasing evidence that exposure to neonicotinoid insecticides at sublethal levels impairs colonies of honeybee and other pollinators. Recently, it was found that sublethal contamination with neonicotinoids also affect growth and behavior of ants. In this study, we exposed red imported fi...

  8. Survival rate of honeybee (Apis mellifera) workers after exposure to sublethal concentrations of imidacloprid

    NARCIS (Netherlands)

    Blacquiere, T.

    2010-01-01

    Imidacloprid is a commonly used systemic insecticide which can induce several sublethal effects. Previous research has not shown any increased mortality in bees that were fed with sublethal doses. However, there is very little research conducted with the focus on survival rate of honeybees in the

  9. Toxicokinetic-toxicodynamic modeling of quantal and graded sublethal endpoints: a brief discussion of concepts

    NARCIS (Netherlands)

    Ashauer, R.; Agatz, A.; Albert, C.; Ducrot, V.; Galic, N.G.; Hendriks, J.; Jager, T.; Kretschmann, A.; O'Connor, I.; Rubach, M.N.; Nyman, M.; Schmitt, W.; Stadnicka, J.; Brink, van den P.J.

    2011-01-01

    We report on the advantages and problems of using toxicokinetic-toxicodynamic (TKTD) models for the analysis, understanding, and simulation of sublethal effects. Only a few toxicodynamic approaches for sublethal effects are available. These differ in their effect mechanism and emphasis on linkages

  10. Sublethal effects of cadmium, manganese, lead, zinc and iron on the ...

    African Journals Online (AJOL)

    This study also evaluated the sublethal effects of cadmium, manganese, lead, zinc and iron in plasma samples utilising plasma electrolyte parameters as a biomarker using an albino mice model, M. musculus. Mice were subjected to sublethal concentrations of the selected heavy metals (1/10th of 96 hrLC50). Blood plasma ...

  11. Sub-lethal radiation enhances anti-tumor immunotherapy in a transgenic mouse model of pancreatic cancer

    Directory of Open Access Journals (Sweden)

    Hanahan Douglas

    2002-05-01

    Full Text Available Abstract Background It is not uncommon to observe circulating tumor antigen-specific T lymphocytes in cancer patients despite a lack of significant infiltration and destruction of their tumors. Thus, an important goal for tumor immunotherapy is to identify ways to modulate in vivo anti-tumor immunity to achieve clinical efficacy. We investigate this proposition in a spontaneous mouse tumor model, Rip1-Tag2. Methods Experimental therapies were carried out in two distinctive trial designs, intended to either intervene in the explosive growth of small tumors, or regress bulky end-stage tumors. Rip1-Tag2 mice received a single transfer of splenocytes from Tag-specific, CD4+ T cell receptor transgenic mice, a single sub-lethal radiation, or a combination therapy in which the lymphocyte transfer was preceded by the sub-lethal radiation. Tumor burden, the extent of lymphocyte infiltration into solid tumors and host survival were used to assess the efficacy of these therapeutic approaches. Results In either intervention or regression, the transfer of Tag-specific T cells alone did not result in significant lymphocyte infiltration into solid tumors, not did it affect tumor growth or host survival. In contrast, the combination therapy resulted in significant reduction in tumor burden, increase in lymphocyte infiltration into solid tumors, and extension of survival. Conclusions The results indicate that certain types of solid tumors may be intrinsically resistant to infiltration and destruction by tumor-specific T lymphocytes. Our data suggest that such resistance can be disrupted by sub-lethal radiation. The combinatorial approach presented here merits consideration in the design of clinical trials aimed to achieve T cell-mediated anti-tumor immunity.

  12. Accumulating microglia phagocytose injured neurons in hippocampal slice cultures: involvement of p38 MAP kinase.

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    Takahiro Katayama

    Full Text Available In this study, microglial migration and phagocytosis were examined in mouse organotypic hippocampal slice cultures, which were treated with N-methyl-D-aspartate (NMDA to selectively injure neuronal cells. Microglial cells were visualized by the expression of enhanced green fluorescent protein. Daily observation revealed microglial accumulation in the pyramidal cell layer, which peaked 5 to 6 days after NMDA treatment. Time-lapse imaging showed that microglia migrated to the pyramidal cell layer from adjacent and/or remote areas. There was no difference in the number of proliferating microglia between control and NMDA-treated slices in both the pyramidal cell layer and stratum radiatum, suggesting that microglial accumulation in the injured areas is mainly due to microglial migration, not to proliferation. Time-lapse imaging also showed that the injured neurons, which were visualized by propidium iodide (PI, disappeared just after being surrounded by microglia. Daily observation revealed that the intensity of PI fluorescence gradually attenuated, and this attenuation was suppressed by pretreatment with clodronate, a microglia toxin. These findings suggest that accumulating microglia phagocytosed injured neurons, and that PI fluorescence could be a useful indicator for microglial phagocytosis. Using this advantage to examine microglial phagocytosis in living slice cultures, we investigated the involvements of mitogen-activated protein (MAP kinases in microglial accumulation and phagocytosis. p38 MAP kinase inhibitor SB203580, but not MAP kinase/extracellular signal-regulated kinase inhibitor PD98059 or c-Jun N-terminal kinase inhibitor SP600125, suppressed the attenuation of PI fluorescence. On the other hand, microglial accumulation in the injured areas was not inhibited by any of these inhibitors. These data suggest that p38 MAP kinase plays an important role in microglial phagocytosis of injured neurons.

  13. Microcurrent electrical neuromuscular stimulation facilitates regeneration of injured skeletal muscle in mice.

    Science.gov (United States)

    Fujiya, Hiroto; Ogura, Yuji; Ohno, Yoshitaka; Goto, Ayumi; Nakamura, Ayane; Ohashi, Kazuya; Uematsu, Daiki; Aoki, Haruhito; Musha, Haruki; Goto, Katsumasa

    2015-06-01

    Conservative therapies, mainly resting care for the damaged muscle, are generally used as a treatment for skeletal muscle injuries (such as muscle fragmentation). Several past studies reported that microcurrent electrical neuromuscular stimulation (MENS) facilitates a repair of injured soft tissues and shortens the recovery period. However, the effects of MENS on the regeneration in injured skeletal muscle are still unclear. The purpose of this study was to investigate the effect of MENS on the regenerative process of injured skeletal muscle and to elucidate whether satellite cells in injured skeletal muscle are activated by MENS by using animal models. Male C57BL/6J mice, aged 7 weeks old, were used (n = 30). Mice were randomly divided into two groups: (1) cardiotoxin (CTX)-injected (CX, n = 15) and (2) CTX-injected with MENS treatment (MX, n=15) groups. CTX was injected into tibialis anterior muscle (TA) of mice in CX and MX groups to initiate the necrosis-regeneration cycle of the muscle. TA was dissected 1, 2, and 3 weeks after the injection. Muscle weight, muscle protein content, the mean cross-sectional areas of muscle fibers, the relative percentage of fibers having central nuclei, and the number of muscle satellite cells were evaluated. MENS facilitated the recovery of the muscle dry weight and protein content relative to body weight, and the mean cross-sectional areas of muscle fibers in CTX-induced injured TA muscle. The number of Pax7-positive muscle satellite cells was increased by MENS during the regenerating period. Decrease in the percentages of fibers with central nuclei after CTX-injection was facilitated by MENS. MENS may facilitate the regeneration of injured skeletal muscles by activating the regenerative potential of skeletal muscles. Key pointsMicrocurrent electrical neuromuscular stimulation (MENS) facilitated the recovery of the relative muscle dry weight, the relative muscle protein content, and the mean cross-sectional areas of muscle

  14. Field-level sublethal effects of approved bee hive chemicals on Honey Bees (Apis mellifera L).

    Science.gov (United States)

    Berry, Jennifer A; Hood, W Michael; Pietravalle, Stéphane; Delaplane, Keith S

    2013-01-01

    In a study replicated across two states and two years, we tested the sublethal effects on honey bees of the miticides Apistan (tau fluvalinate) and Check Mite+ (coumaphos) and the wood preservative copper naphthenate applied at label rates in field conditions. A continuous covariate, a colony Varroa mite index, helped us disambiguate the effects of the chemicals on bees while adjusting for a presumed benefit of controlling mites. Mite levels in colonies treated with Apistan or Check Mite+ were not different from levels in non-treated controls. Experimental chemicals significantly decreased 3-day brood survivorship and increased construction of queen supercedure cells compared to non-treated controls. Bees exposed to Check Mite+ as immatures had higher legacy mortality as adults relative to non-treated controls, whereas bees exposed to Apistan had improved legacy mortality relative to non-treated controls. Relative to non-treated controls, Check Mite+ increased adult emergence weight. Although there was a treatment effect on a test of associative learning, it was not possible to statistically separate the treatment means, but bees treated with Apistan performed comparatively well. And finally, there were no detected effects of bee hive chemical on colony bee population, amount of brood, amount of honey, foraging rate, time required for marked released bees to return to their nest, percentage of released bees that return to the nest, and colony Nosema spore loads. To our knowledge, this is the first study to examine sublethal effects of bee hive chemicals applied at label rates under field conditions while disambiguating the results from mite control benefits realized from the chemicals. Given the poor performance of the miticides at reducing mites and their inconsistent effects on the host, these results defend the use of bee health management practices that minimize use of exotic hive chemicals.

  15. Field-level sublethal effects of approved bee hive chemicals on Honey Bees (Apis mellifera L.

    Directory of Open Access Journals (Sweden)

    Jennifer A Berry

    Full Text Available In a study replicated across two states and two years, we tested the sublethal effects on honey bees of the miticides Apistan (tau fluvalinate and Check Mite+ (coumaphos and the wood preservative copper naphthenate applied at label rates in field conditions. A continuous covariate, a colony Varroa mite index, helped us disambiguate the effects of the chemicals on bees while adjusting for a presumed benefit of controlling mites. Mite levels in colonies treated with Apistan or Check Mite+ were not different from levels in non-treated controls. Experimental chemicals significantly decreased 3-day brood survivorship and increased construction of queen supercedure cells compared to non-treated controls. Bees exposed to Check Mite+ as immatures had higher legacy mortality as adults relative to non-treated controls, whereas bees exposed to Apistan had improved legacy mortality relative to non-treated controls. Relative to non-treated controls, Check Mite+ increased adult emergence weight. Although there was a treatment effect on a test of associative learning, it was not possible to statistically separate the treatment means, but bees treated with Apistan performed comparatively well. And finally, there were no detected effects of bee hive chemical on colony bee population, amount of brood, amount of honey, foraging rate, time required for marked released bees to return to their nest, percentage of released bees that return to the nest, and colony Nosema spore loads. To our knowledge, this is the first study to examine sublethal effects of bee hive chemicals applied at label rates under field conditions while disambiguating the results from mite control benefits realized from the chemicals. Given the poor performance of the miticides at reducing mites and their inconsistent effects on the host, these results defend the use of bee health management practices that minimize use of exotic hive chemicals.

  16. Using a sequential regimen to eliminate bacteria at sublethal antibiotic dosages.

    Directory of Open Access Journals (Sweden)

    Ayari Fuentes-Hernandez

    2015-04-01

    Full Text Available We need to find ways of enhancing the potency of existing antibiotics, and, with this in mind, we begin with an unusual question: how low can antibiotic dosages be and yet bacterial clearance still be observed? Seeking to optimise the simultaneous use of two antibiotics, we use the minimal dose at which clearance is observed in an in vitro experimental model of antibiotic treatment as a criterion to distinguish the best and worst treatments of a bacterium, Escherichia coli. Our aim is to compare a combination treatment consisting of two synergistic antibiotics to so-called sequential treatments in which the choice of antibiotic to administer can change with each round of treatment. Using mathematical predictions validated by the E. coli treatment model, we show that clearance of the bacterium can be achieved using sequential treatments at antibiotic dosages so low that the equivalent two-drug combination treatments are ineffective. Seeking to treat the bacterium in testing circumstances, we purposefully study an E. coli strain that has a multidrug pump encoded in its chromosome that effluxes both antibiotics. Genomic amplifications that increase the number of pumps expressed per cell can cause the failure of high-dose combination treatments, yet, as we show, sequentially treated populations can still collapse. However, dual resistance due to the pump means that the antibiotics must be carefully deployed and not all sublethal sequential treatments succeed. A screen of 136 96-h-long sequential treatments determined five of these that could clear the bacterium at sublethal dosages in all replicate populations, even though none had done so by 24 h. These successes can be attributed to a collateral sensitivity whereby cross-resistance due to the duplicated pump proves insufficient to stop a reduction in E. coli growth rate following drug exchanges, a reduction that proves large enough for appropriately chosen drug switches to clear the bacterium.

  17. Sub-lethal effects of the neurotoxic pyrethroid insecticide Fastac 50EC on the general motor and locomotor activities of the non-targeted beneficial carabid beetle Platynus assimilis (Coleoptera: Carabidae).

    Science.gov (United States)

    Tooming, Ene; Merivee, Enno; Must, Anne; Sibul, Ivar; Williams, Ingrid

    2014-06-01

    Sub-lethal effects of pesticides on behavioural endpoints are poorly studied in carabids (Coleoptera: Carabidae) though changes in behaviour caused by chemical stress may affect populations of these non-targeted beneficial insects. General motor activity and locomotion are inherent in many behavioural patterns, and changes in these activities that result from xenobiotic influence mirror an integrated response of the insect to pesticides. Influence of pyrethroid insecticides over a wide range of sub-lethal doses on the motor activities of carabids still remains unclear. Video tracking of Platynus assimilis showed that brief exposure to alpha-cypermethrin at sub-lethal concentrations ranged from 0.01 to 100 mg L(-1) caused initial short-term (24 h) locomotor hypo-activity. In addition, significant short- and long-term concentration and time-dependent changes occurred in general motor activity patterns and rates. Conspicuous changes in motor activity of Platynus assimilis beetles treated at alpha-cypermethrin concentrations up to 75,000-fold lower than maximum field recommended concentration (MFRC) suggest that many, basic fitness-related behaviours might be severely injured as well. These changes may negatively affect carabid populations in agro-ecosystems. Long-term hypo-activity could directly contribute to decreased trap captures of carabids frequently observed after insecticide application in the field. © 2013 Society of Chemical Industry.

  18. Injured pedestrians in Cape Town - the role of alcohol | Peden ...

    African Journals Online (AJOL)

    Objective. To establish a profile of injured adult pedestrians and attempt to define the role which alcohol plays in this regard. Design. Prospective survey of injured pedestrians who presented consecutively over 9 weeks to Groote Schuur Hospital. Data on fatally injured pedestrians were retrospectively collected from the ...

  19. Cadaverous Particles and Infection in Injured Man,

    Science.gov (United States)

    1993-01-01

    A D-A 277 941 E J Surg 159: 515-520, 1993 CADAVEROUS PARTICLES AND INFECTION IN INJURED MAN*D T IC Clinical Review based on the Semmelweis Lecture WM...systemic hematogencus dissemination of bacteria versary year of the publication of his findings in Die which had invaded tissue from an infected wound...surgical infections to evaluate therapeutic interventions (35). The effec- and the importance of " cadaverous particles" in the tiveness of topical

  20. Sublethal effects of waterborne herbicides in tropical freshwater fish.

    Science.gov (United States)

    Rossi, Stéfani Cibele; Dreyer da Silva, Manuela; Piancini, Laercio Dante Stein; Oliveira Ribeiro, Ciro Alberto; Cestari, Marta Margarete; Silva de Assis, Helena Cristina

    2011-12-01

    The study evaluated the sublethal effects of the herbicides glyphosate (Roundup) and diuron (Hexaron) and the mixture of them, used extremely in agriculture, through biomarkers in fish. The glutathione S-transferase activity increased (74%) and catalase activity decreased (37%) at the higher exposure concentration of Hexaron in comparison to the control group, suggesting an activation of this metabolism route. Membrane damage was observed at the higher exposure of Roundup and in the mixture group compared to the control group, which can be related to the nuclear alterations observed in these exposed groups. The cholinesterase activity was also inhibited (37%) in mixture group compared to the control group and no gill morphology damage was found. The results suggested a potential synergic effect in some analysed parameters.

  1. A longitudinal study of the mechanical properties of injured brain tissue in a mouse model.

    Science.gov (United States)

    Feng, Yuan; Gao, Yuan; Wang, Tao; Tao, Luyang; Qiu, Suhao; Zhao, Xuefeng

    2017-07-01

    Mechanical properties of brain tissue are crucial to understand the mechanism of traumatic brain injury (TBI). Over the past several decades, most of the studies focused on healthy brain tissues, while few of them are about the injured tissues. Therefore, limited knowledge is known about the mechanical properties of the injured brain tissues. In this study, we used an in vivo mouse model with a weight drop device to study injured brain tissues. Around the injury site, mechanical properties of the injured, neighboring, and the corresponding contralateral regions of interest (ROIs) were measured over five temporal points by indentation. Longitudinal and regional comparisons of the mechanical properties revealed that the ROI of the injured tissue had a higher elastic modulus than the contralateral counterpart one-hour post-injury. However, the elastic modulus decreased one-day post-injury and recovered to be close to the contralateral ROI in 7 days. The elastic modulus curves of the injured and the contralateral counterpart ROIs crossed at time points of 12h and 1 day post-injury, where two significant increases of glial fibrillary acidic protein (GFAP) positive cells were observed. Biological staining results indicated that both the astrocytic responses and the morphological structure could affect the mechanical properties of the injured tissue. The observed longitudinal changes of the mechanical properties at the tissue level and the morphological and biological changes at the cellular level provide insights into understanding the mechanism of TBI. Results are also meaningful for applying emerging in vivo diagnostic tools such as magnetic resonance elastography (MRE) in TBI detection. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Microcurrent Electrical Neuromuscular Stimulation Facilitates Regeneration of Injured Skeletal Muscle in Mice

    Directory of Open Access Journals (Sweden)

    Hiroto Fujiya, Yuji Ogura, Yoshitaka Ohno, Ayumi Goto, Ayane Nakamura, Kazuya Ohashi, Daiki Uematsu, Haruhito Aoki, Haruki Musha, Katsumasa Goto

    2015-06-01

    Full Text Available Conservative therapies, mainly resting care for the damaged muscle, are generally used as a treatment for skeletal muscle injuries (such as muscle fragmentation. Several past studies reported that microcurrent electrical neuromuscular stimulation (MENS facilitates a repair of injured soft tissues and shortens the recovery period. However, the effects of MENS on the regeneration in injured skeletal muscle are still unclear. The purpose of this study was to investigate the effect of MENS on the regenerative process of injured skeletal muscle and to elucidate whether satellite cells in injured skeletal muscle are activated by MENS by using animal models. Male C57BL/6J mice, aged 7 weeks old, were used (n = 30. Mice were randomly divided into two groups: (1 cardiotoxin (CTX-injected (CX, n = 15 and (2 CTX-injected with MENS treatment (MX, n=15 groups. CTX was injected into tibialis anterior muscle (TA of mice in CX and MX groups to initiate the necrosis-regeneration cycle of the muscle. TA was dissected 1, 2, and 3 weeks after the injection. Muscle weight, muscle protein content, the mean cross-sectional areas of muscle fibers, the relative percentage of fibers having central nuclei, and the number of muscle satellite cells were evaluated. MENS facilitated the recovery of the muscle dry weight and protein content relative to body weight, and the mean cross-sectional areas of muscle fibers in CTX-induced injured TA muscle. The number of Pax7-positive muscle satellite cells was increased by MENS during the regenerating period. Decrease in the percentages of fibers with central nuclei after CTX-injection was facilitated by MENS. MENS may facilitate the regeneration of injured skeletal muscles by activating the regenerative potential of skeletal muscles.

  3. Lethal and sublethal toxicity of the industrial chemical epichlorohydrin on Rhinella arenarum (Anura, Bufonidae) embryos and larvae.

    Science.gov (United States)

    Hutler Wolkowicz, Ianina R; Aronzon, Carolina M; Pérez Coll, Cristina S

    2013-12-15

    Lethal and sublethal toxicity of the major chemical used in epoxide compounds, epichlorohydrin (ECH) was evaluated on the early life cycle of the common South American toad, Rhinella arenarum (Anura, Bufonidae). The stages evaluated were (according to Del Conte and Sirlin): early blastula (S.3-S.4), gastrula (S.10-S.12), rotation (S.15), tail bud (S.17), muscular response (S.18), gill circulation (S.20), open mouth (S.21), opercular folds (S.23) and complete operculum (S.25). The LC50 and EC50 values for lethal and sublethal effects were calculated. The early blastula was the most sensitive stage to ECH both for continuously and pulse-exposures (LC50-24h=50.9 mg L(-1)), while S.20 was the most resistant (LC50-24h=104.9 mg L(-1)). Among sublethal effects, early blastula was also the most sensitive stage (LOEC-48 h=20 mg L(-1)) and it has a Teratogenic Index of 2.5, which indicates the teratogenic potential of the substance. The main abnormalities were persistent yolk plugs, cell dissociation, tumors, hydropsy, oral malformations, axial/tail flexures, delayed development and reduced body size. ECH also caused neurotoxicity including scarce response to stimuli, reduction in the food intake, general weakness, spasms and shortening, erratic or circular swimming. Industrial contamination is considered an important factor on the decline of amphibian populations. Considering the available information about ECH's toxicity and its potential hazard to the environment, this work shows the first results of its developmental toxicity on a native amphibian species, Rhinella arenarum. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Study of human mesenchymal stem cells plasticity into radiation injured tissues in a N.O.D./S.C.I.D. mouse model: therapeutic approach of the multiple organ dysfunction; Etude de la capacite plastique des Cellules Souches Mesenchymateuses humaines (CSM) apres irradiation du tissu receveur: approche therapeutique de l'atteinte multiorgane radio-induite

    Energy Technology Data Exchange (ETDEWEB)

    Francois, S

    2006-01-15

    The therapeutic potential of bone marrow-derived human mesenchymal stem cells (h.M.S.C.) has recently been brought into the spotlight of many fields of research. One possible application of the approach is the repair of injured tissues arising from side effects of radiation treatments and accidents. The first challenge in cell therapy is to assess the quality of the cell and the ability to retain their differentiation potential during the expansion process. Efficient delivery to the sites of intended action is also necessary. We addressed both questions using h.M.S.C. cultured and then infused to Non Obese Diabetes/Severe Combined Immunodeficiency (N.O.D./S.C.I.D.) mice submitted to total body irradiation. Further, we tested the impact of additional local irradiation superimposed to total body irradiation (T.B.I.), as a model of accidental irradiation. Our results showed that the h.M.S.C. used for transplant have been expanded without significant loss in their differentiation capacities. After transplantation into adult unconditioned mice, h.M.S.C. not only migrate in bone marrow but also into other tissues. Total body irradiation increased h.M.S.C. implantation in bone marrow and muscle and further led to engraftment in brain, heart, and liver. Local irradiation, in addition to T.B.I., increased both specific homing of injected cells to the injured tissues and to other tissues outside the local irradiation field. M.S.C. may participate to restoration of intestinal homeostasis 3 days post abdominal irradiation. This study suggests that using the potential of h.M.S.C. to home to various organs in response to tissue injuries could be a promising strategy to repair the radiation induced damages. (author)

  5. The repair of sub-lethal damage and the stimulated repair of potentially lethal damage in Saintpaulia.

    Science.gov (United States)

    Leenhouts, H P; Sijsma, M J; Litwiniszyn, M; Chadwick, K H

    1981-10-01

    The repair of sublethal and potentially lethal damage in stationary resting epidermal cells of Saintpaulia has been investigated. Fractionation experiments reveal an efficient repair of sublethal damage with a half-life of 1.9 hours. No repair of potentially lethal damage was noted when cultivation of the leaves was delayed for 24 hours after irradiation. At delay times of 2, 3 and 4 days some repair of potentially lethal damage has been found. A small pre-dose given 24 hours before a challenging dose improved the cells' chance to regenerate and the improvement has been shown to be compatible with an improved repair of potentially lethal damage induced by X-rays and fast neutrons. It hs been shown that the stimulated repair process takes 12 to 24 hours to develop, is dependent on the size of the pre-dose, has single-hit dose kinetics, and an r.b.e. of 1 for neutrons. With delayed cultivation of 2 days the stimulated repair process leads to an alteration in the shape of the regeneration (survival)-dose relationship which increases the low dose r.b.e. for neutrons from 10 to 35.

  6. Comparative proteomic analysis of Listeria monocytogenes ATCC 7644 exposed to a sublethal concentration of nisin.

    Science.gov (United States)

    Miyamoto, Kendi Nishino; Monteiro, Karina Mariante; da Silva Caumo, Karin; Lorenzatto, Karina Rodrigues; Ferreira, Henrique Bunselmeyer; Brandelli, Adriano

    2015-04-24

    Listeria monocytogenes infections have been frequently reported in many food poisoning outbreaks around the world. In this work, the protein repertoires of L. monocytogenes ATCC 7644 cells treated or not with a 10(-3)mg/mL nisin sublethal concentration, established by antimicrobial susceptibility tests, were analyzed by LC-MS/MS. Overall, 179 proteins were identified, 9 of them more abundant in nisin-treated samples, and 4 more abundant in non-treated control samples. In nisin treated cells, proteins associated to oxidative stress response showed higher abundance. Also, the higher abundance of an enzyme related to the production of cell membrane lipids upon nisin exposure is suggestive of both a failure in conventional cell division mechanism and the activation of an alternative L-form mediated division mechanism. Finally, flagellar and motility proteins' overexpression upon nisin exposure is indicative of increased bacterial motility in response to the bacteriocin. Taken together, these results provide new insights on nisin effects on L. monocytogenes cells and on how this bacterium may overcome a bacteriocin-containing environment. The antimicrobial mechanism of nisin on target bacterial cells has been extensively studied since discovery of this bacteriocin. The nisin pore-forming mechanism is mediated by its binding to the pyrophosphate portion of membrane lipid II [1], but some evidences point out to alternative mechanisms. Results from assays with mutacin 1140 hybrids [2] showed that the portion of nisin that is not involved with lipid II binding could damage the bacterial cell, independently of pore formation [3,4]. Moreover, there are insufficient data to explain how nisin affects the bacterial survival. In this scenario, proteomics is an interesting approach, as a comparison between treated and untreated cells may provide insights of both antimicrobial mechanisms of action and bacterial response mechanisms [5]. Copyright © 2015 Elsevier B.V. All rights

  7. Expression of liver functions following sub-lethal and non-lethal doses of allyl alcohol and acetaminophen in the rat

    DEFF Research Database (Denmark)

    Tygstrup, N; Jensen, S A; Krog, B

    1997-01-01

    synthesis, and for proteins related to liver regeneration. RESULTS: After allyl alcohol, mRNA of "positive" acute phase proteins was higher than after acetaminophen and increased with the dose, whereas after acetaminophen it decreased with the dose. The mRNA of the urea cycle enzymes and glutamine...... cycle enzymes, whereas that of the rate limiting enzymes carbamoylphosphate synthase and argininosuccinate synthetase was increased by the non-lethal but not by the sub-lethal dose of acetaminophen. The expression of the cell-cycle dependent proteins was more suppressed after the sub-lethal than after......BACKGROUND/AIMS: To relate severity of intoxication with allyl alcohol and acetaminophen to modulated hepatic gene expression of liver functions and regeneration. METHODS: Rats fasted for 12 h received acetaminophen 3.5 or 5.6 g per kg body weight, or allyl alcohol 100 or 125 microl by gastric tube...

  8. Profiling of recovery efficiencies for three standard protocols (FDA-BAM, ISO-11290, and modified USDA) on temperature-injured Listeria monocytogenes.

    Science.gov (United States)

    Lee, Hai Yen; Chai, Lay Ching; Pui, Chai Fung; Wong, Woan Chwen; Mustafa, Shuhaimi; Cheah, Yoke Kqueen; Mat Issa, Zuraini; Nishibuchi, Mitsuaki; Radu, Son

    2011-09-01

    There have been a number of studies conducted in order to compare the efficiencies of recovery rates, utilizing different protocols, for the isolation of L. monocytogenes. However, the severity of multiple cell injury has not been included in these studies. In the current study, L. monocytogenes ATCC 19112 was injured by exposure to extreme temperatures (60°C and -20°C) for a one-step injury, and for a two-step injury the cells were transferred directly from a heat treatment to frozen state to induce a severe cell injury (up to 100% injury). The injured cells were then subjected to the US Food and Drug Administration (FDA), the ISO-11290, and the modified United States Department of Agriculture (mUSDA) protocols, and plated on TSAyeast (0.6% yeast), PALCAM agar, and CHROMAgar Listeria for 24 h or 48 h. The evaluation of the total recovery of injured cells was also calculated based on the costs involved in the preparation of media for each protocol. Results indicate that the mUSDA method is best able to aid the recovery of heat-injured, freeze-injured, and heat-freeze-injured cells and was shown to be the most cost effective for heat-freeze-injured cells.

  9. Effects of Sublethal Doses of Imidacloprid on Young Adult Honeybee Behaviour: e0140814

    National Research Council Canada - National Science Library

    Gonalons, Carolina Mengoni; Farina, Walter Marcelo

    2015-01-01

    .... As young adult workers perform in-hive duties that are crucial for colony maintenance and survival, we aimed to assess the effect of sublethal IMI doses on honeybee behaviour during this period...

  10. Effects of Sublethal Doses of Imidacloprid on Young Adult Honeybee Behaviour

    National Research Council Canada - National Science Library

    Mengoni Goñalons, Carolina; Farina, Walter Marcelo

    2015-01-01

    .... As young adult workers perform in-hive duties that are crucial for colony maintenance and survival, we aimed to assess the effect of sublethal IMI doses on honeybee behaviour during this period...

  11. Haematological, blood biochemical and histopathological effects of sublethal cadmium and lead concentrations in common carp

    Directory of Open Access Journals (Sweden)

    M.K.Khalesi

    2017-06-01

    Full Text Available The present research aimed at examining the effects of common carp (Cyprinus carpio exposure to sublethal concentrations of two non-essential heavy metals: cadmium (Cd: 8.4 mg/L and lead (Pb: 6.2 mg/L for 15 days to evaluate occurring biochemical and haematological effects. The examined parameters included haematocrit (Hct, haemoglobin (Hb, lymphocytes (Lym, neutrophils (Neu, total protein (TP, albumin (Alb, immunoglobulin M (IgM, glucose, red and white blood cells counts (RBC & WBC, mean corpuscular volume (MCV, mean corpuscular haemoglobin (MCH, and mean corpuscular haemoglobin concentration (MCHC. Exposure to both metals significantly (P<0.05 reduced the amounts of WBC and MCHC. MCV values decreased (P<0.05 after the Pb treatment but MCV estimates with Cd exposure showed no differences. MCH levels increased in both treatments (P<0.05 whereas Hct, Hb, RBC, Lym, and Neu following both metal exposures were almost similar to those in the control. IgM values were elevated in fish contaminated with both Pb and Cd (P<0.05. The exposed fish showed fusion of gill lamellae, vessel dilatation, hyperaemia, and hyperplasia of gill epithelial cells whereas muscle histology remained unchanged. The observed responses can be secondary to low heavy metals concentrations reflecting the trigger of stress reactions in affected fish

  12. Sublethal impact of short term exposure to the organophosphate pesticide azamethiphos in the marine mollusc Mytilus edulis.

    Science.gov (United States)

    Canty, M N; Hagger, J A; Moore, R T B; Cooper, L; Galloway, T S

    2007-04-01

    Concern has been raised that the increased use of pesticides in intensive aquaculture practices may cause adverse sublethal effects to non-target aquatic species. Azamethiphos is an organophosphate (OP) pesticide used to combat sea lice infestations in farmed salmonids. Here, the sublethal impact on the blue mussel, Mytilus edulis, of short term exposure to azamethiphos was determined. The testing regime included biomarkers of exposure (acetylcholinesterase activity), cytotoxicity (neutral red retention), immune function (phagocytic index) and physiological condition (feeding rate). The distribution and sensitivity of M. edulis acetylcholinesterase to inhibition by azamethiphos was first determined, yielding IC(50) values of 0.736 and 1.30 mg l(-1) for gill and haemolymph, respectively. Exposure of mussels to 0.1 mg l(-1) azamethiphos for periods of up to 24h caused a significant reduction in acetylcholinesterase activity in both the haemolymph (Pgill (P<0.002), alteration in cell viability (P<0.02) and decrease in phagocytic index (P<0.03). The feeding rate remained unaffected. The results support the hypothesis that, in addition to its neurotoxic effects, azamethiphos can modulate haemocyte function and immune defence in M. edulis at environmentally relevant concentrations after only a few hours.

  13. Glycosylation-induced depolarization facilitates subthreshold membrane oscillation in injured primary sensory neurons.

    Science.gov (United States)

    Li, Chen-Xu; Jing, Yan-Ling; Xie, Yi-Kuan

    2007-03-30

    Subthreshold membrane potential oscillations (SMPO) in the injured dorsal root ganglion (DRG) neurons are involved in the generation of spontaneous activity, which can directly evoke neuropathic pain. Nerve injury usually triggers the synthesis of large quantities of membrane protein in nerve injured DRG neurons. Membrane proteins are glycosylated by addition of sugars, especially negatively charged sialic acid residues, which may depolarize the resting membrane potential (Vm), open voltage-gated channels in injured neurons, and cause spontaneous activity. In the present study, we aimed to determine if increased negative charge on the cell surface, carried by the sialic acid residues, could contribute to the generation of SMPO in injured DRG neurons. Intracellular recording was performed in DRG neurons following chronic constrictive injury (CCI) of the sciatic nerve. Results indicated that both A- and C-type injured DRG neurons exhibited a higher incidence of SMPO and more depolarized Vm than those of the control neurons. Ca(2+), Mg(2+), Mn(2+), or poly-lysine, a positively charged organic compound, when topically applied to the DRG, not only reduced SMPO but also caused a rapid hyperpolarizing shift in Vm. Topical application of neuraminidase to selectively remove sialic acid residues on the extracellular membrane normalized the depolarized Vm and inhibited both spontaneous and evoked SMPO. However, application of Ca(2+), Mg(2+), Mn(2+) or neuraminidase had no effect on excitability and Vm in normal neurons. The results demonstrated that the increase in negatively charged sialic acid residues on the extracellular membrane of neuronal somata is a critical factor in the generation of SMPO and hyperexcitability in injured sensory neurons.

  14. Gamma knife irradiation of injured sciatic nerve induces histological and behavioral improvement in the rat neuropathic pain model.

    Directory of Open Access Journals (Sweden)

    Yuki Yagasaki

    Full Text Available We examined the effects of gamma knife (GK irradiation on injured nerves using a rat partial sciatic nerve ligation (PSL model. GK irradiation was performed at one week after ligation and nerve preparations were made three weeks after ligation. GK irradiation is known to induce immune responses such as glial cell activation in the central nervous system. Thus, we determined the effects of GK irradiation on macrophages using immunoblot and histochemical analyses. Expression of Iba-1 protein, a macrophage marker, was further increased in GK-treated injured nerves as compared with non-irradiated injured nerves. Immunohistochemical study of Iba-1 in GK-irradiated injured sciatic nerves demonstrated Iba-1 positive macrophage accumulation to be enhanced in areas distal to the ligation point. In the same area, myelin debris was also more efficiently removed by GK-irradiation. Myelin debris clearance by macrophages is thought to contribute to a permissive environment for axon growth. In the immunoblot study, GK irradiation significantly increased expressions of βIII-tubulin protein and myelin protein zero, which are markers of axon regeneration and re-myelination, respectively. Toluidine blue staining revealed the re-myelinated fiber diameter to be larger at proximal sites and that the re-myelinated fiber number was increased at distal sites in GK-irradiated injured nerves as compared with non-irradiated injured nerves. These results suggest that GK irradiation of injured nerves facilitates regeneration and re-myelination. In a behavior study, early alleviation of allodynia was observed with GK irradiation in PSL rats. When GK-induced alleviation of allodynia was initially detected, the expression of glial cell line-derived neurotrophic factor (GDNF, a potent analgesic factor, was significantly increased by GK irradiation. These results suggested that GK irradiation alleviates allodynia via increased GDNF. This study provides novel evidence that GK

  15. Investigation into the potential of sub-lethal photodynamic antimicrobial chemotherapy (PACT) to reduce susceptibility of meticillin-resistant Staphylococcus aureus (MRSA) to antibiotics

    Science.gov (United States)

    Cassidy, C. M.; Donnelly, R. F.; Tunney, M. M.

    2009-06-01

    In PACT, a combination of a sensitising drug and visible light cause the selective destruction of microbial cells via singlet oxygen production. As singlet oxygen is a non-specific oxidizing agent and is only present during illumination, development of resistance to this treatment is thought to be unlikely. However, in response to oxidative stress, bacteria can up-regulate oxidative stress genes and associated antibiotic resistance genes. The up-regulation of these genes and potential transfer of genetic material may result in a resistant bacterial population. This study determined whether treatment of clinically isolated meticillin resistant Staphylococcus aureus (MRSA) strains with sub-lethal doses of methylene blue (MB) and meso-tetra (N-methyl-4-pyridyl) porphine tetra tosylate (TMP)-PACT resulted in reduced susceptibility to antibiotics and previously lethal PACT. Exposure of strains to sub-lethal doses of photosensitizer in combination with light had no effect on susceptibility to previously lethal photosensitization. Furthermore, exposure to sub-lethal concentrations of both photosensitizers caused no significant changes in the minimum inhibitory concentration (MIC) for each strain tested. Any differences in susceptibility were not significant as they did not cross breakpoints between resistant and susceptible for any organism or antibiotic tested. Therefore, PACT remains an attractive alternative option for treatment of MRSA infections.

  16. Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, Catherine J., E-mail: cjwalsh@mote.org [Marine Immunology Program, Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236 (United States); Butawan, Matthew, E-mail: mattbutawan@outlook.com [Marine Immunology Program, Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236 (United States); Yordy, Jennifer, E-mail: jennifer.e.balmer@gmail.com [Marine Immunology Program, Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236 (United States); Ball, Ray, E-mail: Ray.Ball@lowryparkzoo.com [Lowry Park Zoo, 1101 W Sligh Ave, Tampa, FL 33604 (United States); Flewelling, Leanne, E-mail: Leanne.Flewelling@MyFWC.com [Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission, 100 8th Ave SE, St. Petersburg, FL 33701 (United States); Wit, Martine de, E-mail: Martine.deWit@MyFWC.com [Fish and Wildlife Research Institute, Florida Fish and Wildlife Conservation Commission, 100 8th Ave SE, St. Petersburg, FL 33701 (United States); Bonde, Robert K., E-mail: rbonde@usgs.gov [U.S. Geological Survey, Sirenia Project, 7920 NE 71st Street, Gainesville, FL 32653 (United States)

    2015-04-15

    Highlights: • Sublethal brevetoxin exposure affects manatee immune function. • Plasma brevetoxin levels correlate with oxidative stress in rescued manatees. • Brevetoxin exposure affects lymphocyte proliferation in rescued manatees. • Plasma brevetoxin concentrations ranged from 0 to 19 ng PbTx-3 eq/mL. - Abstract: The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected by exposure to blooms of the toxic dinoflagellate, Karenia brevis. K. brevis blooms are common in manatee habitats of Florida’s southwestern coast and produce a group of cyclic polyether toxins collectively referred to as red tide toxins, or brevetoxins. Although a large number of manatees exposed to significant levels of red tide toxins die, several manatees are rescued from sublethal exposure and are successfully treated and returned to the wild. Sublethal brevetoxin exposure may potentially impact the manatee immune system. Lymphocyte proliferative responses and a suite of immune function parameters in the plasma were used to evaluate effects of brevetoxin exposure on health of manatees rescued from natural exposure to red tide toxins in their habitat. Blood samples were collected from rescued manatees at Lowry Park Zoo in Tampa, FL and from healthy, unexposed manatees in Crystal River, FL. Peripheral blood leukocytes (PBL) isolated from whole blood were stimulated with T-cell mitogens, ConA and PHA. A suite of plasma parameters, including plasma protein electrophoresis profiles, lysozyme activity, superoxide dismutase (SOD) activity, and reactive oxygen/nitrogen (ROS/RNS) species, was also used to assess manatee health. Significant decreases (p < 0.05) in lymphocyte proliferation were observed in ConA and PHA stimulated lymphocytes from rescued animals compared to non-exposed animals. Significant correlations were observed between oxidative stress markers (SOD, ROS/RNS) and plasma brevetoxin concentrations. Sublethal exposure to brevetoxins in the

  17. Multi-ligand nanoparticles for targeted drug delivery to the injured vascular wall

    Science.gov (United States)

    Kona, Soujanya

    Pathological conditions like coronary artery disease, acute myocardial infarction, stroke, and peripheral artery diseases as well as cardiovascular interventions used in the treatment of coronary artery diseases such as angioplasty and stenting damage/injure the blood vessel wall, leading to inflamed or activated endothelial cells that have been implicated in events leading to thrombosis, inflammation, and restenosis. Oral administration of anti-coagulant and anti-inflammatory drugs causes systemic toxicity, bleeding, patient incompliance, and inadequate amounts of drugs at the injured area. Though drug-eluting stents have shown therapeutic benefits, complications such as in-stent restenosis and late thrombosis still remain and are a cause for concern. Rapid growth in the field of nanotechnology and nanoscience in recent years has paved the way for new targeted and controlled drug delivery strategies. In this perspective, the development of biodegradable nanoparticles for targeted intracellular drug delivery to the inflamed endothelial cells may offer an improved avenue for treatment of cardiovascular diseases. The major objective of this research was to develop "novel multi-ligand nanoparticles," as drug carriers that can efficiently target and deliver therapeutic agents to the injured/inflamed vascular cells under dynamic flow conditions. Our approach mimics the natural binding ability of platelets to injured/activated endothelial cells through glycoprotein Ib (GPIb) bound to P-selectin expressed on inflamed endothelial cells and to the subendothelium through GPIb binding to von Willebrand factor (vWF) deposited onto the injured vascular wall. Our design also exploits the natural cell membrane translocation ability of the internalizing cell peptide - trans-activating transcriptor (TAT) to enhance the nanoparticle uptake by the targeted cells. Our hypothesis is that these multi-ligand nanoparticles would show an increased accumulation at the injury site since GPIb

  18. Sublethal effects of atrazine on embryo-larval development of Rhinella arenarum (Anura: Bufonidae).

    Science.gov (United States)

    Svartz, Gabriela V; Herkovits, Jorge; Pérez-Coll, Cristina S

    2012-05-01

    Atrazine (ATR), one of the most widely used herbicides in the world, affects not only target organisms but also the biota in general. Here, the teratogenic and neurotoxic effects of ATR on Rhinella arenarum (South American toad) embryos, and larvae were evaluated by means of standardized bioassays during acute and chronic exposures. The herbicide had a significant incidence of malformations, with a Teratogenic Index (TI) of 3.28. The main effects were delayed development, reduced body size, microcephaly, axial flexures, wavy tail and edema. In addition, delayed development, reduced development of forelimbs, and edema were recorded at metamorphosis stages. Scanning electron microscopy allowed observing different degrees of cellular dissociation and persistent cilliar cells in specific regions like the adhesive structure and tail fin. Results obtained by ATR 24 h pulse exposures at six developmental stages pointed out blastula as the most susceptible developmental stage both for immediate and delayed adverse effects. A noteworthy recovery capacity from acute toxic effects was recorded from the neural plate stage onwards. Regarding neurotoxic effects, abnormal, and erratic swimming and spasmodic contractions were recorded. Both the teratogenic and neurotoxic effects reported in this study demonstrate the importance of evaluating sublethal effects in non-target organisms as they could imply reduced fitness of individuals and eventually a population decline. The Hazard Quotients (HQ) for ATR ranged from 0.14 to 10.80, and the fact that some of these values are above USEPA's level of concern indicate that ATR is likely a risk to R. arenarum.

  19. Sublethal toxicity and biotransformation of pyrene in Lumbriculus variegatus (Oligochaeta)

    Energy Technology Data Exchange (ETDEWEB)

    Maeenpaeae, K. [Faculty of Biosciences, University of Joensuu, FIN-80101 Joensuu (Finland)], E-mail: kimmo.maenpaa@joensuu.fi; Leppaenen, M.T.; Kukkonen, J.V.K. [Faculty of Biosciences, University of Joensuu, FIN-80101 Joensuu (Finland)

    2009-04-01

    The aim of this work was to study the toxicity and biotransformation of polyaromatic hydrocarbon (PAH) pyrene in the oligochaete aquatic worm, Lumbriculus variegatus. PAHs are ubiquitous environmental pollutants that pose a hazard to aquatic organisms, and metabolizing capability is poorly known in the case of many invertebrate species. To study the toxicity and biotransformation of pyrene, the worm was exposed for 15 days to various concentrations of water-borne pyrene. The dorsal blood vessel pulse rate was used as a sublethal endpoint. Pyrene biotransformation by L. variegatus was studied and the critical body residues (CBR) were estimated for pyrene toxicity. The toxicokinetics of pyrene uptake was evaluated. A combination of radiolabeled ({sup 14}C) and nonlabeled pyrene was used in the exposures, and liquid scintillation counting (LSC) and high-pressure liquid chromatography were employed in both water and tissue residue analyses. The results showed that L. variegatus was moderately able to metabolize pyrene to 1-hydroxypyrene (1-HP), thus demonstrating that the phase-I-like oxidizing enzyme system metabolizes pyrene in L. variegatus. The amount of the 1-HP was 1-2% of the amount of pyrene in the worm tissues. The exposure to pyrene reduced the blood vessel pulse rate significantly (p < 0.05), showing that pyrene had a narcotic effect. The estimated CBRs remained constant during the exposure time, varying from 0.120 to 0.174 mmol pyrene/kg worm wet weight. The bioconcentration factors (BCF) decreased as exposure concentration increased. It was suggested that the increased toxicity of pyrene accounted for the decrease in BCFs by lowering the activity of the organism.

  20. Psychoactive substances in seriously injured drivers in Denmark

    DEFF Research Database (Denmark)

    Simonsen, Kirsten Wiese; Steentoft, Anni; Bernhoft, Inger Marie

    2013-01-01

    found to be above the Danish legal limit in 4.9% of injured drivers. Young men (median age 31 years) were over-represented among injured drivers who violated Danish law for alcohol and drugs. Diazepam (4.4%), tramadol (3.2%), and clonazepam (3.0%) were the medicinal drugs most frequently detected...

  1. Injured pedestrians in Cape Town - the role of alcohol

    African Journals Online (AJOL)

    alcohol. Margaret M. Peden, John D. Knotlenbelt,. Johan van der Spuy, Ravi Oodit, Hendrik J. Scholtz,. J. Morris Stokol. Objective. To establish a profile of injured adult pedestrians and attempt to define the role which alcohol plays in this regard. Design. Prospective survey of injured pedestrians who presented consecutively ...

  2. Wound healing process of injured pulp tissues with emdogain gel.

    Science.gov (United States)

    Kaida, Hikaru; Hamachi, Takafumi; Anan, Hisashi; Maeda, Katsumasa

    2008-01-01

    This study aimed to investigate the wound healing process of injured pulp tissues with Emdogain gel (EMD). Pulpotomy was performed for the first molars of the mandibles in rats. EMD or Vitapex (VIT)-containing calcium hydroxide was applied to the exposed pulp tissues. The treated teeth were extracted after 7, 14, and 28 days and prepared for histologic examination. In the VIT-treated group, the number of interleukin-1 beta (IL-1 beta)-expressing macrophages initially increased, followed by that of transforming growth factor-beta1 (TGF-beta1)-expressing macrophages. The number of cells expressing bone morphogenetic proteins (BMPs) gradually increased with reparative dentin formation. Meanwhile, in the EMD-treated group, cells expressing IL-1 beta or TGF-beta1 were few. However, the number of BMP-expressing cells, partly macrophages, increased in the early phase, and large amounts of reparative dentin were observed. This study demonstrated that different healing processes existed for EMD and VIT. BMP-expressing macrophages might play important roles in reparative dentin formation.

  3. Optimizing detection of heat-injured Listeria monocytogenes in pasteurized milk.

    Science.gov (United States)

    Teo, A Y; Ziegler, G R; Knabel, S J

    2001-07-01

    Optimal conditions for the detection of heat-injured cells of Listeria monocytogenes in modified Pennsylvania State University (mPSU) broth were determined using a response surface design generated by a computer program, EChip. Different combinations of incubation temperatures and lithium, magnesium, and D-serine concentrations were evaluated to determine the optimum conditions for the detection of heat-injured L. monocytogenes in filter-sterilized whole milk inoculated with selected problematic background microflora. A concentration of 212 mM lithium chloride completely inhibited the growth of Enterococcus faecium while permitting recovery and detection of L. monocytogenes. A concentration of 15.8 mM MgSO4 was found to be optimum for the recovery and detection of L. monocytogenes. A concentration of 140.2 mM D-serine was found to completely inhibit the germination of Bacillus subtilis var. globii spores but not recovery and detection of L. monocytogenes. Under optimum concentrations of LiCl, MgSO4, and D-serine and in the absence of background microflora, the effect of incubation temperature on percentage detection was described by a second-order polynomial model, and 28 degrees C was determined to be optimal. In the presence of background microflora, the effect of incubation temperature on percentage detection of heat-injured cells was described by a third-order polynomial model, and 30 degrees C was found to be optimal. Optimizing the levels of highly specific and selective agents, nutrients, and incubation temperature in one recovery enrichment system dramatically increased the Listeria/background microflora ratio. This resulting medium, optimized PSU (oPSU) broth, greatly improved the detection of heat-injured and nonheat-injured L. monocytogenes by both conventional and molecular methods (Oxoid's Listeria Rapid Test, Gen-Probe's Accuprobe Listeria monocytogenes Culture Identification Test, and Qualicon's BAX for screening Listeria monocytogenes).

  4. Can molecular motors drive distance measurements in injured neurons?

    Directory of Open Access Journals (Sweden)

    Naaman Kam

    2009-08-01

    Full Text Available Injury to nerve axons induces diverse responses in neuronal cell bodies, some of which are influenced by the distance from the site of injury. This suggests that neurons have the capacity to estimate the distance of the injury site from their cell body. Recent work has shown that the molecular motor dynein transports importin-mediated retrograde signaling complexes from axonal lesion sites to cell bodies, raising the question whether dynein-based mechanisms enable axonal distance estimations in injured neurons? We used computer simulations to examine mechanisms that may provide nerve cells with dynein-dependent distance assessment capabilities. A multiple-signals model was postulated based on the time delay between the arrival of two or more signals produced at the site of injury-a rapid signal carried by action potentials or similar mechanisms and slower signals carried by dynein. The time delay between the arrivals of these two types of signals should reflect the distance traversed, and simulations of this model show that it can indeed provide a basis for distance measurements in the context of nerve injuries. The analyses indicate that the suggested mechanism can allow nerve cells to discriminate between distances differing by 10% or more of their total axon length, and suggest that dynein-based retrograde signaling in neurons can be utilized for this purpose over different scales of nerves and organisms. Moreover, such a mechanism might also function in synapse to nucleus signaling in uninjured neurons. This could potentially allow a neuron to dynamically sense the relative lengths of its processes on an ongoing basis, enabling appropriate metabolic output from cell body to processes.

  5. DUI histories in intoxicated injured bicyclists.

    Science.gov (United States)

    Maximus, Steven; Figueroa, Cesar; Pham, Jacqueline; Kuncir, Eric; Barrios, Cristobal

    2016-10-01

    It has been well documented that the use of alcohol correlates with injury risk, especially in DUI (driving under the influence) and DWI (driving while intoxicated). Consumption of alcohol in patients presenting with bicycle-related injuries is associated with greater injury severity, longer hospitalization, and higher health care costs. We hypothesized that intoxicated patients operating a bicycle with traumatic injuries have previous DUI or DWI convictions and had lost their privilege to drive a motor vehicle, resorted to bicycling, and had continued alcohol consumption despite negative previous consequences. We retrospectively collected data on injured bicyclists older than 18 years with positive blood alcohol content levels treated from the period January 2009 to June 2014 at a large Level 1 urban trauma center. We then matched each patient by name and date of birth and were able to obtain public criminal records through the Superior Court of California for the local of county. A total of 149 injured bicyclists with positive blood alcohol levels were identified. Their average blood alcohol content was 236.0 mg/dL, and their average age was 41 years. Sixty-six (44.2%) of these patients had prior DUI/DWI convictions with suspension of driving privileges. Ninety-five patients in this group (63.8%) had no health insurance, and 51 patients (34.2%) tested positive for other drugs. Intoxicated bicyclists trended toward longer hospital length compared with nonintoxicated bicyclists (4.60 vs. 3.44 days; p = 0.07). Three (0.02%) of 149 patients were charged with bicycling while intoxicated. Intoxicated bicyclists involved in trauma are more likely to have a previous DUI/DWI, have other drug use, tend to have longer hospital stays, and are less likely to have insurance. Bicycle safety education and behavior modification targeting DUI/DWI offenders are warranted. In order to promote injury prevention, resources to increase awareness of this underestimated public health

  6. Supporting data for comparative proteomic analysis of Listeria monocytogenes ATCC 7644 exposed to a sublethal concentration of nisin

    Directory of Open Access Journals (Sweden)

    Kendi Nishino Miyamoto

    2015-06-01

    Full Text Available Here we provide the LC–MS/MS data from a comparative analysis of Listeria monocytogenes ATCC 7644 treated and non-treated with a sublethal concentration of nisin (10−3 mg/mL. Protein samples were analyzed by multidimensional protein identification technology (MudPIT approach, in an off-line configuration. The raw MS/MS data allowed the detection of 49,591 spectra which resulted in 576 protein identifications. After Scaffold validation, 179 proteins were identified with high confidence. A label-free quantitative analysis based of normalized spectral abundance factor (NSAF was used and 13 proteins were found differentially expressed between nisin-treated and non-treated cells. Gene ontology analysis of differentially expressed proteins revealed that most of them are correlated to metabolic process, oxidative stress response mechanisms and molecular binding. A detailed analysis and discussion of these data may be found in Miyamoto et al. [1].

  7. Supporting data for comparative proteomic analysis of Listeria monocytogenes ATCC 7644 exposed to a sublethal concentration of nisin

    Science.gov (United States)

    Miyamoto, Kendi Nishino; Mariante Monteiro, Karina; da Silva Caumo, Karin; Rodrigues Lorenzatto, Karina; Bunselmeyer Ferreira, Henrique; Brandelli, Adriano

    2015-01-01

    Here we provide the LC–MS/MS data from a comparative analysis of Listeria monocytogenes ATCC 7644 treated and non-treated with a sublethal concentration of nisin (10−3 mg/mL). Protein samples were analyzed by multidimensional protein identification technology (MudPIT) approach, in an off-line configuration. The raw MS/MS data allowed the detection of 49,591 spectra which resulted in 576 protein identifications. After Scaffold validation, 179 proteins were identified with high confidence. A label-free quantitative analysis based of normalized spectral abundance factor (NSAF) was used and 13 proteins were found differentially expressed between nisin-treated and non-treated cells. Gene ontology analysis of differentially expressed proteins revealed that most of them are correlated to metabolic process, oxidative stress response mechanisms and molecular binding. A detailed analysis and discussion of these data may be found in Miyamoto et al. [1]. PMID:26217729

  8. Qigong in Injured Military Service Members.

    Science.gov (United States)

    Reb, Anne Marie; Saum, Nancy Seaby; Murphy, Deborah Ann; Breckenridge-Sproat, Sara Todd; Su, Xiaogang; Bormann, Jill Ellen

    2017-03-01

    Wounded, ill, and injured (WII) Military Service members experience significant stress and are at risk for developing chronic conditions including posttraumatic stress disorder and depression. Qigong, a meditative movement practice, may positively affect their ability to engage in successful rehabilitation. We assessed the feasibility of Qigong practice in WII Service members returning from combat; effects on stress, sleep, and somatic symptoms; satisfaction; and participants' experience with the practice. Single-group, pre- and posttest, mixed methods approach. Twenty-six WII were enrolled. The program was designed to include 20 classes over 10 weeks. Participants completed self-report questionnaires, practice logs, and an exit interview. Average attendance was 8.14 classes ( SD = 4.9); mean engagement was 5.7 ( SD = 3.5) weeks. Participants endorsed a high level of satisfaction with the practice. Qualitative themes included coping with stress; feeling more resilient and empowered; improvement in symptoms including sleep and physical function; and factors affecting practice. Participant-reported facilitators included accessibility and portability of the practice; barriers included scheduling conflicts and personal challenges. Participants recommended offering shorter programs with flexible scheduling options, increasing program awareness, and including significant others in future classes. Qigong was safe, portable, and easily adapted for WII Service members.

  9. Sublethal effects of some synthetic and botanical insecticides on Bemisia tabaci (Hemiptera: Aleyrodidae

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    Esmaeily Saeideh

    2014-07-01

    Full Text Available In addition to direct mortality caused by insecticides, some biological traits of insects may also be affected by sublethal insecticide doses. In this study, we used the age-stage, two-sex life table method to evaluate the sublethal effects of the four synthetic insecticides: abamectin, imidacloprid, diazinon, and pymetrozin as well as the botanical insecticide taken from Calotropis procera (Asclepiadaceae extract, on eggs of the cotton whitefly, Bemisia tabaci (Hem.: Aleyrodidae. The lowest and highest survival rates and oviposition periods were observed in whiteflies treated by diazinon and imidacloprid, respectively. We found significant differences in the net reproductive rate (R0, the intrinsic rate of increase (r, the finite rate of increase (?, and the gross reproductive rate (GRR among different insecticides. Altogether, our results showed that pymetrozin and C. procera induced the most sublethal effects, thus they may be suitable candidates for use in integrated pest management programs of B. tabaci.

  10. Sublethal Dosage of Imidacloprid Reduces the Microglomerular Density of Honey Bee Mushroom Bodies

    Science.gov (United States)

    Peng, Yi-Chan; Yang, En-Cheng

    2016-01-01

    The dramatic loss of honey bees is a major concern worldwide. Previous studies have indicated that neonicotinoid insecticides cause behavioural abnormalities and have proven that exposure to sublethal doses of imidacloprid during the larval stage decreases the olfactory learning ability of adults. The present study shows the effect of sublethal doses of imidacloprid on the neural development of the honey bee brain by immunolabelling synaptic units in the calyces of mushroom bodies. We found that the density of the synaptic units in the region of the calyces, which are responsible for olfactory and visual functions, decreased after being exposed to a sublethal dose of imidacloprid. This not only links a decrease in olfactory learning ability to abnormal neural connectivity but also provides evidence that imidacloprid damages the development of the nervous system in regions responsible for both olfaction and vision during the larval stage of the honey bee. PMID:26757950

  11. Sub-lethal antibiotic treatment leads to multidrug resistance via radical-induced mutagenesis

    Science.gov (United States)

    Kohanski, Michael A.; DePristo, Mark A.; Collins, James J.

    2010-01-01

    Summary Antibiotic resistance arises through mechanisms such as selection of naturally occurring resistant mutants and horizontal gene transfer. Recently, oxidative stress has been implicated as one of the mechanisms whereby bactericidal antibiotics kill bacteria. Here we show that sub-lethal levels of bactericidal antibiotics induce mutagenesis, resulting in heterogeneous increases in the minimum inhibitory concentration for a range of antibiotics, irrespective of the drug target. This increase in mutagenesis correlates with an increase in ROS, and is prevented by the ROS scavenger thiourea and by anaerobic conditions, indicating that sub-lethal concentrations of antibiotics induce mutagenesis by stimulating the production of ROS. We demonstrate that these effects can lead to mutant strains that are sensitive to the applied antibiotic but resistant to other antibiotics. This work establishes a radical-based molecular mechanism whereby sub-lethal levels of antibiotics can lead to multidrug resistance, which has important implications for the widespread use and misuse of antibiotics. PMID:20159551

  12. Sublethal concentrations of silver nanoparticles affect the mechanical stability of biofilms.

    Science.gov (United States)

    Grün, Alexandra Y; Meier, Jutta; Metreveli, George; Schaumann, Gabriele E; Manz, Werner

    2016-12-01

    Bacterial biofilms are most likely confronted with silver nanoparticles (Ag NPs) as a pollutant stressor in aquatic systems. In this study, biofilms of Aquabacterium citratiphilum were exposed for 20 h to 30 and 70 nm citrate stabilized Ag NPs in low-dose concentrations ranging from 600 to 2400 μg l(-1), and the Ag NP-mediated effects on descriptive, structural, and functional biofilm characteristics, including viability, protein content, architecture, and mechanical stability, were investigated. Viability, based on the bacterial cell membrane integrity of A. citratiphilum, as determined by epifluorescence microscopy, remained unaffected after Ag NP exposure. Moreover, in contrast to information in the current literature, protein contents of cells and extracellular polymeric substances (EPS) and biofilm architecture, including dry mass, thickness, and density, were not significantly impacted by exposure to Ag NPs. However, the biofilms themselves served as effective sinks for Ag NPs, exhibiting enrichment factors from 5 to 8. Biofilms showed a greater capacity to accumulate 30 nm sized Ag NPs than 70 nm Ag NPs. Furthermore, Ag NPs significantly threatened the mechanical stability of biofilms, as determined by a newly developed assay. For 30 nm Ag NPs, the mechanical stability of biofilms decreased as the Ag NP concentrations applied to them increased. In contrast, 70 nm Ag NPs produced a similar decrease in mechanical stability for each applied concentration. Overall, this finding demonstrates that exposure to Ag NPs triggers remarkable changes in biofilm adhesion and/or cohesiveness. Because of biofilm-mediated ecological services, this response raises environmental concerns regarding Ag NP release into freshwater systems, even in sublethal concentrations.

  13. Metallothionein deficiency in the injured peripheral nerves of complex regional pain syndrome as revealed by proteomics.

    Science.gov (United States)

    Oki, Gosuke; Wada, Takuro; Iba, Kosuke; Aiki, Hikono; Sasaki, Kouichi; Imai, Shin-ichi; Sohma, Hitoshi; Matsumoto, Kayo; Yamaguchi, Mami; Fujimiya, Mineko; Yamashita, Toshihiko; Kokai, Yasuo

    2012-03-01

    Complex regional pain syndrome (CRPS) is characterized by persistent and severe pain after trauma or surgery; however, its molecular mechanisms in the peripheral nervous system are poorly understood. Using proteomics, we investigated whether injured peripheral nerves of CRPS patients have altered protein profiles compared with control nerves. We obtained nerve samples from 3 patients with CRPS-2 who underwent resection of part of an injured peripheral nerve. Sural nerves from fresh cadavers with no history of trauma or neuropathic pain served as controls. Proteomic analysis showed that the number and functional distribution of proteins expressed in CRPS and control nerves was similar. Interestingly, metallothionein was absent in the injured nerves of CRPS-2, although it was readily detected in control nerves. Western blotting further confirmed the absence of metallothionein in CRPS-2 nerves, and immunohistochemistry corroborated the deficiency of metallothionein expression in injured nerves from 5 of 5 CRPS patients and 2 of 2 patients with painful neuromas. In contrast, all control nerves, including 5 sural nerves from fresh cadavers and 41 nerves obtained from surgically resected tumors, expressed MT. Furthermore, expression of S100 as a marker for Schwann cells, and neurofilament M as a marker of axons was comparable in both CRPS-2 and controls. Metallothioneins are zinc-binding proteins that are probably involved in protection against injury and subsequent regeneration after CNS damage. Their absence from the injured peripheral nerves of patients with CRPS-2 suggests a potential pathogenic role in generating pain in the damaged peripheral nerves. Copyright © 2011 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.

  14. Matrix metalloproteinase-9 facilitates glial scar formation in the injured spinal cord

    OpenAIRE

    Hsu, Jung-Yu C.; Bourguignon, Lilly Y.W.; Adams, Christen M.; Peyrollier, Karine; Zhang, Haoqian; Fandel, Thomas; Cun, Christine L.; Werb, Zena; Noble-Haeusslein, Linda J.

    2008-01-01

    In the injured spinal cord, a glial scar forms and becomes a major obstacle to axonal regeneration. Formation of the glial scar involves migration of astrocytes toward the lesion. Matrix metalloproteinases (MMPs), including MMP-9 and MMP-2, govern cell migration through their ability to degrade constituents of the extracellular matrix. Although MMP-9 is expressed in reactive astrocytes, its involvement in astrocyte migration and formation of a glial scar is unknown. Here we found that spinal ...

  15. Vertical transmission of sublethal granulovirus infection in the Indian meal moth, Plodia interpunctella.

    Science.gov (United States)

    Burden, J P; Griffiths, C M; Cory, J S; Smith, P; Sait, S M

    2002-03-01

    Knowledge of the mechanisms of pathogen persistence in relation to fluctuations in host density is crucial to our understanding of disease dynamics. In the case of insect baculoviruses, which are typically transmitted horizontally via a lifestage that can persist outside the host, a key issue that remains to be elucidated is whether the virus can also be transmitted vertically as a sublethal infection. We show that RNA transcripts for the Plodia interpunctella GV granulin gene are present in a high proportion of P. interpunctella insects that survive virus challenge. Granulin is a late-expressed gene that is only transcribed after viral genome replication, its presence thus strongly indicates that viral genome replication has occurred. Almost all insects surviving the virus challenge tested positive for viral RNA in the larval and pupal stage. However, this proportion declined in the emerging adults. Granulin mRNA was also detected in both the ovaries and testes, which may represent a putative mechanism by which reduced fecundity in sublethally affected hosts might be manifested. RNA transcripts were also detected in 60-80% of second-generation larvae that were derived from mating surviving adults, but there was no difference between the sexes, with both males and females capable of transmitting a sublethal infection to their offspring. The data indicate that low-level persistent infection, with at least limited gene expression, can occur in P. interpunctella following survival of a granulovirus challenge. We believe that this is the first demonstration of a persistent, sublethal infection by a baculovirus to be initiated by a sublethal virus dose. We hypothesize that the 'latent' baculovirus infections frequently referred to in the literature may also be low level persistent, sublethal infections resulting from survival from initial baculovirus exposure.

  16. Sublethal Effects of Thiamethoxam on the Demographic Parameters of Myzus persicae (Hemiptera: Aphididae).

    Science.gov (United States)

    Wang, Pan; Zhou, Li-Lin; Yang, Fan; Li, Mang; Liu, Xiao-Ming; Wang, Yong; Lei, Chao-Liang; Si, Sheng-Yun

    2017-08-01

    The green peach aphid, Myzus persicae Sulzer (Hemiptera: Aphididae), is an important sap-sucking pest of many crops, including Chinese cabbage, Brassinca oleracea L. The neonicotinoid insecticide thiamethoxam has been used as an effective insecticide to control M. persicae in cabbage fields. In this study, we assessed the effects of sublethal concentrations of thiamethoxam on demographic parameters of M. persicae. In leaf-dip bioassays, thiamethoxam showed a relatively high toxicity against M. persicae with an LC50 of 6.80 mg liter-1. The duration of the preadult stage was not significantly affected in the sublethal bioassay. Additionally, the longevity and adult preoviposition period were not significantly affected by sublethal thiamethoxam. However, sublethal thiamethoxam significantly increased fecundity (LC10) and prolonged the total preoviposition period (LC40). Consequently, the finite rate of increase (λ) and the intrinsic rate of increase (rm) of aphids exposed to the LC40 were significantly lower than those of control aphids, whereas the net reproductive rate (R0) was higher, and the generation time (T) and the population doubling time (DT) were longer in the treated group. Based on these results, hormesis was induced by sublethal thiamethoxam in M. persicae, with the population growth of M. persicae negatively affected at higher sublethal concentrations of thiamethoxam. Therefore, our study indicated that the possible effects of thiamethoxam on aphids require further study to develop optimized integrated pest management strategies. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. An experimental model for the transplantation of fetal central nervous system cells to the injured spinal cord in rats Modelo experimental de transplante de células do sistema nervoso central fetal para lesão de medula espinal em ratos

    Directory of Open Access Journals (Sweden)

    Tarcísio Eloy Pessoa de Barros Filho

    2002-01-01

    Full Text Available INTRODUCTION: Traumatic spinal cord injury is one of the most disabling conditions occurring in man and thus stimulates a strong interest in its histopathological, biochemical, and functional changes, primarily as we search for preventive and therapeutic methods. PURPOSE: To develop an experimental model for transplantation of cells from the fetal rat central nervous system to the site of an injured spinal cord of an adult rat in which the transplanted cells survive and become integrated. This experimental model will facilitate investigations of factors that promote regeneration and functional recovery after spinal cord trauma. MATERIAL AND METHODS: Fifteen adult Wistar rats underwent laminectomy, and an spinal cord lesion was made with microdissection. Fetal spinal cord tissue was then transplanted to the site of the injury. The rats were monitored over a 48-hour period, and then their vertebral column was completely removed for histological analysis. RESULTS: In 60% of transplanted rats, the fetal tissue at the injured site remained viable in the site of the lesion.INTRODUÇÃO: A lesão traumática da medula espinal consiste numa das mais incapacitantes lesões que o ser humano pode sofrer e tem despertado grande interesse no conhecimento das alterações histopatológicas, bioquímicas, funcionais e principalmente na busca de métodos de prevenção e tratamento. OBJETIVO: Propor um modelo experimental de transplante de células do sistema nervoso fetal de ratos para o sítio de lesão medular de ratos adultos que permitisse sua sobrevivência e integração para possibilitar protocolos de pesquisa para identificar outros fatores de regeneração e recuperação funcional pós trauma raquimedular. MATERIAL E MÉTODOS: Utilizaram-se 15 ratos adultos que foram submetidos a laminectomia e lesão de 5mm de hemimelula realizada com auxílio de microscópio óptico. Os ratos tiveram seu sítio de lesão medular transplantado com células do

  18. Expression and function of Toll-like receptor 9 in severely injured patients prone to sepsis

    Science.gov (United States)

    Baiyee, E E; Flohe, S; Lendemans, S; Bauer, S; Mueller, N; Kreuzfelder, E; Grosse-Wilde, H

    2006-01-01

    The purpose of this prospective study was to enumerate Toll-like receptor 9 (TLR9)+ cells and measure their function using synthetic oligonucleotides enriched in CG dinucleotide motifs (CpG)-induced proliferation within 48 h after trauma in severely injured patients prone to sepsis. Sixteen consecutive trauma patients with an injury severity score (ISS) > 21 and 16 blood donors (controls) were included in this study. Using two-colour flow cytometry, TLR9 expression was detectable intracellularly and also on the surface of B lymphocytes. The surface expression of TLR9 of B lymphocytes from whole blood and peripheral blood mononuclear cells (PBMC) stimulated with CpG was significantly increased in B cells of severely injured patients prone to sepsis compared to controls. No significant differences could be observed between CpG-induced proliferation of PBMC of severely injured patients prone to sepsis and controls. As a measure of immunosuppression, human leucocyte antigen (HLA)-DR expression of monocytes of the trauma patients was significantly diminished compared with controls in PBMC and in whole blood. Immunosuppression in the early phase after trauma seems not to be associated with a disturbed sensing of bacterial DNA. PMID:16907913

  19. Trauma in children injured by physical violence

    Directory of Open Access Journals (Sweden)

    Karina S Solovyova

    2016-09-01

    Full Text Available Introduction. In recent years, legislation has changed to include the rights of children injured because of physical violence. Trauma departments of St. Petersburg outpatient clinics admit children with injuries of varying severity after physical violence. The actions of medical institutions are always aimed at protecting the child. Aims. The aim of the present study was to analyze the cases of children in connection with injuries sustained as a result of physical violence in 2014–2015, and to compare the results with those of previous studies (2007–2008. Material and methods. In 2014–2015, the trauma department of City Children's Outpatient clinic No 62 treated 268 children, who had suffered from physical violence at home, on the street, or in educational institutions, which accounted for 1.6 per 1000 children living in the district, and 1.2% of all children admitted during 2 years. Results. Compared to 2007–2008, the number of children who suffered from physical violence decreased by almost two times in 2014–2015; in addition, the severity of injuries slightly decreased but the frequency of hospital admission of victims remained high (38% in 2007–2008. With regard to the circumstances in which the injury occurred, violence from strangers was lower, but violence among peers was higher. Conclusions. Positive results have been achieved by a complex of measures, including the implementation of the Federal Law “On Basic Guarantees of the Rights of the Child” to improve the care and safety of children, and an investigation of each case of violence is conducted by local authorities for internal affairs.

  20. Extended hypoxia in the alfalfa leafcutting bee, Megachile rotundata, increases survival but causes sub-lethal effects.

    Science.gov (United States)

    Abdelrahman, H; Rinehart, J P; Yocum, G D; Greenlee, K J; Helm, B R; Kemp, W P; Schulz, C H; Bowsher, J H

    2014-05-01

    Many insects are tolerant of hypoxic conditions, but survival may come at a cost to long-term health. The alfalfa leaf-cutting bee, Megachile rotundata, develops in brood cells inside natural cavities, and may be exposed to hypoxic conditions for extended periods of time. Whether M. rotundata is tolerant of hypoxia, and whether exposure results in sub-lethal effects, has never been investigated. Overwintering M. rotundata prepupae were exposed to 10%, 13%, 17%, 21% and 24% O2 for 11 months. Once adults emerged, five indicators of quality - emergence weight, body size, feeding activity, flight performance, and adult longevity, - were measured to determine whether adult bees that survived past exposure to hypoxia were competent pollinators. M. rotundata prepupae are tolerant of hypoxic condition and have higher survival rates in hypoxia, than in normoxia. Under hypoxia, adult emergence rates did not decrease over the 11 months of the experiment. In contrast, bees reared in normoxia had decreased emergence rates by 8 months, and were dead by 11 months. M. rotundata prepupae exposed to extended hypoxic conditions had similar emergence weight, head width, and cross-thorax distance compared to bees reared in standard 21% oxygen. Despite no significant morphological differences, hypoxia-exposed bees had lower feeding rates and shorter adult lifespans. Hypoxia may play a role in post-diapause physiology of M. rotundata, with prepupae showing better survival under hypoxic conditions. Extended exposure to hypoxia, while not fatal, causes sub-lethal effects in feeding rates and longevity in the adults, indicating that hypoxia tolerance comes at a cost. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. Adenovirus vector-mediated ex vivo gene transfer of brain-derived neurotrophic factor (BDNF) tohuman umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) promotescrush-injured rat sciatic nerve regeneration.

    Science.gov (United States)

    Hei, Wei-Hong; Almansoori, Akram A; Sung, Mi-Ae; Ju, Kyung-Won; Seo, Nari; Lee, Sung-Ho; Kim, Bong-Ju; Kim, Soung-Min; Jahng, Jeong Won; He, Hong; Lee, Jong-Ho

    2017-03-16

    This study was designed toinvestigate the efficacy of adenovirus vector-mediated brain-derived neurotrophic factor (BDNF) ex vivo gene transfer to human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) in a rat sciatic nerve crush injury model. BDNF protein and mRNA expression after infection was checked through an enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR). Male Sprague-Dawley rats (200-250g, 6 weeks old) were distributed into threegroups (n=20 each): the control group, UCB-MSC group, and BDNF-adenovirus infected UCB-MSC (BDNF-Ad+UCB-MSC) group. UCB-MSCs (1×10 6 cells/10μl/rat) or BDNF-Ad+UCB-MSCs (1×10 6 cells/10μl/rat)were transplantedinto the rats at the crush site immediately after sciatic nerve injury. Cell tracking was done with PKH26-labeled UCB-MSCs and BDNF-Ad+UCB-MSCs (1×10 6 cells/10μl/rat). The rats were monitored for 4 weeks post-surgery. Results showed that expression of BDNF at both the protein and mRNA levels was higher inthe BDNF-Ad+UCB-MSC group compared to theUCB-MSC group in vitro.Moreover, BDNF mRNA expression was higher in both UCB-MSC group and BDNF-Ad+ UCB-MSC group compared tothe control group, and BDNF mRNA expression in theBDNF-Ad+UCB-MSC group was higher than inboth other groups 5days after surgeryin vivo. Labeled neurons in the dorsal root ganglia (DRG), axon counts, axon density, and sciatic function index were significantly increased in the UCB-MSC and BDNF-Ad+ UCB-MSCgroupscompared to the controlgroup four weeksaftercell transplantation. Importantly,the BDNF-Ad+UCB-MSCgroup exhibited more peripheral nerve regeneration than the other two groups.Our results indicate thatboth UCB-MSCs and BDNF-Ad+UCB-MSCscan improve rat sciatic nerve regeneration, with BDNF-Ad+UCB-MSCsshowing a greater effectthan UCB-MSCs. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Sub-lethal levels of electric current elicit the biosynthesis of plant secondary metabolites.

    Science.gov (United States)

    Kaimoyo, Evans; Farag, Mohamed A; Sumner, Lloyd W; Wasmann, Catherine; Cuello, Joel L; VanEtten, Hans

    2008-01-01

    Many secondary metabolites that are normally undetectable or in low amounts in healthy plant tissue are synthesized in high amounts in response to microbial infection. Various abiotic and biotic agents have been shown to mimic microorganisms and act as elicitors of the synthesis of these plant compounds. In the present study, sub-lethal levels of electric current are shown to elicit the biosynthesis of secondary metabolites in transgenic and non-transgenic plant tissue. The production of the phytoalexin (+)-pisatin by pea was used as the main model system. Non-transgenic pea hairy roots treated with 30-100 mA of electric current produced 13 times higher amounts of (+)-pisatin than did the non-elicited controls. Electrically elicited transgenic pea hairy root cultures blocked at various enzymatic steps in the (+)-pisatin biosynthetic pathway also accumulated intermediates preceding the blocked enzymatic step. Secondary metabolites not usually produced by pea accumulated in some of the transgenic root cultures after electric elicitation due to the diversion of the intermediates into new pathways. The amount of pisatin in the medium bathing the roots of electro-elicited roots of hydroponically cultivated pea plants was 10 times higher 24 h after elicitation than in the medium surrounding the roots of non-elicited control plants, showing not only that the electric current elicited (+)-pisatin biosynthesis but also that the (+)-pisatin was released from the roots. Seedlings, intact roots or cell suspension cultures of fenugreek (Trigonella foenum-graecum), barrel medic, (Medicago truncatula), Arabidopsis thaliana, red clover (Trifolium pratense) and chickpea (Cicer arietinum) also produced increased levels of secondary metabolites in response to electro-elicitation. On the basis of our results, electric current would appear to be a general elicitor of plant secondary metabolites and to have potential for application in both basic and commercial research.

  3. Intramuscular Cobinamide Sulfite in a Rabbit Model of Sub-Lethal Cyanide Toxicity

    Science.gov (United States)

    Brenner, Matthew; Kim, Jae G.; Mahon, Sari B.; Lee, Jangwoen; Kreuter, Kelly A.; Blackledge, William; Mukai, David; Patterson, Steve; Mohammad, Othman; Sharma, Vijay S.; Boss, Gerry R.

    2009-01-01

    Objective To determine the ability of an intramuscular cobinamide sulfite injection to rapidly reverse the physiologic effects of cyanide toxicity. Background Exposure to cyanide in fires and industrial exposures and intentional cyanide poisoning by terrorists leading to mass casualties is an ongoing threat. Current treatments for cyanide poisoning must be administered intravenously, and no rapid treatment methods are available for mass casualty cyanide exposures. Cobinamide is a cobalamin (vitamin B12) analog with an extraordinarily high affinity for cyanide that is more water-soluble than cobalamin. We investigated the use of intramuscular cobinamide sulfite to reverse cyanide toxicity induced physiologic changes in a sublethal cyanide exposure animal model. Methods New Zealand white rabbits were given 10 mg sodium cyanide intravenously over 60 minutes. Quantitative diffuse optical spectroscopy and continuous wave near infrared spectroscopy monitoring of tissue oxy- and deoxyhemoglobin concentrations were performed concurrently with blood cyanide level measurements and cobinamide levels. Immediately after completion of the cyanide infusion, the rabbits were injected intramuscularly with cobinamide sulfite (n=6) or inactive vehicle (controls, n=5). Results Intramuscular administration led to rapid mobilization of cobinamide and was extremely effective at reversing the physiologic effects of cyanide on oxyhemoglobin and deoxyhemoglobin extraction. Recovery time to 63% of their baseline values in the central nervous system was in a mean of 1032 minutes in the control group and 9 minutes in the cobinamide group with a difference of 1023 minutes (95% confidence interval [CI] 116, 1874 minutes). In muscle tissue, recovery times were 76 and 24 minutes with a difference of 52 minutes (95% CI 7, 98min). Red blood cell cyanide levels returned towards normal significantly faster in cobinamide sulfite-treated animals than in control animals. Conclusions Intramuscular

  4. Exenatide promotes regeneration of injured rat sciatic nerve

    Directory of Open Access Journals (Sweden)

    Ersin Kuyucu

    2017-01-01

    Full Text Available Damage to peripheral nerves results in partial or complete dysfunction. After peripheral nerve injuries, a full functional recovery usually cannot be achieved despite the standard surgical repairs. Neurotrophic factors and growth factors stimulate axonal growth and support the viability of nerve cells. The objective of this study is to investigate the neurotrophic effect of exenatide (glucagon like peptide-1 analog in a rat sciatic nerve neurotmesis model. We injected 10 μg/d exenatide for 12 weeks in the experimental group (n = 12 and 0.1 mL/d saline for 12 weeks in the control group (n = 12. We evaluated nerve regeneration by conducting electrophysiological and motor functional tests. Histological changes were evaluated at weeks 1, 3, 6, and 9. Nerve regeneration was monitored using stereomicroscopy. The electrophysiological and motor functions in rats treated with exenatide were improved at 12 weeks after surgery. Histological examination revealed a significant increase in the number of axons in injured sciatic nerve following exenatide treatment confirmed by stereomicroscopy. In an experimentally induced neurotmesis model in rats, exenatide had a positive effect on nerve regeneration evidenced by electromyography, functional motor tests, histological and stereomicroscopic findings.

  5. Rewiring the injured CNS: lessons from the optic nerve

    Science.gov (United States)

    Benowitz, Larry; Yin, Yuqin

    2008-01-01

    The optic nerve offers a number of advantages for investigating mechanisms that govern axon regeneration in the CNS. Although mature retinal ganglion cells (RGCs) normally show no ability to regenerate injured axons through the optic nerve, this situation can be partially reversed by inducing an inflammatory response in the eye. The secretion of a previously unknown growth factor, oncomodulin, along with co-factors, causes RGCs to undergo dramatic changes in gene expression and regenerate lengthy axons into the highly myelinated optic nerve. By themselves, strategies that counteract inhibitory signals associated with myelin and the glial scar are insufficient to promote extensive regeneration in this system. However, combinatorial treatments that activate neurons' intrinsic growth state and overcome inhibitory signals result in dramatic axon regeneration in vivo. Because of the ease of introducing trophic factors, soluble receptors, drugs, or viruses expressing any gene or small interfering RNA of interest into RGCs, this system is ideal for identifying intracellular signaling pathways, transcriptional cascades, and ligand-receptor interactions that enable axon regeneration to occur in the CNS and to develop means to augment this process. PMID:17610877

  6. Histological and functional benefit following transplantation of motor neuron progenitors to the injured rat spinal cord.

    Directory of Open Access Journals (Sweden)

    Sharyn L Rossi

    2010-07-01

    Full Text Available Motor neuron loss is characteristic of cervical spinal cord injury (SCI and contributes to functional deficit.In order to investigate the amenability of the injured adult spinal cord to motor neuron differentiation, we transplanted spinal cord injured animals with a high purity population of human motor neuron progenitors (hMNP derived from human embryonic stem cells (hESCs. In vitro, hMNPs displayed characteristic motor neuron-specific markers, a typical electrophysiological profile, functionally innervated human or rodent muscle, and secreted physiologically active growth factors that caused neurite branching and neuronal survival. hMNP transplantation into cervical SCI sites in adult rats resulted in suppression of intracellular signaling pathways associated with SCI pathogenesis, which correlated with greater endogenous neuronal survival and neurite branching. These neurotrophic effects were accompanied by significantly enhanced performance on all parameters of the balance beam task, as compared to controls. Interestingly, hMNP transplantation resulted in survival, differentiation, and site-specific integration of hMNPs distal to the SCI site within ventral horns, but hMNPs near the SCI site reverted to a neuronal progenitor state, suggesting an environmental deficiency for neuronal maturation associated with SCI.These findings underscore the barriers imposed on neuronal differentiation of transplanted cells by the gliogenic nature of the injured spinal cord, and the physiological relevance of transplant-derived neurotrophic support to functional recovery.

  7. Biodegradable biomatrices and bridging the injured spinal cord: the corticospinal tract as a proof of principle.

    Science.gov (United States)

    Joosten, Elbert A J

    2012-07-01

    Important advances in the development of smart biodegradable implants for axonal regeneration after spinal cord injury have recently been reported. These advances are evaluated in this review with special emphasis on the regeneration of the corticospinal tract. The corticospinal tract is often considered the ultimate challenge in demonstrating whether a repair strategy has been successful in the regeneration of the injured mammalian spinal cord. The extensive know-how of factors and cells involved in the development of the corticospinal tract, and the advances made in material science and tissue engineering technology, have provided the foundations for the optimization of the biomatrices needed for repair. Based on the findings summarized in this review, the future development of smart biodegradable bridges for CST regrowth and regeneration in the injured spinal cord is discussed.

  8. Toxicity bioassay and effects of sub-lethal exposure of malathion on ...

    African Journals Online (AJOL)

    Clarias gariepinus were exposed to different concentrations of malathion to determine the 96 h LC50 value and its sub-lethal effects on haematological parameters and biochemical composition were also investigated. The 96 h LC50 value concluded was 8.22 mg/L. Specimens of C. gariepinus were exposed to sub-lethal ...

  9. Sublethal effects of imidacloprid on interactions in a tritrophic system of non-target species.

    Science.gov (United States)

    Uhl, Philipp; Bucher, Roman; Schäfer, Ralf B; Entling, Martin H

    2015-08-01

    Imidacloprid is one of the most used insecticides worldwide, but is highly toxic to non-target arthropods. Effects of sublethal imidacloprid intoxication can potentially propagate in food webs, yet little is known about the impact on non-target populations and communities. We investigated short-term sublethal toxicity of imidacloprid in a tritrophic model system of wild strawberry Fragaria vesca, wood cricket Nemobius sylvestris and nursery web spider Pisaura mirabilis. Strawberries were treated two times with 0mg (control), 1mg (low rate) and 10mg (high rate) of Confidor® WG 70 and crickets were allowed to feed on them. In four lab experiments, we quantified the impact of imidacloprid on leaf damage, growth, behaviour and survival of crickets. The high imidacloprid rate reduced feeding, mass gain, thorax growth and mobility in crickets compared to the control, while mortality was similarly low in all treatments. The low rate reduced mass gain only. Cricket survival of spider predation was higher in the low rate treatment than in the control. Overall, herbivory and predation were reduced at sublethal imidacloprid rates in a non-target organism, three-level food chain, which demonstrates possible propagation of sublethal effects through trophic interactions. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Assessment of chronic sublethal effects of imidacloprid on honey bee colony health

    Science.gov (United States)

    Here we present results of a three-year study to determine the fate of imidacloprid residues in hive matrices and to assess chronic sublethal effects on whole honey bee colonies fed supplemental pollen diet containing imidacloprid at 5, 20 and 100 µg/kg over multiple brood cycles. Various endpoints ...

  11. Effect of sublethal preculturing on the survival of probiotics and metabolite formation in set-yoghurt

    NARCIS (Netherlands)

    Settachaimongkon, S.; Valenberg, van H.J.F.; Winata, V.; Wang, X.; Nout, M.J.R.; Hooijdonk, van A.C.M.; Zwietering, M.H.; Smid, E.J.

    2015-01-01

    The objective of this study was to investigate the effect of preculturing of Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB12 under sublethal stress conditions on their survival and metabolite formation in set-yoghurt. Prior to co-cultivation with yoghurt starters in milk,

  12. Sub-lethal effects of neonicitinoids on the alfalfa leafcutter bee, Megachile rotundata

    Science.gov (United States)

    Neonicotinoids are commonly used pesticides in U.S. agriculture. For many beneficial insect species, lethal effects of neonicotinoids are well-documented; however, much less is known about sublethal exposure. The alfalfa leaf cutter bee Megachile rotundata is a managed pollinator that constructs com...

  13. Monitoring colony-level effects of sublethal pesticide exposure on honey bees

    Science.gov (United States)

    The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment methods. Here we describe methods to measure the quantities of adult bees, brood and food resources by weighing hives and hive parts, by photogra...

  14. Physiological stress and ethanol accumulation in tree stems and woody tissues at sublethal temperatures from fire

    Science.gov (United States)

    Rick G. Kelsey; Douglas J. Westlind

    2017-01-01

    The lethal temperature limit is 60 degrees Celsius (°C) for plant tissues, including trees, with lower temperatures causing heat stress. As fire injury increases on tree stems, there is an accompanying rise in tissue ethanol concentrations, physiologically linked to impaired mitochondrial oxidative phosphorylation energy production. We theorize that sublethal tissue...

  15. Sublethal Triclosan Exposure Decreases Susceptibility to Gentamicin and Other Aminoglycosides in Listeria monocytogenes

    DEFF Research Database (Denmark)

    Christensen, Ellen Gerd; Gram, Lone; Kastbjerg, Vicky Gaedt

    2011-01-01

    The human food-borne pathogen Listeria monocytogenes is capable of persisting in food processing plants despite cleaning and sanitation and is likely exposed to sublethal biocide concentrations. This could potentially affect susceptibility of the bacterium to biocides and other antimicrobial agents...

  16. Lethal and sublethal effects of an insect growth regulator, pyriproxyfen, on obliquebanded leafroller (Lepidoptera: Tortricidae).

    Science.gov (United States)

    Sial, Ashfaq A; Brunner, Jay F

    2010-04-01

    The obliquebanded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae), is one of the most destructive pests of tree fruit in Washington. The development of insecticide resistance in C. rosaceana has led us to explore new management tactics. The use of very low doses of insecticides that have strong sublethal effects represents an environmentally friendly option to improve existing integrated pest management (IPM) strategies. We tested the insect growth regulator pyriproxyfen to determine its lethal and sublethal effects on growth and development of C. rosaceana. A leaf-disk bioassay was used to test seven concentrations of pyriproxyfen ranging from 0 to 30 ppm on fifth-instar C. rosaceana. Male and female larvae were assessed separately for mortality as well as other parameters of growth and development. The LC, values for males and females were 2.4 and 4.8 ppm, respectively. The response to pyriproxyfen was concentration-dependent: only 5-6% of the larvae treated with the highest concentration emerged as morphologically normal adults compared with 86% emergence in the controls. The pupation and adult emergence was significantly delayed at concentrations higher than 1 ppm. The weights of C. rosaceana pupae and adults were significantly increased, whereas fecundity and fertility were significantly reduced at a sublethal concentration of 0.3 ppm. We conclude that both lethal and sublethal effects might exhibit significant impacts on the population dynamics of C. rosaceana in tree fruit orchards treated with low concentrations of pyriproxyfen.

  17. Effects of sub-lethal concentrations of a vegetable oil mill effluent on ...

    African Journals Online (AJOL)

    Effects of sub-lethal concentrations of a vegetable oil mill effluent on growth of Clarius gariepinus. JA Adakole, E Alabi. Abstract. No Abstract. Nigerian Journal of Fisheries Vol. 4 (2) 2007: pp. 182-189. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  18. Effects of IL-10 on systemic inflammatory responses during sublethal primate endotoxemia

    NARCIS (Netherlands)

    van der Poll, T.; Jansen, P. M.; Montegut, W. J.; Braxton, C. C.; Calvano, S. E.; Stackpole, S. A.; Smith, S. R.; Swanson, S. W.; Hack, C. E.; Lowry, S. F.; Moldawer, L. L.

    1997-01-01

    IL-10 protects mice from LPS-induced lethality. To determine the effects of IL-10 on LPS-induced inflammatory responses, six Papio anubis baboons were i.v. injected with a sublethal dose of LPS (Salmonella typhimurium; 500 microg/kg) directly preceded by either human rIL-10 (n = 3, 500 microg/kg) or

  19. Effects of sublethal concentrations of formalin on weight gain in the ...

    African Journals Online (AJOL)

    The African Catfish, Clarias gariepinus, was exposed to various sublethal concentrations (25.0, 12.50, 6.25, 3.125, 1.56 and 0.0 mgl-1) of formalin to investigate their effects on the weight gain of the fish. Decrease in weight gain, directly proportional to the toxicant concentration, was observed in fish exposed to ...

  20. Effects of in vivo applications of peripheral blood-derived mesenchymal stromal cells (PB-MSCs) and platlet-rich plasma (PRP) on experimentally injured deep digital flexor tendons of sheep.

    Science.gov (United States)

    Martinello, Tiziana; Bronzini, Ilaria; Perazzi, Anna; Testoni, Stefania; De Benedictis, Gulia Maria; Negro, Alessandro; Caporale, Giovanni; Mascarello, Francesco; Iacopetti, Ilaria; Patruno, Marco

    2013-02-01

    Tendon injuries, degenerative tendinopathies, and overuse tendinitis are common in races horses. Novel therapies aim to restore tendon functionality by means of cell-based therapy, growth factor delivery, and tissue engineering approaches. This study examined the use of autologous mesenchymal stromal cells derived from peripheral blood (PB-MSCs), platelet-rich plasma (PRP) and a combination of both for ameliorating experimental lesions on deep digital flexor tendons (DDFT) of Bergamasca sheep. In particular, testing the combination of blood-derived MSCs and PRP in an experimental animal model represents one of the few studies exploring a putative synergistic action of these treatments. Effectiveness of treatments was evaluated at 30 and 120 days comparing clinical, ultrasonographic, and histological features together with immunohistochemical expression of collagen types 1 and 3, and cartilage oligomeric matrix protein (COMP). Significant differences were found between treated groups and their corresponding controls (placebo) regarding tendon morphology and extracellular matrix (ECM) composition. However, our results indicate that the combined use of PRP and MSCs did not produce an additive or synergistic regenerative response and highlighted the predominant effect of MSCs on tendon healing, enhanced tissue remodeling and improved structural organization. Copyright © 2012 Orthopaedic Research Society.

  1. RFID tracking of sublethal effects of two neonicotinoid insecticides on the foraging behavior of Apis mellifera.

    Directory of Open Access Journals (Sweden)

    Christof W Schneider

    Full Text Available The development of insecticides requires valid risk assessment procedures to avoid causing harm to beneficial insects and especially to pollinators such as the honeybee Apis mellifera. In addition to testing according to current guidelines designed to detect bee mortality, tests are needed to determine possible sublethal effects interfering with the animal's vitality and behavioral performance. Several methods have been used to detect sublethal effects of different insecticides under laboratory conditions using olfactory conditioning. Furthermore, studies have been conducted on the influence insecticides have on foraging activity and homing ability which require time-consuming visual observation. We tested an experimental design using the radiofrequency identification (RFID method to monitor the influence of sublethal doses of insecticides on individual honeybee foragers on an automated basis. With electronic readers positioned at the hive entrance and at an artificial food source, we obtained quantifiable data on honeybee foraging behavior. This enabled us to efficiently retrieve detailed information on flight parameters. We compared several groups of bees, fed simultaneously with different dosages of a tested substance. With this experimental approach we monitored the acute effects of sublethal doses of the neonicotinoids imidacloprid (0.15-6 ng/bee and clothianidin (0.05-2 ng/bee under field-like circumstances. At field-relevant doses for nectar and pollen no adverse effects were observed for either substance. Both substances led to a significant reduction of foraging activity and to longer foraging flights at doses of ≥0.5 ng/bee (clothianidin and ≥1.5 ng/bee (imidacloprid during the first three hours after treatment. This study demonstrates that the RFID-method is an effective way to record short-term alterations in foraging activity after insecticides have been administered once, orally, to individual bees. We contribute further

  2. Equivalency of the quality of sublethal lesions after photons and high-linear energy transfer ion beams.

    Science.gov (United States)

    Furusawa, Yoshiya; Nakano-Aoki, Mizuho; Matsumoto, Yoshitaka; Hirayama, Ryoichi; Kobayashi, Alisa; Konishi, Teruaki

    2017-11-01

    The quality of the sublethal damage (SLD) after irradiation with high-linear energy transfer (LET) ion beams was investigated with low-LET photons. Chinese hamster V79 cells and human squamous carcinoma SAS cells were first exposed to a priming dose of different ion beams at different LETs at the Heavy Ion Medical Accelerator in the Chiba facility. The cells were kept at room temperature and then exposed to a secondary test dose of X-rays. Based on the repair kinetics study, the surviving fraction of cells quickly increased with the repair time, and reached a plateau in 2-3 h, even when cells had received priming monoenergetic high-LET beams or spread-out Bragg peak beams as well as X-ray irradiation. The shapes of the cell survival curves from the secondary test X-rays, after repair of the damage caused by the high-LET irradiation, were similar to those obtained from cells exposed to primary X-rays only. Complete SLD repairs were observed, even when the LET of the primary ion beams was very high. These results suggest that the SLD caused by high-LET irradiation was repaired well, and likewise, the damage caused by the X-rays. In cells where the ion beam had made a direct hit in the core region in an ion track, lethal damage to the domain was produced, resulting in cell death. On the other hand, in domains that had received a glancing hit in the low-LET penumbra region, the SLD produced was completely repaired. © The Author 2017. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.

  3. Temporal dynamics of epidermal responses of guppies Poecilia reticulata to a sublethal range of waterborne zinc concentrations.

    Science.gov (United States)

    Gheorghiu, C; Marcogliese, D J; Scott, M E

    2009-12-01

    This study assessed the histological changes in the epidermis of guppies Poecilia reticulata induced by waterborne zinc (Zn). Laboratory-reared P. reticulata fry were maintained individually in separate vessels containing artificial water (8 microg l(-1) Zn) to which 0, 15, 30, 60 or 120 microg l(-1) Zn was added. Their epidermal response to Zn was monitored regularly over 4 weeks. Compared with controls, mucus was rapidly released and mucous cell numbers decreased at all concentrations. Thereafter mucous release, epidermal thickness, numbers and size of mucous cells fluctuated at a rate that varied with Zn concentration, but fluctuations declined after day 18. Results clearly highlight the dynamic nature of the epidermal response to sublethal concentrations of waterborne Zn. In general, low concentrations of Zn induced a rapid response with reduced numbers and size of mucous cells and shift in mucin composition, and a subsequent thickening of the epidermis. Epidermal thickness and mucous cell area fluctuated over time but were normal after a month of exposure to low Zn concentrations. The number of mucous cells, however, remained low. Virtually all mucous cells from fish maintained in 15 and 60 microg l(-1) Zn contained acidic mucins throughout the month, whereas fish maintained at 30 microg l(-1) Zn responded by production of neutral mucins during the first 12 days followed by a mixture of neutral and acidic mucins. At 120 microg l(-1) Zn, the most dramatic effects were the gradual but sustained decrease in numbers and area of mucous cells, and the shift to acidic mucins in these cells. Thus, as concentration of Zn increased, the epidermal responses indicated a disturbed host response (dramatic decline in mucous cell numbers, with mixed composition of mucins), which may have been less effective in preventing Zn uptake across the epithelium.

  4. Sublethal red tide toxin exposure in free-ranging manatees (Trichechus manatus) affects the immune system through reduced lymphocyte proliferation responses, inflammation, and oxidative stress.

    Science.gov (United States)

    Walsh, Catherine J; Butawan, Matthew; Yordy, Jennifer; Ball, Ray; Flewelling, Leanne; de Wit, Martine; Bonde, Robert K

    2015-04-01

    The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected by exposure to blooms of the toxic dinoflagellate, Karenia brevis. K. brevis blooms are common in manatee habitats of Florida's southwestern coast and produce a group of cyclic polyether toxins collectively referred to as red tide toxins, or brevetoxins. Although a large number of manatees exposed to significant levels of red tide toxins die, several manatees are rescued from sublethal exposure and are successfully treated and returned to the wild. Sublethal brevetoxin exposure may potentially impact the manatee immune system. Lymphocyte proliferative responses and a suite of immune function parameters in the plasma were used to evaluate effects of brevetoxin exposure on health of manatees rescued from natural exposure to red tide toxins in their habitat. Blood samples were collected from rescued manatees at Lowry Park Zoo in Tampa, FL and from healthy, unexposed manatees in Crystal River, FL. Peripheral blood leukocytes (PBL) isolated from whole blood were stimulated with T-cell mitogens, ConA and PHA. A suite of plasma parameters, including plasma protein electrophoresis profiles, lysozyme activity, superoxide dismutase (SOD) activity, and reactive oxygen/nitrogen (ROS/RNS) species, was also used to assess manatee health. Significant decreases (p<0.05) in lymphocyte proliferation were observed in ConA and PHA stimulated lymphocytes from rescued animals compared to non-exposed animals. Significant correlations were observed between oxidative stress markers (SOD, ROS/RNS) and plasma brevetoxin concentrations. Sublethal exposure to brevetoxins in the wild impacts some immune function components, and thus, overall health, in the Florida manatee. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Buyang Huanwu decoction up-regulates Notch1 gene expression in injured spinal cord

    Directory of Open Access Journals (Sweden)

    Zhan-peng Guo

    2015-01-01

    Full Text Available Expression of genes in the Notch signaling pathway is altered in the injured spinal cord, which indicates that Notch participates in repair after spinal cord injury. Buyang Huanwu decoction, a traditional Chinese herbal preparation, can promote the growth of nerve cells and nerve fibers; however, it is unclear whether Buyang Huanwu decoction affects the Notch signaling pathway in injured spinal cord. In this study, a rat model was established by injuring the T 10 spinal cord. At 2 days after injury, rats were intragastrically administered 2 mL of 0.8 g/mL Buyang Huanwu decoction daily until sacrifice. Real-time reverse transcription polymerase chain reaction analysis demonstrated that at 7, 14 and 28 days after injury, the expression of Notch1 was increased in the Buyang Huanwu decoction group compared with controls. These findings confirm that Buyang Huanwu decoction can promote the expression of Notch1 in rats with incomplete spinal cord injury, and may indicate a mechanism to promote the repair of spinal cord injury.

  6. Chronic exposure to sublethal hexavalent chromium affects organ histopathology and serum cortisol profile of a teleost, Channa punctatus (Bloch)

    Energy Technology Data Exchange (ETDEWEB)

    Mishra, Ashish K. [Department of Zoology, University of Allahabad, Allahabad-211002 (India); Mohanty, Banalata, E-mail: drbana_mohanty@rediffmail.com [Department of Zoology, University of Allahabad, Allahabad-211002 (India)

    2009-09-01

    Effects of chronic exposures (one and two months) to sublethal doses of hexavalent chromium (2 and 4 mg/L potassium dichromate) on organ histopathology and serum cortisol profile were investigated and their overall impact on growth and behavior of a teleost fish, Channa punctatus was elucidated. Histopathological lesions were distinct in the vital organs gill, kidney and liver. The gill lamellae became lifted, fused, and showed oedema. Hyperplasia and hypertrophy of lamellar epithelial cells were distinct with desquamation. Hypertrophy of epithelial cells of renal tubules and reduction in tubular lumens were observed in the trunk kidney. The atrophy of the head kidney interrenal cells and decreased serum cortisol level indicated exhaustion of interrenal activity. Hepatocyte vacuolization and shrinkage, nuclear pyknosis and increase of sinusoidal spaces were observed in the liver. Abnormal behavioral patterns and reduced growth rate were also noticed in the exposed fish. The chronic hexavalent chromium exposure thus by affecting histopathology of gill, kidney (including interrenal tissue) and liver could impair the vital functions of respiration, excretion, metabolic regulation and maintenance of stress homeostasis which in the long-run may pose serious threat to fish health and affect their population.

  7. Histological and histochemical alterations in the liver following intramuscular injection with a sublethal dose of the Egyptian cobra venom.

    Science.gov (United States)

    Rahmy, T R; Hemmaid, K Z

    2000-02-01

    In the present study, the effects of intramuscular (i.m.) injection of a sublethal dose (0.015 microgram/gm b.wt.) of Naja haje venom were histologically and histochemically examined in the hepatic tissues of rabbits after 3, 6, and 12 hr. of envenomation. Three hours after venom injection, the hepatic cells showed a generalized cytoplasmic granulation and cellular swelling accompanied with narrowing of the sinusoidal spaces. Occurrence of inflammatory cells and hypertrophy of Kupffer cells were also noticed. After 6 hr. of envenomation, the hepatic tissues revealed severe cellular swelling, cytoplasmic deterioration, nuclear pyknosis, and appearance of numerous basophilic granules. The central veins were engorged with hemolyzed blood. Hepatic tissues investigated after 12 hr. of envenomation exhibited highly damaged hepatic cells to the extent that the individual cell cannot be identified and remnants of cell debris were seen intermixed together. Some hepatic cells were intensively swollen and their contents were dissoluted except for a few dusty cytoplasmic granules and pyknotic nuclei. The histochemical observations showed a time dependent depletion in polysaccharide, lipid, and protein contents in the hepatic cells of the envenomed groups. As for the nucleic acids, slight depletion of RNA together with no changes in DNA contents were observed by 3 hr. of envenomation. Nevertheless, severe degrees of RNA depletion and moderate contents of DNA were recorded in 6 hr. envenomed tissues. Highly obvious depletion of RNA and DNA were demonstrated by 12 hr. after venom injection. From the above results, it is obvious that cobra venom induces a hepatotoxic action reflected by alterations in the histological and histochemical patterns of the hepatic tissues. These alterations are initiated at early stages of envenomation and could indicate a disturbance in the functional activities of the liver during envenomation.

  8. Cellular origin of pro-coagulant and (anti)-fibrinolytic factors in bleomycin-injured lungs.

    Science.gov (United States)

    Wygrecka, M; Markart, P; Ruppert, C; Petri, K; Preissner, K T; Seeger, W; Guenther, A

    2007-06-01

    Excessive pro-coagulant and decreased fibrinolytic activities in the alveolar compartment have been repeatedly documented for inflammatory and fibrotic lung diseases. The current authors determined the contribution of different resident lung cells to the altered local production of coagulation- and fibrinolysis-system components in bleomycin-injured mouse lungs via cell-specific and quantitative assessment of mRNA levels of various pro-coagulant and (anti)-fibrinolytic factors. Laser-assisted microdissection technology was used to sample specific cell populations in combination with subsequent mRNA analysis by real-time quantitative reverse transcriptase-PCR. Additionally, western blot analysis, immunohistochemistry and activity assays were performed. Following bleomycin challenge, the strongest induction of tissue factor and plasminogen activator inhibitor (PAI)-1 mRNA expression was observed in alveolar macrophages (approximately 250- and 60-fold induction, respectively). These factors were also upregulated in alveolar type II cells, but to an approximately six-fold lesser extent. In contrast, PAI-2 expression was induced exclusively in alveolar macrophages. A slight increase of urokinase-type plasminogen activator (uPA) expression was also observed in alveolar macrophages (two-fold induction), but uPA activity was reduced due to a disproportionate increase of PAI production. Alveolar macrophages and, to a lesser extent, alveolar type II cells are the main sources of locally produced pro-coagulant and anti-fibrinolytic factors in bleomycin-injured lungs.

  9. Effects of sub-lethal concentrations of thyme and oregano essential oils, carvacrol, thymol, citral and trans-2-hexenal on membrane fatty acid composition and volatile molecule profile of Listeria monocytogenes, Escherichia coli and Salmonella enteritidis.

    Science.gov (United States)

    Siroli, Lorenzo; Patrignani, Francesca; Gardini, Fausto; Lanciotti, Rosalba

    2015-09-01

    The aim of this work was to investigate the modifications of cell membrane fatty acid composition and volatile molecule profiles of Listeria monocytogenes, Salmonella enteritidis, Escherichia coli, during growth in the presence of different sub-lethal concentrations of thyme and oregano essential oils as well as carvacrol, thymol, trans-2-hexenal and citral. The results evidenced that the tested molecules induced noticeable modifications of membrane fatty acid profiles and volatile compounds produced during the growth. Although specific differences in relation to the species considered were identified, the tested compounds induced a marked increase of some membrane associated fatty acids, particularly unsaturated fatty acids, trans-isomers, and specific released free fatty acids. These findings can contribute to the comprehension of the stress response mechanisms used by different pathogenic microorganisms often involved in food-borne diseases in relation to the exposure to sub-lethal concentrations of natural antimicrobials. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Effects of sub-lethal heroin administration on thyroid stimulating hormone (TSH), thyroid hormones (T3, T4) and thyroid gland of Mus norvegicus.

    Science.gov (United States)

    Bhoir, Kaminidevi K; Suryawanshi, S A; Pandey, A K

    2009-11-01

    Serum TSH level of control Mus norvegicus fluctuated between 498.20 +/- 21.92 and 506.80 +/- 22.35 ng ml(-1), thyroxine (T4) between 68.17 +/- 3.46 and 69.03 +/- 4.12 microg dl(-1) and triiodothyronine (T3) between 4.76 +/- 0.52 and 5.00 +/- 0.66 microg dl(-1). Sub-lethal heroin administration induced a significant decline in the levels of all the three hormones at 24 hr and 15 days post-administration. Decline in the levels of these hormones registered the lowest values (pThyroid gland of control rat consisted of spherical, round follicles lined with low cuboidal and columnar epithelial cells and lumina filled with eosinophilic colloid. Ultrastructurally, the thyroid follicular cells showed the presence of round nuclei, polymorphic mitochondria, Golgi complex as well as lysosomes located on the apical side of the nucleus and cytoplasm with different sizes of lipid droplets and smooth along with rough endoplasmic reticulum. Basal lamina of the follicular cells was often in association with the endothelium of the capillaries. Sub-lethal heroin administration for 30 days elicited degenerative changes in the follicular epithelial cells as evident by the vacuolization of cytoplasm, pycnotic nuclei and reduced colloidal content. Ultrastructurally, the thyroid follicular cells showed indented nuclei with heavy deposition of chromatin material on the inner membrane of nucleus and dilated rough endoplasmic reticulum. Along with RBC infiltration, vesiculated mitochondria owing to the loss of cristae were also seen. Diffused electron-dense material was seen at the periphery of the cell body. Heroin treatment caused cellular necrosis as revealed by the fragmentation of cytoplasmic materials in follicular epithelial cells of the gland.

  11. Electrical stimulation promotes regeneration of injured oculomotor nerves in dogs

    Directory of Open Access Journals (Sweden)

    Lei Du

    2016-01-01

    Full Text Available Functional recovery after oculomotor nerve injury is very poor. Electrical stimulation has been shown to promote regeneration of injured nerves. We hypothesized that electrical stimulation would improve the functional recovery of injured oculomotor nerves. Oculomotor nerve injury models were created by crushing the right oculomotor nerves of adult dogs. Stimulating electrodes were positioned in both proximal and distal locations of the lesion, and non-continuous rectangular, biphasic current pulses (0.7 V, 5 Hz were administered 1 hour daily for 2 consecutive weeks. Analysis of the results showed that electrophysiological and morphological recovery of the injured oculomotor nerve was enhanced, indicating that electrical stimulation improved neural regeneration. Thus, this therapy has the potential to promote the recovery of oculomotor nerve dysfunction.

  12. Perceptions of Supervision Among Injured and Non-Injured Teens Working in the Retail or Service Industry.

    Science.gov (United States)

    Zierold, Kristina M

    2016-04-01

    According to the National Institute for Occupational Safety and Health (NIOSH), a teen is injured every 9 minutes at work. Workplace supervision may affect whether teens are injured on the job. Because research on workplace supervision among teens is limited, the objectives of this study were to characterize the perceptions of supervision among injured and non-injured teen workers and assess the characteristics and perceptions of supervisors that may be associated with work-related injuries. In 2011, a cross-sectional survey was conducted among high school students. Teens who worked in retail or service industries (n= 270) were included in the sample. Non-injured teens were more likely to have reported that their supervisors cared about their safety, were helpful, listened well, and ensured that teen workers understood workplace safety. Most teens (70%) did not feel comfortable talking about safety issues with their supervisors. The importance of supervision and how supervisors are perceived in the workplace may be significant in creating a safety culture that leaves a lasting impression. © 2015 The Author(s).

  13. Locomotion Inhibition of Cimex lectularius L. Following Topical, Sublethal Dose Application of the Chitin Synthesis Inhibitor Lufenuron

    Directory of Open Access Journals (Sweden)

    Brittany Campbell

    2017-09-01

    Full Text Available To date, few studies have evaluated chitin synthesis inhibitors against bed bugs, although they would provide an alternative mode of action to circumvent insecticide resistance. Acute and sublethal effects of lufenuron were evaluated against two strains of the common bed bug. Combined acute and sublethal effects were used to calculate effective doses. The dose that was effective against 50% of Harlan strain bed bugs was 0.0081% (w/v, and was much higher against Bradenton strain bed bugs (1.11% w/v. Sublethal doses were chosen to determine the effect that leg abnormalities had on pulling force. Both Harlan and Bradenton strain bed bugs had significantly lower locomotion ability (p < 0.0001 following topical application of lufenuron. The observed sublethal effects that limit locomotion could prevent bed bugs from moving within a domicile and taking a blood meal, subsequently reducing a bed bug population over time.

  14. Comparison of postural stability between injured and uninjured ballet dancers.

    Science.gov (United States)

    Lin, Cheng-Feng; Lee, I-Jung; Liao, Jung-Hsien; Wu, Hong-Wen; Su, Fong-Chin

    2011-06-01

    Ballet movements require a limited base of support; thus, ballet dancers require a high level of postural control. However, postural stability in ballet dancers is still unclear and needs to be understood. To evaluate ballet dancers' postural stability in performing single-leg standing, the en pointe task, and the first and fifth positions and to determine differences in task performance among healthy nondancers, healthy dancers, and dancers with ankle sprains. Controlled laboratory study. Injured dancers, uninjured dancers, and nondancers were recruited for this study (N = 33 age-matched participants; n= 11 per group). The tasks tested were single-leg standing with eyes open and closed, first position, fifth position, and en pointe. Center of pressure parameters were calculated from the ground-reaction force collected with 1 force plate. Analysis of variance was used to assess the differences of center of pressure parameters among 3 groups in single-leg standing; independent t test was used to examine the differences of center of pressure parameters between injured and uninjured dancers. During single-leg standing, injured dancers had significantly greater maximum displacement in the medial-lateral direction and total trajectory of center of pressure, compared with the uninjured dancers and nondancers. During the first and fifth positions, the injured dancers demonstrated significantly greater standard deviation of center of pressure position in the medial-lateral and anterior-posterior directions, compared with the uninjured dancers. During en pointe, the injured dancers had significantly greater maximum displacement in the medial-lateral direction and the anterior-posterior direction, compared with the uninjured dancers. The injured and uninjured dancers demonstrated differences in postural stability in the medial-lateral direction during single-leg standing and the ballet postures. Although the injured dancers received ballet training, their postural stability

  15. Combination Treatment with Sublethal Ionizing Radiation and the Proteasome Inhibitor, Bortezomib, Enhances Death-Receptor Mediated Apoptosis and Anti-Tumor Immune Attack

    Directory of Open Access Journals (Sweden)

    Ercan Cacan

    2015-12-01

    Full Text Available Sub-lethal doses of radiation can modulate gene expression, making tumor cells more susceptible to T-cell-mediated immune attack. Proteasome inhibitors demonstrate broad anti-tumor activity in clinical and pre-clinical cancer models. Here, we use a combination treatment of proteasome inhibition and irradiation to further induce immunomodulation of tumor cells that could enhance tumor-specific immune responses. We investigate the effects of the 26S proteasome inhibitor, bortezomib, alone or in combination with radiotherapy, on the expression of immunogenic genes in normal colon and colorectal cancer cell lines. We examined cells for changes in the expression of several death receptors (DR4, DR5 and Fas commonly used by T cells for killing of target cells. Our results indicate that the combination treatment resulted in increased cell surface expression of death receptors by increasing their transcript levels. The combination treatment further increases the sensitivity of carcinoma cells to apoptosis through FAS and TRAIL receptors but does not change the sensitivity of normal non-malignant epithelial cells. Furthermore, the combination treatment significantly enhances tumor cell killing by tumor specific CD8+ T cells. This study suggests that combining radiotherapy and proteasome inhibition may simultaneously enhance tumor immunogenicity and the induction of antitumor immunity by enhancing tumor-specific T-cell activity.

  16. Effects of sublethal exposure to metofluthrin on the fitness of Aedes aegypti in a domestic setting in Cairns, Queensland

    OpenAIRE

    Buhagiar, Tamara S.; Gregor J Devine; Scott A. Ritchie

    2017-01-01

    Background Metofluthrin is highly effective at reducing biting activity in Aedes aegypti. Its efficacy lies in the rapid onset of confusion, knockdown, and subsequent kill of a mosquito. In the field, there are a variety of scenarios that might result in sublethal exposure to metofluthrin, including mosquitoes that are active at the margins of the chemical?s lethal range, brief exposure as mosquitoes fly in and out of treated spaces or decreasing efficacy of the emanators with time. Sublethal...

  17. Sublethal doses of imidacloprid disrupt sexual communication and host finding in a parasitoid wasp

    Science.gov (United States)

    Tappert, Lars; Pokorny, Tamara; Hofferberth, John; Ruther, Joachim

    2017-02-01

    Neonicotinoids are widely used insecticides, but their use is subject of debate because of their detrimental effects on pollinators. Little is known about the effect of neonicotinoids on other beneficial insects such as parasitoid wasps, which serve as natural enemies and are crucial for ecosystem functioning. Here we show that sublethal doses of the neonicotinoid imidacloprid impair sexual communication and host finding in the parasitoid wasp Nasonia vitripennis. Depending on the dose, treated females were less responsive to the male sex pheromone or unable to use it as a cue at all. Courtship behaviour of treated couples was also impeded resulting in a reduction of mating rates by up to 80%. Moreover, treated females were no longer able to locate hosts by using olfactory cues. Olfaction is crucial for the reproductive success of parasitoid wasps. Hence, sublethal doses of neonicotinoids might compromise the function of parasitoid wasps as natural enemies with potentially dire consequences for ecosystem services.

  18. Both genome and cytosol dynamics change in E. coli challenged with sublethal rifampicin

    Science.gov (United States)

    Wlodarski, Michal; Raciti, Bianca; Kotar, Jurij; Cosentino Lagomarsino, Marco; Fraser, Gillian M.; Cicuta, Pietro

    2017-02-01

    While the action of many antimicrobial drugs is well understood at the molecular level, a systems-level physiological response to antibiotics remains largely unexplored. This work considers fluctuation dynamics of both the chromosome and cytosol in Escherichia coli, and their response to sublethal treatments of a clinically important antibiotic, rifampicin. We precisely quantify the changes in dynamics of chromosomal loci and cytosolic aggregates (a rheovirus nonstructural protein known as μNS-GFP), measuring short time-scale displacements across several hours of drug exposure. To achieve this we develop an empirical method correcting for photo-bleaching and loci size effects. This procedure allows us to characterize the dynamic response to rifampicin in different growth conditions, including a customised microfluidic device. We find that sub-lethal doses of rifampicin cause a small but consistent increase in motility of both the chromosomal loci and cytosolic aggregates. Chromosomal and cytosolic responses are consistent with each other and between different growth conditions.

  19. Sublethal effects of spinetoram on the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae).

    Science.gov (United States)

    Wang, Ling; Zhang, Youjun; Xie, Wen; Wu, Qingjun; Wang, Shaoli

    2016-09-01

    The two-spotted spider mite Tetranychus urticae is a serious pest of many agricultural crops and ornamental plants. The sublethal effects of a new chemical, spinetoram, on T. urticae were investigated by treating adult females and eggs with LC10 and LC20 in the laboratory. The data were assessed based on age-stage, two-sex life table analysis. The results showed that T. urticae developmental time from egg to adult was reduced and that fecundity was increased by treatment with LC10 and LC20 of spinetoram. The LC10 and LC20 of spinetoram also increased the intrinsic and finite rate of increase and the net reproductive rate and reduced the mean generation time, egg duration, and larval duration whether eggs or adult females were treated. These laboratory results suggest that sublethal or lethal doses of spinetoram may cause outbreaks of T. urticae. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Sublethal effect of nanosilver on the structure of gill of Caspian roach (Rutilus rutilus caspicus fingerlings

    Directory of Open Access Journals (Sweden)

    Maryam Sharifian

    2013-04-01

    Full Text Available Widespread use of nanosilver can be led the contamination of aquatic environment and impact on living organisms such as fishes. We investigated histopathological changes in the gills tissue of Caspian roach fingerlings after two weeks exposure to sublethal concentrations of nanosilver. Following one and two weeks exposure, necrosis, shortening of secondary lamellae, edema, destruction of epithelial lamella, shortening of secondary lamellae, epithelial lifting and curling of secondary lamellae were observed in gill tissues. This observation showed that exposure to sub-lethal concentrations of nanosilver is caused damages in the gill tissues of Caspian roach. The results demonstrated direct correlation of gill tissue damage and toxin exposure i.e. increasing nanosilver concentration is caused more tissue damage. Hence, histopathological changes of gill can considered as a proper indicator for nanosilver contamination of aquatic environments.

  1. Unbiased high-throughput characterization of mussel transcriptomic responses to sublethal concentrations of the biotoxin okadaic acid

    Science.gov (United States)

    Suarez-Ulloa, Victoria; Fernandez-Tajes, Juan; Aguiar-Pulido, Vanessa; Prego-Faraldo, M. Veronica; Florez-Barros, Fernanda; Sexto-Iglesias, Alexia; Mendez, Josefina

    2015-01-01

    Background. Harmful Algal Blooms (HABs) responsible for Diarrhetic Shellfish Poisoning (DSP) represent a major threat for human consumers of shellfish. The biotoxin Okadaic Acid (OA), a well-known phosphatase inhibitor and tumor promoter, is the primary cause of acute DSP intoxications. Although several studies have described the molecular effects of high OA concentrations on sentinel organisms (e.g., bivalve molluscs), the effect of prolonged exposures to low (sublethal) OA concentrations is still unknown. In order to fill this gap, this work combines Next-Generation sequencing and custom-made microarray technologies to develop an unbiased characterization of the transcriptomic response of mussels during early stages of a DSP bloom. Methods. Mussel specimens were exposed to a HAB episode simulating an early stage DSP bloom (200 cells/L of the dinoflagellate Prorocentrum lima for 24 h). The unbiased characterization of the transcriptomic responses triggered by OA was carried out using two complementary methods of cDNA library preparation: normalized and Suppression Subtractive Hybridization (SSH). Libraries were sequenced and read datasets were mapped to Gene Ontology and KEGG databases. A custom-made oligonucleotide microarray was developed based on these data, completing the expression analysis of digestive gland and gill tissues. Results. Our findings show that exposure to sublethal concentrations of OA is enough to induce gene expression modifications in the mussel Mytilus. Transcriptomic analyses revealed an increase in proteasomal activity, molecular transport, cell cycle regulation, energy production and immune activity in mussels. Oppositely, a number of transcripts hypothesized to be responsive to OA (notably the Serine/Threonine phosphatases PP1 and PP2A) failed to show substantial modifications. Both digestive gland and gill tissues responded similarly to OA, although expression modifications were more dramatic in the former, supporting the choice of

  2. Unbiased high-throughput characterization of mussel transcriptomic responses to sublethal concentrations of the biotoxin okadaic acid

    Directory of Open Access Journals (Sweden)

    Victoria Suarez-Ulloa

    2015-11-01

    Full Text Available Background. Harmful Algal Blooms (HABs responsible for Diarrhetic Shellfish Poisoning (DSP represent a major threat for human consumers of shellfish. The biotoxin Okadaic Acid (OA, a well-known phosphatase inhibitor and tumor promoter, is the primary cause of acute DSP intoxications. Although several studies have described the molecular effects of high OA concentrations on sentinel organisms (e.g., bivalve molluscs, the effect of prolonged exposures to low (sublethal OA concentrations is still unknown. In order to fill this gap, this work combines Next-Generation sequencing and custom-made microarray technologies to develop an unbiased characterization of the transcriptomic response of mussels during early stages of a DSP bloom.Methods. Mussel specimens were exposed to a HAB episode simulating an early stage DSP bloom (200 cells/L of the dinoflagellate Prorocentrum lima for 24 h. The unbiased characterization of the transcriptomic responses triggered by OA was carried out using two complementary methods of cDNA library preparation: normalized and Suppression Subtractive Hybridization (SSH. Libraries were sequenced and read datasets were mapped to Gene Ontology and KEGG databases. A custom-made oligonucleotide microarray was developed based on these data, completing the expression analysis of digestive gland and gill tissues.Results. Our findings show that exposure to sublethal concentrations of OA is enough to induce gene expression modifications in the mussel Mytilus. Transcriptomic analyses revealed an increase in proteasomal activity, molecular transport, cell cycle regulation, energy production and immune activity in mussels. Oppositely, a number of transcripts hypothesized to be responsive to OA (notably the Serine/Threonine phosphatases PP1 and PP2A failed to show substantial modifications. Both digestive gland and gill tissues responded similarly to OA, although expression modifications were more dramatic in the former, supporting the

  3. The Protective Role of Thiourea on Leuciscus cephalus Exposed to Sublethal Doses of Pendigan 330EC (Pendimethalin) Herbicide.

    Science.gov (United States)

    Paunescu, Alina; Zgurschi, Gabriela; Soare, Liliana Cristina; Man, George Mihail; Brinzea, Gheorghita; Fierascu, Radu Claudiu; Fierascu, Irina; Ponepal, Maria Cristina

    2016-08-01

    The aim of the study was to investigate the protective role of thiourea on the physiological, hematological, biochemical and histopathological parameters of Leuciscus cephalus exposed to sublethal concentration of Pendigan 330 EC herbicide. The animals were divided in four experimental groups (control, animals subjected to 1 ‰ thiourea, animals subjected to 4 × 10(-4) mL/L herbicide and, respectively, animals subjected to 4 × 10(-4) mL/L herbicide and 1 ‰ thiourea). Exposure of European chub to herbicide administered in water for 2 weeks determined installation of pathological changes in the liver and gills tissues. Also, were observed a decrease in the number of white blood cells and oxygen consumption, breathing frequency, and an increase in the number of red blood cells and glycaemia values. Thiourea counteracts the toxic action, describing itself as normal liver parenchyma and normal gills in animals intoxicated with herbicide, without lesion, and a return to normal values of the studied markers.

  4. Injured astrocytes are repaired by Synaptotagmin XI-regulated lysosome exocytosis.

    Science.gov (United States)

    Sreetama, S C; Takano, T; Nedergaard, M; Simon, S M; Jaiswal, J K

    2016-04-01

    Astrocytes are known to facilitate repair following brain injury; however, little is known about how injured astrocytes repair themselves. Repair of cell membrane injury requires Ca(2+)-triggered vesicle exocytosis. In astrocytes, lysosomes are the main Ca(2+)-regulated exocytic vesicles. Here we show that astrocyte cell membrane injury results in a large and rapid calcium increase. This triggers robust lysosome exocytosis where the fusing lysosomes release all luminal contents and merge fully with the plasma membrane. In contrast to this, receptor stimulation produces a small sustained calcium increase, which is associated with partial release of the lysosomal luminal content, and the lysosome membrane does not merge into the plasma membrane. In most cells, lysosomes express the synaptotagmin (Syt) isoform Syt VII; however, this isoform is not present on astrocyte lysosomes and exogenous expression of Syt VII on lysosome inhibits their exocytosis. Deletion of one of the most abundant Syt isoform in astrocyte--Syt XI--suppresses astrocyte lysosome exocytosis. This identifies lysosome as Syt XI-regulated exocytic vesicle in astrocytes. Further, inhibition of lysosome exocytosis (by Syt XI depletion or Syt VII expression) prevents repair of injured astrocytes. These results identify the lysosomes and Syt XI as the sub-cellular and molecular regulators, respectively of astrocyte cell membrane repair.

  5. Sublethal triclosan exposure decreases susceptibility to gentamicin and other aminoglycosides in Listeria monocytogenes.

    Science.gov (United States)

    Christensen, Ellen G; Gram, Lone; Kastbjerg, Vicky G

    2011-09-01

    The human food-borne pathogen Listeria monocytogenes is capable of persisting in food processing plants despite cleaning and sanitation and is likely exposed to sublethal biocide concentrations. This could potentially affect susceptibility of the bacterium to biocides and other antimicrobial agents. The purpose of the present study was to determine if sublethal biocide concentrations affected antibiotic susceptibility in L. monocytogenes. Exposure of L. monocytogenes strains EGD and N53-1 to sublethal concentrations of Incimaxx DES (containing peroxy acids and hydrogen peroxide) and Triquart Super (containing quaternary ammonium compound) in four consecutive cultures did not alter the frequency of antibiotic-tolerant isolates, as determined by plating on 2× the MIC for a range of antibiotics. Exposure of eight strains of L. monocytogenes to 1 and 4 μg/ml triclosan did not alter triclosan sensitivity. However, all eight strains became resistant to gentamicin (up to 16-fold increase in MIC) after exposure to sublethal triclosan concentrations. Gentamicin-resistant isolates of strains N53-1 and 4446 were also resistant to other aminoglycosides, such as kanamycin, streptomycin, and tobramycin. Gentamicin resistance remained at a high level also after five subcultures without triclosan or gentamicin. Aminoglycoside resistance can be caused by mutations in the target site, the 16S rRNA gene. However, such mutations were not detected in the N53-1-resistant isolates. A combination of gentamicin and ampicillin is commonly used in listeriosis treatment. The triclosan-induced resistance is, hence, of great concern. Further investigations are needed to determine the molecular mechanisms underlying the effect of triclosan.

  6. Sublethal Triclosan Exposure Decreases Susceptibility to Gentamicin and Other Aminoglycosides in Listeria monocytogenes▿

    Science.gov (United States)

    Christensen, Ellen G.; Gram, Lone; Kastbjerg, Vicky G.

    2011-01-01

    The human food-borne pathogen Listeria monocytogenes is capable of persisting in food processing plants despite cleaning and sanitation and is likely exposed to sublethal biocide concentrations. This could potentially affect susceptibility of the bacterium to biocides and other antimicrobial agents. The purpose of the present study was to determine if sublethal biocide concentrations affected antibiotic susceptibility in L. monocytogenes. Exposure of L. monocytogenes strains EGD and N53-1 to sublethal concentrations of Incimaxx DES (containing peroxy acids and hydrogen peroxide) and Triquart Super (containing quaternary ammonium compound) in four consecutive cultures did not alter the frequency of antibiotic-tolerant isolates, as determined by plating on 2× the MIC for a range of antibiotics. Exposure of eight strains of L. monocytogenes to 1 and 4 μg/ml triclosan did not alter triclosan sensitivity. However, all eight strains became resistant to gentamicin (up to 16-fold increase in MIC) after exposure to sublethal triclosan concentrations. Gentamicin-resistant isolates of strains N53-1 and 4446 were also resistant to other aminoglycosides, such as kanamycin, streptomycin, and tobramycin. Gentamicin resistance remained at a high level also after five subcultures without triclosan or gentamicin. Aminoglycoside resistance can be caused by mutations in the target site, the 16S rRNA gene. However, such mutations were not detected in the N53-1-resistant isolates. A combination of gentamicin and ampicillin is commonly used in listeriosis treatment. The triclosan-induced resistance is, hence, of great concern. Further investigations are needed to determine the molecular mechanisms underlying the effect of triclosan. PMID:21746948

  7. Assessment of sublethal effects of methoxyfenozide on oriental fruit Moth (Lepidoptera: Tortricidae).

    Science.gov (United States)

    Borchert, Daniel M; Walgenbach, James F; Kennedy, George G

    2005-06-01

    Sublethal effects of the insect growth regulator methoxyfenozide were examined in oriental fruit moth, Grapholita molesta (Busck), in laboratory and field studies. In laboratory studies, oriental fruit moth larvae reared on diet amended with 0.1 ppm methoxyfenozide developed at the same rate as larvae reared on untreated diet, and paired moths reared as larvae from the same treated or untreated diets exhibited similar fecundity and fertility. Population growth differences over multiple generations were used to examine sublethal effects of methoxyfenozide on population dynamics in the field. Multiple single-tree cages were placed over apple (Malus spp.) trees treated with two applications of methoxyfenozide (70 g [AI] /ha) and nontreated trees. Cages were infested at a single time point with virgin male and female oriental fruit moth adults, and population growth was evaluated by egg counts, shoot infestation, fruit damage, and larval counts over a 12-wk period. Significantly fewer eggs, larvae, and damaged fruit were found on methoxyfenozide-treated compared with nontreated trees in 2001. Observed population differences may have been a result of direct mortality to eggs and larvae of the first generation rather than sublethal effects. In 2002, no differences were observed between treatments, but a heavy rain event shortly after the early infestation impacted the experiment. A late moth release treatment was tested in 2002 to examine the effects of residual methoxyfenozide 55 d after initial application. Significantly fewer eggs were found in the methoxyfenozide treatment compared with the control, but no differences existed among treatments in shoot infestation, percentage of damaged fruit, or larval populations. It was concluded direct mortality of eggs and larvae exposed to methoxyfenozide rather than sublethal effects were most important in reduction of subsequent generations.

  8. Lethal and sub-lethal effects of copper to the african catfish ( clarias ...

    African Journals Online (AJOL)

    Lethal and sub-lethal effects of copper on Clarias gariepinus were studied using a 96-hour static bioassay. Copper (as copper chloride, CuCl2 . H2O) was used to prepare the stock solution from which five standard concentrations 0.0, 1.8, 3.2, 5.6, and 10.0 mg/L were prepared (coded A – E). 15 juvenile C. gariepinus fish ...

  9. Wood anatomical analysis of Alnus incana and Betula pendula injured by a debris-flow event.

    Science.gov (United States)

    Arbellay, Estelle; Stoffel, Markus; Bollschweiler, Michelle

    2010-10-01

    Vessel chronologies in ring-porous species have been successfully employed in the past to extract the climate signal from tree rings. Environmental signals recorded in vessels of ring-porous species have also been used in previous studies to reconstruct discrete events of drought, flooding and insect defoliation. However, very little is known about the ability of diffuse-porous species to record environmental signals in their xylem cells. Moreover, time series of wood anatomical features have only rarely been used to reconstruct former geomorphic events. This study was therefore undertaken to characterize the wood anatomical response of diffuse-porous Alnus incana (L.) Moench and Betula pendula Roth to debris-flow-induced wounding. Tree microscopic response to wounding was assessed through the analysis of wood anatomical differences between injured rings formed in the debris-flow event year and uninjured rings formed in the previous year. The two ring types were examined close and opposite to the injury in order to determine whether wound effects on xylem cells decrease with increasing tangential distance from the injury. Image analysis was used to measure vessel parameters as well as fiber and parenchyma cell (FPC) parameters. The results of this study indicate that injured rings are characterized by smaller vessels as compared with uninjured rings. By contrast, FPC parameters were not found to significantly differ between injured and uninjured rings. Vessel and FPC parameters mainly remained constant with increasing tangential distance from the injury, except for a higher proportion of vessel lumen area opposite to the injury within A. incana. This study highlights the existence of anatomical tree-ring signatures-in the form of smaller vessels-related to past debris-flow activity and addresses a new methodological approach to date injuries inflicted on trees by geomorphic processes.

  10. HGF mediates the anti-inflammatory effects of PRP on injured tendons.

    Directory of Open Access Journals (Sweden)

    Jianying Zhang

    Full Text Available Platelet-rich plasma (PRP containing hepatocyte growth factor (HGF and other growth factors are widely used in orthopaedic/sports medicine to repair injured tendons. While PRP treatment is reported to decrease pain in patients with tendon injury, the mechanism of this effect is not clear. Tendon pain is often associated with tendon inflammation, and HGF is known to protect tissues from inflammatory damages. Therefore, we hypothesized that HGF in PRP causes the anti-inflammatory effects. To test this hypothesis, we performed in vitro experiments on rabbit tendon cells and in vivo experiments on a mouse Achilles tendon injury model. We found that addition of PRP or HGF decreased gene expression of COX-1, COX-2, and mPGES-1, induced by the treatment of tendon cells in vitro with IL-1β. Further, the treatment of tendon cell cultures with HGF antibodies reduced the suppressive effects of PRP or HGF on IL-1β-induced COX-1, COX-2, and mPGES-1 gene expressions. Treatment with PRP or HGF almost completely blocked the cellular production of PGE2 and the expression of COX proteins. Finally, injection of PRP or HGF into wounded mouse Achilles tendons in vivo decreased PGE2 production in the tendinous tissues. Injection of platelet-poor plasma (PPP however, did not reduce PGE2 levels in the wounded tendons, but the injection of HGF antibody inhibited the effects of PRP and HGF. Further, injection of PRP or HGF also decreased COX-1 and COX-2 proteins. These results indicate that PRP exerts anti-inflammatory effects on injured tendons through HGF. This study provides basic scientific evidence to support the use of PRP to treat injured tendons because PRP can reduce inflammation and thereby reduce the associated pain caused by high levels of PGE2.

  11. Sublethal dose of phoxim and Bombyx mori nucleopolyhedrovirus interact to elevate silkworm mortality.

    Science.gov (United States)

    Gu, ZhiYa; Li, FanChi; Hu, JingSheng; Ding, Chao; Wang, Chaoqian; Tian, JiangHai; Xue, Bin; Xu, KaiZun; Shen, WeiDe; Li, Bing

    2017-03-01

    Silkworm (Bombyx mori) is an economically important insect. It is relatively less resistant to certain chemicals and environment exposures such as pesticides and pathogens. After pesticide exposures, the silkworms are more susceptible to microbial infections. The mechanism underlying the susceptibility might be related to immune response and oxidative stress. A sublethal dose of phoxim combined with Bombyx mori nucleopolyhedrovirus (BmNPV) elevated the silkworm mortality at 96 h. We found a higher content of H2 O2 and increased levels of genes related to oxidative stress and immune response after treatment with a sublethal dose of phoxim for 24 h or 48 h. However, such response decreased with longer pesticide treatment. Mortality increased by 44% when B. mori was exposed to combined treatment with BmNPV and phoxim rather than BmNPV alone. The level of examined immune-related and oxidative-stress-related genes significantly decreased in the combined treatment group compared with the BmNPV group. Our results indicated that, with long-term exposure to pesticides such as OPs, even at sublethal dose, the oxidative stress response and immune responses in silkworm were inhibited, which may lead to further immune impairment and accumulation of oxidative stress, resulting in susceptibility to the virus and harm to the silkworm. Our study provided insights for understanding the susceptibility to pathogen after pesticide exposures, which may promote the development of better pesticide controls to avoid significant economic losses. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  12. Acute toxicity and sublethal effects of fipronil on detoxification enzymes in juvenile zebrafish (Danio rerio).

    Science.gov (United States)

    Wu, Haihua; Gao, Cuie; Guo, Yaping; Zhang, Yuping; Zhang, Jianzhen; Ma, Enbo

    2014-10-01

    The acute toxicity of fipronil and its sublethal effects on detoxification enzymes (carboxylesterases (CarEs), glutathione S-transferases (GSTs), and 7-ethoxycoumarin O-deethylase (ECOD)) in zebrafish (Danio rerio) were investigated. The results indicated that the 24-h LC50 of fipronil for zebrafish was 220.4 μg/L (95% CI: 173.7-272.4 μg/L). Sublethal concentrations of fipronil did not cause significant changes in CarEs activities. In the liver and muscle tissues, GST activities at the tested concentrations did not significantly differ from those in the control. In the brain and gill tissues, GST activities at a concentration of 4 μg/L were significantly lower than those at a concentration of 2 μg/L. The results suggest that CarEs and GSTs were not suitable biomarkers for fipronil effects in D. rerio. A significant induction in the ECOD activities in the brain, gill, liver, and muscle tissues was observed compared with the control. Moreover, the dose-dependent responses of the ECOD activity were observed after treatment with sublethal concentrations of fipronil in the range of 2-20 μg/L. The results suggested that ECOD could be a suitable biomarker of fipronil effects in D. rerio. Copyright © 2014. Published by Elsevier Inc.

  13. Fumigant Toxicity and Sublethal Effects of Artemisia khorassanica and Artemisia sieberi on Sitotroga cerealella (Lepidoptera: Gelechiidae).

    Science.gov (United States)

    Naseri, B; Abedi, Z; Abdolmaleki, A; Jafary-Jahed, M; Borzoui, E; Mozaffar Mansouri, Seyed

    2017-09-01

    Fumigant toxicity and sublethal effects of essential oils from Artemisia khorassanica Podl. and Artemisia sieberi Bess were investigated against adults of Sitotroga cerealella Olivier. To assess the sublethal effects, adult moths were exposed to the LC30 of each essential oil, and life table parameters of the surviving S. cerealella were studied. Higher fumigant toxicity of A. khorassanica (LC50: 7.38 µl/liter air) than A. sieberi (LC50: 9.26 µl/liter air) was observed against S. cerealella. Also, the insecticidal effects of A. khorassanica (LT50: 9.01 h) were faster than A. sieberi (LT50: 14.37 h). A significant extension was observed in the developmental time (egg to adult) of S. cerealella treated with the essential oils. In addition, fecundity of S. cerealella reduced by 25.29 and 35.78% following exposure to sublethal concentrations of A. sieberi and A. khorassanica, respectively. Both tested essential oils caused a significant reduction in the gross and net reproductive rates, intrinsic rate of increase (rm), and finite rate of increase of S. cerealella. The rm values following exposure to A. sieberi, A. khorassanica, and control were 0.098, 0.094, and 0.107 d-1, respectively. The results of this study suggest that tested essential oils have a good potential to apply in integrated pest management of S. cerealella. © The Author 2017. Published by Oxford University Press on behalf of Entomological Society of America.

  14. Fumigant Toxicity and Sublethal Effects of Artemisia khorassanica and Artemisia sieberi on Sitotroga cerealella (Lepidoptera: Gelechiidae)

    Science.gov (United States)

    Abedi, Z; Abdolmaleki, A; Jafary-Jahed, M; Borzoui, E; Mozaffar Mansouri, Seyed

    2017-01-01

    Abstract Fumigant toxicity and sublethal effects of essential oils from Artemisia khorassanica Podl. and Artemisia sieberi Bess were investigated against adults of Sitotroga cerealella Olivier. To assess the sublethal effects, adult moths were exposed to the LC30 of each essential oil, and life table parameters of the surviving S. cerealella were studied. Higher fumigant toxicity of A. khorassanica (LC50: 7.38 µl/liter air) than A. sieberi (LC50: 9.26 µl/liter air) was observed against S. cerealella. Also, the insecticidal effects of A. khorassanica (LT50: 9.01 h) were faster than A. sieberi (LT50: 14.37 h). A significant extension was observed in the developmental time (egg to adult) of S. cerealella treated with the essential oils. In addition, fecundity of S. cerealella reduced by 25.29 and 35.78% following exposure to sublethal concentrations of A. sieberi and A. khorassanica, respectively. Both tested essential oils caused a significant reduction in the gross and net reproductive rates, intrinsic rate of increase (rm), and finite rate of increase of S. cerealella. The rm values following exposure to A. sieberi, A. khorassanica, and control were 0.098, 0.094, and 0.107 d−1, respectively. The results of this study suggest that tested essential oils have a good potential to apply in integrated pest management of S. cerealella. PMID:29117375

  15. Sublethal effects of some botanical and chemical insecticides on the cotton whitefly, Bemisia tabaci (Hem: Aleyrodidae

    Directory of Open Access Journals (Sweden)

    Fatemeh Jafarbeigi

    2014-09-01

    Full Text Available In addition to direct mortalities caused by acute concentrations of insecticides, some biological traits of target pests may be also affected by sublethal doses. The cotton whitefly, Bemisia tabaci (Hem: Aleyrodidae is an important pest of a wide variety of agricultural crops across the world. The control of B. tabaci largely relies on wide application of chemical insecticides. In this study, we analyzed the life table parameters to evaluate the sublethal effect of three plant-derived insecticides (Fumaria parviflora (Fumariaceae, Teucrium polium (Lamiaceae, and Thymus vulgaris (Lamiaceae and two chemical insecticides (pymetrozin and neemarin on B. tabaci. The whiteflies were allowed to oviposit on plants infected with each of the five insecticides using leaf-dip method. The data were analyzed using the age-stage two-sex life table. We found significant differences in the gross reproductive rate (GRR, the net reproductive rat (R0, the intrinsic rate of increase (r and the finite rate of increase (λ of treated whiteflies compared to control. Our results showed that some biological traits of B. tabaci are affected by sub-lethal doses of the plant-derived extracts and that these effects are comparable to those of chemical insecticides. Given the detrimental effects of chemical insecticides on human, environment and non-target organisms, plant-derived insecticides may provide valuable environmentally friendly tools for pest management programs.

  16. Sexuality in Spinal-Cord-Injured High School Students.

    Science.gov (United States)

    Dailey, Anne Louise

    1982-01-01

    Presents facts, literature, and organization sources on the sexual functioning of spinal-cord-injured persons. Suggests using this information in counseling situations. Urges counselors to help their clients reach sexual maturity by speaking frankly about sexuality and expressing affection, and by accepting their sexual behavior as normal. (JAC)

  17. Psychoactive substances in seriously injured drivers in Denmark

    DEFF Research Database (Denmark)

    Simonsen, Kirsten Wiese; Steentoft, Anni; Bernhoft, IM

    2013-01-01

    , which is 0.53 g/l in Denmark. The percentage of seriously injured drivers testing positive for medicinal drugs at levels above the Danish legal limit was 6.8%. Benzodiazepines and Z-drugs (6.4%) comprised the majority of this group. One or more illegal drugs (primarily amphetamines and cannabis) were...

  18. Prehospital transport of spinal cord-injured patients in Nigeria ...

    African Journals Online (AJOL)

    Background. Well-organised and efficient prehospital transport is associated with an improved outcome in trauma patients. In Nigeria there is a paucity of information on prehospital transport of spinal cord-injured patients and its relation to mortality. Objective. To determine whether prehospital transportation is a predictor of ...

  19. Prehospital transport of spinal cord-injured patients in Nigeria

    African Journals Online (AJOL)

    that the means of transport are generally not optimal for those with spinal cord injury. The aim of the present study was therefore to highlight the importance of prehospital transport of spinal cord-injured patients and the contribution of these injuries to mortality in Nigeria. Patients, materials and methods. The records of spinal ...

  20. Outcome in elderly injured patients : injury severity versus host factors

    NARCIS (Netherlands)

    van der Sluis, CK; Timmer, HW; Eisma, WH; ten Duis, HJ

    1997-01-01

    To evaluate the differences between the outcome of elderly patients with severe injuries and that of their contemporaries with a less severe injury, we reviewed 42 severely injured elderly patients and compared them with 76 patients with a femoral neck fracture. We analysed the influence of injury

  1. Addressing the Needs of Children and Families of Combat Injured

    Science.gov (United States)

    2012-04-01

    subsequent treatments. His death was not related to participation in the current study. Participant death , specifically the combat-injured service member...connected her with bereavement services. One participant (service member receiving treatment at WRNMMC) voluntarily withdrew from the study at

  2. Prevent Tipping Furniture from Injuring or Killing Young Children

    Science.gov (United States)

    ... this! Home » Health Tips » Child Emergencies Prevent Tipping Furniture from Injuring or Killing Young Children The nation’s ... a child — killed by a piece of a furniture, appliance or a television falling on them. “It ...

  3. Effect of sildenafil on erectile dysfunction in spinal Cord injured ...

    African Journals Online (AJOL)

    Background: Erectile dysfunction is a preoccupying issue, just like motor and bladder disability, in spinal cord injured (SCI) patients. This is particularly so because of the increasing prevalence of paraplegic and tetraplegic subjects and the fact that these patients are younger, and sexually active. Objective: To determine the ...

  4. Repair and Enumeration of Injured Coliforms by a Plating Procedure

    Science.gov (United States)

    Speck, M. L.; Ray, B.; Read, R. B.

    1975-01-01

    Surface plating of coliforms on Trypticase soy agar, followed by 1 to 2 h of incubation at 25 C and subsequent overlay with violet red bile agar, was found to be a useful method for the repair and enumeration of coliforms injured by freezing. PMID:1092266

  5. Efficacy of a metalloproteinase inhibitor in spinal cord injured dogs.

    Science.gov (United States)

    Levine, Jonathan M; Cohen, Noah D; Heller, Michael; Fajt, Virginia R; Levine, Gwendolyn J; Kerwin, Sharon C; Trivedi, Alpa A; Fandel, Thomas M; Werb, Zena; Modestino, Augusta; Noble-Haeusslein, Linda J

    2014-01-01

    Matrix metalloproteinase-9 is elevated within the acutely injured murine spinal cord and blockade of this early proteolytic activity with GM6001, a broad-spectrum matrix metalloproteinase inhibitor, results in improved recovery after spinal cord injury. As matrix metalloproteinase-9 is likewise acutely elevated in dogs with naturally occurring spinal cord injuries, we evaluated efficacy of GM6001 solubilized in dimethyl sulfoxide in this second species. Safety and pharmacokinetic studies were conducted in naïve dogs. After confirming safety, subsequent pharmacokinetic analyses demonstrated that a 100 mg/kg subcutaneous dose of GM6001 resulted in plasma concentrations that peaked shortly after administration and were sustained for at least 4 days at levels that produced robust in vitro inhibition of matrix metalloproteinase-9. A randomized, blinded, placebo-controlled study was then conducted to assess efficacy of GM6001 given within 48 hours of spinal cord injury. Dogs were enrolled in 3 groups: GM6001 dissolved in dimethyl sulfoxide (n = 35), dimethyl sulfoxide (n = 37), or saline (n = 41). Matrix metalloproteinase activity was increased in the serum of injured dogs and GM6001 reduced this serum protease activity compared to the other two groups. To assess recovery, dogs were a priori stratified into a severely injured group and a mild-to-moderate injured group, using a Modified Frankel Scale. The Texas Spinal Cord Injury Score was then used to assess long-term motor/sensory function. In dogs with severe spinal cord injuries, those treated with saline had a mean motor score of 2 (95% CI 0-4.0) that was significantly (Pdogs receiving dimethyl sulfoxide (mean, 5; 95% CI 2.0-8.0) or GM6001 (mean, 5; 95% CI 2.0-8.0). As there was no independent effect of GM6001, we attribute improved neurological outcomes to dimethyl sulfoxide, a pleotropic agent that may target diverse secondary pathogenic events that emerge in the acutely injured cord.

  6. Altered hematological and immunological parameters in silver catfish (Rhamdia quelen) following short term exposure to sublethal concentration of glyphosate.

    Science.gov (United States)

    Kreutz, Luiz Carlos; Gil Barcellos, Leonardo José; de Faria Valle, Stella; de Oliveira Silva, Tális; Anziliero, Deniz; Davi dos Santos, Ezequiel; Pivato, Mateus; Zanatta, Rafael

    2011-01-01

    Using agrichemicals to control unwanted species has become a necessary and common worldwide practice to improve crop production. Although most currently used agrichemicals are considered relatively safe, continuous usage contributes for soil and water contamination and collateral toxic effects on aquatic species. Few studies correlated the presence of agrichemicals on fish blood cells and natural immune system. Thus, in this study, silver catfish (Rhamdia quelen) were exposed to sublethal concentrations (10% of the LC(50-96 h)) of a glyphosate based herbicide and hematological and natural immune system parameters were evaluated. Silver catfish fingerlings exposed to glyphosate for 96 h had a significant reduction on blood erythrocytes, thrombocytes, lymphocytes and total leukocytes in contrast to a significant increase in the number of immature circulating cells. The effect of glyphosate on natural immune system was evaluated after 24h or 10 days exposure by measuring the phagocytic index of coelomic cells, and lysozyme, total peroxidase, bacteria agglutination, bactericidal activity and natural complement hemolytic activity in the serum of fingerlings. A significant reduction on phagocytic index, serum bacteria agglutination and total peroxidase was observed only after 24h exposure to glyphosate. In contrast, fingerlings exposed to glyphosate for 10 days had a significant lower serum bacteria agglutination and lysozyme activity. Glyphosate had no effect on serum bactericidal and complement natural hemolytic activity after 24h or 10 days exposure. Nonetheless, the information obtained in this study indicates that glyphosate contaminated water contributes to alter blood cells parameters and to reduce the activity of natural immune components important to mediate fish resistance to infecting microorganisms. Copyright © 2010 Elsevier Ltd. All rights reserved.

  7. Characteristics of injured children attending the emergency department : patients potentially in need of rehabilitation

    NARCIS (Netherlands)

    Sturms, L.M.; van der Sluis, C.K.; Groothoff, J.W.; Ten Duis, H.J.; Eisma, W.H.

    Objective: To provide an epidemiological overview of the characteristics of injured children and to compare hospitalized and nonhospitalized injured children to identify predictors of hospitalization and, with that, possible predictors of disablement. Design: Retrospective analysis of data obtained

  8. Risk factors for subsequent impaired driving by injured passengers.

    Science.gov (United States)

    Purssell, Roy A; Chan, Herbert; Brown, Douglas; Fang, Ming; Brubacher, Jeffrey R

    2014-01-01

    The purpose of this study was to evaluate the rate of, and risk factors for, subsequent impaired driving activity (IDA) in a cohort of injured passengers who were treated for injuries in a Canadian trauma center. We studied adult passengers who were occupants in vehicles involved in motor vehicle crashes (MVCs) and either included in the British Columbia (BC) trauma registry (January 1, 1992-December 31, 2004) or treated in the emergency department (ED) of Vancouver General Hospital (VGH; January 1, 1999-December 31, 2003). Passengers were linked to their driver's license and hence to their driving record using personal health number and demographic information. Injured passengers were stratified into 3 groups based on their blood alcohol concentration (BAC) at time of ED presentation: group 1: BAC = 0, group 2: 0 17.3 mM (0.08%). Two outcome variables were studied: involvement in a subsequent IDA and time to their first subsequent IDA. IDA was defined as a criminal code conviction for impaired driving, a 24-h or 90-day license suspension for impaired driving, and/or involvement in an MVC where police cited alcohol as a factor. Time to first IDA following the index event among passenger BAC groups was compared with Kaplan-Meier survival analysis. Cox proportional hazards models were employed to examine the effect of various potential risk factors on time to engage in first IDA. Injured passengers with any BAC at the time of ED visit were more likely to engage in IDA and had their first IDA sooner after the index event than those with zero BAC. Among this cohort of injured passengers, 12.1 percent with BAC = 0, 29.9 percent with 0 17.3 mM (0.08%) engaged in IDA. Compared to passengers with BAC = 0, group 3 passengers and group 2 passengers were 2.06 times and 1.79 times more likely to engage in future IDA. Twenty-five percent of injured passengers engaged their first IDA by 57 and 38 months in groups 2 and 3, respectively. Previous IDA and being male were also

  9. Sialic acid accelerates the electrophoretic velocity of injured dorsal root ganglion neurons

    OpenAIRE

    Chen-xu Li; Guo-ying Ma; Min-fang Guo; Ying Liu

    2015-01-01

    Peripheral nerve injury has been shown to result in ectopic spontaneous discharges on soma and injured sites of sensory neurons, thereby inducing neuropathic pain. With the increase of membrane proteins on soma and injured site neurons, the negatively charged sialic acids bind to the external domains of membrane proteins, resulting in an increase of this charge. We therefore speculate that the electrophoretic velocity of injured neurons may be faster than non-injured neurons. The present stud...

  10. Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice.

    Science.gov (United States)

    Ieronimakis, Nicholas; Pantoja, Mario; Hays, Aislinn L; Dosey, Timothy L; Qi, Junlin; Fischer, Karin A; Hoofnagle, Andrew N; Sadilek, Martin; Chamberlain, Jeffrey S; Ruohola-Baker, Hannele; Reyes, Morayma

    2013-08-01

    Presently, there is no effective treatment for the lethal muscle wasting disease Duchenne muscular dystrophy (DMD). Here we show that increased sphingosine-1-phoshate (S1P) through direct injection or via the administration of the small molecule 2-acetyl-4(5)-tetrahydroxybutyl imidazole (THI), an S1P lyase inhibitor, has beneficial effects in acutely injured dystrophic muscles of mdx mice. We treated mdx mice with and without acute injury and characterized the histopathological and functional effects of increasing S1P levels. We also tested exogenous and direct administration of S1P on mdx muscles to examine the molecular pathways under which S1P promotes regeneration in dystrophic muscles. Short-term treatment with THI significantly increased muscle fiber size and extensor digitorum longus (EDL) muscle specific force in acutely injured mdx limb muscles. In addition, the accumulation of fibrosis and fat deposition, hallmarks of DMD pathology and impaired muscle regeneration, were lower in the injured muscles of THI-treated mdx mice. Furthermore, increased muscle force was observed in uninjured EDL muscles with a longer-term treatment of THI. Such regenerative effects were linked to the response of myogenic cells, since intramuscular injection of S1P increased the number of Myf5nlacz/+ positive myogenic cells and newly regenerated myofibers in injured mdx muscles. Intramuscular injection of biotinylated-S1P localized to muscle fibers, including newly regenerated fibers, which also stained positive for S1P receptor 1 (S1PR1). Importantly, plasma membrane and perinuclear localization of phosphorylated S1PR1 was observed in regenerating muscle fibers of mdx muscles. Intramuscular increases of S1P levels, S1PR1 and phosphorylated ribosomal protein S6 (P-rpS6), and elevated EDL muscle specific force, suggest S1P promoted the upregulation of anabolic pathways that mediate skeletal muscle mass and function. These data show that S1P is beneficial for muscle regeneration and

  11. Fast revascularization of the injured area is essential to support zebrafish heart regeneration.

    Science.gov (United States)

    Marín-Juez, Rubén; Marass, Michele; Gauvrit, Sebastien; Rossi, Andrea; Lai, Shih-Lei; Materna, Stefan C; Black, Brian L; Stainier, Didier Y R

    2016-10-04

    Zebrafish have a remarkable capacity to regenerate their heart. Efficient replenishment of lost tissues requires the activation of different cell types including the epicardium and endocardium. A complex set of processes is subsequently needed to support cardiomyocyte repopulation. Previous studies have identified important determinants of heart regeneration; however, to date, how revascularization of the damaged area happens remains unknown. Here, we show that angiogenic sprouting into the injured area starts as early as 15 h after injury. To analyze the role of vegfaa in heart regeneration, we used vegfaa mutants rescued to adulthood by vegfaa mRNA injections at the one-cell stage. Surprisingly, vegfaa mutants develop coronaries and revascularize after injury. As a possible explanation for these observations, we find that vegfaa mutant hearts up-regulate the expression of potentially compensating genes. Therefore, to overcome the lack of a revascularization phenotype in vegfaa mutants, we generated fish expressing inducible dominant negative Vegfaa. These fish displayed minimal revascularization of the damaged area. In the absence of fast angiogenic revascularization, cardiomyocyte proliferation did not occur, and the heart failed to regenerate, retaining a fibrotic scar. Hence, our data show that a fast endothelial invasion allows efficient revascularization of the injured area, which is necessary to support replenishment of new tissue and achieve efficient heart regeneration. These findings revisit the model where neovascularization is considered to happen concomitant with the formation of new muscle. Our work also paves the way for future studies designed to understand the molecular mechanisms that regulate fast revascularization.

  12. Histologic evaluation of the effects of Emdogain gel on injured root apex in rats.

    Science.gov (United States)

    Matsumoto, Noriyoshi; Minakami, Masahiko; Hatakeyama, Junko; Haruna, Chieko; Morotomi, Takahiko; Izumi, Toshio; Anan, Hisashi

    2014-12-01

    This study investigated the effects of Emdogain gel (EMD) on the injured open apex within periapical lesions. Periapical lesions were induced in rats by opening the pulp chambers of the mandibular first molars and filing the apical foramen through the distal root canal with #25 K-files to make an open apex. The teeth were exposed to the oral environment for 7 days. Then we irrigated the distal root canals and divided them into EMD-treated and propylene glycol alginate-treated groups. The rats were killed 7, 14, and 28 days after treatment and examined histochemically. In the EMD-treated rats, more cells expressed transforming growth factor-β1 or bone morphogenetic protein-2 at 7 days after treatment, and the regeneration of cementum and bone was observed around the root apex at 14 days after treatment. Conversely, in the propylene glycol alginate-treated group, few cells expressed transforming growth factor-β1 or bone morphogenetic protein-2, and apical periodontal tissue recovery was rarely seen within the periapical lesions throughout the experiment. These results suggest that EMD does not irritate injured periapical tissue and may create a favorable environment that promotes the healing of destroyed periapical tissues. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  13. Acute and Perioperative Care of the Burn-Injured Patient

    Science.gov (United States)

    Bittner, Edward A.; Shank, Erik; Woodson, Lee; Martyn, J.A. Jeevendra

    2016-01-01

    Care of burn-injured patients requires knowledge of the pathophysiologic changes affecting virtually all organs from the onset of injury until wounds are healed. Massive airway and/or lung edema can occur rapidly and unpredictably after burn and/or inhalation injury. Hemodynamics in the early phase of severe burn injury are characterized by a reduction in cardiac output, increased systemic and pulmonary vascular resistance. Approximately 2–5 days after major burn injury, a hyperdynamic and hypermetabolic state develops. Electrical burns result in morbidity much higher than expected based on burn size alone. Formulae for fluid resuscitation should serve only as guideline; fluids should be titrated to physiologic end points. Burn injury is associated basal and procedural pain requiring higher than normal opioid and sedative doses. Operating room concerns for the burn-injured patient include airway abnormalities, impaired lung function, vascular access, deceptively large and rapid blood loss, hypothermia and altered pharmacology. PMID:25485468

  14. Imagery use by injured athletes: a qualitative analysis.

    Science.gov (United States)

    Driediger, Molly; Hall, Craig; Callow, Nichola

    2006-03-01

    The purpose of this study was to expand our knowledge and increase our understanding of imagery use by athletes in sport-injury rehabilitation using a qualitative approach. The participants were 10 injured athletes who were receiving physiotherapy at the time they were interviewed. During the interviews, the athletes provided extensive information about their use of imagery during injury rehabilitation and it was clear that they believed imagery served cognitive, motivational and healing purposes in effectively rehabilitating an injury. Cognitive imagery was used to learn and properly perform the rehabilitation exercises. They employed motivational imagery for goal setting (e.g. imagined being fully recovered) and to enhance mental toughness, help maintain concentration and foster a positive attitude. Imagery was used to manage pain. The methods they employed for controlling pain included using imagery to practise dealing with expected pain, using imagery as a distraction, imagining the pain dispersing, and using imagery to block the pain. With respect to what they imaged (i.e. the content of their imagery), they employed both visual and kinaesthetic imagery and their images tended to be positive and accurate. It was concluded that the implementation of imagery alongside physical rehabilitation should enhance the rehabilitation experience and, therefore, facilitate the recovery rates of injured athletes. Moreover, it was recommended that those responsible for the treatment of injured athletes (e.g. medical doctors, physiotherapists) should understand the benefits of imagery in athletic injury rehabilitation, since it is these practitioners who are in the best position to encourage injured athletes to use imagery.

  15. Psychiatric morbidity among physically injured Syrian refugees in Turkey.

    Science.gov (United States)

    Al-Nuaimi, Saleem; Aldandashi, Samer; Easa, Abdul Kadir Saed; Saqqur, Maher

    2018-01-01

    To the best of our knowledge, the mental health status of physically injured Syrian refugees has not yet been investigated. The aim of this study was to examine the prevalence of psychiatric morbidity among physically injured Syrian refugees in Turkey receiving treatment at the main rehabilitation centre near the Syrian border. This is a cross sectional study. Information was collected from consenting injured Syrian refugees at Dar-el-Shefa'a Hospital in Reyhanlı (Turkey) during a one week period in December 2012 and another one week period in August 2013. A clinical psychiatric interview was conducted to determine a diagnosis according to the diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM) IV-TR. A total of 40 refugees consented and completed a clinical psychiatric interview. All refugees in this study did not have a significant past psychiatric history. The most prevalent current diagnosis was major depressive disorder (22.5%), adjustment disorder (20%), and post-traumatic stress disorder (15%). Five (12.5%) patients had no evidence of a psychiatric disorder. The prevalence of psychiatric morbidity among injured Syrian refugees in our study was extremely high. This may help guide the treatment and management of this select population. This study had a low number of participants. The method of assessment was not standardized with a validated tool. This study may help guide the treatment and management of this select population, both in neighbouring countries and as resettled refugees in Western host countries. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Early diagnostics of infectiouscomplications in injured persons with heavy traumas

    Directory of Open Access Journals (Sweden)

    S. A. Svistunov

    2012-01-01

    Full Text Available In this study specific features of the etiological structure of infectiouscomplications occurring early in treatment of heavily injured patients is established and the need is demonstrated for bacteriological examination of clinicalmaterials according to a developed algorithm on the second or third day after admission to hospital, which contributes totimely isolation of etiological agents of infectiouscomplications, rational etiotropic antibacterial treatment and reduced mortality.

  17. Injured pedestrians in Cape Town - the role of alcohol

    African Journals Online (AJOL)

    average, 35.6 years old. They were most frequently injured at night and over weekends. The BAC was positive in. 62.1 % of pedestrians, and the mean BAC was 0.19 g/dl. Most pedestrians had at least one lower limb injury and nearly half had a head injury; however, BAC-positive pedestrians were 2.6 times more likely to ...

  18. Contact irritant responses of Aedes aegypti Using sublethal concentration and focal application of pyrethroid chemicals.

    Directory of Open Access Journals (Sweden)

    Hortance Manda

    Full Text Available BACKGROUND: Previous studies have demonstrated contact irritant and spatial repellent behaviors in Aedes aegypti following exposure to sublethal concentrations of chemicals. These sublethal actions are currently being evaluated in the development of a push-pull strategy for Ae. aegypti control. This study reports on mosquito escape responses after exposure to candidate chemicals for a contact irritant focused push-pull strategy using varying concentrations and focal application. METHODS: Contact irritancy (escape behavior, knockdown and 24 hour mortality rates were quantified in populations of female Ae. aegypti under laboratory conditions and validated in the field (Thailand and Peru using experimental huts. Evaluations were conducted using varying concentrations and treatment surface area coverage (SAC of three pyrethroid insecticides: alphacypermethrin, lambacyhalothrin and deltamethrin. RESULTS: Under laboratory conditions, exposure of Ae. aegypti to alphacypermethrin using the standard field application rate (FAR resulted in escape responses at 25% and 50% SAC that were comparable with escape responses at 100% SAC. Significant escape responses were also observed at <100% SAC using ½FAR of all test compounds. In most trials, KD and 24 hour mortality rates were higher in mosquitoes that did not escape than in those that escaped. In Thailand, field validation studies indicated an early time of exit (by four hours and 40% increase in escape using ½FAR of alphacypermethrin at 75% SAC compared to a matched chemical-free control. In Peru, however, the maximum increase in Ae. aegypti escape from alphacypermethrin-treated huts was 11%. CONCLUSIONS/SIGNIFICANCE: Results presented here suggest a potential role for sublethal and focal application of contact irritant chemicals in an Ae. aegypti push-pull strategy to reduce human-vector contact inside treated homes. However, the impact of an increase in escape response on dengue virus transmission is

  19. Identification of sublethal toxicants in a BC coastal pulp and paper mill effluent

    Energy Technology Data Exchange (ETDEWEB)

    Eickhoff, C.V.; Pickard, J.; Kinnee, K. [BC Research Inc., Vancouver, BC (Canada); Dwernychuk, W. [Hatfield Consultants Ltd., West Vancouver, BC (Canada); Birkholz, D. [EnviroTest Lab., Edmonton, AB (Canada); Kilback, D. [Pacifica Papers, Powell River, BC (Canada)

    2001-06-01

    BC Research Inc. conducted a toxicity identification evaluation to identify the different compounds comprised in the mill Outfall number 1 effluent. The Environmental Effects Monitoring program had determined that these compounds were responsible for sublethal effects to organisms. Echinoderm species like the sand dollar, Dendraster excentricus Eshscholtz, the purple sea urchin, Stronglyocentrotus purpuratus Stimpson, and the marine algae, Champia parvula had suffered toxicity caused by the mill effluent. The last several Environmental Effects Monitoring testing periods had shown the sublethal toxicity of the Outfall number 1 effluent to echinoderms was very consistent. Based on the high cost and shipping associated with the Champia bioassays, toxicity tests conducted during the peak spawning season of the sea urchin and the non significant difference between the sensitivity of the sand dollar and the purple sea urchin, the purple sea urchin was selected to evaluate the toxicity of the manipulated samples for the tests. The tests conducted were: a baseline toxicity test performed immediately upon receipt of the effluent sample, the pH adjustment filtration test to determine if the toxic compound can be removed using filtration, the pH adjustment aeration test to determine if volatile compounds in the sample are toxic, the pH adjustment solid phase extraction test to determine the level of toxicity from organic compounds and metal chelates that can be removed by solid phase extraction. The results indicated that it seems high molecular weight molecules were responsible for the sublethal toxicity observed. Two different sources could be responsible: lignin derived macromolecules, and polymer compounds used as flocculants and sizing agents. Further testing of the pulp mill effluent to identify the source of the toxic high molecular weight compounds was recommended. 22 refs., 4 tabs., 6 figs.

  20. Manure source and age affect survival of zoonotic pathogens during aerobic composting at sublethal temperatures.

    Science.gov (United States)

    Erickson, Marilyn C; Smith, Chris; Jiang, Xiuping; Flitcroft, Ian D; Doyle, Michael P

    2015-02-01

    Heat is the primary mechanism by which aerobic composting inactivates zoonotic bacterial pathogens residing within animal manures, but at sublethal temperatures, the time necessary to hold the compost materials to ensure pathogen inactivation is uncertain. To determine the influence of the type of nitrogen amendment on inactivation of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in compost mixtures stored at sublethal temperatures, specific variables investigated in these studies included the animal source of the manure, the initial carbon/nitrogen (C:N) ratio of the compost mixture, and the age of the manure. Salmonella and L. monocytogenes were both inactivated more rapidly in chicken and swine compost mixtures stored at 20°C when formulated to an initial C:N ratio of 20:1 compared with 40:1, whereas a C:N ratio did not have an effect on inactivation of these pathogens in cow compost mixtures. Pathogen inactivation was related to the elevated pH of the samples that likely arises from ammonia produced by the indigenous microflora in the compost mixtures. Indigenous microbial activity was reduced when compost mixtures were stored at 30°C and drier conditions (compost mixtures prepared with aged chicken litter compared with fresh chicken litter, whereas E. coli O157:H7 survived to similar extents in compost mixtures prepared with either fresh or aged cow manure. The different responses observed when different sources of manure were used in compost mixtures reveal that guidelines with times required for pathogen inactivation in compost mixtures stored at sublethal temperatures should be dependent on the source of nitrogen, i.e., type of animal manure, present.

  1. Toxicity and Sublethal Effects of Cantharidin on Musca domestica (Diptera: Muscidae).

    Science.gov (United States)

    Yasoob, Hassan; Ali Khan, Hafiz Azhar; Zhang, Yalin

    2017-09-27

    The house fly, Musca domestica L. (Diptera: Muscidae), is a major pest of medical and veterinary importance all over the world. Management efforts for house flies are usually compromised owing to their resistance to many groups of conventional insecticides. Cantharidin, a natural toxin produced by meloid beetles, is a biopesticide with a reported toxicity to some insect pests including house flies. However, the effects of cantharidin on biological and fitness parameters of house flies have not yet been investigated. In the present study, we investigated the toxicity and sublethal effects of cantharidin on biological parameters of house flies for two consecutive generations. The results revealed that the values of LC50, LC25, LC10, and LC2 against house flies were to be 2.45, 1.23, 0.66, and 0.30 mg/liter, respectively. Sublethal effects of these concentrations on the development and reproduction parameters of house flies revealed that cantharidin reduced population growth by affecting pupation rate, adult emergence, and by lengthening developmental time. The female ratio, fecundity, egg hatching, and survival of adult flies were significantly reduced at LC2, LC10, LC25, and LC50 of cantharidin when compared with the control group. Furthermore, the increase in concentration of cantharidin had a significant effect on reducing the mean values of mean relative growth rate, net reproductive rate (Ro), intrinsic rate of natural increase (rm), and biotic potential (bp). In conclusion, the results of this study revealed the toxicity of cantharidin against house flies and the adverse effects of sublethal concentrations on biological parameters which may have positive implications for effective management of house flies. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  2. Sublethal effects on wood frogs chronically exposed to environmentally relevant concentrations of two neonicotinoid insecticides.

    Science.gov (United States)

    Robinson, Stacey A; Richardson, Sarah D; Dalton, Rebecca L; Maisonneuve, France; Trudeau, Vance L; Pauli, Bruce D; Lee-Jenkins, Stacey S Y

    2017-04-01

    Neonicotinoids are prophylactically used globally on a variety of crops, and there is concern for the potential impacts of neonicotinoids on aquatic ecosystems. The intensive use of pesticides on crops has been identified as a contributor to population declines of amphibians, but currently little is known regarding the sublethal effects of chronic neonicotinoid exposure on amphibians. The objective of the present study was to characterize the sublethal effect(s) of exposure to 3 environmentally relevant concentrations (1 μg/L, 10 μg/L, and 100 μg/L) of 2 neonicotinoids on larval wood frogs (Lithobates sylvaticus) using outdoor mesocosms. We exposed tadpoles to solutions of 2 commercial formulations containing imidacloprid and thiamethoxam, and assessed survival, growth, and development. Exposure to imidacloprid at 10 μg/L and 100 μg/L increased survival and delayed completion of metamorphosis compared with controls. Exposure to thiamethoxam did not influence amphibian responses. There was no significant effect of any treatment on body mass or size of the metamorphs. The results suggest that current usage of imidacloprid and thiamethoxam does not pose a threat to wood frogs. However, further assessment of both direct and indirect effects on subtle sublethal endpoints, and the influence of multiple interacting stressors at various life stages, is needed to fully understand the effects of neonicotinoids on amphibians. Environ Toxicol Chem 2017;36:1101-1109. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

  3. Understanding contextual influences of community reintegration among injured servicemembers.

    Science.gov (United States)

    Hawkins, Brent L; McGuire, Francis A; Linder, Sandra M; Britt, Thomas W

    2015-01-01

    As part of a larger mixed-methods research project investigating the influence of contextual factors on community reintegration (CR), this qualitative study sought to understand the subjective experiences of injured servicemembers and their perception of how contextual factors influenced their CR. More specifically, this article addresses how the influences of contextual factors differ between injured servicemembers with different levels of CR. Using a phenomenological framework, semistructured interviews were conducted with nine injured, community-dwelling servicemembers with low, moderate, and high levels of CR (three per category). Participants provided in-depth descriptions of the contextual barriers and facilitators of CR. Thematic analysis indicated the importance of social support and personal factors (e.g., self-efficacy, personal motivation) as the primary means for being reintegrated into their homes and communities. Other themes indicated factors that had an indirect but important influence on CR, including adapted sports, recreation, and other social programs; rehabilitation programs and therapists; school, work, and volunteering; and organizations and policies in developing social supports and personal factors. Comparisons between servicemembers indicated participants with low CR described many more contextual barriers and far fewer contextual facilitators to reintegration than those with high CR. Those with moderate CR were unique in that they described many facilitators and barriers to reintegration.

  4. Decreased mortality after prehospital interventions in severely injured trauma patients.

    Science.gov (United States)

    Meizoso, Jonathan P; Valle, Evan J; Allen, Casey J; Ray, Juliet J; Jouria, Jassin M; Teisch, Laura F; Shatz, David V; Namias, Nicholas; Schulman, Carl I; Proctor, Kenneth G

    2015-08-01

    We test the hypothesis that prehospital interventions (PHIs) performed by skilled emergency medical service providers during ground or air transport adversely affect outcome in severely injured trauma patients. Consecutive trauma activations (March 2012 to June 2013) transported from the scene by air or ground emergency medical service providers were reviewed. PHI was defined as intubation, needle decompression, tourniquet, cricothyroidotomy, or advanced cardiac life support. In 3,733 consecutive trauma activations (71% blunt, 25% penetrating, 4% burns), age was 39 years, 74% were male, Injury Severity Score (ISS) was 5, and Glasgow Coma Score (GCS) was 15, with 32% traumatic brain injury (TBI) and 7% overall mortality. Those who received PHI (n = 130, 3.5% of the trauma activations) were more severely injured: ISS (26 vs. 5), GCS (3 vs. 15), TBI (57% vs. 31%), Revised Trauma Score (RTS, 5.45 vs. 7.84), Trauma and Injury Severity Score (TRISS, 1.32 vs. 4.89), and mortality (56% vs. 5%) were different (all p blunt injury, high ISS, and long prehospital times (all p blunt trauma, and air transport were similar, but mortality was significantly lower (43% vs. 23%, p= 0.021). In our urban trauma system, PHIs are associated with a lower incidence of mortality in severely injured trauma patients and do not delay transport to definitive care. Prognostic/epidemiologic study, level III; therapeutic study, level IV.

  5. Response to and recovery from acute sublethal gamma radiation in the Amazon molly, Poecilia formosa

    Energy Technology Data Exchange (ETDEWEB)

    Woodhead, A.D.; Setlow, R.B.

    1979-05-01

    Acute irradiation of the Amazon molly with a sublethal dose of 1,000 rad caused some damage to the intestinal tract and to the haematopoietic system. Histologically, the intestine appeared to have regenerated by the end of a week; damage to the haematopoietic tissue appeared more slowly, but repair was almost complete some two months later. Nevertheless, recovery to the intestine cannot have been entirely completed in seven days, since the fish did not feed well for the following two weeks. After this, there were no obvious deleterious effects upon the survival and viability of the fish, although irradiated fish weighed less at the termination of the experiment.

  6. A translational murine model of sub-lethal intoxication with Shiga toxin 2 reveals novel ultrastructural findings in the brain striatum.

    Directory of Open Access Journals (Sweden)

    Carla Tironi-Farinati

    Full Text Available Infection by Shiga toxin-producing Escherichia coli causes hemorrhagic colitis, hemolytic uremic syndrome (HUS, acute renal failure, and also central nervous system complications in around 30% of the children affected. Besides, neurological deficits are one of the most unrepairable and untreatable outcomes of HUS. Study of the striatum is relevant because basal ganglia are one of the brain areas most commonly affected in patients that have suffered from HUS and since the deleterious effects of a sub-lethal dose of Shiga toxin have never been studied in the striatum, the purpose of this study was to attempt to simulate an infection by Shiga toxin-producing E. coli in a murine model. To this end, intravenous administration of a sub-lethal dose of Shiga toxin 2 (0.5 ηg per mouse was used and the correlation between neurological manifestations and ultrastructural changes in striatal brain cells was studied in detail. Neurological manifestations included significant motor behavior abnormalities in spontaneous motor activity, gait, pelvic elevation and hind limb activity eight days after administration of the toxin. Transmission electron microscopy revealed that the toxin caused early perivascular edema two days after administration, as well as significant damage in astrocytes four days after administration and significant damage in neurons and oligodendrocytes eight days after administration. Interrupted synapses and mast cell extravasation were also found eight days after administration of the toxin. We thus conclude that the chronological order of events observed in the striatum could explain the neurological disorders found eight days after administration of the toxin.

  7. Exposure to a sublethal concentration of imidacloprid and the side effects on target and nontarget organs of Apis mellifera (Hymenoptera, Apidae).

    Science.gov (United States)

    Catae, Aline Fernanda; Roat, Thaisa Cristina; Pratavieira, Marcel; Silva Menegasso, Anally Ribeiro da; Palma, Mario Sergio; Malaspina, Osmar

    2018-03-01

    The use of insecticides has become increasingly frequent, and studies indicate that these compounds are involved in the intoxication of bees. Imidacloprid is a widely used neonicotinoid; thus, we have highlighted the importance of assessing its oral toxicity to Africanized bees and used transmission electron microscopy to investigate the sublethal effects in the brain, the target organ, and the midgut, responsible for the digestion/absorption of food. In addition, the distribution of proteins involved in important biological processes in the brain were evaluated on the 1st day of exposure by MALDI-imaging analysis. Bioassays were performed to determine the Median Lethal Concentration (LC 50 ) of imidacloprid to bees, and the value obtained was 1.4651 ng imidacloprid/μL diet. Based on this result, the sublethal concentration to be administered at 1, 4 and 8 days was established as a hundredth (1/100) of the LC 50 . The results obtained from the ultrastructural analysis showed alterations in the midgut cells of bees as nuclear and mitochondrial damage and an increase of vacuoles. The insecticide caused spacing among the Kenyon cells in the mushroom bodies, chromatin condensation and loss of mitochondrial cristae. The MALDI-imaging analysis showed an increase in the expression of such proteins as vascular endothelial growth factor receptor, amyloid protein precursor and protein kinase C, which are related to oxygen supply, neuronal degeneration and memory/learning, and a decrease in the expression of the nicotinic acetylcholine receptor alpha 1, which is fundamental to the synapses. These alterations demonstrated that imidacloprid could compromise the viability of the midgut epithelium, as well as inhibiting important cognitive processes in individuals, and may be reflected in losses of the colony.

  8. Premature induction of aging in sublethally H2O2-treated young MRC5 fibroblasts correlates with increased glutathione peroxidase levels and resistance to DNA breakage.

    Science.gov (United States)

    Caldini, R; Chevanne, M; Mocali, A; Tombaccini, D; Paoletti, F

    1998-09-15

    Human MRC5 fibroblasts, at different passages in cultures, were used as an in vitro model to assess variations and/or induction of aging parameters under basal conditions or following sublethal oxidative stress by H2O2. DNA sensitivities to oxidatively-induced breakage, rather than basal levels of damaged DNA, were significantly different between cultures at low and high population doubling level (PDL): old cells maintained most of their DNA integrity even at high concentrations of H2O2, while young cells showed more extensive DNA damage which developed in a dose-dependent fashion. However, young cells pretreated with low doses of H2O2 exhibited increased resistance against further oxidative damage to DNA thus reproducing a senescent-like profile of sensitivity. In turn, DNA from old cultures incubated in a NAD precursor-free medium was more prone to H2O2-induced strand breaks mimicking DNA sensitivity of young cells. The extent of oxidatively-induced DNA damage in MRC5 populations correlated inversely with the levels of glutathione peroxidase (GPx) activity that almost doubled when cells passed from the young to the senescent stage. In addition, H2O2-pretreatment of young cells induced an increase in GPx expression approaching old cell values and promoted also the premature appearance of neutral beta-galactosidase activity and decreased c-fos expression upon serum stimulation, both of which were assumed to be characteristic traits of the senescent phenotype.

  9. Neurotrauma: The Crosstalk between Neurotrophins and Inflammation in the Acutely Injured Brain

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    Lindolfo da Silva Meirelles

    2017-05-01

    Full Text Available Traumatic brain injury (TBI is a major cause of morbidity and mortality among young individuals worldwide. Understanding the pathophysiology of neurotrauma is crucial for the development of more effective therapeutic strategies. After the trauma occurs, immediate neurologic damage is produced by the traumatic forces; this primary injury triggers a secondary wave of biochemical cascades together with metabolic and cellular changes, called secondary neural injury. In the scenario of the acutely injured brain, the ongoing secondary injury results in ischemia and edema culminating in an uncontrollable increase in intracranial pressure. These areas of secondary injury progression, or areas of “traumatic penumbra”, represent crucial targets for therapeutic interventions. Neurotrophins are a class of signaling molecules that promote survival and/or maintenance of neurons. They also stimulate axonal growth, synaptic plasticity, and neurotransmitter synthesis and release. Therefore, this review focuses on the role of neurotrophins in the acute post-injury response. Here, we discuss possible endogenous neuroprotective mechanisms of neurotrophins in the prevailing environment surrounding the injured areas, and highlight the crosstalk between neurotrophins and inflammation with focus on neurovascular unit cells, particularly pericytes. The perspective is that neurotrophins may represent promising targets for research on neuroprotective and neurorestorative processes in the short-term following TBI.

  10. Sublethal and hormesis effects of imidacloprid on the soybean aphid Aphis glycines.

    Science.gov (United States)

    Qu, Yanyan; Xiao, Da; Li, Jinyu; Chen, Zhou; Biondi, Antonio; Desneux, Nicolas; Gao, Xiwu; Song, Dunlun

    2015-04-01

    The soybean aphid, Aphis glycines Matsumura, is a major pest in soybean crop. Current management of this pest relies mainly on insecticides applications, and the neonicotinoid imidacloprid has been proposed as an effective insecticide to control A. glycines in soybean field. Imidacloprid at lethal concentrations not only exerts acute toxicity to A. glycines, but also cause various biological changes when aphids are chronically exposed to lower concentrations. In this study, we assessed the effects of a low-lethal (0.20 mg L(-1)) and two sublethal (0.05 and 0.10 mg L(-1)) imidacloprid concentrations on various A. glycines life history traits. Aphid exposure to 0.20 mg L(-1) imidacloprid caused slower juvenile development, shorter reproductive period, and reduced adult longevity, fecundity and total lifespan. Stimulatory effects, i.e. hormesis, on reproduction and immature development duration were observed in aphids exposed to the lower sublethal imidacloprid concentrations. Consequently, the net reproduction rate (R 0) was significantly higher than in the control aphids. These findings stress the importance of the actual imidacloprid concentration in its toxicological properties on A. glycines. Therefore, our results would be useful for assessing the overall effects of imidacloprid on A. glycines and for optimizing integrated pest management programs targeting this pest.

  11. Gene expression changes in honey bees induced by sublethal imidacloprid exposure during the larval stage.

    Science.gov (United States)

    Wu, Ming-Cheng; Chang, Yu-Wen; Lu, Kuang-Hui; Yang, En-Cheng

    2017-09-01

    Honey bee larvae exposed to sublethal doses of imidacloprid show behavioural abnormalities as adult insects. Previous studies have demonstrated that this phenomenon originates from abnormal neural development in response to imidacloprid exposure. Here, we further investigated the global gene expression changes in the heads of newly emerged adults and observed that 578 genes showed more than 2-fold changes in gene expression after imidacloprid exposure. This information might aid in understanding the effects of pesticides on the health of pollinators. For example, the genes encoding major royal jelly proteins (MRJPs), a group of multifunctional proteins with significant roles in the sustainable development of bee colonies, were strongly downregulated. These downregulation patterns were further confirmed through analyses using quantitative reverse transcription-polymerase chain reaction on the heads of 6-day-old nurse bees. To our knowledge, this study is the first to demonstrate that sublethal doses of imidacloprid affect mrjp expression and likely weaken bee colonies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Macroalgal extracts induce bacterial assemblage shifts and sublethal tissue stress in Caribbean corals.

    Science.gov (United States)

    Morrow, Kathleen M; Ritson-Williams, Raphael; Ross, Cliff; Liles, Mark R; Paul, Valerie J

    2012-01-01

    Benthic macroalgae can be abundant on present-day coral reefs, especially where rates of herbivory are low and/or dissolved nutrients are high. This study investigated the impact of macroalgal extracts on both coral-associated bacterial assemblages and sublethal stress response of corals. Crude extracts and live algal thalli from common Caribbean macroalgae were applied onto the surface of Montastraea faveolata and Porites astreoides corals on reefs in both Florida and Belize. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene amplicons was used to examine changes in the surface mucus layer (SML) bacteria in both coral species. Some of the extracts and live algae induced detectable shifts in coral-associated bacterial assemblages. However, one aqueous extract caused the bacterial assemblages to shift to an entirely new state (Lobophora variegata), whereas other organic extracts had little to no impact (e.g. Dictyota sp.). Macroalgal extracts more frequently induced sublethal stress responses in M. faveolata than in P. astreoides corals, suggesting that cellular integrity can be negatively impacted in selected corals when comparing co-occurring species. As modern reefs experience phase-shifts to a higher abundance of macroalgae with potent chemical defenses, these macroalgae are likely impacting the composition of microbial assemblages associated with corals and affecting overall reef health in unpredicted and unprecedented ways.

  13. Histopathological effect of sub-lethal concentration of aluminum phosphide (phostoxin on Clarias gariepinus juveniles

    Directory of Open Access Journals (Sweden)

    Kayode B. Olurin

    Full Text Available Abtsract: The study evaluated the effect of sub-lethal concentration of phostoxin on Clarias gariepinus juveniles. C. gariepinus juveniles belonging to the same cohort (40.1±1.2g; 18.1±1.1cm from a commercial fish farm were randomly placed ten in each of 15 plastic tanks containing 15 liters of water. They were exposed for 96 hrs to three sub-lethal concentrations (treatments of phostoxin (0.125, 0.250, 0.5mg L-1 and a phostoxin free control. At the end of 96 hrs exposure, they were dissected and the tissues need for histopathology removed and fixed in Bouin's fluid. The gill filament exhibited fusion at the secondary lamella that was progressive with concentration. At the highest concentration of exposure, the secondary lamellae showed marked pyknotic and necrotic changes characterized by epithelia detachment. The hepatic tissue showed mild inflammatory changes at lower concentrations while at the highest concentration of exposure there was marked inflammation with observed hydropic degeneration. In the kidney, an inflammatory change was only observed in the interstices at the highest dose of exposure with the convoluted tubules showing partial shrinkage. Phostoxin showed to have significantly caused alterations in cyto-architecture of the gills and to a considerable extent liver and kidney of C. gariepinus.

  14. Effects of Sublethal Concentrations of Insecticides on the Functional Response of Two Mirid Generalist Predators.

    Directory of Open Access Journals (Sweden)

    Angeliki F Martinou

    Full Text Available The use of agrochemicals particularly pesticides, can hamper the effectiveness of natural enemies, causing disruption in the ecosystem service of biological control. In the current study, the effects of the insecticides thiacloprid and chlorantraniliprole on the functional response curves were assessed for two mirid predator nymphs, Macrolophus pygmaeus Rambur and Nesidiocoris tenuis Reuter. In the absence of insecticides, both predators exhibited a type II functional response when feeding on eggs of the moth Ephestia kuehniella. N. tenuis seems to be a more efficient predator than M. pygmaeus, as model estimated handling time was significantly lower for the former than for the latter. Residual exposure of M. pygmaeus to sublethal concentrations of either insecticide was associated with a change in the asymptote but not the type of the functional response curve. Thiacloprid seems to be the least compatible with M. pygmaeus, as it led to both a significant reduction of the attack rate and an increase in handling time. In contrast, chlorantraniliprole exposure significantly increased the handling time, but not the attack rate of the predator. Residual exposure of N. tenuis to sublethal concentrations of either insecticide did not have a significant effect on the type nor the parameters of the functional response model. The results show that pesticide residues that do not have lethal effects on beneficial arthropods can reduce prey consumption depending on predator species and on likely risks associated with toxicity.

  15. Using photopigment biomarkers to quantify sub-lethal effects of petroleum pollution on natural phytoplankton assemblages

    Energy Technology Data Exchange (ETDEWEB)

    Swistak, J.; Pinckney, J.; Piehler, M.; Paerl, H. [Univ. of North Carolina, Morehead City, NC (United States). Inst. of Marine Sciences

    1995-12-31

    Although much work has been undertaken to determine the toxicity of petroleum pollutants to phytoplankton, most studies have used pure cultures to monitor growth of selected phytoplankton species. Fewer have considered the net effect on entire microalgal communities. Using high performance liquid chromatography (HPLC) to characterize diagnostic microalgal pigments, the authors were able to simultaneously assess sub-lethal pollutant effects on entire communities as well as on individual phytoplankton functional groups. Incubations of natural water samples with diesel fuel, an important contributor to coastal petroleum pollution, revealed significant changes in photopigments and relative abundance of taxonomic groups at sub-lethal concentrations. Differential rates of change of indicator pigment concentrations suggest a range of sensitivity among phytoplankton groups. In preliminary experiments, cyanobacteria exhibited the greatest overall tolerance to the diesel fuel concentrations tested, while cryptomonads displayed the most sensitivity. The authors are currently evaluating the responses of seasonal phytoplankton populations from 3 sites exposed to varied levels of petroleum pollution. HPLC will be used to characterize phytoplankton populations and to determine if the most abundant groups are also the most tolerant of diesel fuel. Preliminary experiments indicate that diesel fuel pollution may modify the structure and function of phytoplankton communities and subsequently alter the trophodynamics of impacted systems.

  16. Chronic exposure of corals to fine sediments: lethal and sub-lethal impacts.

    Science.gov (United States)

    Flores, Florita; Hoogenboom, Mia O; Smith, Luke D; Cooper, Timothy F; Abrego, David; Negri, Andrew P

    2012-01-01

    Understanding the sedimentation and turbidity thresholds for corals is critical in assessing the potential impacts of dredging projects in tropical marine systems. In this study, we exposed two species of coral sampled from offshore locations to six levels of total suspended solids (TSS) for 16 weeks in the laboratory, including a 4 week recovery period. Dose-response relationships were developed to quantify the lethal and sub-lethal thresholds of sedimentation and turbidity for the corals. The sediment treatments affected the horizontal foliaceous species (Montipora aequituberculata) more than the upright branching species (Acropora millepora). The lowest sediment treatments that caused full colony mortality were 30 mg l(-1) TSS (25 mg cm(-2) day(-1)) for M. aequituberculata and 100 mg l(-1) TSS (83 mg cm(-2) day(-1)) for A. millepora after 12 weeks. Coral mortality generally took longer than 4 weeks and was closely related to sediment accumulation on the surface of the corals. While measurements of damage to photosystem II in the symbionts and reductions in lipid content and growth indicated sub-lethal responses in surviving corals, the most reliable predictor of coral mortality in this experiment was long-term sediment accumulation on coral tissue.

  17. Chronic exposure of corals to fine sediments: lethal and sub-lethal impacts.

    Directory of Open Access Journals (Sweden)

    Florita Flores

    Full Text Available Understanding the sedimentation and turbidity thresholds for corals is critical in assessing the potential impacts of dredging projects in tropical marine systems. In this study, we exposed two species of coral sampled from offshore locations to six levels of total suspended solids (TSS for 16 weeks in the laboratory, including a 4 week recovery period. Dose-response relationships were developed to quantify the lethal and sub-lethal thresholds of sedimentation and turbidity for the corals. The sediment treatments affected the horizontal foliaceous species (Montipora aequituberculata more than the upright branching species (Acropora millepora. The lowest sediment treatments that caused full colony mortality were 30 mg l(-1 TSS (25 mg cm(-2 day(-1 for M. aequituberculata and 100 mg l(-1 TSS (83 mg cm(-2 day(-1 for A. millepora after 12 weeks. Coral mortality generally took longer than 4 weeks and was closely related to sediment accumulation on the surface of the corals. While measurements of damage to photosystem II in the symbionts and reductions in lipid content and growth indicated sub-lethal responses in surviving corals, the most reliable predictor of coral mortality in this experiment was long-term sediment accumulation on coral tissue.

  18. Lethal and sublethal effects of cyantraniliprole on Bactrocera dorsalis (Hendel) (Diptera: Tephritidae).

    Science.gov (United States)

    Zhang, Ruimin; Jang, Eric B; He, Shiyu; Chen, Jiahua

    2015-02-01

    The oriental fruit fly, Bactrocera dorsalis (Hendel), is one of the most globally important insect pests. Studies were conducted with the novel anthranilic diamide insecticide cyantraniliprole to determine its lethal and sublethal effects on B. dorsalis. An ingestion toxicity bioassay showed that cyantraniliprole was active against B. dorsalis, and the 72 h feeding LC50 was 3.22 µg g(-1) in adult diet for a susceptible strain. Sublethal doses of cyantraniliprole (1.30 µg g(-1) adult diet) induced a hormesis effect on B. dorsalis. The mating competitiveness of B. dorsalis treated with cyantraniliprole at 3.27 µg g(-1) adult diet was significantly lower when compared with the controls. The lower dose (1.30 µg g(-1) adult diet) of cyantraniliprole improved the total mating times of both mating pairs in treated groups and also the mating competitiveness of the treated males when compared with the higher dose and controls. Cyantraniliprole-treated females of the mated pairs with the lower dose laid more eggs. On the fifth day, female receptivity in the treated group was significantly reduced when compared with the controls. These results indicate that cyantraniliprole is effective against B. dorsalis. The inhibition and stimulation effect of cyantraniliprole on the adult's mating performance at different concentrations was proved. © 2014 Society of Chemical Industry.

  19. Development and application of a sublethal toxicity test to PAH using marine harpacticoid copepods. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Fleeger, J.W.; Lotufo, G.R.

    1999-01-01

    This research project was designed to improve the understanding of the acute and sublethal effects of PAHs to benthic invertebrates. Sublethal bioassay protocols for benthic harpacticoid copepods were developed, and two species of harpacticoids were exposed to a range of concentrations of sediment-amended PAHs; the single compounds fluoranthene and phenanthrene as well as a complex mixture (diesel fuel). The harpacticoid copepods Schizopera knabeni and Nitocra lacustris were tested using several bioassay approaches. Reproductive assays, feeding assays and avoidance tests were conducted in addition to lethal tests for S. knabeni. Species-specific differences in sensitivity were detected. Early life history stages were much more sensitive than adults in one species but not in the other. Concentrations of PAH as low as 26 micrograms PAH decreased copepod offspring production, egg hatching success, and embryonic and early-stage development, demonstrating the high sensitivity of life history-related endpoints. In addition, grazing on microalgae was significantly impaired at concentrations as low as 20 micrograms/g PAH after short exposures (<30 h). Finally it was demonstrated that harpacticoids can actively avoid contamination.

  20. Effects of Sublethal Doses of Imidacloprid on Young Adult Honeybee Behaviour

    Science.gov (United States)

    Mengoni Goñalons, Carolina; Farina, Walter Marcelo

    2015-01-01

    Imidacloprid (IMI), a neonicotinoid used for its high selective toxicity to insects, is one of the most commonly used pesticides. However, its effect on beneficial insects such as the honeybee Apis mellifera L is still controversial. As young adult workers perform in-hive duties that are crucial for colony maintenance and survival, we aimed to assess the effect of sublethal IMI doses on honeybee behaviour during this period. Also, because this insecticide acts as a cholinergic-nicotinic agonist and these pathways take part in insect learning and memory processes; we used IMI to assess their role and the changes they suffer along early adulthood. We focused on appetitive behaviours based on the proboscis extension response. Laboratory reared adults of 2 to 10 days of age were exposed to sublethal IMI doses (0.25 or 0.50ng) administered orally or topically prior to behavioural assessment. Modification of gustatory responsiveness and impairment of learning and memory were found as a result of IMI exposure. These outcomes differed depending on age of evaluation, type of exposure and IMI dose, being the youngest bees more sensitive and the highest oral dose more toxic. Altogether, these results imply that IMI administered at levels found in agroecosystems can reduce sensitivity to reward and impair associative learning in young honeybees. Therefore, once a nectar inflow with IMI traces is distributed within the hive, it could impair in-door duties with negative consequences on colony performance. PMID:26488410

  1. Sub-lethal toxicity of chlorpyrifos on Common carp, Cyprinus carpio (Linnaeus, 1758: Biochemical response

    Directory of Open Access Journals (Sweden)

    Mahdi Banaee

    2014-01-01

    Full Text Available Chlorpyrifos, an organophosphate pesticide, is widely used to control pests in agriculture farms and orchards of fruit trees. In this study, the fish were exposed to sub-lethal concentrations of chlorpyrifos which were determined based on numerical value of 96 h LC50. Blood was sampled after 10, 20 and 30 days and biochemical parameters including glucose, total protein, albumin, globulin, triglyceride and cholesterol levels, and aspartate aminotransferase (AST, alanine aminotransferase (ALT, lactate dehydrogenase (LDH, creatine kinase (CK, alkaline phosphatase (ALP and acetylcholinsetrase (AChE activities were measured. Behavioral changes in the fish were also recorded during the experiment. Unbalanced swimming, swimming in the surface water and hyperglycemia, increased blood triglyceride, and increased levels of AST, LDH and CK activities as well as decreased levels of AChE activity were important changes that were observed in the specimens exposed to chlorpyrifos during experimental periods. The most important alterations in the blood biochemical parameters were measured in the specimens exposed to 40 µg/L chlorpyrifos on the 20th and 30th day of the trial. In conclusion, results of the present study indicated that exposure to sub-lethal concentrations of chlorpyrifos as low as 40 µg/L may cause biochemical and behavioral changes in Cyprinus carpio.

  2. The role of sublethal effects in evaluating earthworm responses to soil contamination

    Energy Technology Data Exchange (ETDEWEB)

    Wilborn, D.; Bollman, M.; Linder, G. [ManTech Environmental Technology, Inc., Corvallis, OR (United States)

    1994-12-31

    Frequently, standard test methods rely upon relatively straightforward, easily interpreted endpoints to evaluate biological effects, like growth inhibition, gross morbidity or death. In soil contamination evaluations, for example, earthworm toxicity tests are routinely completed in order to consider adverse biological effects associated with exposures to soil samples in the laboratory or field. Here, the toxicity endpoint measured in the standard test using Eisenia foetida is death; however, if chronic effects are more appropriate to the questions being asked within a risk assessment context, then alternative test endpoints must be developed and standardized. Prior evaluations have relied upon sublethal endpoints, most frequently behavioral and morphological observations, for evaluating chronic effects associated with contaminant exposures. The authors applied these behavioral and morphological endpoints in analyzing potential chronic effects in earthworms exposed to heavy metal-contaminated soils in both the laboratory and field. In using a relatively standard set of these sublethal endpoints the authors found that these endpoints could be used to evaluate chronic effects associated with soil exposures, but that selection of the specific end-points had to be adequately standardized and that observer bias had to be adequately characterized in order for these measures of chronic effects to be unequivocally applied within an ecological risk assessment.

  3. Avoidance behaviour and anxiety in rats irradiated with a sublethal dose of gamma-rays.

    Science.gov (United States)

    Tomášová, Lenka; Smajda, B; Bona, M

    2011-12-01

    The aim of this study was to assess, whether a sublethal dose of gamma-rays will influence the avoidance behaviour and anxiety in rats and whether the response to radiation depends on time of day of its application. Adult male Wistar rats were tested in elevated plus-maze, in hot plate test and in the light/dark box in 4 regular intervals during a day. After two weeks the animals were irradiated with a whole-body dose 6 Gy of gamma-rays. One day after irradiation the animals were repeatedly tested in the same way, as before irradiation. In the plus-maze test an increased level of anxiety was established. The irradiation significantly decreased the locomotor activity of rats, but the extent of exploratory and comfortable behaviour were not altered. After irradiation, an elevated aversion to the thermal stimulus was observed in the hot plate test. The effects of radiation were more pronounced in the light period of the day, than in the dark one. No significant differences in aversion to light were detected after irradiation. The obtained results indicate, that sublethal doses of ionizing radiation can markedly influence the reactivity of animals to adverse stimuli, their motoric activity and emotional status, as well.

  4. Lethal and sublethal effects of four essential oils on the egg parasitoids Trissolcus basalis.

    Science.gov (United States)

    Werdin González, Jorge Omar; Laumann, Raúl Alberto; da Silveira, Samantha; Moraes, Maria Carolina Blassioli; Borges, Miguel; Ferrero, Adriana Alicia

    2013-07-01

    The essential oils from leaves of Schinus molle var. areira, Aloysia citriodora, Origanum vulgare and Thymus vulgaris have showed potential as phytoinsecticides against the green stink bug, Nezara viridula. In this work were evaluated their toxicological and behavioral effects on the parasitoid Trissolcus basalis, a biological control agent of this pest insect. Essential oils were obtained via hydrodestillation from fresh leaves. Insecticide activity in T. basalis females was evaluated in direct contact and fumigation bioassays. Behavioral effects were evaluated in olfactometer bioassays. To evaluate the residual toxicity, females of the parasitoids were exposed to oil residues; in these insects, the sublethal effects were evaluated (potential parasitism and survivorship of immature stages). The essential oils from O. vulgare and T. vulgaris proved to be highly selective when used as fumigant and did not change parasitoid behavior. After one week, the residues of these oils were harmless and did not show sublethal effects against T. basalis. According with these results, essential oils have potential applications for the integrated management of N. viridula. Published by Elsevier Ltd.

  5. Valsartan attenuates intimal hyperplasia in balloon-injured rat aortic arteries through modulating the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor axis.

    Science.gov (United States)

    Li, Yonghong; Cai, Shanglang; Wang, Qixin; Zhou, Jingwei; Hou, Bo; Yu, Haichu; Ge, Zhiming; Guan, Renyan; Liu, Xu

    2016-05-15

    The role of the Mas receptor in the activity of valsartan against intimal hyperplasia is unclear. Herein, we investigated the role of the angiotensin-converting enzyme 2 (ACE2)-angiotensin-(1-7)-Mas receptor axis on the activity of valsartan against intimal hyperplasiain balloon-injured rat aortic arteries. Wistar rats were randomized equally into the sham control group, injured group, and injured plus valsartan (20 mg/kg/d)-treated group. Valsartan significantly attenuated the vascular smooth muscle cell proliferation and intimal and medial thickening on days 14 and 28 after injury. The angiotensin-(1-7) levels as well as ACE2 and Mas receptor mRNA/protein expression were significantly decreased in the injured rats, compared to the uninjured rats; meanwhile, the angiotensin II level as well as the ACE and AT1 receptor mRNA/protein expression were increased (all P Mas receptor mRNA/protein expression but decreased the angiotensin II level, ACE and AT1 receptor mRNA/protein expression, as well as the p-ERK protein expression, compared to the injured group (all P Mas axis as well as inhibition of the ACE-angiotensin II-AT1 and p-ERK pathways. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Alexithymia, impulsiveness, and psychopathology in nonsuicidal self-injured adolescents

    Science.gov (United States)

    Gatta, Michela; Dal Santo, Francesco; Rago, Alessio; Spoto, Andrea; Battistella, Pier Antonio

    2016-01-01

    Introduction Nonsuicidal self-injury (NSSI) is a multifaceted phenomenon and a major health issue among adolescents. A better understanding of self-injury comorbidities is crucial to improve our ability to assess, treat, and prevent NSSI. Purpose This study aimed at analyzing some of the psychobehavioral correlates of NSSI: psychological problems, alexithymia, impulsiveness, and sociorelational aspects. Patients and methods This was a case–control study. The clinical sample (n=33) included adolescents attending our unit for NSSI and other issues; the controls (n=79) were high-school students. Data were collected using six questionnaires: Youth Self-Report, Barratt’s Impulsiveness Scale, Toronto Alexithymia Scale, Children’s Depression Inventory, Symptom Checklist-90-R, and Child Behavior Checklist. Results Cases scored significantly higher in all questionnaires. Habitual self-injurers scored higher on impulsiveness and alexithymia. The gesture’s repetition seems relevant to the global clinical picture: habitual self-injurers appear more likely to seek help from the sociosanitary services. We found a difference between the self-injurers’ and their parents’ awareness of the disorder. Conclusion Habitual self-injurers show signs of having difficulty with assessing the consequences of their actions (nonplanning impulsiveness) and the inability to manage their feelings. Given the significantly higher scores found for cases than for controls on all the psychopathological scales, NSSI can be seen as a cross-category psychiatric disorder, supporting the Diagnostic and Statistical Manual of Mental Disorders decision to include it as a pathological entity in its own right. PMID:27672324

  7. In-hospital mortality pattern of severely injured children

    DEFF Research Database (Denmark)

    Do, Hien Quoc; Steinmetz, Jacob; Rasmussen, Lars S

    2012-01-01

    BACKGROUND: Although trauma remains a major cause of morbidity and mortality in children, less attention has been directed to this group of patients. Whilst there is considerable literature on trauma in adults, only few studies describe paediatric trauma. The aim of this study was to describe...... the mortality pattern of severely injured children admitted to a Danish level I trauma centre. METHODS: We included trauma patients aged 15 years or less, who subsequent a trauma team activation were admitted during the 9-year period 1999-2007. Data were collected prospectively for subjects who had a length...

  8. Toxicity, sublethal effects, and potential modes of action of select fungicides on freshwater fish and invertebrates

    Science.gov (United States)

    Elskus, Adria A.

    2012-01-01

    Despite decades of agricultural and urban use of fungicides and widespread detection of these pesticides in surface waters, relatively few data are available on the effects of fungicides on fish and invertebrates in the aquatic environment. Nine fungicides are reviewed in this report: azoxystrobin, boscalid, chlorothalonil, fludioxonil, myclobutanil, fenarimol, pyraclostrobin, pyrimethanil, and zoxamide. These fungicides were identified as emerging chemicals of concern because of their high or increasing global use rates, detection frequency in surface waters, or likely persistence in the environment. A review of the literature revealed significant sublethal effects of fungicides on fish, aquatic invertebrates, and ecosystems, including zooplankton and fish reproduction, fish immune function, zooplankton community composition, metabolic enzymes, and ecosystem processes, such as leaf decomposition in streams, among other biological effects. Some of these effects can occur at fungicide concentrations well below single-species acute lethality values (48- or 96-hour concentration that effects a response in 50 percent of the organisms, that is, effective concentration killing 50 percent of the organisms in 48 or 96 hours) and chronic sublethal values (for example, 21-day no observed adverse effects concentration), indicating that single-species toxicity values may dramatically underestimate the toxic potency of some fungicides. Fungicide modes of toxic action in fungi can sometimes reflect the biochemical and (or) physiological effects of fungicides observed in vertebrates and invertebrates; however, far more studies are needed to explore the potential to predict effects in nontarget organisms based on specific fungicide modes of toxic action. Fungicides can also have additive and (or) synergistic effects when used with other fungicides and insecticides, highlighting the need to study pesticide mixtures that occur in surface waters. For fungicides that partition to

  9. A fish of many scales: extrapolating sublethal pesticide exposures to the productivity of wild salmon populations.

    Science.gov (United States)

    Baldwin, David H; Spromberg, Julann A; Collier, Tracy K; Scholz, Nathaniel L

    2009-12-01

    For more than a decade, numerous pesticides have been detected in river systems of the western United States that support anadromous species of Pacific salmon and steelhead. Over the same interval, several declining wild salmon populations have been listed as either threatened or endangered under the U.S. Endangered Species Act (ESA). Because pesticides occur in surface waters that provide critical habitat for ESA-listed stocks, they are an ongoing concern for salmon conservation and recovery throughout California and the Pacific Northwest. Because pesticide exposures are typically sublethal, a key question is whether toxicological effects at (or below) the scale of the individual animal ultimately reduce the productivity and recovery potential of wild populations. In this study we evaluate how the sublethal impacts of pesticides on physiology and behavior can reduce the somatic growth of juvenile chinook salmon (Oncorhynchus tshawytscha) and, by extension, subsequent size-dependent survival when animals migrate to the ocean and overwinter in their first year. Our analyses focused on the organophosphate and carbamate classes of insecticides. These neurotoxic chemicals have been widely detected in aquatic environments. They inhibit acetylcholinesterase, an enzyme in the salmon nervous system that regulates neurotransmitter-mediated signaling at synapses. Based on empirical data, we developed a model that explicitly links sublethal reductions in acetylcholinesterase activity to reductions in feeding behavior, food ration, growth, and size at migration. Individual size was then used to estimate size-dependent survival during migration and transition to the sea. Individual survival estimates were then integrated into a life-history population projection matrix and used to calculate population productivity and growth rate. Our results indicate that short-term (i.e., four-day) exposures that are representative of seasonal pesticide use may be sufficient to reduce the

  10. A comparison of the sublethal and lethal toxicity of four pesticides in Hyalella azteca and Chironomus dilutus.

    Science.gov (United States)

    Hasenbein, Simone; Connon, Richard E; Lawler, Sharon P; Geist, Juergen

    2015-08-01

    Laboratory toxicity testing is the primary tool used for surface water environmental risk assessment; however, there are critical information gaps regarding the sublethal effects of pesticides. In 10-day exposures, we assessed the lethal and sublethal (motility and growth) toxicities of four commonly used pesticides, bifenthrin, permethrin, cyfluthrin, and chlorpyrifos, on two freshwater invertebrates, Chironomus dilutus and Hyalella azteca. Pyrethroids were more toxic than the organophosphate chlorpyrifos in both species. Bifenthrin was most toxic to H. azteca survival and growth. Cyfluthrin was most toxic to C. dilutus. However, cyfluthrin had the greatest effect on motility on both H. azteca and C. dilutus. The evaluated concentrations of chlorpyrifos did not affect C. dilutus motility or growth, but significantly impacted H. azteca growth. Motility served as the most sensitive endpoint in assessing sublethal effects at low concentrations for both species, while growth was a good indicator of toxicity for all four pesticides for H. azteca. The integration of sublethal endpoints in ambient water monitoring and pesticide regulation efforts could improve identification of low-level pesticide concentrations that may eventually cause negative effects on food webs and community structure in aquatic environments.

  11. Evaluating sub-lethal effects of orchard-applied pyrethroids using video-tracking software to quantify honey bee behaviors.

    Science.gov (United States)

    Ingram, Erin M; Augustin, Julie; Ellis, Marion D; Siegfried, Blair D

    2015-09-01

    Managed honey bee, Apis mellifera L., colonies are contracted to pollinate fruit and nut orchards improving crop quality and yield. Colonies placed in orchards are potentially exposed to pyrethroid insecticides used for broad-spectrum pest control. Pyrethroids have been reported to pose minimal risk to bees due to their low application rates in the field and putative repellent properties. This repellency is believed to alter foraging behavior with the benefit of preventing bees from encountering a lethal dose in the field. However, sub-lethal exposure to pyrethroids may adversely impact bee behavior potentially resulting in social dysfunction or disruption of foraging. This study quantified behaviors associated with sub-lethal exposure to orchard-applied pyrethroids including, lambda-cyhalothrin, esfenvalerate, and permethrin, using video tracking software, Ethovision XT (Noldus Information Technologies). Bee locomotion, social interaction, and time spent near a food source were measured over a 24-h period. Bees treated with a pyrethroid traveled 30-71% less than control bees. Social interaction time decreased by 43% for bees treated with a high sub-lethal dose of esfenvalerate. Bees exposed to a high sub-lethal dose of permethrin spent 67% less time in social interaction and spent more than 5 times as long in the food zone compared to control bees. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Lethal and sub-lethal responses of native freshwater mussels exposed to granular Bayluscide®, a sea lamprey larvicide

    Science.gov (United States)

    Newton, Teresa; Boogaard, Michael A.; Gray, Brian R.; Hubert, Terrance D.; Schloesser, Nicholas

    2017-01-01

    The invasive sea lamprey (Petromyzon marinus) poses a substantial threat to fish communities in the Great Lakes. Efforts to control sea lamprey populations typically involve treating tributary streams with lampricides on a recurring cycle. The presence of a substantial population of larval sea lampreys in the aquatic corridor between Lakes Huron and Erie prompted managers to propose a treatment using the granular formulation of Bayluscide® that targets larval sea lampreys that reside in sediments. However, these treatments could cause adverse effects on native freshwater mussels—imperiled animals that also reside in sediments. We estimated the risk of mortality and sub-lethal effects among eight species of adult and sub-adult mussels exposed to Bayluscide® for durations up to 8 h to mimic field applications. Mortality was appreciable in some species, especially in sub-adults (range, 23–51%). The lethal and sub-lethal effects were positively associated with the duration of exposure in most species and life stage combinations. Estimates of the median time of exposure that resulted in lethal and sub-lethal effects suggest that sub-adults were often affected by Bayluscide® earlier than adults. Siphoning activity and burrowing position of mussels during exposure may have moderated the uptake of Bayluscide® and may have influenced lethal and sub-lethal responses. Given that the various species and life stages were differentially affected, it will be difficult to predict the effects of Bayluscide® treatments on mussels.

  13. Toxicity and sub-lethal effect of endemic plants from family Anacardiaceae on oviposition behavior of Aedes albopictus

    Directory of Open Access Journals (Sweden)

    Wan Fatma Zuharah

    2015-08-01

    Conclusions: These results clearly indicate that the acetone extract of G. renghas could be served as potential larvicide, whereas M. fasciculiflora has better sub-lethal effect for oviposition deterrence and against Ae. albopictus as an oviciding agent.

  14. Uptake and elimination of 1,2,4-trichlorobenzene in the guppy (Poecilia reticulata) at sublethal and lethal aqueous concentrations.

    NARCIS (Netherlands)

    Eck, van J.M.C.; Koelmans, A.A.; Deneer, J.W.

    1997-01-01

    The kinetics of 1,2,4-trichlorobenzene in guppies (Poecilia reticulata) have been studied in an accumulation and elimination experiment. At a sublethal exposure, uptake and elimination rate constants of 1,2,4-trichlorobenzene were determined, employing a first-order one-compartment model. The

  15. (1)H NMR-based metabolomics of Daphnia magna responses after sub-lethal exposure to triclosan, carbamazepine and ibuprofen.

    Science.gov (United States)

    Kovacevic, Vera; Simpson, André J; Simpson, Myrna J

    2016-09-01

    Pharmaceuticals and personal care products are a class of emerging contaminants that are present in wastewater effluents, surface water, and groundwater around the world. There is a need to determine rapid and reliable bioindicators of exposure and the toxic mode of action of these contaminants to aquatic organisms. (1)H nuclear magnetic resonance (NMR)-based metabolomics in combination with multivariate statistical analysis was used to determine the metabolic profile of Daphnia magna after exposure to a range of sub-lethal concentrations of triclosan (6.25-100μg/L), carbamazepine (1.75-14mg/L) and ibuprofen (1.75-14mg/L) for 48h. Sub-lethal triclosan exposure suggested a general oxidative stress condition and the branched-chain amino acids, glutamine, glutamate, and methionine emerged as potential bioindicators. The aromatic amino acids, serine, glycine and alanine are potential bioindicators for sub-lethal carbamazepine exposure that may have altered energy metabolism. The potential bioindicators for sub-lethal ibuprofen exposure are serine, methionine, lysine, arginine and leucine, which showed a concentration-dependent response. The differences in the metabolic changes were related to the dissimilar modes of toxicity of triclosan, carbamazepine and ibuprofen. (1)H NMR-based metabolomics gave an improved understanding of how these emerging contaminants impact the keystone species D. magna. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Lethal and sub-lethal effects of five pesticides used in rice farming on the earthworm Eisenia fetida

    NARCIS (Netherlands)

    Rico, Andreu; Sabater, Consuelo; Castillo, María Ángeles

    2016-01-01

    The toxicity of five pesticides typically used in rice farming (trichlorfon, dimethoate, carbendazim, tebuconazole and prochloraz) was evaluated on different lethal and sub-lethal endpoints of the earthworm Eisenia fetida. The evaluated endpoints included: avoidance behaviour after an exposure

  17. Sublethal effects of Imidacloprid on honey bee colony growth and activity at three sites in the U.S.

    Science.gov (United States)

    Field experiments in southern Arizona, central Arkansas and southern Mississippi were conducted to evaluate the effects of sublethal concentrations (0, 5, 20 and 100 ppb) of imidacloprid in sugar syrup on honey bee colony growth and activity. Response variables included discrete data from hive inspe...

  18. Reinitialization of evolutionary clocks during sublethal environmental stress in some invertebrates

    Science.gov (United States)

    Guex, Jean

    2006-02-01

    This paper describes the influence of high environmental stress on evolutionary trends in some selected Mesozoic ammonite lineages and some protists. During extinction periods, many ammonoids are affected by drastic simplifications of their shell geometry, ornamentation and suture line. We observe that relatively tightly coiled ammonites can give rise to highly evolute forms or uncoiled heteromorphs with simple ornamentation and almost ceratitic suture line—a phenomenon called "proteromorphosis". Such simplifications often correspond to a reappearance of ancestral geometries (primitive ornamentation, evolute coiling or uncoiling) which suggest that the evolutionary clock of these organisms can be reinitialized by extreme, sublethal, environmental stress such as giant volcanism (including its consequences on diverse pollutions and on climatic changes) and major regressive events.

  19. The growth and development of Schistosoma mansoni in mice exposed to sublethal doses of radiation

    Energy Technology Data Exchange (ETDEWEB)

    Aitken, R.; Wilson, R.A. (Univ. of York, Heslington (England))

    1989-12-01

    The maturation of Schistosoma mansoni was studied in mice exposed to various sublethal doses of radiation. Although the treatment of mice with 500 rads of radiation prior to infection did not alter parasite maturation, doses in excess of 500 rads led to a reduction in worm burden. This could not be attributed to a delay in the arrival of parasites in the hepatic portal system. Worms developing in mice treated with 800 rads commenced egg-laying about 1 wk later than worms in intact mice, and the rate of egg deposition appeared to be lower in irradiated hosts. The data demonstrate that exposure of C57BL/6 mice to doses of radiation in excess of 500 rads impairs their ability to carry infections of S. mansoni. The findings do not support the hypothesis that primary worm burdens in the mouse are controlled by a host immune response.

  20. Feeding Inhibition: the Ups and Downs of Sublethal Effects on Grazers and Detritivores

    Science.gov (United States)

    Alexander, A.; Culp, J.; Liber, K.; Baird, D.

    2005-05-01

    Sublethal impacts are likely the primary mechanism of exposure for the aquatic community in the case of soluble agricultural pesticides. This study examines the effects of pulsed exposures of the common insecticide, imidacloprid, on the feeding and growth of the mayfly Epeorus longimanus, and the oligochaete, Lumbriculus variegatus. Examining the effects of pulsed exposures of imidacloprid is particularly relevant due to the soluble (0.51g/L) nature of this compound. Recovery experiments were conducted by exposing mayflies and oligochaetes to an environmentally realistic range (0, 0.1, 0.5, 1, 5, 10 ppb) of concentrations for a short period. Effects on feeding were measured by quantifying the foodstuffs consumed by mayflies and egested by oligochaetes. In tandem with the feeding experiments, a series of artificial stream experiments were undertaken that demonstrate the changes in growth and abundance of adult mayflies in response to this common insecticide stressor.

  1. Effects of Glyphosate-Based Herbicide Sub-Lethal Concentrations on Fish Feeding Behavior.

    Science.gov (United States)

    Giaquinto, Percilia Cardoso; de Sá, Marina Borges; Sugihara, Vanessa Seiko; Gonçalves, Bruno Bastos; Delício, Helton Carlos; Barki, Assaf

    2017-04-01

    Glyphosate-based herbicides are widely used in agricultural systems. Although the target organism are particularly plant organisms, there are numerous studies showing adverse effects in aquatic animals, such as inhibition of acetyl cholinesterase, effects on kidney, liver, and gill and stressors effects. This study analyzed the effects of commercial formulation of glyphosate on feeding behavior in Pacu (Piaractus mesopotamicus). Fish were exposed to three glyphosate concentrations (0.2, 0.6, and 1.8 ppm) for 15 days. At concentrations of 0.2 and 0.6 ppm, food intake decreased on day 13 and then returned to normal on day 15. At the highest glyphosate-based herbicide concentration, 1.8 ppm, food consumption decreased dramatically and did not recover on day 15. This study showed that glyphosate-based herbicide at sub-lethal concentrations can affect feed intake in pacu and consequently inhibits its growth.

  2. Influence of biorhythms on sensitivity of Nerita to pollutants at Sublethal levels

    Energy Technology Data Exchange (ETDEWEB)

    Battershill, C.N.; Bergquist, P.R.

    1984-01-01

    Intrinsic rhythmic activity of Nerita (Melanerita) atramentosa melanotragus was assessed under constant laboratory conditions. Activity proved to be a sensitive indicator of toxicity, and was affected by low levels of a relatively new oil dispersing agent, Shell SD LTX. How the state of activity influenced animal sensitivity to Shell SD LTX and to an oil, Maui Condensate, was investigated using short-term recovery experiments. Nerita were most sensitive during their active phase, and results during this period differed significantly from tests carried out during the inactive phase of the animal. Dispersant/oil mixture proved to be highly toxic. These findings have ecological implications and permit comment relating to the design of sublethal toxicity tests. These subjects are discussed. 37 references, 3 figures, 2 tables.

  3. Response of larval fish, Leiostomus xanthurus, to environmental stress following sublethal cadmium exposure

    Energy Technology Data Exchange (ETDEWEB)

    Middaugh, D.P.; Davis, W.R.; Yoakum, R.L.

    1975-08-01

    The toxicity of cadmium to larval fish, Leiostomus xanthurus, was studied. An incipient LC/sub 50/ concentration of approximately 0.2 to 0.3 mg/l cadmium was first estimated. Subsequent short-term sublethal tests were conducted to determine the relationship of cadmium exposure and accumulated whole body residues of the metal on the response of larvae to thermal stress and low dissolved oxygen. Results of this study indicated a significant decrease (..cap alpha.. = 0.05, t-Test) in the critical thermal maximum (CTM) for larvae exposed to 0.5 and 0.8 mg/l cadmium for 96 hours at 20/sup 0/C. Significant decreases (..cap alpha.. = 0.05, chi/sup 2/) in survival of larvae subjected to a dissolved oxygen (DO) level of 1.6 mg/l after exposure to 0.5 and 0.8 mg/l cadmium were also observed.

  4. Effects of sublethal entrainment stresses on the vulnerability of juvenile bluegill sunfish to predation

    Energy Technology Data Exchange (ETDEWEB)

    Cada, G.F.; Solomon, J.A.; Loar, J.M.

    1981-07-01

    This report provides a review of literature concerning the effects of sublethal stresses on predator-prey interactions in aquatic systems. In addition, the results of a preliminary laboratory study of the susceptibility of entrainment-stressed juvenile bluegill to striped bass predation are presented. Juvenile bluegill were exposed to thermal and physical entrainment stresses in the ORNL Power Plant Simulator and subsequently to predation by juvenile striped bass in a susceptibility to predation experimental design. None of the entrainment stresses tested (thermal shock, physical effects of pump and condenser passage, and combination of thermal and physical shock) was found to significantly increase predation rates as compared to controls, and no significant interactions between thermal and physical stresses were detected. The validity of laboratory predator-prey studies and the application of indirect mortality information for setting protective standards and predicting environmental impacts are discussed.

  5. Sublethal gamma irradiation affects reproductive impairment and elevates antioxidant enzyme and DNA repair activities in the monogonont rotifer Brachionus koreanus

    Energy Technology Data Exchange (ETDEWEB)

    Han, Jeonghoon; Won, Eun-Ji [Department of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Kim, Il-Chan; Yim, Joung Han [Division of Life Sciences, Korea Polar Research Institute, Incheon 406-840 (Korea, Republic of); Lee, Su-Jae [Department of Life Science, College of Natural Sciences, Hanyang University, Seoul 133-791 (Korea, Republic of); Lee, Jae-Seong, E-mail: jslee2@skku.edu [Department of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of)

    2014-10-15

    Highlights: • No mortality within 96 h even at a high intensity (1200 Gy). • A reduced fecundity of Brachionus koreanus at over 150 Gy with a decrease in lifespan. • Dose-dependent ROS increase with GST enzyme activity at sub-lethal doses. • Significant impact on life table parameters, particularly fecundity. • Significant up-regulation of DNA repair-associated genes at sublethal doses. - Abstract: To examine the effects of gamma radiation on marine organisms, we irradiated several doses of gamma ray to the microzooplankton Brachionus koreanus, and measured in vivo and in vitro endpoints including the survival rate, lifespan, fecundity, population growth, gamma ray-induced oxidative stress, and modulated patterns of enzyme activities and gene expressions after DNA damage. After gamma radiation, no individuals showed any mortality within 96 h even at a high intensity (1200 Gy). However, a reduced fecundity (e.g. cumulated number of offspring) of B. koreanus at over 150 Gy was observed along with a slight decrease in lifespan. At 150 Gy and 200 Gy, the reduced fecundity of the rotifers led to a significant decrease in population growth, although in the second generation the population growth pattern was not affected even at 200 Gy when compared to the control group. At sub-lethal doses, reactive oxygen species (ROS) levels dose-dependently increased with GST enzyme activity. In addition, up-regulations of the antioxidant and chaperoning genes in response to gamma radiation were able to recover cellular damages, and life table parameters were significantly influenced, particularly with regard to fecundity. DNA repair-associated genes showed significantly up-regulated expression patterns in response to sublethal doses (150 and 200 Gy), as shown in the expression of the gamma-irradiated B. koreanus p53 gene, suggesting that these sublethal doses were not significantly fatal to B. koreanus but induced DNA damages leading to a decrease of the population size.

  6. Mortality, Temporary Sterilization, and Maternal Effects of Sublethal Heat in Bed Bugs

    Science.gov (United States)

    Rukke, Bjørn Arne; Aak, Anders; Edgar, Kristin Skarsfjord

    2015-01-01

    Adult bed bugs were exposed to the sublethal temperatures 34.0°C, 35.5°C, 37.0°C, 38.5°C, or 40.0°C for 3, 6, or 9 days. The two uppermost temperatures induced 100% mortality within 9 and 2 days, respectively, whereas 34.0°C had no observable effect. The intermediate temperatures interacted with time to induce a limited level of mortality but had distinct effects on fecundity, reflected by decreases in the number of eggs produced and hatching success. Adult fecundity remained low for up to 40 days after heat exposure, and the time until fertility was restored correlated with the temperature-sum experienced during heat exposure. Three or 6 days of parental exposure to 38.5°C significantly lowered their offspring’s feeding and moulting ability, which consequently led to a failure to continue beyond the third instar. Eggs that were deposited at 22.0°C before being exposed to 37.0°C for 3 or 6 days died, whereas eggs that were exposed to lower temperatures were not significantly affected. Eggs that were deposited during heat treatment exhibited high levels of mortality also at 34.0°C and 35.5°C. The observed negative effects of temperatures between 34.0°C and 40.0°C may be utilized in pest management, and sublethal temperature exposure ought to be further investigated as an additional tool to decimate or potentially eradicate bed bug populations. The effect of parental heat exposure on progeny demonstrates the importance of including maternal considerations when studying bed bug environmental stress reactions. PMID:25996999

  7. Use of sublethal endpoints in sediment toxicity tests with the amphipod Hyalella azteca

    Science.gov (United States)

    Ingersoll, Chris G.; Brunson, Eric L.; Dwyer, F. James; Hardesty, Douglas K.; Kemble, Nile E.

    1998-01-01

    Short-term sediment toxicity tests that only measure effects on survival can be used to identify high levels of contamination but may not be able to identify marginally contaminated sediments. The objective of the present study was to develop a method for determining the potential sublethal effects of contaminants associated with sediment on the amphipod Hyalella azteca (e.g., reproduction). Exposures to sediment were started with 7- to 8-d-old amphipods. On day 28, amphipods were isolated from the sediment and placed in water-only chambers where reproduction was measured on day 35 and 42. Typically, amphipods were first in amplexus at about day 21 to 28 with release of the first brood between day 28 to 42. Endpoints measured included survival (day 28, 35, and 42), growth (as length and weight on day 28 and 42), and reproduction (number of young/female produced from day 28 to 42). This method was used to evaluate a formulated sediment and field-collected sediments with low to moderate concentrations of contaminants. Survival of amphipods in these sediments was typically >85% after the 28-d sediment exposures and the 14-d holding period in water to measure reproduction. Reproduction was more variable than growth; hence, more replicates might be needed to establish statistical differences among treatments. Previous studies have demonstrated that growth of H. azteca in sediment tests often provides unique information that can be used to discriminate toxic effects of exposure to contaminants. Either length or weight can be measured in sediment tests with H. azteca. However, additional statistical options are available if length is measured on individual amphipods, such as nested analysis of variance that can account for variance in length within replicates. Ongoing water-only studies testing select contaminants will provide additional data on the relative sensitivity and variability of sublethal endpoints in toxicity tests with H. azteca.

  8. Intra- and intergenerational persistence of an insect nucleopolyhedrovirus: adverse effects of sublethal disease on host development, reproduction, and susceptibility to superinfection.

    Science.gov (United States)

    Cabodevilla, Oihana; Villar, Eduardo; Virto, Cristina; Murillo, Rosa; Williams, Trevor; Caballero, Primitivo

    2011-05-01

    Sublethal infections by Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) are common in field populations of the beet armyworm (S. exigua, Hübner) in the Almerian horticultural region of Spain. Inoculation of second, third, and fourth instars with occlusion bodies (OBs) of an isolate (VT-SeAl1) associated with vertically transmitted infections resulted in 15 to 100% of sublethal infection in adult survivors, as determined by reverse transcription-PCR (RT-PCR) detection of viral DNA polymerase transcripts, and quantitative PCR (qPCR) targeted at the DNA polymerase gene. The prevalence of adult sublethal infection was positively related to the inoculum OB concentration consumed during the larval stage. Sublethal infections persisted in OB-treated insects for at least five generations. Viral transcripts were more frequently detected in adult insects than in third instars. qPCR analysis indicated a consistently higher prevalence of sublethal infection than RT-PCR. Sublethal infection was associated with significant reductions in pupal weight, adult emergence, fecundity, and fertility (egg hatch) and significant increases in larval development time and duration of the preoviposition period. Insects taken from a persistently infected experimental population were significantly more susceptible to the OB inoculum than control insects that originated from the same virus-free colony as the persistently infected insects. We conclude that OB treatment results in rapid establishment of sublethal infections that persist between generations and which incur costs in the development and reproductive capacity of the host insect.

  9. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells.

    Science.gov (United States)

    Branco, Patrícia; Viana, Tiago; Albergaria, Helena; Arneborg, Nils

    2015-07-16

    Saccharomyces cerevisiae produces antimicrobial peptides (AMPs) during alcoholic fermentation that are active against several wine-related yeasts (e.g. Hanseniaspora guilliermondii) and bacteria (e.g. Oenococcus oeni). In the present study, the physiological changes induced by those AMPs on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20 min of exposure to inhibitory concentrations of AMPs, and after 24 h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h of exposure to AMPs, PI-stained (dead) cells increased from 0% to 77.7% and the number of viable cells fell from 1×10(5) to 10 CFU/ml. This means that virtually all cells (99.99%) became unculturable but that a sub-population of 22.3% of the cells remained viable (as determined by PI staining). Besides, pHi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0<∆pH<3). Taken together, these results indicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals that these AMPs compromise the plasma membrane integrity (and possibly also the vacuole membrane) of H. guilliermondii cells, disturbing the pHi homeostasis and inducing a loss of culturability. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Distribution of the amelogenin protein in developing, injured and carious human teeth

    Directory of Open Access Journals (Sweden)

    Thimios eMitsiadis

    2014-12-01

    Full Text Available Amelogenin is the major enamel matrix protein with key roles in amelogenesis. Although for many decades amelogenin was considered to be exclusively expressed by ameloblasts, more recent studies have shown that amelogenin is also expressed in other dental and no-dental cells. However, amelogenin expression in human tissues remains unclear. Here, we show that amelogenin protein is not only expressed during human embryonic development but also in pathological conditions such as carious lesions and injuries after dental cavity preparation. In developing embryonic teeth, amelogenin stage-specific expression is found in all dental epithelia cell populations but with different instensities. In the different layers of enamel matrix, waves of positive versus negative immunostaining for amelogenin are detected suggesting that the secretion of amelogenin protein is orchestreted by a biological clock. Amelogenin is also expressed transiently in differentiating odontoblasts during predentin formation, but was absent in mature functional odontoblasts. In intact adult teeth, amelogenin was not present in dental pulp, odontoblasts, and dentin. However, in injured and carious adult human teeth amelogenin is strongly re-expressed in newly differentiated odontoblasts and is distributed in the dentinal tubuli under the lesion site. In an in vitro culture system, amelogenin is expressed preferentially in human dental pulp cells that start differentiating into odontoblast-like cells and form mineralization nodules. These data suggest that amelogenin plays important roles not only during cytodifferentiation, but also during tooth repair processes in humans.

  11. Thymol accelerates the recovery of the skeletal muscle of mice injured with cardiotoxin.

    Science.gov (United States)

    Cardoso, Eroneide S B; Santana, Tayse A; Diniz, Polyana Borges França; Montalvão, Monalisa M; Bani, Cristiane C; Thomazzi, Sara M

    2016-03-01

    The aim of this study was to investigate the preventive effect of thymol in in vivo muscle inflammation and regeneration on cardiotoxin-induced injury. Mice were pretreated (p.o.) with thymol (10-100 mg/kg), and after 1 h, cardiotoxin (25 μM, 40 μl) was administrated into the gastrocnemius muscle. The quantification of the areas of inflammation and regeneration of muscle tissue (3, 7 and 10 days) in HE-stained slides as well as the count of total mast cells and different phenotypes of mast cells were made. Sirius red staining was used to analyse total collagen expression. The pretreatment with thymol significantly reduced the area of inflammation (30 and 100 mg/kg) and increased the area of regeneration (100 mg/kg) 3 days after the cardiotoxin injection. Thymol at 30 and 100 mg/kg increased the area of collagen in 3 days and also decreased this area in 7 and 10 days, compared to the injured group. The pretreatment with thymol did not affect the number of total mast cells; however, it was able to change the number of mucosal mast cells within 10 days. This study suggests that thymol ameliorates inflammatory response and accelerates regeneration in cardiotoxin-induced muscle injury. © 2016 Royal Pharmaceutical Society, Journal of Pharmacy and Pharmacology.

  12. Compensatory strategies in the language of closed head injured patients.

    Science.gov (United States)

    Penn, C; Cleary, J

    1988-01-01

    The study reported here examined the nature and effectiveness of compensatory strategies appearing in the conversational discourse of six closed head injured patients. Subject performance on a range of communication measures was compared with their judged abilities on a taxonomy of 32 compensatory strategies. Results indicated that all subjects employed a wide range of strategies but to differential effect. The overall effectiveness of these strategies correlated strongly with performance on the oral language subtest of the Western Aphasia Battery, the Communicative Abilities in Daily Living Test and a pragmatic protocol. The development of compensation following brain injury is viewed as a process of equilibration, determined in part by neurological and subject variables, and in part by ecologic variables. The implications for therapeutic management are discussed.

  13. The acutely ACL injured knee assessed by MRI

    DEFF Research Database (Denmark)

    Frobell, R B; Roos, H P; Roos, E M

    2008-01-01

    depression fractures, meniscal injuries and patient characteristics. METHODS: One hundred and twenty-one subjects (26% women, mean age 26 years) with an ACL rupture to a previously un-injured knee were studied using a 1.5T MR imager within 3 weeks from trauma. Meniscal injuries and fractures were classified...... by type, size and location. BML location and volume were quantified using a multi-spectral image data set analyzed by computer software, edited by an expert radiologist. RESULTS: Fractures were found in 73 (60%) knees. In 67 (92%) of these knees at least one cortical depression fracture was found. Uni......-compartmental meniscal tears were found in 44 (36%) subjects and bi-compartmental in 24 (20%). One hundred and nineteen (98%) knees had at least one BML, all but four (97%) located in the lateral compartment. Knees with a cortical depression fracture had larger BML volumes (P

  14. Resuscitation and coagulation in the severely injured trauma patient

    Science.gov (United States)

    Midwinter, Mark J.; Woolley, Tom

    2011-01-01

    Developments in the resuscitation of the severely injured trauma patient in the last decade have been through the increased understanding of the early pathophysiological consequences of injury together with some observations and experiences of recent casualties of conflict. In particular, the recognition of early derangements of haemostasis with hypocoagulopathy being associated with increased mortality and morbidity and the prime importance of tissue hypoperfusion as a central driver to this process in this population of patients has led to new resuscitation strategies. These strategies have focused on haemostatic resuscitation and the development of the ideas of damage control resuscitation and damage control surgery continuum. This in turn has led to a requirement to be able to more closely monitor the physiological status, of major trauma patients, including their coagulation status, and react in an anticipatory fashion. PMID:21149355

  15. Effects of hypertonic dextrose on injured rat skeletal muscles.

    Science.gov (United States)

    Kunduracioglu, Burak; Ulkar, Bulent; Sabuncuoglu, Bizden T; Can, Belgin; Bayrakci, Kenan

    2006-04-01

    Histological examination of proliferative therapy effects on the healing process of muscular injury. We performed this study between March and August 2002 at Ankara University, School of Medicine, Laboratory of Animal Experiments, Ankara, Turkey. We used an experimental animal model by conducting a standardized cut injury of the gastrocnemius muscle in 30 adult male albino rats, which we divided into 2 groups; proliferative therapy group and control group. We evaluated the injured rat muscles by light microscopy on the fifth, eight, and twelfth day of injury. The muscular regeneration process began at day 5 in both the control and proliferative therapy groups. The proliferative therapy group revealed a prominent inflammatory reaction, fibroblast migration, and necrosis with accompanying regeneration and excessive connective tissue formation. We cannot consider proliferative therapy an appropriate treatment modality for muscular injuries, unless there is evidence of normal muscle physiology and biomechanics post traumatically.

  16. Evaluating Age in the Field Triage of Injured Persons

    Science.gov (United States)

    Nakamura, Yoko; Daya, Mohamud; Bulger, Eileen M.; Schreiber, Martin; Mackersie, Robert; Hsia, Renee Y.; Mann, N. Clay; Holmes, James F.; Staudenmayer, Kristan; Sturges, Zachary; Liao, Michael; Haukoos, Jason; Kuppermann, Nathan; Barton, Erik D.; Newgard, Craig D.

    2012-01-01

    Study Objective In this study, we evaluated (1) trauma under-triage by age group; (2) the association between age and serious injury after accounting for other field triage criteria and confounders; and (3) the potential impact of a mandatory age triage criterion for field triage. Methods This was a retrospective cohort study of injured children and adults transported by 48 EMS agencies to 105 hospitals in 6 regions of the Western U.S. from 2006 through 2008. We used probabilistic linkage to match EMS records to hospital records, including: trauma registries, state discharge databases and emergency department databases. The primary outcome measure was serious injury, as measured by an Injury Severity Score (ISS) ≥ 16. We assessed under-triage (ISS ≥ 16 and triage-negative or transport to a non-trauma center) by age decile and used multivariable logistic regression models to estimate the association (linear and non-linear) between age and ISS ≥ 16, adjusted for important confounders. We also evaluated the potential impact of age on triage efficiency and trauma center volume. Results 260,027 injured patients were evaluated and transported by EMS over the 3-year study period. Under-triage increased for patients over 60 years of age, reaching approximately 60% for those older than 90 years. There was a strong non-linear association between age and ISS ≥ 16. For patients not meeting other triage criteria, the probability of serious injury was most notable after 60 years. A mandatory age triage criterion would have decreased under-triage at the expense of over-triage, with one ISS ≥ 16 patient identified for every 60–65 additional patients transported to major trauma centers. Conclusion Trauma under-triage increases in patients older than 60 years. While the probability of serious injury increases among triage-negative patients with increasing age, the use of a mandatory age triage criterion appears inefficient for improving field triage. PMID:22633339

  17. Accuracy of Prehospital Triage in Selecting Severely Injured Trauma Patients.

    Science.gov (United States)

    Voskens, Frank J; van Rein, Eveline A J; van der Sluijs, Rogier; Houwert, Roderick M; Lichtveld, Robert Anton; Verleisdonk, Egbert J; Segers, Michiel; van Olden, Ger; Dijkgraaf, Marcel; Leenen, Luke P H; van Heijl, Mark

    2017-11-01

    A major component of trauma care is adequate prehospital triage. To optimize the prehospital triage system, it is essential to gain insight in the quality of prehospital triage of the entire trauma system. To prospectively evaluate the quality of the field triage system to identify severely injured adult trauma patients. Prehospital and hospital data of all adult trauma patients during 2012 to 2014 transported with the highest priority by emergency medical services professionals to 10 hospitals in Central Netherlands were prospectively collected. Prehospital data collected by the emergency medical services professionals were matched to hospital data collected in the trauma registry. An Injury Severity Score of 16 or more was used to determine severe injury. The quality and diagnostic accuracy of the field triage protocol and compliance of emergency medical services professionals to the protocol. A total of 4950 trauma patients were evaluated of which 436 (8.8%) patients were severely injured. The undertriage rate based on actual destination facility was 21.6% (95% CI, 18.0-25.7) with an overtriage rate of 30.6% (95% CI, 29.3-32.0). Analysis of the protocol itself, regardless of destination facility, resulted in an undertriage of 63.8% (95% CI, 59.2-68.1) and overtriage of 7.4% (95% CI, 6.7-8.2). The compliance to the field triage trauma protocol was 73% for patients with a level 1 indication. More than 20% of the patients with severe injuries were not transported to a level I trauma center. These patients are at risk for preventable morbidity and mortality. This finding indicates the need for improvement of the prehospital triage protocol.

  18. Homologous stress adaptation, antibiotic resistance, and biofilm forming ability of Salmonella enterica serovar Heidelberg ATCC8326 on different food-contact surfaces following exposure to sublethal chlorine concentrations.

    Science.gov (United States)

    Obe, Tomi; Nannapaneni, Rama; Sharma, Chander Shekhar; Kiess, Aaron

    2018-01-13

    Salmonella enterica serovar Heidelberg (American Type Culture Collection; ATCC 8326) was examined for the ability to adapt to the homologous stress of chlorine through exposure to increasing chlorine concentrations (25 ppm daily increments) in tryptic soy broth (TSB). The tested strain exhibited an acquired tolerance to chlorine in TSB with the tolerant cells growing in concentrations up to 400 ppm. In addition, the chlorine stressed cells displayed rugose morphology on tryptic soy agar (TSA) plates at 37°C. The minimum inhibitory concentration (MIC) of chlorine for adapted (rugose and smooth) cells was determined to be 550 ppm and 500 ppm, respectively whereas the MIC for the control was 450 ppm. The biofilm forming ability of the adapted and control cells were examined on both plastic and stainless steel surface at room temperature and 37°C. The rugose variant, in contrast to the smooth (adapted and control) showed the ability to form strong biofilms (P ≤ 0.05) on a plastic surface at room temperature and 37°C. Rugose cells compared to smooth and control attached more (P ≤ 0.05) to steel surfaces as well. The possibility of cross-adaptation was examined by exposing the adapted and control cells to different antibiotics according to the Clinical & Laboratory Standards Institute guidelines. Adapted cells exhibited reduced susceptibility to some of the antibiotics tested as compared to control. The findings of this study suggest that exposure to sublethal chlorine concentration during the sanitization procedure can result in tolerant Salmonella cells. Chlorine may confer cross-protection that aids in the survival of the tolerant population to other environmental stresses. © 2018 Poultry Science Association Inc.

  19. Matrix metalloproteinase-9 facilitates glial scar formation in the injured spinal cord.

    Science.gov (United States)

    Hsu, Jung-Yu C; Bourguignon, Lilly Y W; Adams, Christen M; Peyrollier, Karine; Zhang, Haoqian; Fandel, Thomas; Cun, Christine L; Werb, Zena; Noble-Haeusslein, Linda J

    2008-12-10

    In the injured spinal cord, a glial scar forms and becomes a major obstacle to axonal regeneration. Formation of the glial scar involves migration of astrocytes toward the lesion. Matrix metalloproteinases (MMPs), including MMP-9 and MMP-2, govern cell migration through their ability to degrade constituents of the extracellular matrix. Although MMP-9 is expressed in reactive astrocytes, its involvement in astrocyte migration and formation of a glial scar is unknown. Here we found that spinal cord injured, wild-type mice expressing MMPs developed a more severe glial scar and enhanced expression of chondroitin sulfate proteoglycans, indicative of a more inhibitory environment for axonal regeneration/plasticity, than MMP-9 null mice. To determine whether MMP-9 mediates astrocyte migration, we conducted a scratch wound assay using astrocytes cultured from MMP-9 null, MMP-2 null, and wild-type mice. Gelatin zymography confirmed the expression of MMP-9 and MMP-2 in wild-type cultures. MMP-9 null astrocytes and wild-type astrocytes, treated with an MMP-9 inhibitor, exhibited impaired migration relative to untreated wild-type controls. MMP-9 null astrocytes showed abnormalities in the actin cytoskeletal organization and function but no detectable untoward effects on proliferation, cellular viability, or adhesion. Interestingly, MMP-2 null astrocytes showed increased migration, which could be attenuated in the presence of an MMP-9 inhibitor. Collectively, our studies provide explicit evidence that MMP-9 is integral to the formation of an inhibitory glial scar and cytoskeleton-mediated astrocyte migration. MMP-9 may thus be a promising therapeutic target to reduce glial scarring during wound healing after spinal cord injury.

  20. Process of tight junction recovery in the injured vocal fold epithelium: Morphological and paracellular permeability analysis.

    Science.gov (United States)

    Suzuki, Ryo; Katsuno, Tatsuya; Kishimoto, Yo; Nakamura, Ryosuke; Mizuta, Masanobu; Suehiro, Atsushi; Yamashita, Masaru; Nakamura, Tatsuo; Tateya, Ichiro; Omori, Koichi

    2017-10-31

    The vocal fold epithelium that includes tight junction (TJ)-based barrier function protects underlying connective tissues from external insults. TJs play an important role to control paracellular permeability of not only solutes but also ions, and preserve the vocal fold homeostasis. However, the distribution of TJs and paracellular diffusion barrier across the entire vocal fold epithelium are still unknown. The aim of this study was to identify the distribution of TJs in the vocal fold epithelium and to characterize the recovery process of TJ-based paracellular diffusion barrier in a rat model of vocal fold injury. Animal experiments with controls. Normal and vocal fold-injured rats were used. Larynges were harvested for immunohistochemical examination of TJ proteins. For functional analysis, a tracer permeability assay was performed using EZ-Link Sulfo-NHS-LC-Biotin. TJ proteins occludin and zonula occludens 1 signals were localized to the junctional regions of the most luminal cell layers of the vocal fold epithelium. The injured region had been recovered with epithelium at 5 days postinjury, but the paracellular diffusion barrier assays revealed that biotinylation reagents diffused into the lamina propria at 5 days postinjury, and were blocked at the epithelium at 14 and 28 days postinjury. It was strongly suggested that TJs in the vocal fold epithelium exist at the junctional regions of the first layer of stratified squamous epithelium. TJ-based paracellular diffusion barrier following vocal fold injury is recovered by 14 days postinjury, and this period corresponds with the time course of structural changes in the regenerating epithelium layer. NA Laryngoscope, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  1. Development of a New Technique to Assess Susceptibility to Predation Resulting from Sublethal Stresses (Indirect Mortality)

    Energy Technology Data Exchange (ETDEWEB)

    Cada, G.F.

    2003-08-25

    Fish that pass through a hydroelectric turbine may not be killed directly, but may nonetheless experience sublethal stresses that will increase their susceptibility to predators (indirect mortality). There is a need to develop reliable tests for indirect mortality so that the full consequences of passage through turbines (and other routes around a hydroelectric dam) can be assessed. We evaluated a new technique for assessing indirect mortality, based on a behavioral response to a startling stimulus (akin to perceiving an approaching predator). We compare this technique to the standard predator preference test. The behavioral response is a rapid movement commonly referred to as a startle response, escape response, or C-shape, based on the characteristic body position assumed by the fish. When viewed from above, a startled fish bends into a C-shape, then springs back and swims away in a direction different from its original orientation. This predator avoidance (escape) behavior can be compromised by sublethal stresses that temporarily stun or disorient the fish. We subjected striped shiners and fathead minnows to varying intensities of either turbulence (10-, 20- or 30-min) or 2-min exposures to a fish anesthetic (100 or 200 mg/L of tricaine methanesulfonate), and evaluated their subsequent behavior. Individual fish were given a startle stimulus and filmed with a high-speed video camera. Each fish was startled and filmed twice before being stressed, and then at 1-, 5-, 15-, and 30-min post-exposure. The resulting image files were analyzed for a variety of behavioral measures including: presence of a response, time to first reaction, duration of reaction, time to formation of maximum C-shape, time to completion of C-shape, and completeness of C-shape. The most immediate measure of potential changes in fish behavior was whether stressed fish exhibited a startle response. For striped shiners, the number of fish not responding to the stimulus was significantly different

  2. Employment among Spinal Cord Injured Patients Living in Turkey: A Cross-Sectional Study

    Science.gov (United States)

    Gunduz, Berrin; Erhan, Belgin; Bardak, Ayse Nur

    2010-01-01

    The aim of this study was to determine the rate of employment and to establish the factors affecting vocational status in spinal cord injured patients living in Turkey. One hundred and fifty-two traumatic spinal cord injured patients older than 18 years with injury duration of at least 1 year and living in the community were included in the study;…

  3. The experience of being a partner to a spinal cord injured person:

    DEFF Research Database (Denmark)

    Angel, Sanne; Buus, Niels

    2011-01-01

    supporting the injured partner and the demanding tasks of everyday life outside the institution. After discharge, partners struggled for the injured partner to regain a well-functioning everyday life and for reestablishing life as a couple. The partner struggled to manage the overwhelming amount of everyday...

  4. Prevalence of alcohol and other psychoactive substances in injured drivers : comparison between Belgium and The Netherlands.

    NARCIS (Netherlands)

    Legrand, S.-A. Houwing, S. Hagenzieker, M. & Verstraete, A.G.

    2013-01-01

    The study objective was to compare the prevalence of alcohol and (il)licit drugs in seriously injured drivers in Belgium (BE) and the Netherlands (NL). Injured car and van drivers admitted to the emergency departments of five hospitals in Belgium and three in the Netherlands from January 2008 to May

  5. Alcohol and drugs in seriously injured drivers in six European countries

    DEFF Research Database (Denmark)

    Legrand, Sara‐Ann; Isalberti, Cristina; Linden, Trudy Van

    2013-01-01

    The objective of this study was to determine the presence of alcohol and drugs in drivers severely injured in traffic crashes in six European countries. Data were collected from 2492 seriously injured drivers of cars and vans in Belgium, Denmark, Finland, Italy, Lithuania, and the Netherlands...

  6. Alcohol and drugs in seriously injured drivers in 6 European countries

    DEFF Research Database (Denmark)

    Legrand, Sara-Ann; Isalberti, Cristina; Linden, Trudy Van der

    2012-01-01

    Objective: To determine the presence of alcohol and drugs in drivers severely injured in traffic crashes in 6 European countries. Methods: Data were collected from 2492 seriously injured drivers of cars and vans in Belgium, Denmark, Finland, Italy, Lithuania and The Netherlands, between 2007...

  7. 9 CFR 311.27 - Injured animals slaughtered at unusual hours.

    Science.gov (United States)

    2010-01-01

    ... slaughter an injured animal at night or on Sunday or a holiday when the inspector cannot be obtained, the... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Injured animals slaughtered at unusual hours. 311.27 Section 311.27 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT...

  8. Differences in kinetic asymmetry between injured and noninjured novice runners : A prospective cohort study

    NARCIS (Netherlands)

    Bredeweg, S. W.; Buist, I.; Kluitenberg, B.

    Purpose: The purpose of this prospective study was to describe natural levels of asymmetry in running, compare levels of asymmetry between injured and noninjured novice runners and compare kinetic variables between the injured and noninjured lower limb within the novice runners with an injury.

  9. Early propranolol administration to severely injured patients can improve bone marrow dysfunction.

    Science.gov (United States)

    Bible, Letitia E; Pasupuleti, Latha V; Alzate, Walter D; Gore, Amy V; Song, Kim J; Sifri, Ziad C; Livingston, David H; Mohr, Alicia M

    2014-07-01

    Bone marrow (BM) dysfunction is common in severely injured trauma patients, resulting from elevated catecholamines and plasma granulocyte colony-stimulating factor (G-CSF) as well as prolonged mobilization of hematopoietic progenitor cells (HPCs). We have previously shown that propranolol (β-blocker [BB]) reduces HPC mobilization in a rodent model of injury and hemorrhagic shock. We hypothesize that BB would prevent BM dysfunction in humans following severe injury. Forty-five severely injured trauma patients were studied in a prospective, randomized pilot trial. Twenty-five patients received BB, and 20 served as untreated controls. The dose of propranolol was adjusted to decrease the heart rate by 10% to 20% from baseline. Blood was analyzed for the presence of HPC (blast-forming unit erythroid cells [BFU-E] and colony-forming unit erythroid cells) and G-CSF. Demographic data, Injury Severity Score (ISS), hemoglobin, reticulocyte number, and outcome data were obtained. The mean age of the study population was 33 years; 87% were male, with a mean ISS of 29. There is a significant increase in BFU-E in peripheral blood immediately following traumatic injury, and this mobilization persists for 30 days. The use of BB significantly decreases BFU-E and colony-forming unit erythroid cells at all time points. G-CSF is significantly elevated in both groups on admission; the use of BB decreases G-CSF levels by 51% as compared with 37% for controls. The average hemoglobin is nearly 1 g higher on the day of discharge with propranolol treatment (BB, 9.9 ± 0.4 g/dL vs. no BB, 9.1 ± 0.6 g/dL). Following severe trauma, early treatment with propranolol following resuscitation is safe. The use of propranolol blunts early tachycardia, reduces HPC mobilization, and results in a faster return to baseline of the G-CSF peak seen after injury. There is also a trend toward faster recovery and resolution of anemia. Propranolol may be the first therapeutic agent to show improved BM

  10. A novel strictly anaerobic recovery and enrichment system incorporating lithium for detection of heat-injured Listeria monocytogenes in pasteurized milk containing background microflora.

    Science.gov (United States)

    Mendonca, A F; Knabel, S J

    1994-11-01

    Heat-injured cells of Listeria monocytogenes were recovered from heated raw milk containing noninjured Enterococcus faecium by combining a simple method for obtaining strict anaerobiosis with a novel enrichment broth, Penn State University broth (PSU broth). Strictly anaerobic conditions were rapidly achieved by adding 0.5 g of filter-sterilized cysteine per liter to PSU broth and then purging the preparation with N2 gas. Little resuscitation or growth occurred in strictly anaerobic PSU broth without lithium chloride because of overgrowth by E. faecium. The growth of E. faecium decreased dramatically with increasing LiCl concentration; LiCl concentrations of 8 and 10 g/liter were completely bacteriostatic. The mechanism of inhibition by LiCl appeared to involve competition with the divalent cations Ca2+ and Mg2+. Heat-injured L. monocytogenes consistently recovered and grew rapidly in strictly anaerobic PSU broth containing 4, 6, or 7 g of LiCl per liter. The use of strictly anaerobic PSU broth containing 7 g of LiCl per liter permitted detection of severely heat-injured L. monocytogenes in one simple recovery-enrichment step by eliminating oxygen toxicity and inhibiting the growth of background microflora, without preventing the resuscitation and subsequent growth of heat-injured L. monocytogenes. L. monocytogenes heated in raw milk at 62.8 degrees C for 10, 15, and 20 min could be consistently recovered from strictly anaerobic PSU broth enrichment cultures at 30 degrees C after 48, 96, and 144 h, respectively, and hence, use of PSU broth may result in better recovery of both injured and noninjured cells from foods than currently used U.S. Department of Agriculture and Food and Drug Administration preenrichment procedures.

  11. Transplantation of embryonic spinal cord neurons to the injured distal nerve promotes axonal regeneration after delayed nerve repair.

    Science.gov (United States)

    Zhang, Wenming; Fang, Xinyu; Zhang, Chaofan; Li, Wen; Wong, Wai Man; Xu, Yejun; Wu, Wutian; Lin, Jianhua

    2017-03-01

    Peripheral nerve injury (PNI) usually results in poor functional recovery. Nerve repair is the common clinical treatment for PNI but is always obstructed by the chronic degeneration of the distal stump and muscle. Cell transplantation can alleviate the muscle atrophy after PNI, but the subsequent recovery of the locomotive function is seldom described. In this study, we combined cell transplantation and nerve repair to investigate whether the transplantation of embryonic spinal cord cells could benefit the delayed nerve repair. The experiment consisted of 3 stages: transection of the tibial nerve to induce 'pre-degeneration', a second surgery performed 2 weeks later for transplantation of E14 embryonic spinal cord cells or vehicle (culture medium) at the distal end of the injured nerve, and, 3 months later, the removal of the grafted cells and the cross-suturing of the residual distal end to the proximal end of a freshly cut ipsilateral common peroneal (CP) nerve. Cell survival and fate after the transplantation were investigated, and the functional recovery after the cross-suturing was compared between the groups. The grafted cells could survive and generate motor neurons, extending axons that were subsequently myelinated and forming synapses with the muscle. After the cross-suturing, the axonal regeneration from the proximal stump of the injured CP nerve and the functional recovery of the denervated gastrocnemius muscle were significantly promoted in the group receiving the cells. Our study presents a new perspective indicating that the transplantation of embryonic spinal cord neurons may be a valuable therapeutic strategy for PNI. © 2016 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  12. Sublethal pesticide doses negatively affect survival and the cellular responses in American foulbrood-infected honeybee larvae

    Science.gov (United States)

    López, Javier Hernández; Krainer, Sophie; Engert, Antonia; Schuehly, Wolfgang; Riessberger-Gallé, Ulrike; Crailsheim, Karl

    2017-02-01

    Disclosing interactions between pesticides and bee infections is of most interest to understand challenges that pollinators are facing and to which extent bee health is compromised. Here, we address the individual and combined effect that three different pesticides (dimethoate, clothianidin and fluvalinate) and an American foulbrood (AFB) infection have on mortality and the cellular immune response of honeybee larvae. We demonstrate for the first time a synergistic interaction when larvae are exposed to sublethal doses of dimethoate or clothianidin in combination with Paenibacillus larvae, the causative agent of AFB. A significantly higher mortality than the expected sum of the effects of each individual stressor was observed in co-exposed larvae, which was in parallel with a drastic reduction of the total and differential hemocyte counts. Our results underline that characterizing the cellular response of larvae to individual and combined stressors allows unmasking previously undetected sublethal effects of pesticides in colony health.

  13. Evaluation of Pyrethrin Formulations on Dengue/Dengue Haemorrhagic Fever Vectors in the Laboratory and Sublethal Effects

    Directory of Open Access Journals (Sweden)

    S Sulaiman

    2007-12-01

    Full Text Available In Southeast Asia, Aedes aegypti (L. has been incriminated as principal vector of dengue viruses and Ae. albopictus as the secondary vector of dengue fever. Therefore, the aim of this study was to investigate the effectiveness of three for-mula¬tions of pyrethrin derived from Tanacetum cinerariaefolium against the dengue/dengue haemorrhagic fever vectors Aedes aegypti and Ae. albopictus in the laboratory. The testings employed 2 methodologies: the WHO Larval Bioassay and WHO Adult Bioassay. The results showed that all the three pyrethrin formulations had larvicidal and adulticidal activi-ties. The impact of the sublethal doses of pyrethrin formulations on Aedes spp. larvae resulted in 4-6% of alive adult emergence compared to 90% of Ae. aegypti emerging adults and 96% Ae. albopictus alive adult emergence in the control. The impact of sublethal doses of the pyrethrin formulations caused very low fecundity on both Aedes spp. compared to the control (P< 0.05.

  14. Physicochemical Evidence on Sublethal Neonicotinoid Imidacloprid Interacting with an Odorant-Binding Protein from the Tea Geometrid Moth, Ectropis obliqua.

    Science.gov (United States)

    Li, Hongliang; Zhao, Lei; Fu, Xiaobin; Song, Xinmi; Wu, Fan; Tang, Mingzhu; Cui, Hongchun; Yu, Jizhong

    2017-04-26

    Nowadays the excessive usage of neonicotinoid insecticides always results in residues in Chinese tea fields. It is not clear whether the insecticide residue at the sublethal level influences the physiological processes of tea pests. Here, we provide evidence of interaction between the neonicotinoid imidacloprid and a general odorant-binding protein, EoblGOBP2, from the tea geometrid moth, Ectropis obliqua. The interacting process was demonstrated through multiple fluorescence spectra, UV absorption spectra, circular dichroism (CD) spectra, molecular docking, etc. The binding mode was determined to be static (from 300 to 310 K) and dynamic quenching (from 290 to 300 K). The binding distance was calculated to be 6.9 nm on the basis of FRET theory. According to the thermodynamic analysis, the process was mainly driven by enthalpy (ΔH neonicotinoid insecticide at sublethal level may still affect the olfactory cognition of the tea geometrid moth to volatile compounds from tea leaves.

  15. Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum.

    Science.gov (United States)

    Amaradasa, B Sajeewa; Everhart, Sydney E

    2016-01-01

    Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor), iprodione (unclear mode of action), thiophanate methyl (inhibition of microtubulin synthesis) and azoxystrobin and pyraclostrobin (quinone outside inhibitors). Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50-100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR) loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs). SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each). Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed) generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA) and discriminant analysis of principal components (DAPC) identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001). Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the experiment, and

  16. Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum.

    Directory of Open Access Journals (Sweden)

    B Sajeewa Amaradasa

    Full Text Available Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor, iprodione (unclear mode of action, thiophanate methyl (inhibition of microtubulin synthesis and azoxystrobin and pyraclostrobin (quinone outside inhibitors. Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50-100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs. SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each. Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA and discriminant analysis of principal components (DAPC identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001. Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the

  17. Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum

    Science.gov (United States)

    Amaradasa, B. Sajeewa

    2016-01-01

    Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor), iprodione (unclear mode of action), thiophanate methyl (inhibition of microtubulin synthesis) and azoxystrobin and pyraclostrobin (quinone outside inhibitors). Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50–100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR) loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs). SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each). Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed) generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA) and discriminant analysis of principal components (DAPC) identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001). Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the experiment

  18. Exposure to sublethal blast overpressure reduces the food intake and exercise performance of rats.

    Science.gov (United States)

    Bauman, R A; Elsayed, N; Petras, J M; Widholm, J

    1997-07-25

    Exposure to blast overpressure can typically inflict generalized damage on major organ systems, especially gas-containing organs such as the lungs and the gastrointestinal tract. The purpose of the present study was to use rat's food intake and exercise wheel running as behavioral correlates of the perhaps more subtle damage to these organ systems induced by sublethal blast overpressure. Toward this end, all rats were exposed to a 12-h light/dark cycle and food was available only in the dark period. Prior to exposure, rats in the (E)xercise group were required to execute five rotations of an activity wheel for a food pellet; wheel turns that occurred at times other than when a rat was feeding were recorded separately and labeled exercise running. In the (S)edentary and (A)nesthesia groups, wheel running was not possible and rats were required to execute five leverpresses for a single pellet. A compressed air-driven shock tube was used to expose rats to a supra-atmospheric wave of air pressure. The tube was separated into two sections by a polyester membrane, the thickness of which determined peak and duration of overpressure. All rats were anesthetized with 50 mg/kg of phenobarbital. After reaching a deep plane of anesthesia, they were individually tied in a stockinet across one end of the shock tube. In preliminary tests, the membrane thickness was 1000 (A)ngstroms and rats in Group L(ethality) were exposed to a 129 kPa (peak amplitude) wave of overpressure. Three of six rats survived exposure to this peak pressure; pathology was evident in the lungs and gastrointestinal tract of all non-survivors. Rats in Groups E and S were tested with a 500 A membrane, which resulted in an 83 kPa peak amplitude. All rats survived exposure to this lower peak pressure. On the day of exposure to blast, the relative reduction of intake during the first 3 h of the dark period was significantly greater for Group E than for Groups S and A; the intake of Groups E and S remained reduced

  19. Basal cell skin cancer

    Science.gov (United States)

    Basal cell skin cancer almost never spreads. If it is left untreated, it may spread into surrounding areas and nearby tissues and bone. In these cases, treatment can injure the appearance of the skin.

  20. Multi-OMIC profiling of survival and metabolic signaling networks in cells subjected to photodynamic therapy.

    Science.gov (United States)

    Weijer, Ruud; Clavier, Séverine; Zaal, Esther A; Pijls, Maud M E; van Kooten, Robert T; Vermaas, Klaas; Leen, René; Jongejan, Aldo; Moerland, Perry D; van Kampen, Antoine H C; van Kuilenburg, André B P; Berkers, Celia R; Lemeer, Simone; Heger, Michal

    2017-03-01

    Photodynamic therapy (PDT) is an established palliative treatment for perihilar cholangiocarcinoma that is clinically promising. However, tumors tend to regrow after PDT, which may result from the PDT-induced activation of survival pathways in sublethally afflicted tumor cells. In this study, tumor-comprising cells (i.e., vascular endothelial cells, macrophages, perihilar cholangiocarcinoma cells, and EGFR-overexpressing epidermoid cancer cells) were treated with the photosensitizer zinc phthalocyanine that was encapsulated in cationic liposomes (ZPCLs). The post-PDT survival pathways and metabolism were studied following sublethal (LC50) and supralethal (LC90) PDT. Sublethal PDT induced survival signaling in perihilar cholangiocarcinoma (SK-ChA-1) cells via mainly HIF-1-, NF-кB-, AP-1-, and heat shock factor (HSF)-mediated pathways. In contrast, supralethal PDT damage was associated with a dampened survival response. PDT-subjected SK-ChA-1 cells downregulated proteins associated with EGFR signaling, particularly at LC90. PDT also affected various components of glycolysis and the tricarboxylic acid cycle as well as metabolites involved in redox signaling. In conclusion, sublethal PDT activates multiple pathways in tumor-associated cell types that transcriptionally regulate cell survival, proliferation, energy metabolism, detoxification, inflammation/angiogenesis, and metastasis. Accordingly, tumor cells sublethally afflicted by PDT are a major therapeutic culprit. Our multi-omic analysis further unveiled multiple druggable targets for pharmacological co-intervention.

  1. Do biopesticides affect the demographic traits of a parasitoid wasp and its biocontrol services through sublethal effects?

    Directory of Open Access Journals (Sweden)

    Antonio Biondi

    Full Text Available Pesticide risk assessments are usually based on short-term acute toxicity tests, while longer-term population dynamic related traits, critical to the success of biological control and Integrated Pest Management (IPM programs, are often overlooked. This is increasingly important with respect to new biopesticides that frequently cause no short-term acute effects, but that can induce multiple physiological and behavioral sublethal effects, leading to a decrease in population growth and ecosystem services. In this study we assessed the lethal and sublethal effects of six biopesticides [abamectin, azadirachtin, Bacillus thuringiensis, borax plus citrus oil (Prev-Am®, emamectin benzoate, and spinosad], used in tomato crops to control the invasive pest Tuta absoluta (Lepidoptera: Gelechiidae, on adults and pupae of the parasitoid Bracon nigricans (Hymenoptera: Braconidae. Data on female survival and production of female offspring were used to calculate population growth indexes as a measure of population recovery after pesticide exposure. Spinosad caused 100% and 80% mortality in exposed adults (even 10 d after the treatment and pupae, respectively. Although most of the biopesticides had low levels of acute toxicity, multiple sublethal effects were observed. The biocontrol activity of both females that survived 1-h and 10-d old residues, and females that emerged from topically treated pupae was significantly affected by the application of the neurotoxic insecticides emamectin benzoate and abamectin. Furthermore, very low B. nigricans demographic growth indices were estimated for these two insecticides, indicating potential local extinction of the wasp populations. Among the tested products, Bt proved to be the safest for B. nigricans adults and pupae. Our findings emphasize that acute toxicity assessment alone cannot fully predict the actual impact of pesticides on non-target parasitoids. Thus, sublethal effects related to the species specific life

  2. Do Biopesticides Affect the Demographic Traits of a Parasitoid Wasp and Its Biocontrol Services through Sublethal Effects?

    Science.gov (United States)

    Biondi, Antonio; Zappalà, Lucia; Stark, John D.; Desneux, Nicolas

    2013-01-01

    Pesticide risk assessments are usually based on short-term acute toxicity tests, while longer-term population dynamic related traits, critical to the success of biological control and Integrated Pest Management (IPM) programs, are often overlooked. This is increasingly important with respect to new biopesticides that frequently cause no short-term acute effects, but that can induce multiple physiological and behavioral sublethal effects, leading to a decrease in population growth and ecosystem services. In this study we assessed the lethal and sublethal effects of six biopesticides [abamectin, azadirachtin, Bacillus thuringiensis, borax plus citrus oil (Prev-Am®), emamectin benzoate, and spinosad], used in tomato crops to control the invasive pest Tuta absoluta (Lepidoptera: Gelechiidae), on adults and pupae of the parasitoid Bracon nigricans (Hymenoptera: Braconidae). Data on female survival and production of female offspring were used to calculate population growth indexes as a measure of population recovery after pesticide exposure. Spinosad caused 100% and 80% mortality in exposed adults (even 10 d after the treatment) and pupae, respectively. Although most of the biopesticides had low levels of acute toxicity, multiple sublethal effects were observed. The biocontrol activity of both females that survived 1-h and 10-d old residues, and females that emerged from topically treated pupae was significantly affected by the application of the neurotoxic insecticides emamectin benzoate and abamectin. Furthermore, very low B. nigricans demographic growth indices were estimated for these two insecticides, indicating potential local extinction of the wasp populations. Among the tested products, Bt proved to be the safest for B. nigricans adults and pupae. Our findings emphasize that acute toxicity assessment alone cannot fully predict the actual impact of pesticides on non-target parasitoids. Thus, sublethal effects related to the species specific life-history variables

  3. Sublethal doses of the pesticide imidacloprid alter honey bee (Apis mellifera) response threshold and navigation, potentially affecting colony health

    OpenAIRE

    Eiri, Daren

    2011-01-01

    Much attention on honey bee declines has focused on the sublethal effects the pesticide, imidacloprid, has on honey bee behavior. How it affects individual foragers and their preference for nectar or their ability to navigate to communicated food sources is unknown. We use the proboscis extension reflex (PER) assay to test an individual's response threshold. Bees treated with the pesticide have higher response thresholds and respond less often to high concentrations of sucrose than control be...

  4. Sub-lethal doses of photodynamic therapy affect biofilm formation ability and metabolic activity of Enterococcus faecalis.

    Science.gov (United States)

    Pourhajibagher, M; Chiniforush, N; Shahabi, S; Ghorbanzadeh, R; Bahador, A

    2016-09-01

    During photodynamic therapy (PDT) in the treatment of a primary endodontic infection, it is extremely likely that microorganisms would be exposed to sub-lethal doses of PDT (sPDT). Although sPDT cannot kill microorganisms, it can considerably influence microbial virulence. This study was conducted to characterize the effect of sPDT using toluidine blue O (TBO), methylene blue (MB), and indocyanine green (ICG) on biofilm formation ability and metabolic activity of Enterococcus faecalis. The antimetabolic and antibiofilm potential of ICG-, TBO-, and MB-sPDT against E. faecalis was analyzed at sub-lethal doses (1/2-1/64 minimum inhibitory concentration) using the XTT reduction assay, crystal violet assay, and scanning electron microscopy. Higher doses of sPDT adversely affected biofilm formation ability and metabolic activity. ICG-, TBO-, and MB-PDT at a maximum sub-lethal dose markedly reduced the formation of biofilm up to 42.8%, 22.6%, and 19.5%, respectively. ICG-, TBO-, and MB-sPDT showed a marked reduction in bacterial metabolic activity by 98%, 94%, and 82%, respectively. ICG-PDT showed a stronger inhibitory effect on biofilm formation in E. faecalis than MB- and TBO-PDT at sub-lethal levels. Interestingly, a gradual increase in metabolic activity and biofilm formation upon exposure to a lower dose of test sPDT were observed. sPDT showed dual effect on biofilm formation ability and metabolic activity of E. faecalis. High doses revealed antimetabolic and antibiofilm potential activity, whereas lower doses had conflicting results. Hence, when PDT is prescribed in clinical settings, the dose of PDT used in vivo should be taken into consideration. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Sublethal Exposure to Diatomaceous Earth Increases Net Fecundity of Flour Beetles (Tribolium confusum) by Inhibiting Egg Cannibalism

    OpenAIRE

    Shostak, Allen W.

    2014-01-01

    Population regulation results from an interplay of numerous intrinsic and external factors, and for many insects cannibalism is such a factor. This study confirms a previously-reported observation that sublethal exposure to the fossilized remains of diatoms (i.e. diatomaceous earth) increases net fecundity (eggs produced minus eggs destroyed/day) of flour beetles, Tribolium confusum. The aim was to experimentally test two non-mutually-exclusive ecological mechanisms potentially responsible fo...

  6. Linking mechanistic and behavioral responses to sublethal esfenvalerate exposure in the endangered delta smelt; Hypomesus transpacificus (Fam. Osmeridae).

    Science.gov (United States)

    Connon, Richard E; Geist, Juergen; Pfeiff, Janice; Loguinov, Alexander V; D'Abronzo, Leandro S; Wintz, Henri; Vulpe, Christopher D; Werner, Inge

    2009-12-15

    The delta smelt (Hypomesus transpacificus) is a pelagic fish species listed as endangered under both the USA Federal and Californian State Endangered Species Acts and considered an indicator of ecosystem health in its habitat range, which is limited to the Sacramento-San Joaquin estuary in California, USA. Anthropogenic contaminants are one of multiple stressors affecting this system, and among them, current-use insecticides are of major concern. Interrogative tools are required to successfully monitor effects of contaminants on the delta smelt, and to research potential causes of population decline in this species. We have created a microarray to investigate genome-wide effects of potentially causative stressors, and applied this tool to assess effects of the pyrethroid insecticide esfenvalerate on larval delta smelt. Selected genes were further investigated as molecular biomarkers using quantitative PCR analyses. Exposure to esfenvalerate affected swimming behavior of larval delta smelt at concentrations as low as 0.0625 mug.L-1, and significant differences in expression were measured in genes involved in neuromuscular activity. Alterations in the expression of genes associated with immune responses, along with apoptosis, redox, osmotic stress, detoxification, and growth and development appear to have been invoked by esfenvalerate exposure. Swimming impairment correlated significantly with expression of aspartoacylase (ASPA), an enzyme involved in brain cell function and associated with numerous human diseases. Selected genes were investigated for their use as molecular biomarkers, and strong links were determined between measured downregulation in ASPA and observed behavioral responses in fish exposed to environmentally relevant pyrethroid concentrations. The results of this study show that microarray technology is a useful approach in screening for, and generation of molecular biomarkers in endangered, non-model organisms, identifying specific genes that can be

  7. Linking mechanistic and behavioral responses to sublethal esfenvalerate exposure in the endangered delta smelt; Hypomesus transpacificus (Fam. Osmeridae

    Directory of Open Access Journals (Sweden)

    Wintz Henri

    2009-12-01

    Full Text Available Abstract Background The delta smelt (Hypomesus transpacificus is a pelagic fish species listed as endangered under both the USA Federal and Californian State Endangered Species Acts and considered an indicator of ecosystem health in its habitat range, which is limited to the Sacramento-San Joaquin estuary in California, USA. Anthropogenic contaminants are one of multiple stressors affecting this system, and among them, current-use insecticides are of major concern. Interrogative tools are required to successfully monitor effects of contaminants on the delta smelt, and to research potential causes of population decline in this species. We have created a microarray to investigate genome-wide effects of potentially causative stressors, and applied this tool to assess effects of the pyrethroid insecticide esfenvalerate on larval delta smelt. Selected genes were further investigated as molecular biomarkers using quantitative PCR analyses. Results Exposure to esfenvalerate affected swimming behavior of larval delta smelt at concentrations as low as 0.0625 μg.L-1, and significant differences in expression were measured in genes involved in neuromuscular activity. Alterations in the expression of genes associated with immune responses, along with apoptosis, redox, osmotic stress, detoxification, and growth and development appear to have been invoked by esfenvalerate exposure. Swimming impairment correlated significantly with expression of aspartoacylase (ASPA, an enzyme involved in brain cell function and associated with numerous human diseases. Selected genes were investigated for their use as molecular biomarkers, and strong links were determined between measured downregulation in ASPA and observed behavioral responses in fish exposed to environmentally relevant pyrethroid concentrations. Conclusions The results of this study show that microarray technology is a useful approach in screening for, and generation of molecular biomarkers in endangered

  8. Acute Toxicity and Sublethal Effects of Terpenoids and Essential Oils on the Predator Chrysoperla externa (Neuroptera: Chrysopidae).

    Science.gov (United States)

    Castilhos, R V; Grützmacher, A D; Coats, J R

    2017-07-31

    The search for new safer insecticides has increased in recent agriculture. Botanical compounds such as terpenoids and plant essential oils with insecticidal activity could represent important tools in pest management, and their risk assessment against non-target organisms is necessary since they may serve as a precursor for the synthesis of new insecticide active ingredients. For this study, the acute toxicity and sublethal effects of seven terpenoids and three essential oils with recognized insecticidal activity were evaluated on the predator Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) in laboratory bioassays. Results indicate that these compounds feature relative selectivity to the predator C. externa; however, sublethal effects on reproduction were recorded for some compounds. The phenolic monoterpenoids carvacrol and thymol were more acutely toxic than other terpenoids screened, with LD 50 <20,000 μg/g; however, they were less toxic than natural pyrethrins (toxicity standard) in these bioassays. Sublethal effects on fecundity and fertility were observed for R-(+)-limonene, while oregano oil only affected fecundity. The compounds evaluated here have potential to be used as insecticides and can serve as backbone for future synthesis of selective active ingredients; however, a complete risk assessment to C. externa and other non-target organisms is necessary for their incorporation in future crop protection paradigms.

  9. Lethal and Sublethal Effects of Some Chemical and Biological Insecticides on Tuta absoluta (Lepidoptera: Gelechiidae) Eggs and Neonates.

    Science.gov (United States)

    Nozad-Bonab, Z; Hejazi, M J; Iranipour, Sh; Arzanlou, M

    2017-06-01

    Tuta absoluta (Meyrick) is one of the serious pests of tomatoes. This study was conducted to evaluate the effects of some chemical and biological insecticides on T. absoluta. The insecticides tested were diazinon, dichlorvos, chlorantraniliprole, deltamethrin, acetamiprid, imidacloprid, spinosad, abamectin, indoxacarb, Bacillus thuringiensis Berliner, Beauveria bassiana (Balsamo) Vuillemin, and Metarhizium anisopliae (Metschnikoff) Sorokin. Sublethal effects for the most effective insecticides tested, namely, abamectin, spinosad, chlorantraniliprole, and indoxacarb were assessed, and life table parameters were calculated. Chlorantraniliprole had the highest lethal effect on T. absoluta followed by spinosad, abamectin, and indoxacarb. On the other hand, imidacloprid was not considerably effective on T. absoluta eggs. Metarhizium anisopliae was 11 and 518 times more effective on the eggs and newly hatched larvae than B. bassiana and B. thuringiensis, respectively. Chlorantraniliprole, spinosad, abamectin, and indoxacarb affected life table parameters of T. absoluta significantly (α = 0.05). Spinosad had the highest sublethal effect on T. absoluta followed by abamectin, chlorantraniliprole, and indoxacarb. The results revealed that chlorantraniliprole, spinosad, abamectin, and indoxacarb had considerable lethal and sublethal effects on T. absoluta, and if they perform similarly in commercial greenhouses and fields, they would be suitable candidates to be considered in IPM programs for this pest. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. The Wiggle Index: An Open Source Bioassay to Assess Sub-Lethal Insecticide Response in Drosophila melanogaster.

    Science.gov (United States)

    Denecke, Shane; Nowell, Cameron J; Fournier-Level, Alexandre; Perry, Trent; Batterham, Phil

    2015-01-01

    Toxicological assays measuring mortality are routinely used to describe insecticide response, but sub-lethal exposures to insecticides can select for resistance and yield additional biological information describing the ways in which an insecticide impacts the insect. Here we present the Wiggle Index (WI), a high-throughput method to quantify insecticide response by measuring the reduction in motility during sub-lethal exposures in larvae of the vinegar fly Drosophila melanogaster. A susceptible wild type strain was exposed to the insecticides chlorantraniliprole, imidacloprid, spinosad, and ivermectin. Each insecticide reduced larval motility, but response times and profiles differed among insecticides. Two sets of target site mutants previously identified in mortality studies on the basis of imidacloprid or spinosad resistance phenotypes were tested. In each case the resistant mutant responded significantly less than the control. The WI was also able to detect a spinosad response in the absence of the primary spinosad target site. This response was not detected in mortality assays suggesting that spinosad, like many other insecticides, may have secondary targets affecting behaviour. The ability of the WI to detect changes in insecticide metabolism was confirmed by overexpressing the imidacloprid metabolizing Cyp6g1 gene in digestive tissues or the central nervous system. The data presented here validate the WI as an inexpensive, generic, sub-lethal assay that can complement information gained from mortality assays, extending our understanding of the genetic basis of insecticide response in D. melanogaster.

  11. Does selective logging stress tropical forest invertebrates? Using fat stores to examine sublethal responses in dung beetles.

    Science.gov (United States)

    França, Filipe; Barlow, Jos; Araújo, Bárbara; Louzada, Julio

    2016-12-01

    The increased global demand for tropical timber has driven vast expanses of tropical forests to be selectively logged worldwide. While logging impacts on wildlife are predicted to change species distribution and abundance, the underlying physiological responses are poorly understood. Although there is a growing consensus that selective logging impacts on natural populations start with individual stress-induced sublethal responses, this literature is dominated by investigations conducted with vertebrates from temperate zones. Moreover, the sublethal effects of human-induced forest disturbance on tropical invertebrates have never been examined. To help address this knowledge gap, we examined the body fat content and relative abundance of three dung beetle species (Coleoptera: Scarabaeinae) with minimum abundance of 40 individuals within each examined treatment level. These were sampled across 34 plots in a before-after control-impact design (BACI) in a timber concession area of the Brazilian Amazon. For the first time, we present evidence of logging-induced physiological stress responses in tropical invertebrates. Selective logging increased the individual levels of fat storage and reduced the relative abundance of two dung beetle species. Given this qualitative similarity, we support the measurement of body fat content as reliable biomarker to assess stress-induced sublethal effects on dung beetles. Understanding how environmental modification impacts the wildlife has never been more important. Our novel approach provides new insights into the mechanisms through which forest disturbances impose population-level impacts on tropical invertebrates.

  12. {sup 1}H NMR metabolomics of earthworm exposure to sub-lethal concentrations of phenanthrene in soil

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Sarah A.E.; McKelvie, Jennifer R.; Simpson, Andre J. [Department of Physical and Environmental Sciences, University of Toronto, 1265 Military Trail Toronto, Ontario, M1C 1A4 (Canada); Simpson, Myrna J., E-mail: myrna.simpson@utoronto.c [Department of Physical and Environmental Sciences, University of Toronto, 1265 Military Trail Toronto, Ontario, M1C 1A4 (Canada)

    2010-06-15

    {sup 1}H NMR metabolomics was used to monitor earthworm responses to sub-lethal (50-1500 mg/kg) phenanthrene exposure in soil. Total phenanthrene was analyzed via soxhlet extraction, bioavailable phenanthrene was estimated by hydroxypropyl-beta-cyclodextrin (HPCD) and 1-butanol extractions and sorption to soil was assessed by batch equilibration. Bioavailable phenanthrene (HPCD-extracted) comprised approx65-97% of total phenanthrene added to the soil. Principal component analysis (PCA) showed differences in responses between exposed earthworms and controls after 48 h exposure. The metabolites that varied with exposure included amino acids (isoleucine, alanine and glutamine) and maltose. PLS models indicated that earthworm response is positively correlated to both total phenanthrene concentration and bioavailable (HPCD-extracted) phenanthrene in a freshly spiked, unaged soil. These results show that metabolomics is a powerful, direct technique that may be used to monitor contaminant bioavailability and toxicity of sub-lethal concentrations of contaminants in the environment. These initial findings warrant further metabolomic studies with aged contaminated soils. - {sup 1}H NMR metabolomics is used to directly monitor metabolic responses of Eisenia fetida after 48 h of exposure to sub-lethal concentrations of phenanthrene in soil.

  13. The Effect of Acclimation to Sublethal Temperature on Subsequent Susceptibility of Sitophilus zeamais Mostchulsky (Coleoptera: Curculionidae) to High Temperatures.

    Science.gov (United States)

    Lü, Jianhua; Zhang, Huina

    2016-01-01

    Heat treatment is a popular alternative to synthetic pesticides in disinfesting food-processing facilities and empty grain storages. Sitophilus zeamais Mostchulsky is one of the most cosmopolitan and destructive insects found in empty grain storage facilities and processing facilities. The effect of acclimation in S. zeamais adults to sublethal high temperature on their subsequent susceptibility to high temperatures was investigated. S. zeamais adults were acclimated to 36°C for 0 (as a control), 1, 3, and 5 h, and then were exposed at 43, 47, 51, and 55°C for different time intervals respectively. Acclimation to sublethal high temperature significantly reduced subsequent susceptibility of S. zeamais adults to lethal high temperatures of 43, 47, 51, and 55°C, although the mortality of S. zeamais adults significantly increased with increasing exposure time at lethal high temperatures. The mortality of S. zeamais adults with 1, 3, and 5 h of acclimation to 36°C was significantly lower than that of S. zeamais adults without acclimation when exposed to the same lethal high temperatures. The present results suggest that the whole facility should be heated to target lethal high temperature as soon as possible, avoiding decreasing the control effectiveness of heat treatment due to the acclimation in stored product insects to sublethal temperature.

  14. The sub-lethal effects of repeated freezing in the woolly bear caterpillar Pyrrharctia isabella.

    Science.gov (United States)

    Marshall, Katie E; Sinclair, Brent J

    2011-04-01

    Repeated freeze-thaw cycles are common and are increasing in frequency with climate change in many temperate locations, yet understanding of their impact on freeze-tolerant insects is extremely limited. We investigated the effects of repeated freezing and thawing on the freeze-tolerant final instar caterpillars of the moth Pyrrharctia isabella (Lepidoptera: Arctiidae) by subjecting individuals to either a single sustained 35 h freeze or five 7 h freezes. Sub-lethal effects were quantified with changes in three broad groups of measures: (1) cold hardiness, (2) metabolic rate and energy reserves and (3) survival after challenge with fungal spores. Repeated freeze-thaw cycles increased mortality to almost 30% and increased tissue damage in Malpighian tubules and hemocytes. Repeated freezing increased caterpillar glycerol concentration by 0.82 mol l(-1). There were no changes in metabolic rate or energy reserves with repeated freezing. For the first time, we report increased survival after immune challenge in caterpillars after freezing and suggest that this may be linked to wounding during freezing. We suggest that little repair of freezing damage is possible in P. isabella caterpillars and repeated freeze-thaw cycles may present significant challenges to survival in this species.

  15. Sublethal Effects of Spirodiclofen on Tetranychus urticae Koch_Pre-Ovipositional Females After Different Exposure Times

    Directory of Open Access Journals (Sweden)

    Dejan Marčić

    2006-01-01

    Full Text Available Sublethal effect of spirodiclofen on Tetranychus urticae females that survived different exposure times in the pre-ovipositional period was evaluated calculating two parameters - instantaneous rate of increase and net fertility - after six days of reproduction. The females were exposed to four concentrations/doses of the acaricide: 96 mg /L (0.24 μg/cm2, 48 mg/L(0.12 μg/cm2, 24 mg/L (0.06 μg/cm2 and 12 mg./L (0.03 μg/cm2 for 2, 6 and 24h in a leaf disc bioassay. After 24h exposure to 12 mg/L, instantaneous rate of increase was significantlyreduced (0.545; 0.634 in control, while significant reduction in net fertility (20.61; 28.57 in the control was recorded even after 2h exposure to the same concentration. The effect of all tested concentrations of spirodiclofen on both parameters increased with exposure time. The lowest values of instantaneous rate of increase (0.268 and net fertility (2.58 were recorded after 24h exposure to 96 mg/L. After 24h exposure, the concentration increase from 12 to 24 mg/L significantly reduced both parameters, while a further increase from 24 to 96 mg/L significantly reduced instantaneous rate of increase, but not net-fertility. The results regarding T. urticae population management are discussed.

  16. Haematological changes in Bufo maculatus treated with sublethal concentrations of Cadmium

    Directory of Open Access Journals (Sweden)

    Lawrence Ikechukwu Ezemonye

    2011-12-01

    Full Text Available Adult Bufo maculatus was exposed to sublethal cadmium concentrations of 0.25, 0.50, 1.00 and 2.00 mg/L. The toxicant from which the cadmium concentrations were prepared was cadmium chloride (CdCl2.H2O. There were three replicate tanks per treatment and three individuals per tank including control groups. The hematologic alterations based on the examination of blood indices during the 28 days of exposure showed that total erythrocyte count (TEC, hematocrit (Hct and hemoglobin (Hb concentration decreased (P<0.05 relative to controls. The decline was concentration- dependent as concentration of cadmium increased. The decline in hemoglobin and hematocrit in the experimental organism could be due to a decrease in the synthesis or release of erythrocytes into the circulation or an increase in the rate of erythrocyte destruction inflicted by cadmium toxicity. There was significant (P<0.05 elevation in total leuko- leukocyte count (TLC with increase in the concen- cyte concentration of cadmium. The increase in total leukocyte count observed in this study could be attributed to a stimulation of the immune system in response to tissue damage caused by cadmium toxicity. The study has shown that the exposure of the Bufo maculatus toad to cadmium can inflict alterations in the hematologic indices, which could induce unfavorable physiological changes in the amphibian, which may lead to death. There is, therefore, the need to protect amphibians in order to sustain the biodiversity in the Nigerian Niger Delta ecological zone.

  17. Sublethal Effects of Neonicotinoid Insecticide on Calling Behavior and Pheromone Production of Tortricid Moths.

    Science.gov (United States)

    Navarro-Roldán, Miguel A; Gemeno, César

    2017-09-01

    In moths, sexual behavior combines female sex pheromone production and calling behavior. The normal functioning of these periodic events requires an intact nervous system. Neurotoxic insecticide residues in the agroecosystem could impact the normal functioning of pheromone communication through alteration of the nervous system. In this study we assess whether sublethal concentrations of the neonicotinoid insecticide thiacloprid, that competitively modulates nicotinic acetylcholine receptors at the dendrite, affect pheromone production and calling behavior in adults of three economically important tortricid moth pests; Cydia pomonella (L.), Grapholita molesta (Busck), and Lobesia botrana (Denis & Schiffermüller). Thiacloprid significantly reduced the amount of calling in C. pomonella females at LC 0.001 (a lethal concentration that kills only 1 in 10 5 individuals), and altered its calling period at LC 1 , and in both cases the effect was dose-dependent. In the other two species the effect was similar but started at higher LCs, and the effect was relatively small in L. botrana. Pheromone production was altered only in C. pomonella, with a reduction of the major compound, codlemone, and one minor component, starting at LC 10 . Since sex pheromones and neonicotinoids are used together in the management of these three species, our results could have implications regarding the interaction between these two pest control methods.

  18. Sublethal Effects of the Neonicotinoid Insecticide Thiamethoxam on the Transcriptome of the Honey Bees (Hymenoptera: Apidae).

    Science.gov (United States)

    Shi, Teng-Fei; Wang, Yu-Fei; Liu, Fang; Qi, Lei; Yu, Lin-Sheng

    2017-12-05

    Neonicotinoid insecticides are now the most widely used insecticides in the world. Previous studies have indicated that sublethal doses of neonicotinoids impair learning, memory capacity, foraging, and immunocompetence in honey bees (Apis mellifera, Linnaeus) (Hymenoptera: Apidae). Despite these, few studies have been carried out on the molecular effects of neonicotinoids. In this study, we focus on the second-generation neonicotinoid thiamethoxam, which is currently widely used in agriculture to protect crops. Using high-throughput RNA-Seq, we investigated the transcriptome profile of honey bees after subchronic exposure to 10 ppb thiamethoxam over 10 d. In total, 609 differentially expressed genes (DEGs) were identified, of which 225 were upregulated and 384 were downregulated. Several genes, including vitellogenin, CSP3, defensin-1, Mrjp1, and Cyp6as5 were selected and further validated using real-time quantitative polymerase chain reaction assays. The functions of some DEGs were identified, and Gene Ontology-enrichment analysis showed that the enriched DEGs were mainly linked to metabolism, biosynthesis, and translation. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that thiamethoxam affected biological processes including ribosomes, the oxidative phosphorylation pathway, tyrosine metabolism pathway, pentose and glucuronate interconversions, and drug metabolism. Overall, our results provide a basis for understanding the molecular mechanisms of the complex interactions between neonicotinoid insecticides and honey bees. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Sublethal Effects in Pest Management: A Surrogate Species Perspective on Fruit Fly Control

    Directory of Open Access Journals (Sweden)

    John E. Banks

    2017-07-01

    Full Text Available Tephritid fruit flies are economically important orchard pests globally. While much effort has focused on controlling individual species with a combination of pesticides and biological control, less attention has been paid to managing assemblages of species. Although several tephritid species may co-occur in orchards/cultivated areas, especially in mixed-cropping schemes, their responses to pesticides may be highly variable. Furthermore, predictive efforts about toxicant effects are generally based on acute toxicity, with little or no regard to long-term population effects. Using a simple matrix model parameterized with life history data, we quantified the responses of several tephritid species to the sublethal effects of a toxicant acting on fecundity. Using a critical threshold to determine levels of fecundity reduction below which species are driven to local extinction, we determined that threshold levels vary widely for the three tephritid species. In particular, Bactrocera dorsalis was the most robust of the three species, followed by Ceratitis capitata, and then B. cucurbitae, suggesting individual species responses should be taken into account when planning for area-wide pest control. The rank-order of susceptibility contrasts with results from several field/lab studies testing the same species, suggesting that considering a combination of life history traits and individual species susceptibility is necessary for understanding population responses of species assemblages to toxicant exposure.

  20. Assessment of Mercury Bioaccumulation in Zebra Cichlid (Cichlasoma Nigrofasciatum Exposed to Sublethal Concentrations of Permethrin

    Directory of Open Access Journals (Sweden)

    Mahdi Banaee

    2014-12-01

    Full Text Available Background: Aquatic ecosystems are frequently subjected to contamination by toxic heavy metals and pesticides, yet very little is known about the influence of pesticides on bioaccumulation of heavy metals in aquatic organisms. Mercury is a toxic metal with no known biological benefit to organisms. Bioavailability of mercury in aquatic environments depends on biological and non-biological parameters including other pollutants. Therefore, the objectives of this research were to determine the effects of permethrin on bioaccumulation of mercury in zebra cichlid. Methods: Acute toxicity (LC50 of permethrin and mercury chloride was evaluated by estimating mortality in Probit Model in SPSS (version 19.0 IBM. In sub-lethal toxicity, zebra cichlid (Cichlasoma nigrofasciatum was exposed to various concentrations of permethrin (0.0, 0.40, 0.80, 1.20 and 1.60 µg.L-1 combined with 20 µg.L-1 mercury chloride for 15 days. At the end of the experiment, mercury concentrations were measured using ICP-OES-Perkin elmer (optima 7300-DV. Results: 96 h LC50 values of permethrin and mercury for C. nigrofasciatum were calculated to be 17.55 µg.L-1 and 140.38 µg.L-1, respectively. Our results clearly showed that the bioaccumulation of mercury in the specimens increased with increasing concentrations of permethrin to 1.20 and 1.60 µg.L-1. Conclusion: Increasing the concentration of permethrin had synergistic effects on the bioaccumulation of mercury in fish.

  1. Evaluating sublethal indicators of stress in Asiatic clams (Corbicula fluminea) caged in an urban stream

    Energy Technology Data Exchange (ETDEWEB)

    Black, M.C.; Belin, J.I. [Univ. of Georgia, Athens, GA (United States). College of Agricultural and Environmental Sciences

    1998-12-31

    Freshwater bivalves have been used extensively to monitor chemical accumulation in field exposures, although little information is available on the use of biomarker measurements in field exposures with bivalves. DNA strand breakage, growth rate, condition index and percentage tissue water were measured in freshwater Asiatic clams (Corbicula fluminea) exposed in-situ in a stream that receives urban and industrial stormwater runoff and in a non-impacted reference stream. After 4 weeks exposure, DNA strand lengths in foot tissue from Trail Creek-exposed clams were significantly shorter than DNA from reference clams. These results suggest a reduction in DNA integrity in Trail Creek-exposed clams, possibly indicating exposure to genotoxic chemicals. No significant differences were observed in the growth rates of clams. However, a significant inverse relationship was detected between condition index and % tissue water for all clams. Furthermore, site-specific differences in percentage tissue water and condition indices were observed after 2 and 10 weeks exposure. For this study DNA strand breakage, condition indices, and tissue hydration appear to be more sensitive indicators of sublethal toxicity than growth.

  2. Integrated microfluidic technology for sub-lethal and behavioral marine ecotoxicity biotests

    Science.gov (United States)

    Huang, Yushi; Reyes Aldasoro, Constantino Carlos; Persoone, Guido; Wlodkowic, Donald

    2015-06-01

    Changes in behavioral traits exhibited by small aquatic invertebrates are increasingly postulated as ethically acceptable and more sensitive endpoints for detection of water-born ecotoxicity than conventional mortality assays. Despite importance of such behavioral biotests, their implementation is profoundly limited by the lack of appropriate biocompatible automation, integrated optoelectronic sensors, and the associated electronics and analysis algorithms. This work outlines development of a proof-of-concept miniaturized Lab-on-a-Chip (LOC) platform for rapid water toxicity tests based on changes in swimming patterns exhibited by Artemia franciscana (Artoxkit M™) nauplii. In contrast to conventionally performed end-point analysis based on counting numbers of dead/immobile specimens we performed a time-resolved video data analysis to dynamically assess impact of a reference toxicant on swimming pattern of A. franciscana. Our system design combined: (i) innovative microfluidic device keeping free swimming Artemia sp. nauplii under continuous microperfusion as a mean of toxin delivery; (ii) mechatronic interface for user-friendly fluidic actuation of the chip; and (iii) miniaturized video acquisition for movement analysis of test specimens. The system was capable of performing fully programmable time-lapse and video-microscopy of multiple samples for rapid ecotoxicity analysis. It enabled development of a user-friendly and inexpensive test protocol to dynamically detect sub-lethal behavioral end-points such as changes in speed of movement or distance traveled by each animal.

  3. The concerted action of lactoferrin and bacteriophages in the clearance of bacteria in sublethally infected mice.

    Science.gov (United States)

    Zimecki, Michał; Artym, Jolanta; Kocieba, Maja; Weber-Dabrowska, Beata; Lusiak-Szelachowska, Marzena; Górski, Andrzej

    2008-02-07

    Both lactoferrin (LF) and bacteriophages are potent antibacterial agents. LF is contained in the secretory fluids of mammals and bacteriophages are specific bacterial viruses. The aim of this investigation was to determine whether combined treatment of infected mice may allow lowering the therapeutic dose of specific bacteriophages for Escherichia coli and Staphylococcus aureus. CBA mice were infected intravenously (i.v.) with sublethal doses of E. coli or S. aureus and the specific T4 or A5 bacteriophages, respectively, were administered intraperitoneally (i.p.) or per os one hour following infection. The numbers of colony-forming units (CFUs) were determined in the livers after 24 hours. Comparative administration of bacteriophages i.p. or per os showed that both routes of administration were equally efficacious in the protective action of bacteriophages. The bacteriophages were still very potent in reducing CFU numbers in the liver at a dose of 10(5)/mouse. Application of bovine lactoferrin (LF), 10 mg i.v., 24 h before infection, was also very effective in reducing CFU numbers. Using suboptimal (10(3)-10(4)) doses of bacteriophages and administration of LF, a more potent protective effect in reducing the CFU numbers in the infected mice was demonstrated. The combined effect of LF and bacteriophages in reducing CFU numbers was significantly higher than the effects of either agent alone. The study demonstrated that the combined application of LF and bacteriophages can significantly lower (1000 times) the effective dose of bacteriophages in reducing CFU numbers in infected mice.

  4. Inhibition of acetylcholinesterase in guppies (Poecilia reticulata) by chlorpyrifos at sublethal concentrations: Methodological aspects

    Energy Technology Data Exchange (ETDEWEB)

    van der Wel, H.; Welling, W.

    1989-04-01

    Acetylcholinesterase activity is a potential biochemical indicator of toxic stress in fish and a sensitive parameter for testing water for the presence of organophosphates. A number of methodological aspects regarding the determination of the in vivo effect of chlorpyrifos on acetylcholinesterase in guppies have been investigated. It was found that with acetylthiocholine as a substrate, the contribution of pseudocholinesterase to the total cholinesterase activity can be neglected. Protection of acetylcholinesterase of guppies exposed to chlorpyrifos from additional, artifactual in vitro enzyme inhibition during homogenization is necessary. Very low concentrations of acetone in the exposure medium, resulting from dilution of the stock solution of chlorpyrifos in acetone, can result in large decreases in the oxygen content of this medium. This may affect the uptake rate of the toxic compound and, thereby, cholinesterase inhibition. Very low, sublethal concentrations of chlorpyrifos result in high inhibition levels of acetylcholinesterase (80-90%) in guppies within 2 weeks of continuous exposure. Recovery of the enzyme activity occurs after the exposed animals are kept in clean medium for 4 days, but the rate of recovery is considerably lower than the rate of inhibition.

  5. Sub-lethal effects of herbicides penoxsulam, imazamox, fluridone and glyphosate on Delta Smelt (Hypomesus transpacificus).

    Science.gov (United States)

    Jin, Jiali; Kurobe, Tomofumi; Ramírez-Duarte, Wilson F; Bolotaolo, Melissa B; Lam, Chelsea H; Pandey, Pramod K; Hung, Tien-Chieh; Stillway, Marie E; Zweig, Leanna; Caudill, Jeffrey; Lin, Li; Teh, Swee J

    2018-02-01

    Concerns regarding non-target toxicity of new herbicides used to control invasive aquatic weeds in the San Francisco Estuary led us to compare sub-lethal toxicity of four herbicides (penoxsulam, imazamox, fluridone, and glyphosate) on an endangered fish species Delta Smelt (Hypomesus transpacificus). We measured 17β-estradiol (E2) and glutathione (GSH) concentrations in liver, and acetylcholinesterase (AChE) activity in brain of female and male fish after 6 h of exposure to each of the four herbicides. Our results indicate that fluridone and glyphosate disrupted the E2 concentration and decreased glutathione concentration in liver, whereas penoxsulam, imazamox, and fluridone inhibited brain AChE activity. E2 concentrations were significantly increased in female and male fish exposed to 0.21 μM of fluridone and in male fish exposed to 0.46, 4.2, and 5300 μM of glyphosate. GSH concentrations decreased in males exposed to fluridone at 2.8 μM and higher, and glyphosate at 4.2 μM. AChE activity was significantly inhibited in both sexes exposed to penoxsulam, imazamox, and fluridone, and more pronounced inhibition was observed in females. The present study demonstrates the potential detrimental effects of these commonly used herbicides on Delta Smelt. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. Sublethal toxicity of quinalphos on oxidative stress and antioxidant responses in a freshwater fish Cyprinus carpio.

    Science.gov (United States)

    Hemalatha, Devan; Amala, Antony; Rangasamy, Basuvannan; Nataraj, Bojan; Ramesh, Mathan

    2016-11-01

    Extensive use of quinalphos, an organophosphorus pesticide, is likely to reach the aquatic environment and thereby posing a health concern for aquatic organisms. Oxidative stress and antioxidant responses may be good indicators of pesticide contamination in aquatic organisms. The data on quinalphos induced oxidative stress and antioxidant responses in carps are scanty. This study is aimed to assess the two sublethal concentrations of quinalphos (1.09 and 2.18 μL L -1 ) on oxidative stress and antioxidant responses of Cyprinus carpio for a period of 20 days. In liver, the malondialdehyde level was found to be significantly increased in both the concentrations. The results of the antioxidant parameters obtained show a significant increase in superoxide dismutase, catalase, and glutathione-S-transferase activity in liver of fish. These results demonstrate that environmentally relevant levels of the insecticide quinalphos can cause oxidative damage and increase the antioxidant scavenging capacity in C. carpio. This may reflect the potential role of these parameters as useful biomarkers for the assessment of pesticide contamination. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1399-1406, 2016. © 2015 Wiley Periodicals, Inc.

  7. Analysis of lethal and sublethal impacts of environmental disasters on sperm whales using stochastic modeling.

    Science.gov (United States)

    Ackleh, Azmy S; Chiquet, Ross A; Ma, Baoling; Tang, Tingting; Caswell, Hal; Veprauskas, Amy; Sidorovskaia, Natalia

    2017-08-01

    Mathematical models are essential for combining data from multiple sources to quantify population endpoints. This is especially true for species, such as marine mammals, for which data on vital rates are difficult to obtain. Since the effects of an environmental disaster are not fixed, we develop time-varying (nonautonomous) matrix population models that account for the eventual recovery of the environment to the pre-disaster state. We use these models to investigate how lethal and sublethal impacts (in the form of reductions in the survival and fecundity, respectively) affect the population's recovery process. We explore two scenarios of the environmental recovery process and include the effect of demographic stochasticity. Our results provide insights into the relationship between the magnitude of the disaster, the duration of the disaster, and the probability that the population recovers to pre-disaster levels or a biologically relevant threshold level. To illustrate this modeling methodology, we provide an application to a sperm whale population. This application was motivated by the 2010 Deepwater Horizon oil rig explosion in the Gulf of Mexico that has impacted a wide variety of species populations including oysters, fish, corals, and whales.

  8. Sub-lethal cadmium exposure increases phytochelatin concentrations in the aquatic snail Lymnaea stagnalis

    Energy Technology Data Exchange (ETDEWEB)

    SF, Gonçalves [Department of Biology & CESAM, Centre for Environmental and Marine Studies, University of Aveiro, 3810-193 Aveiro (Portugal); SK, Davies [Department of Surgery and Cancer, Imperial College London, Sir Alexander Fleming Building, London SW7 2AZ (United Kingdom); Bennett, M. [Department of Life Sciences, Imperial College London, Sir Alexander Fleming Building, London SW7 2AZ (United Kingdom); Raab, A.; Feldmann, J. [TESLA, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen AB24 3UE, Scotland (United Kingdom); Kille, P. [Cardiff School of Biosciences, Cardiff University, Park Place, Cardiff CF10 3US (United Kingdom); Loureiro, S. [Department of Biology & CESAM, Centre for Environmental and Marine Studies, University of Aveiro, 3810-193 Aveiro (Portugal); DJ, Spurgeon [Centre for Ecology and Hydrology, Maclean Building, Benson Lane, Wallingford OX10 8BB (United Kingdom); JG, Bundy, E-mail: j.bundy@imperial.ac.uk [Department of Surgery and Cancer, Imperial College London, Sir Alexander Fleming Building, London SW7 2AZ (United Kingdom)

    2016-10-15

    Phytochelatins are metal-binding metabolites found in almost all plant species and some animal groups, including nematodes and annelids, where they can play an important role in detoxifying metals such as cadmium. Species from several other taxa contain a phytochelatin synthase (PCS) gene orthologue, including molluscs, indicating they may have the potential to synthesize phytochelatins. However, the presence of a gene alone does not demonstrate that it plays a functional role in metal detoxification. In the present study, we show that the aquatic snail Lymnaea stagnalis produced both penta- and heptapeptide phytochelatins (i.e. phytochelatin-2 and phytochelatin-3), and their levels increased in response to sub-lethal levels of cadmium. - Highlights: • Little is known about the role of phytochelatins in metal detoxification in animals. • We detected phytochelatins (PC{sub 2} and PC{sub 3}) in a mollusc species, Lymnaea stagnalis. • Phytochelatins increased in Lymnaea stagnalis when exposed to cadmium. • Future research on phytochelatin responses in molluscs would be valuable.

  9. Lethal and sub-lethal effects of UVB on juvenile Biomphalaria glabrata (Mollusca: Pulmonata).

    Science.gov (United States)

    Ruelas, Debbie S; Karentz, Deneb; Sullivan, John T

    2006-11-01

    Although Schistosoma mansoni occurs mainly in the tropics, where intense levels of solar radiation are present, the impact of ultraviolet (UV) light on schistosome transmission is not known. The purpose of this study was to investigate potential effects of UVB (290-320nm) on juvenile Biomphalaria glabrata, the snail intermediate host of S. mansoni. Albino and wild-type snails were exposed to doses of UVB from UV-fluorescent lamps, and the following were measured: survival, photoreactivation (light-mediated DNA repair), effects on feeding behavior, and morphological tissue abnormalities. Irradiation with UVB is lethal to B. glabrata in a dose-dependent manner. Exposure to white light subsequent to UVB irradiation enhances survival, probably by photoreactivation. The shell offers some, but not complete, protection. Experiments in which UVB transmittance through the shell was blocked with black nail polish suggest that injury to both exposed (headfoot) and shell-enclosed (mantle and visceral mass) tissues contributes to mortality in lethally irradiated snails. Wild-type (pigmented) snails are less susceptible to lethal effects of UVB than albino snails, and they may be more capable of photoreactivation. UVB exposure inhibits snail feeding behavior, and causes tentacle forks and growths on the headfoot. Thus, UVB may influence the life cycle of S. mansoni by both lethal and sub-lethal damage to the snail intermediate host. However, the ability of snails to photoreactivate may mitigate these effects.

  10. Pediatric specific shock index accurately identifies severely injured children.

    Science.gov (United States)

    Acker, Shannon N; Ross, James T; Partrick, David A; Tong, Suhong; Bensard, Denis D

    2015-02-01

    Shock index (SI) (heart rate/systolic blood pressure)>0.9 predicts mortality in adult trauma patients. We hypothesized that age adjusted SI could more accurately predict outcomes in children. Retrospective review of children age 4-16 years admitted to two trauma centers between 1/07 and 6/13 following blunt trauma with an injury severity score (ISS)>15 was performed. We evaluated the ability of SI>0.9 at emergency department presentation and elevated shock index, pediatric age adjusted (SIPA) to predict outcomes. SIPA was defined by maximum normal HR and minimum normal SBP by age. Cutoffs included SI>1.22 (age 4-6), >1.0 (7-12), and >0.9 (13-16). Among 543 children, 50% of children had an SI>0.9 but this fell to 28% using age adjusted SI (SIPA). SIPA demonstrated improved discrimination of severe injury relative to SI: ISS>30: 37% vs 26%; blood transfusion within the first 24 hours: 27% vs 20%; Grade III liver/spleen laceration requiring blood transfusion: 41% vs 26%; and in-hospital mortality: 11% vs 7%. A pediatric specific shock index (SIPA) more accurately identifies children who are most severely injured, have intraabdominal injury requiring transfusion, and are at highest risk of death when compared to shock index unadjusted for age. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. The role of resilience in rebuilding lives of injured veterans.

    Science.gov (United States)

    Edward, K-L; Ousey, K

    2016-10-02

    The aim of this commentary is to discuss potential clinical implications of introducing resilience-building interventions into care for veterans who are living with a war wound. Some war veterans are expected to live with a wound upon discharge from an active military role and also to fit into civilian life. These lifestyle adjustments can tax the person's coping abilities and in that context may hinder successful adaptation. The experience of living with a wound or wounds, either acute or chronic, is connected to losses, including loss of mobility, loss of financial capacity (unable to work at times) and losses attached to changed social roles. Psychological stress is also a common experience for veterans returning to civilian life. Psychological stress is associated with impaired wound healing. Modern health practice is centred on symptom reduction and working with pathology, however, working with people's adaptive behaviours such as resilience has not been considered. Using the resilience model as a conceptual framework health-care professionals can engage with veterans towards resilience within the context of their personal experience of ill health. Using this contemporary framework for considering these aspects of care has the potential to facilitate resistance to stressors associated with being injured, possibly preventing quality-of-life impairments. There are no declarations of interest.

  12. Relationship between stress ankle radiographs and injured ligaments on MRI

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Kyoung Min; Chung, Chin Youb; Chung, Myung Ki; Won, Sung Hun; Lee, Seung Yeol; Park, Moon Seok [Seoul National University Bundang Hospital, Department of Orthopaedic Surgery, Kyungki (Korea, Republic of); Kwon, Soon-Sun [Seoul National University Bundang Hospital, Biomedical Research Institute, Kyungki (Korea, Republic of)

    2013-11-15

    This study was performed to investigate the relationship between the injured lateral ankle ligaments on MRI and stress ankle radiographs. Two hundred and twenty-nine consecutive patients (mean age 35.5 years, SD 14.6 years; 136 males and 93 females) that underwent ankle stress radiographs and MRI for lateral ankle instability were included. Tibiotalar tilt angle and anterior translation of talus were measured on stress ankle radiographs. Degree of lateral ligaments (anterior talofibular, calcaneofibular, and posterior talofibular) and deltoid ligament injuries were evaluated and scored as intact (0), partial injury (1), and complete injury (2) on MR images. Effusion of ankle joint was also recorded. The effects of gender, age, injuries of ligaments, and ankle joint effusion on stress radiographs were statistically analyzed. Gender (p = 0.010), age (p = 0.020), and anterior talofibular ligament (ATFL) injury (p < 0.001) were the factors significantly affecting tibiotalar tilt angle. Posterior talofibular ligament (PTFL) injury (p = 0.014) was found to be the only significant factor affecting the anterior translation on the anterior drawer radiographs. ATFL injury and PTFL injury on MRI significantly affected tibiotalar tilt angle and anterior drawer on stress radiographs. Other factors, such as age and gender, need to be considered in evaluating radiographic lateral ankle instability. (orig.)

  13. Lactate and the injured brain: friend or foe?

    Science.gov (United States)

    Bouzat, Pierre; Oddo, Mauro

    2014-04-01

    Energy metabolism is increasingly recognized as a key factor in the pathogenesis of acute brain injury (ABI). We review the role of cerebral lactate metabolism and summarize evidence showing that lactate may act as supplemental fuel after ABI. The role of cerebral lactate has shifted from a waste product to a potentially preferential fuel and signaling molecule. According to the astrocyte-neuron lactate shuttle model, glycolytic lactate might act as glucose-sparing substrate. Lactate also is emerging as a key signal to regulate cerebral blood flow (CBF) and a neuroprotective agent after experimental ABI. Clinical investigation using cerebral microdialysis shows the existence of two main lactate patterns, ischemic - from anaerobic metabolism - and nonischemic, from activated glycolysis, whereby lactate can be used as supplemental energy fuel. Preliminary clinical data suggests hypertonic lactate solutions improve cerebral energy metabolism and are an effective treatment for elevated intracranial pressure (ICP) after ABI. Lactate can be a supplemental fuel for the injured brain and is important to regulate glucose metabolism and CBF. Exogenous lactate supplementation may be neuroprotective after experimental ABI. Recent clinical data from ABI patients suggest hypertonic lactate solutions may be a valid therapeutic option for secondary energy dysfunction and elevated ICP.

  14. Relationship between stress ankle radiographs and injured ligaments on MRI.

    Science.gov (United States)

    Lee, Kyoung Min; Chung, Chin Youb; Kwon, Soon-Sun; Chung, Myung Ki; Won, Sung Hun; Lee, Seung Yeol; Park, Moon Seok

    2013-11-01

    This study was performed to investigate the relationship between the injured lateral ankle ligaments on MRI and stress ankle radiographs. Two hundred and twenty-nine consecutive patients (mean age 35.5 years, SD 14.6 years; 136 males and 93 females) that underwent ankle stress radiographs and MRI for lateral ankle instability were included. Tibiotalar tilt angle and anterior translation of talus were measured on stress ankle radiographs. Degree of lateral ligaments (anterior talofibular, calcaneofibular, and posterior talofibular) and deltoid ligament injuries were evaluated and scored as intact (0), partial injury (1), and complete injury (2) on MR images. Effusion of ankle joint was also recorded. The effects of gender, age, injuries of ligaments, and ankle joint effusion on stress radiographs were statistically analyzed. Gender (p = 0.010), age (p = 0.020), and anterior talofibular ligament (ATFL) injury (p ligament (PTFL) injury (p = 0.014) was found to be the only significant factor affecting the anterior translation on the anterior drawer radiographs. ATFL injury and PTFL injury on MRI significantly affected tibiotalar tilt angle and anterior drawer on stress radiographs. Other factors, such as age and gender, need to be considered in evaluating radiographic lateral ankle instability.

  15. Social reintegration of traumatic brain-injured: the French experience.

    Science.gov (United States)

    Truelle, J-L; Wild, K Von; Onillon, M; Montreuil, M

    2010-01-01

    Traumatic Brain Injury (TBI) may lead to specific handicap, often hidden, mainly due to cognitive and behavioural sequelae. Social re-entry is a long-term, fluctuant and precarious process. The French experience will be illustrated by 6 initiatives answering to 6 challenges to do with TBI specificities:1. bridging the gap, between initial rehabilitation and community re-entry, via transitional units dealing with assessment, retraining, social/vocational orientation and follow-up. Today, there are 30 such units based on multidisciplinary teams.2. assessing recovery by TBI-specific and validated evaluation tools: EBIS holistic document, BNI Screening of higher cerebral functions, Glasgow outcome extended, and QOLIBRI, a TBI-specific quality of life tool.3. promoting specific re-entry programmes founded on limited medication, ecological neuro-psychological rehabilitation, exchange groups and workshops, violence prevention, continuity of care, environmental structuration, and "resocialisation".4. taking into account the "head injured family"5. facilitating recovery after sports-related concussion6. facing medico-legal consequences and compensation: In that perspective, we developed guidelines for TBI-specific expert appraisal, including mandatory neuro-psychological assessment, family interview and an annual forum gathering lawyers and health professionals.

  16. Retaliatory attitudes and violent behaviors among assault-injured youth.

    Science.gov (United States)

    Copeland-Linder, Nikeea; Johnson, Sara B; Haynie, Denise L; Chung, Shang-en; Cheng, Tina L

    2012-03-01

    To examine the effect of retaliatory attitudes on subsequent violent behavior and fight-related injuries among youth who presented to the emergency department with assault injuries. Assault-injured youth were interviewed at baseline, 6 months, and 18 months to assess fighting behavior, retaliatory attitudes, weapon carrying, and injury history as part of a larger randomized control trial. Two emergency departments in urban areas were selected for the study. A total of 129 adolescents aged 10-15 years were included in the study. Fighting behavior, assault injury, weapon carrying, and aggressive behavior. Higher retaliatory attitudes at baseline were associated with more aggression and a higher frequency of fighting over time. Retaliatory attitudes may fuel cycles of violence among youth. Medical professionals in acute care settings have an opportunity to identify youths at risk of future assault injury by assessing retaliation, providing anticipatory guidance, and referring to intervention programs. Copyright © 2012 Society for Adolescent Health and Medicine. All rights reserved.

  17. Management of Sexual Disorders in Spinal Cord Injured Patients

    Directory of Open Access Journals (Sweden)

    Alexander R Vaccaro

    2012-05-01

    Full Text Available Spinal cord injured (SCI patients have sexual disorders including erectile dysfunction (ED, impotence, priapism, ejaculatory dysfunction and infertility. Treatments for erectile dysfunction include four steps. Step 1 involves smoking cessation, weight loss, and increasing physical activity. Step 2 is phosphodiesterase type 5 inhibitors (PDE5I such as Sildenafil (Viagra, intracavernous injections of Papaverine or prostaglandins, and vacuum constriction devices. Step 3 is a penile prosthesis, and Step 4 is sacral neuromodulation (SNM. Priapism can be resolved spontaneously if there is no ischemia found on blood gas measurement or by Phenylephrine. For anejaculatory dysfunction, massage, vibrator, electrical stimulation and direct surgical biopsy can be used to obtain sperm which can then be used for intra-uterine or in-vitro fertilization. Infertility treatment in male SCI patients involves a combination of the above treatments for erectile and anejaculatory dysfunctions. The basic approach to and management of sexual dysfunction in female SCI patients are similar as for men but do not require treatment for erectile or ejaculatory problems.

  18. Plasma fibronectin promotes thrombus growth and stability in injured arterioles

    Science.gov (United States)

    Ni, Heyu; Yuen, Peter S. T.; Papalia, Jessie M.; Trevithick, Jane E.; Sakai, Takao; Fässler, Reinhard; Hynes, Richard O.; Wagner, Denisa D.

    2003-01-01

    Mice lacking both of the best-known platelet ligands, von Willebrand factor and fibrinogen, can still form occlusive thrombi in injured arterioles. The platelets of these animals accumulate excessive amounts of fibronectin (FN). These observations led us to examine the contribution of plasma FN (pFN) to thrombus formation. Inactivation of the FN gene in FN conditional knockout mice reduced pFN levels to <2% and platelet FN to ≈20% of the levels in similarly treated control mice. The mice were then observed in a model of arterial injury to evaluate their capacity to form thrombi. The deficiency of pFN did not affect the initial platelet adhesion, but a delay of several minutes in thrombus formation was observed in the arterioles of pFN-deficient mice as compared with control mice. The thrombi that formed in the absence of pFN were stably anchored to the vessel wall but continuously shed platelets or small platelet clumps, thus slowing their growth significantly; the platelet/platelet cohesion was apparently diminished. Consequently the occlusion of pFN-deficient vessels was delayed, with the majority of vessels remaining patent at the end of the 40-min observation period. We conclude that, in addition to von Willebrand factor and fibrinogen, FN plays a significant role in thrombus initiation, growth, and stability at arterial shear rates and that deficiency in each of the three platelet ligands has its own specific impact on platelet plug formation. PMID:12606706

  19. Neuronal glycosylation differentials in normal, injured and chondroitinase-treated environments

    Energy Technology Data Exchange (ETDEWEB)

    Kilcoyne, Michelle; Sharma, Shashank [Glycoscience Group, National Centre for Biomedical Engineering Science, National University of Ireland, Galway (Ireland); McDevitt, Niamh; O' Leary, Claire [Anatomy, School of Medicine, National University of Ireland, Galway (Ireland); Joshi, Lokesh [Glycoscience Group, National Centre for Biomedical Engineering Science, National University of Ireland, Galway (Ireland); McMahon, Siobhan S., E-mail: siobhan.mcmahon@nuigalway.ie [Anatomy, School of Medicine, National University of Ireland, Galway (Ireland)

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer Carbohydrates are important in the CNS and ChABC has been used for spinal cord injury (SCI) treatment. Black-Right-Pointing-Pointer Neuronal glycosylation in injury and after ChABC treatment is unknown. Black-Right-Pointing-Pointer In silico mining verified that glyco-related genes were differentially regulated after SCI. Black-Right-Pointing-Pointer In vitro model system revealed abnormal sialylation in an injured environment. Black-Right-Pointing-Pointer The model indicated a return to normal neuronal glycosylation after ChABC treatment. -- Abstract: Glycosylation is found ubiquitously throughout the central nervous system (CNS). Chondroitin sulphate proteoglycans (CSPGs) are a group of molecules heavily substituted with glycosaminoglycans (GAGs) and are found in the extracellular matrix (ECM) and cell surfaces. Upon CNS injury, a glial scar is formed, which is inhibitory for axon regeneration. Several CSPGs are up-regulated within the glial scar, including NG2, and these CSPGs are key inhibitory molecules of axonal regeneration. Treatment with chondroitinase ABC (ChABC) can neutralise the inhibitory nature of NG2. A gene expression dataset was mined in silico to verify differentially regulated glycosylation-related genes in neurons after spinal cord injury and identify potential targets for further investigation. To establish the glycosylation differential of neurons that grow in a healthy, inhibitory and ChABC-treated environment, we established an indirect co-culture system where PC12 neurons were grown with primary astrocytes, Neu7 astrocytes (which overexpress NG2) and Neu7 astrocytes treated with ChABC. After 1, 4 and 8 days culture, lectin cytochemistry of the neurons was performed using five fluorescently-labelled lectins (ECA MAA, PNA, SNA-I and WFA). Usually {alpha}-(2,6)-linked sialylation scarcely occurs in the CNS but this motif was observed on the neurons in the injured environment only at day 8. Treatment

  20. Rearing effect of biofloc on antioxidant and antimicrobial transcriptional response in Litopenaeus stylirostris shrimp facing an experimental sub-lethal hydrogen peroxide stress.

    Science.gov (United States)

    Cardona, Emilie; Saulnier, Denis; Lorgeoux, Bénédicte; Chim, Liet; Gueguen, Yannick

    2015-08-01

    This study compares the antioxidant and antimicrobial transcriptional expression of blue shrimps reared according to two different systems, BioFloc Technology (BFT) and Clear sea Water (CW) and their differential responses when facing an experimental sublethal hydrogen peroxide stress. After 30 days of rearing, juvenile shrimps were exposed to H2O2 stress at a concentration of 30 ppm during 6 h. The oxidative stress caused by H2O2 was examined in the digestive glands of the shrimp, in which antioxidant enzyme (AOE) and antimicrobial peptide (AMP) gene expression were analysed by quantitative real-time PCR. Results showed that rearing conditions did not affect the expression of genes encoding AOEs or AMPs. However, H2O2 stress induced a differential response in expression between shrimps from the two rearing treatments (BFT and CW). Comparative analysis of the expression profiles indicates that catalase transcripts were significantly upregulated by H2O2 stress for BFT shrimps while no change was observed for CW shrimps. In contrast, H2O2 caused down-regulation of superoxide dismutase and glutathione transferase transcripts and of the three AMP transcripts studied (penaeidin 2 and 3, and crustin) for CW shrimps, while no effect was observed on BFT shrimp transcript levels. These results suggested that BFT shrimps maintained antioxidant and AMP responses after stress and therefore can effectively protect their cells against oxidative stress, while CW shrimp immune competence seems to decrease after stress. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Effects of chronic exposure to sublethal concentrations of lead acetate on heme synthesis and immune function in red-tailed hawks.

    Science.gov (United States)

    Redig, P T; Lawler, E M; Schwartz, S; Dunnette, J L; Stephenson, B; Duke, G E

    1991-07-01

    Red-tailed hawks were exposed to sublethal levels of lead acetate for periods of 3 or 11 weeks. Alterations in the heme biosynthetic pathway were demonstrated after the first week of exposure to 0.82 mg lead per kilogram body weight per day. Activity of erythrocyte porphobilinogen synthase (aminolevulinic acid dehydratase) was depressed significantly and did not return to normal levels until 5 weeks after the termination of lead treatments. A rapid and relatively brief increase in erythrocyte free protoporphyrin and a slower but more prolonged increase in its zinc complex were also demonstrated with exposure to this dose of lead for 3 weeks. Less substantial decreases in hematocrit and hemoglobin levels occurred but only in the longer experiment with exposure to higher lead levels. Short term, low level lead exposure did not effect immune function significantly in the hawks, as measured by antibody titers to foreign red blood cells or by the mitogenic stimulation of T-lymphocytes. Increased lead exposure produced a significant decrease in the mitogenic response but had no effect on antibody titers.

  2. A biomarker model of sublethal genotoxicity (DNA single-strand breaks and adducts) using the sentinel organism Aporrectodea longa in spiked soil

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Francis L. [Department of Biological Sciences, Institute of Environmental and Natural Sciences, Lancaster University, Lancaster LA1 4YQ (United Kingdom)]. E-mail: f.martin@lancaster.ac.uk; Piearce, Trevor G. [Department of Biological Sciences, Institute of Environmental and Natural Sciences, Lancaster University, Lancaster LA1 4YQ (United Kingdom); Hewer, Alan [Institute of Cancer Research, Brookes Lawley Building, Cotswold Road, Sutton, Surrey SM2 5NG (United Kingdom); Phillips, David H. [Institute of Cancer Research, Brookes Lawley Building, Cotswold Road, Sutton, Surrey SM2 5NG (United Kingdom); Semple, Kirk T. [Department of Environmental Science, Institute of Environmental and Natural Sciences, Lancaster University, Lancaster LA1 4YQ (United Kingdom)

    2005-11-15

    There is a need to develop risk biomarkers during the remediation of contaminated land. We employed the earthworm, Aporrectodea longa (Ude), to determine whether genotoxicity measures could be applied to this organism's intestinal tissues. Earthworms were added, for 24 h or 7 days, to soil samples spiked with benzo[a]pyrene (B[a]P) and/or lindane. After exposure, intestinal tissues (crop/gizzard or intestine) were removed prior to the measurement in disaggregated cells of DNA single-strand breaks (SSBs) by the alkaline comet assay. Damage was quantified by comet tail length (CTL, {mu}m). B[a]P 24-h exposure induced dose-related increases (P<0.0001) in SSBs. Earthworm intestine was significantly (P<0.0001) more susceptible than crop/gizzard to B[a]P and/or lindane. However, both tissues appeared to acquire resistance following 7-day exposure. B[a]P-DNA adducts, measured by {sup 32}P-postlabelling, showed a two-adduct-spot pattern. This preliminary investigation suggests that earthworm tissues may be incorporated into genotoxicity assays to facilitate hazard identification within terrestrial ecosystems. - Sublethal genotoxicity in the sentinel organism A. longa can be used to monitor the effects of contaminants in soil.

  3. Muscle injury in rats induces upregulation of inflammatory cytokines in injured muscle and calcitonin gene-related peptide in dorsal root ganglia innervating the injured muscle.

    Science.gov (United States)

    Sakuma, Yoshihiro; Miyagi, Masayuki; Inoue, Gen; Ishikawa, Tetsuhiro; Kamoda, Hiroto; Yamauchi, Kazuyo; Orita, Sumihisa; Suzuki, Miyako; Oikawa, Yasuhiro; Inage, Kazuhide; Kubota, Go; Sainoh, Takeshi; Sato, Jun; Takahashi, Kazuhisa; Ohtori, Seiji

    2016-10-01

    In this study we evaluated the relationships among the behavioral changes after muscle injury, histological changes, changes in inflammatory cytokines in the injured muscle, and changes in the sensory nervous system innervating the muscle in rats. We established a model of muscle injury in rats using a dropped weight. Behavior was assessed using the CatWalk system. Subsequently, bilateral gastrocnemius muscles and dorsal root ganglia (DRGs) were resected. Muscles were stained with hematoxylin and eosin, and inflammatory cytokines in injured muscles were assayed. DRGs were immunostained for calcitonin gene-related peptide (CGRP). Changes of behavior and upregulation of inflammatory cytokines in injured muscles subsided within 2 days of injury. Repaired tissue was observed 3 weeks after injury. However, upregulation of CGRP in DRG neurons continued for 2 weeks after injury. These findings may explain in part the pathological mechanism of persistent muscle pain. Muscle Nerve 54: 776-782, 2016. © 2016 The Authors Muscle & Nerve Published by Wiley Periodicals, Inc.

  4. Experimental and numerical study of heterogeneous pressure-temperature-induced lethal and sublethal injury of Lactococcus lactis in a medium scale high-pressure autoclave.

    Science.gov (United States)

    Kilimann, K V; Kitsubun, P; Delgado, A; Gänzle, M G; Chapleau, N; Le Bail, A; Hartmann, C

    2006-07-05

    The present contribution is dedicated to experimental and theoretical assessment of microbiological process heterogeneities of the high-pressure (HP) inactivation of Lactococcus lactis ssp. cremoris MG 1363. The inactivation kinetics are determined in dependence of pressure, process time, temperature and absence or presence of co-solutes in the buffer system namely 4 M sodium chloride and 1.5 M sucrose. The kinetic analysis is carried out in a 0.1-L autoclave in order to minimise thermal and convective effects. Upon these data, a deterministic inactivation model is formulated with the logistic equation. Its independent variables represent the counts of viable cells (viable but injured) and of the stress-resistant cells (viable and not injured). This model is then coupled to a thermo-fluiddynamical simulation method, high-pressure computer fluid dynamics technique (HP-CFD), which yields spatiotemporal temperature and flow fields occurring during the HP application inside any considered autoclave. Besides the thermo-fluiddynamic quantities, the coupled model predicts also the spatiotemporal distribution of both viable (VC) and stress-resistant cell counts (SRC). In order to assess the process non-uniformity of the microbial inactivation in a 3.3-L autoclave experimentally, microbial samples are placed at two distinct locations and are exposed to various process conditions. It can be shown with both, experimental and theoretical models that thermal heterogeneities induce process non-uniformities of more than one decimal power in the counts of the viable cells at the end of the treatment. (c) 2006 Wiley Periodicals, Inc.

  5. Differential Gene Expression in Explanted Human Retinal Pigment Epithelial Cells 24-Hours Post-Exposure to 532 nm, 3.0 ns Pulsed Laser Light and 1064 nm, 170 ps Pulsed Laser Light 12-Hours Post-Exposure: Results Compendium

    National Research Council Canada - National Science Library

    Obringer, John

    2004-01-01

    .... We assessed the sublethal insult to human retinal pigment epithelial cells using a cadaver organ donor explant system for genes differentially expressed 12 and 24 hours post- exposure using gene...

  6. Sagittal plane biomechanics cannot injure the ACL during sidestep cutting.

    Science.gov (United States)

    McLean, Scott G; Huang, Xuemei; Su, Anne; Van Den Bogert, Antonie J

    2004-10-01

    Knee joint sagittal plane forces are a proposed mechanism of anterior cruciate ligament injury during sport movements such as sidestep cutting. Ligament force magnitudes for these movements however, remain unknown. The need to examine injury-causing events suggests elucidation via model-based investigations is possible. Using this approach, the current study determined whether sagittal plane knee loading during sidestep cutting could in isolation injure the anterior cruciate ligament. Experiments were performed on subject-specific forward dynamic musculoskeletal models, generated from data obtained from 10 male and 10 female athletes. Models were optimized to simulate subject-specific cutting movements. Random perturbations (n=5000) were applied to initial contact conditions and quadriceps/hamstrings activation levels to simulate their effect on peak 3D knee loads. Injury via the sagittal plane mechanism was based on the criterion of an anterior drawer force greater than 2000 N. Realistic neuromuscular perturbations produced significant increases in external knee anterior force and valgus and internal rotation moments. Peak anterior drawer force never exceeded 2000 N in any model, and thus failed to cause anterior cruciate ligament injuries. Valgus loads reached values that were high enough to rupture the ligament, occurring more frequently in females than in males. Sagittal plane knee joint forces cannot rupture the anterior cruciate ligament during sidestep cutting. The interaction between muscle and joint mechanics and external ground reaction forces in this plane, places a ceiling on ligament loads. Valgus loading is a more likely injury mechanism, especially in females. Modifying sagittal plane biomechanics will thus unlikely contribute to the prevention of anterior cruciate ligament injuries.

  7. Benchmarking outcomes in the critically injured burn patient.

    Science.gov (United States)

    Klein, Matthew B; Goverman, Jeremy; Hayden, Douglas L; Fagan, Shawn P; McDonald-Smith, Grace P; Alexander, Andrew K; Gamelli, Richard L; Gibran, Nicole S; Finnerty, Celeste C; Jeschke, Marc G; Arnoldo, Brett; Wispelwey, Bram; Mindrinos, Michael N; Xiao, Wenzhong; Honari, Shari E; Mason, Philip H; Schoenfeld, David A; Herndon, David N; Tompkins, Ronald G

    2014-05-01

    To determine and compare outcomes with accepted benchmarks in burn care at 6 academic burn centers. Since the 1960s, US morbidity and mortality rates have declined tremendously for burn patients, likely related to improvements in surgical and critical care treatment. We describe the baseline patient characteristics and well-defined outcomes for major burn injuries. We followed 300 adults and 241 children from 2003 to 2009 through hospitalization, using standard operating procedures developed at study onset. We created an extensive database on patient and injury characteristics, anatomic and physiological derangement, clinical treatment, and outcomes. These data were compared with existing benchmarks in burn care. Study patients were critically injured, as demonstrated by mean % total body surface area (TBSA) (41.2 ± 18.3 for adults and 57.8 ± 18.2 for children) and presence of inhalation injury in 38% of the adults and 54.8% of the children. Mortality in adults was 14.1% for those younger than 55 years and 38.5% for those aged 55 years and older. Mortality in patients younger than 17 years was 7.9%. Overall, the multiple organ failure rate was 27%. When controlling for age and % TBSA, presence of inhalation injury continues to be significant. This study provides the current benchmark for major burn patients. Mortality rates, notwithstanding significant % TBSA and presence of inhalation injury, have significantly declined compared with previous benchmarks. Modern day surgical and medically intensive management has markedly improved to the point where we can expect patients younger than 55 years with severe burn injuries and inhalation injury to survive these devastating conditions.

  8. Nursing challenges with a severely injured patient in critical care.

    Science.gov (United States)

    Crossan, Lisa; Cole, Elaine

    2013-09-01

    Patients with severe, multiple, traumatic injuries are challenging to manage in critical care. Early identification of injuries and optimal resuscitation is essential for favourable outcomes. Trauma-related haemorrhage can lead to the lethal triad of hypothermia, coagulopathy and acidosis. Many trauma patients require urgent haemorrhage control and structural fixation through operative intervention. However, metabolic derangement and cardiovascular instability may delay surgery, resulting in an ongoing cycle of deterioration. Damage control surgery (DCS) may be used as a temporizing measure until the patient is stabilized in critical care. The aim of this case study is to discuss the complex issues faced in the critical care management of a severely injured patient. We conducted a patient case study, with analysis of care using published evidence. The key terms used to search for evidence were trauma, injury, damage control surgery, spinal fixation, critical/intensive care and nurse. We report the care of a trauma patient with complex, conflicting injuries requiring management of the lethal triad and DCS. The delay in subsequent definitive repair of spinal column fractures provided many challenges for critical care nurses including restricted patient mobilization, positioning and pressure ulcer prevention. A review of contemporary evidence relating to DCS reveals that whilst this technique is used increasingly in trauma, the research focuses on single system injuries. Evidence and guidelines are required to support DCS for critical care patients with multiple, conflicting injuries including spinal fractures. For patients with delayed surgical intervention, rotational bed therapy may assist critical care nurses in meeting needs. © 2013 The Authors. Nursing in Critical Care © 2013 British Association of Critical Care Nurses.

  9. Dose-dependent hepatic transcriptional responses in Atlantic salmon (Salmo salar) exposed to sublethal doses of gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Song, You, E-mail: you.song@niva.no [Norwegian University of Life Sciences (NMBU), Faculty of Environmental Science and Technology, Department of Environmental Sciences (IMV), Centre for Environmental Radioactivity - CERAD, P.O. Box 5003, N-1432 Ås (Norway); Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, N-0349 Oslo (Norway); Salbu, Brit; Teien, Hans-Christian; Heier, Lene Sørlie [Norwegian University of Life Sciences (NMBU), Faculty of Environmental Science and Technology, Department of Environmental Sciences (IMV), Centre for Environmental Radioactivity - CERAD, P.O. Box 5003, N-1432 Ås (Norway); Rosseland, Bjørn Olav [Norwegian University of Life Sciences (NMBU), Faculty of Environmental Science and Technology, Department of Environmental Sciences (IMV), Centre for Environmental Radioactivity - CERAD, P.O. Box 5003, N-1432 Ås (Norway); Norwegian University of Life Sciences (NMBU), Department of Ecology and Natural Resource Management, P.O. Box 5003, N-1432 Ås (Norway); Tollefsen, Knut Erik [Norwegian University of Life Sciences (NMBU), Faculty of Environmental Science and Technology, Department of Environmental Sciences (IMV), Centre for Environmental Radioactivity - CERAD, P.O. Box 5003, N-1432 Ås (Norway); Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, N-0349 Oslo (Norway)

    2014-11-15

    affected DEGs associated with cellular signaling and immune response; 70 mGy radiation affected cell cycle regulation and DNA damage repair, cellular energy production; and 280 mGy radiation affected pathways related to cell cycle regulation and DNA repair, mitochondrial dysfunction and immune functions. Twelve genes representative of key pathways found in this study were verified by qPCR. Potential common MoAs of low-dose gamma radiation may include induction of oxidative stress, DNA damage and disturbance of oxidative phosphorylation (OXPHOS). Although common MoAs were proposed, a number of DEGs and pathways were still found to be dose-specific, potentially indicating multiple mechanisms of action (MOAs) of low-dose gamma radiation in fish. In addition, plasma glucose displayed an apparent increase with increasing radiation doses, although the results were not significantly different from the control. These findings suggested that sublethal doses of gamma radiation may cause dose-dependent transcriptional changes in the liver of Atlantic salmon after short-term exposure. The current study predicted multiple MoA for gamma radiation and may aid future impact assessment of environmental radioactivity in fish.

  10. Responses of bay anchovy (Anchoa mitchilli) larvae under lethal and sublethal scenarios of crude oil exposure.

    Science.gov (United States)

    Duffy, Tara A; Childress, William; Portier, Ralph; Chesney, Edward J

    2016-12-01

    Bay anchovy (Anchoa mitchilli) is an ecologically important zooplanktivorous fish inhabiting estuaries of the Gulf of Mexico and eastern North America from Maine to Florida. Because they have a protracted spawning season (spring through fall) and are abundant at all life stages in coastal estuaries, their eggs and larvae likely encountered oil that reached the coast during the Deepwater Horizon oil spill. We compared responses to oil exposure at different life stages and at lethal and sublethal conditions using acute, 24h exposures. In a series of experiments, bay anchovy larvae were exposed to high energy water accommodated fractions (HEWAF) and chemically-enhanced WAF (CEWAF) at two stages of larval development (5 and 21 days post hatch, dph). HEWAF oil exposures induced significantly greater life stage dependent sensitivity at 5 dph than at 21 dph but chemically dispersed (CEWAF) exposure mortality was more variable and LC50s were not significantly different between 5 and 21dph larvae. Acute exposure to two low-level concentrations of CEWAF did not result in significant mortality over 24h, but resulted in a 25-77% reduction in larval survival and a 12-34% reduction in weight specific growth after six days of post-exposure growth following the initial 24h exposure. These results show that younger (5 dph) bay anchovy larvae are more vulnerable to acute oil exposure than older (21 dph) larvae, and that acute responses do not accurately reflect potential population level mortality and impacts to growth and development. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Lethal and Sublethal Effects of Insecticides Used on Citrus, on the Ectoparasitoid Tamarixia radiata.

    Science.gov (United States)

    Beloti, Vitor Hugo; Alves, Gustavo Rodrigues; Araújo, Diogo Feliciano Dias; Picoli, Mateus Manara; Moral, Rafael de Andrade; Demétrio, Clarice Garcia Borges; Yamamoto, Pedro Takao

    2015-01-01

    Huanglongbing (HLB) is a disease associated with the bacteria "Candidatus Liberibacter spp." and has been devastating citrus orchards around the world. Its management involves control of the insect vector, the Asian citrus psyllid Diaphorina citri Kuwayama. However, the indiscriminate use of chemicals has caused pest outbreaks and eliminated the natural enemies of the vector, such as the parasitoid Tamarixia radiata (Waterston), the main agent for biological control of D. citri. This study assessed the lethal and sublethal effects of insecticides recommended for integrated production of citrus on the parasitoid T. radiata. When adult parasitoids were exposed to residues of 25 insecticides, 20% of them, i.e., gamma-cyhalothrin, etofenprox, azadirachtin, tebufenozide and pyriproxyfen, were considered as harmless (Class 1), 12% as slightly harmful (Class 2), 12% as moderately harmful (Class 3) and 56% as harmful (Class 4), according to the classification proposed by the IOBC/WPRS. Afterward, 14 insecticides (5 harmless and 9 harmful) were sprayed on the parasitoid pupae. Of the 14 insecticides tested, only the organophosphates dimethoate and chlorpyrifos affected the parasitoid emergence. The effects of insecticides on the parasitism capacity of adults exposed to residues of azadirachtin, etofenprox, gamma-cyhalothrin, pyriproxyfen and tebufenozide (harmless) were also evaluated. Tebufenozide and gamma-cyhalothrin affected the parasitism of the F0 generation, but did not affect the emergence of the F1 and F2 generations. Therefore, for an effective IPM program, selective insecticides or harmful pesticides to adult parasitoids could be used in the field, provided that the adults do not occur naturally and the chemical applications do not coincide with parasitoid releases.

  12. Metalloporphyrin Co(III)TMPyP ameliorates acute, sublethal cyanide toxicity in mice.

    Science.gov (United States)

    Benz, Oscar S; Yuan, Quan; Amoscato, Andrew A; Pearce, Linda L; Peterson, Jim

    2012-12-17

    The formation of Co(III)TMPyP(CN)(2) at pH 7.4 has been shown to be completely cooperative (α(H) = 2) with an association constant of 2.1 (±0.2) × 10(11). The kinetics were investigated by stopped-flow spectrophotometry and revealed a complicated net reaction exhibiting 4 phases at pH 7.4 under conditions where cyanide was in excess. The data suggest molecular HCN (rather than CN(-)) to be the attacking nucleophile around neutrality. The two slower phases do not seem to be present when cyanide is not in excess, and the other two phases have rates comparable to that observed for cobalamin, a known effective cyanide scavenger. Addition of bovine serum albumin (BSA) did not affect the cooperativity of cyanide binding to Co(III)TMPyP, only lowered the equilibrium constant slightly to 1.2 (±0.2) × 10(11) and had an insignificant effect on the observed rate. A sublethal mouse model was used to assess the effectiveness of Co(III)TMPyP as a potential cyanide antidote. The administration of Co(III)TMPyP to sodium cyanide intoxicated mice resulted in the time required for the surviving mice to right themselves from a supine position being significantly decreased (9 ± 2 min) compared to that of the controls (33 ± 2 min). All observations were consistent with the demonstrated antidotal activity of Co(III)TMPyP operating through a cyanide-binding (i.e., scavenging) mechanism.

  13. Sublethal effects of insecticide seed treatments on two nearctic lady beetles (Coleoptera: Coccinellidae).

    Science.gov (United States)

    Moscardini, Valéria Fonseca; Gontijo, Pablo Costa; Michaud, J P; Carvalho, Geraldo Andrade

    2015-07-01

    Predatory insects often feed on plants or use plant products to supplement their diet, creating a potential route of exposure to systemic insecticides used as seed treatments. This study examined whether chlorantraniliprole or thiamethoxam might negatively impact Coleomegilla maculata and Hippodamia convergens when the beetles consumed the extrafloral nectar of sunflowers grown from treated seed. We reared both species on eggs of Ephestia kuehniella and then switched adult H. convergens to a diet of greenbugs, Schizaphis graminum, in order to induce oviposition in this species. Excised sunflower stems, either treated or control and refreshed every 48 h, were provided throughout larval development, or for the first week of adult life. Exposure of C. maculata larvae to chlorantraniliprole and thiamethoxam applied as seed treatments delayed adult emergence by prolonging the pupal period. When adults were exposed, thiamethoxam reduced the preoviposition period compared to chlorantraniliprole, whereas the latter treatment cause females to produce fewer clutches during the observation period. Larvae of C. maculata did not appear to obtain sufficient hydration from the sunflower stems and their subsequent fecundity and fertility were compromised in comparison to the adult exposure experiment where larvae received supplemental water during development. Exposure of H. convergens larvae to thiamethoxam skewed the sex ratio in favor of females; both materials reduced the egg viability of resulting adults and increased the period required for eclosion. Exposure of H. convergens adults to chlorantraniliprole reduced egg eclosion times compared to thiamethoxam and exposure to both insecticides reduced pupation times in progeny. The results indicate that both insecticides have negative, sublethal impacts on the biology of these predators when they feed on extrafloral nectar of sunflower plants grown from treated seed.

  14. Lethal and Sublethal Effects of Insecticides Used on Citrus, on the Ectoparasitoid Tamarixia radiata.

    Directory of Open Access Journals (Sweden)

    Vitor Hugo Beloti

    Full Text Available Huanglongbing (HLB is a disease associated with the bacteria "Candidatus Liberibacter spp." and has been devastating citrus orchards around the world. Its management involves control of the insect vector, the Asian citrus psyllid Diaphorina citri Kuwayama. However, the indiscriminate use of chemicals has caused pest outbreaks and eliminated the natural enemies of the vector, such as the parasitoid Tamarixia radiata (Waterston, the main agent for biological control of D. citri. This study assessed the lethal and sublethal effects of insecticides recommended for integrated production of citrus on the parasitoid T. radiata. When adult parasitoids were exposed to residues of 25 insecticides, 20% of them, i.e., gamma-cyhalothrin, etofenprox, azadirachtin, tebufenozide and pyriproxyfen, were considered as harmless (Class 1, 12% as slightly harmful (Class 2, 12% as moderately harmful (Class 3 and 56% as harmful (Class 4, according to the classification proposed by the IOBC/WPRS. Afterward, 14 insecticides (5 harmless and 9 harmful were sprayed on the parasitoid pupae. Of the 14 insecticides tested, only the organophosphates dimethoate and chlorpyrifos affected the parasitoid emergence. The effects of insecticides on the parasitism capacity of adults exposed to residues of azadirachtin, etofenprox, gamma-cyhalothrin, pyriproxyfen and tebufenozide (harmless were also evaluated. Tebufenozide and gamma-cyhalothrin affected the parasitism of the F0 generation, but did not affect the emergence of the F1 and F2 generations. Therefore, for an effective IPM program, selective insecticides or harmful pesticides to adult parasitoids could be used in the field, provided that the adults do not occur naturally and the chemical applications do not coincide with parasitoid releases.

  15. BAC and crash responsibility of injured older drivers : an analysis of Trauma Center data.

    Science.gov (United States)

    2014-09-01

    This study examined the distribution of blood alcohol concentrations (BACs) in injured drivers 65 and older and the relationship of older-driver BAC to driving record and crash responsibility. Researchers conducted a retrospective examination of 11 y...

  16. Sialic acid accelerates the electrophoretic velocity of injured dorsal root ganglion neurons

    National Research Council Canada - National Science Library

    Chen-xu Li Guo-ying Ma Min-fang Guo Ying Liu

    2015-01-01

    .... With the increase of membrane proteins on soma and injured site neurons, the negatively charged sialic acids bind to the external domains of membrane proteins, resulting in an increase of this charge...

  17. Perceived spasticity in chronic spinal cord injured patients: associations with psychological factors.

    NARCIS (Netherlands)

    Voerman, G.E.; Erren-Wolters, C.V.; Fleuren, J.F.; Hermens, H.J.; Geurts, A.C.H.

    2010-01-01

    PURPOSE: To explore the association between perceived spasticity and psychological factors (pain sensations, coping strategies, and illness cognitions) in chronic spinal cord injured (SCI) patients. METHODS: Cross-sectional study using a set of questionnaires was designed for chronic complete

  18. Perceived spasticity in chronic spinal cord injured patients: Associations with psychological factors

    NARCIS (Netherlands)

    Voerman, Gerlienke; Erren-Wolters, Cathelijne V.; Fleuren, Judith F.M.; Fleuren, J.F.M.; Hermens, Hermanus J.; Geurts, Alexander C.H.

    Purpose: To explore the association between perceived spasticity and psychological factors (pain sensations, coping strategies, and illness cognitions) in chronic spinal cord injured (SCI) patients. - Methods: Cross-sectional study using a set of questionnaires was designed for chronic complete

  19. Differences in kinetic variables between injured and noninjured novice runners : A prospective cohort study

    NARCIS (Netherlands)

    Bredeweg, Steef W.; Kluitenberg, Bas; Bessem, Bram; Buist, Ida

    Objectives: This prospective study examined differences in kinetic variables between injured and noninjured novice female and male runners and their potential contribution to RRIs. Design: A prospective cohort study. Methods: At baseline vertical ground reaction forces were assessed with an

  20. Protective effects of deferasirox and N-acetyl-L-cysteine on iron overload-injured bone marrow

    Directory of Open Access Journals (Sweden)

    J.C. Shen

    2017-10-01

    Full Text Available Using an iron overload mouse model, we explored the protective effect of deferasirox (DFX and N-acetyl-L-cysteine (NAC on injured bone marrow hematopoietic stem/progenitor cells (HSPC induced by iron overload. Mice were intraperitoneally injected with 25 mg iron dextran every 3 days for 4 weeks to establish an iron overload (Fe model. DFX or NAC were co-administered with iron dextran in two groups of mice (Fe+DFX and Fe+NAC, and the function of HSPCs was then examined. Iron overload markedly decreased the number of murine HSPCs in bone marrow. Subsequent colony-forming cell assays showed that iron overload also decreased the colony forming capacity of HSPCs, the effect of which could be reversed by DFX and NAC. The bone marrow hematopoiesis damage caused by iron overload could be alleviated by DFX and NAC.

  1. Vibration modes of injured spine at resonant frequencies under vertical vibration.

    Science.gov (United States)

    Guo, Li-Xin; Zhang, Ming; Zhang, Yi-Min; Teo, Ee-Chon

    2009-09-01

    A detailed three-dimensional finite element model of the spine segment T12-Pelvis was developed to investigate dynamic characteristics of whole lumbar spine with injured cases. This study investigates the motion mechanism of the human lumbar spine and the effect of component injuries on adjacent spinal components under whole body vibration. Several investigations have analyzed the influence of injured spines on adjacent spinal components under static loadings. However, it is not clear how the spine injury affects dynamic characteristics of whole lumbar spine and adjacent components of the injured segment under vibration. The T12-Pelvis model was used to obtain the modal vibration modes of the spine at resonant frequencies. Injury conditions of the spine were simulated and tested, including denucleation and/or facetectomy with removal of capsular ligaments. The results indicate the first-order vertical resonant frequency of the intact model is 7.21 Hz. After the denucleation at L4-L5, it decreases by more than 4% compared with the intact condition. All the injured conditions including disc injury and ligament injury decrease the resonant frequency of the spine. Due to the denucleation at L4-L5 the anteroposterior displacements of the vertebrae from L2 to L5 decrease and the vertical displacements of the vertebrae from L1 to L4 increase under vibration. The denucleation also decreases the rotational deformations of the vertebrae from L1 to L5. The material property sensitivity analysis shows intervertebral discs have a dominating effect on variation of vertical resonant frequency of the spine. The denucleation may decrease cushioning effects of adjacent motion segments at the injured level under vibration. The injured condition may increase the vertical displacement amplitudes of the spine above the injured level. All the injured conditions may decrease the resonant frequency of the spine system.

  2. Sphincter (ring muscle) gymnastics for spinal cord injured and spinal cord transected patients.

    Science.gov (United States)

    Yom-Tov, S

    2000-07-01

    Sphincter gymnastics seem to be able to recruit central pattern generators--networks of neurons that generate motor patterns--through peripheral rhythmic stimuli and to activate them without supraspinal signals. This physical therapy method is thus suitable for treating spinal cord injured and spinal cord transected patients, for it can reach below the injured segment and feed signals into the central nervous system, thus activating vital organs. This article presents some observations and discusses a possible mechanism.

  3. Mechanisms and Treatment of Deployment-Related Lung Injury: Repair of the Injured Epithelium

    Science.gov (United States)

    2017-10-01

    AWARD NUMBER: W81XWH-16-2-0018 TITLE: Mechanisms and Treatment of Deployment-Related Lung Injury: Repair of the Injured Epithelium PRINCIPAL...AND SUBTITLE Mechanisms and Treatment of Deployment-Related Lung Injury: Repair of the Injured Epithelium 5a. CONTRACT NUMBER 5b. GRANT NUMBER...initial experiments confirm our preliminary data that pretreatment with PM from Iraq or Afghanistan (5- 10 µg/cm2) significantly delay wound closure in

  4. 〈Review〉Psychological Response to Sport Injury and Psychological Skills Interventions for Injured Athletes

    OpenAIRE

    Naoi, Airi; Deaner, Heather. R.

    2006-01-01

    [Abstract] This article reviews previous research studies that have examined the psychological responses of injured athletes and the effectiveness of psychological skills interventions for injured athletes. In order to understand psychological response to injury, two models, a cognitive appraisal model of psychological adjustment to athletic injury and an integrated model of psychological response to the sport injury and rehabilitation process, are described. Additionally, the relationship am...

  5. Inhibition of hydrogen peroxide induced injuring on human skin fibroblast by Ulva prolifera polysaccharide.

    Science.gov (United States)

    Cai, Chuner; Guo, Ziye; Yang, Yayun; Geng, Zhonglei; Tang, Langlang; Zhao, Minglin; Qiu, Yuyan; Chen, Yifan; He, Peimin

    2016-10-01

    Ulva prolifera can protect human skin fibroblast from being injured by hydrogen peroxide. This work studied the composition of Ulva prolifera polysaccharide and identified its physicochemical properties. The results showed that the cell proliferation of 0.5mg/mL crude polysaccharide was 154.4% of that in negative control group. Moreover, ROS detection indices, including DCFH-DA, GSH-PX, MDA and CAT, indicated that crude polysaccharide could improve cellular ability to scavenge free radical and decrease the injury on human skin fibroblast by hydrogen peroxide. In purified polysaccharide, the activity of fraction P1-1 was the highest, with 174.6% of that in negative control group. The average molecular weight of P1-1 was 137kD with 18.0% of sulfate content. This work showed the inhibition of hydrogen peroxide induced injuries on human skin fibroblast by Ulva prolifera polysaccharide, which may further evaluate the application of U. prolifera on cosmetics. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Endothelial glycocalyx shedding and vascular permeability in severely injured trauma patients

    DEFF Research Database (Denmark)

    Rahbar, Elaheh; Cardenas, Jessica C; Baimukanova, Gyulnar

    2015-01-01

    of trauma patients. METHODS: Plasma samples were collected from 5 healthy consented volunteers and 22 severely injured trauma patients upon admission to the emergency department. ELISA assays were performed to quantify shed HA, HS, CS and syndecan-1 in plasma. A colloid osmometer and Electric Cell......-parametric statistical tests were used to compare differences between groups. RESULTS: We observed increased shedding of all four proteins in trauma patient plasma compared to healthy controls: 31.7 vs. 21.2 U/L of CS, 175.8 vs. 121.9 ng/ml of HS, 946.7 vs. 618.6 ng/ml of HA and 245.8 vs. 31.6 ng/ml of syndecan-1 (all p...... generation of less than 250 nM on average (ptrauma patients compared to healthy controls in this cohort. However, only syndecan-1 and HA shedding were significantly higher in patients with reduced plasma COP. Thrombin generation was impaired...

  7. Shrimp Protein Hydrolysate Modulates the Timing of Proinflammatory Macrophages in Bupivacaine-Injured Skeletal Muscles in Rats

    Directory of Open Access Journals (Sweden)

    Junio Dort

    2016-01-01

    Full Text Available This study was designed to determine whether marine-derived proteins other than cod could have beneficial effects on inflammation following muscle injury. Macrophage and neutrophil densities were measured from bupivacaine-injured tibialis anterior muscle of rats fed isoenergetic diets containing either shrimp hydrolysate (Shr, casein hydrolysate (CaH, or whole casein (Ca. In this study, Shr reduced ED1+-macrophages at day 2 (p=0.013, day 5 (p=0.006, and day 14 after injury (p=0.038 compared with Ca, indicating faster resolution of inflammation in Shr. Except for day 2 after injury where Shr led to lower ED1+-macrophages compared with CaH (p=0.006, both Shr and CaH responded similarly at days 5, 14, and 28 after injury. This findings suggest that beneficial effects of Shr on ED1+-cells might be related to generation of anti-inflammatory peptides through the hydrolysis process, in addition to its high content of anti-inflammatory amino acids. However, while increasing myofiber cross-sectional area in noninjured muscles compared with both Ca and CaH, Shr failed to have a positive effect in corresponding injured muscles. These data indicate that shrimp hydrolysate can facilitate resolution of inflammation after muscle injury mainly through modulating proinflammatory macrophage accumulation but have less effect on optimal recovery in terms of muscle mass and fiber size.

  8. Shrimp Protein Hydrolysate Modulates the Timing of Proinflammatory Macrophages in Bupivacaine-Injured Skeletal Muscles in Rats.

    Science.gov (United States)

    Dort, Junio; Leblanc, Nadine; Bryl, Piotr; Fortin, Marie-Gil; Carbonneau, Marie-Elise; Lavigne, Charles; Jacques, Hélène

    2016-01-01

    This study was designed to determine whether marine-derived proteins other than cod could have beneficial effects on inflammation following muscle injury. Macrophage and neutrophil densities were measured from bupivacaine-injured tibialis anterior muscle of rats fed isoenergetic diets containing either shrimp hydrolysate (Shr), casein hydrolysate (CaH), or whole casein (Ca). In this study, Shr reduced ED(1+)-macrophages at day 2 (p = 0.013), day 5 (p = 0.006), and day 14 after injury (p = 0.038) compared with Ca, indicating faster resolution of inflammation in Shr. Except for day 2 after injury where Shr led to lower ED(1+)-macrophages compared with CaH (p = 0.006), both Shr and CaH responded similarly at days 5, 14, and 28 after injury. This findings suggest that beneficial effects of Shr on ED(1+)-cells might be related to generation of anti-inflammatory peptides through the hydrolysis process, in addition to its high content of anti-inflammatory amino acids. However, while increasing myofiber cross-sectional area in noninjured muscles compared with both Ca and CaH, Shr failed to have a positive effect in corresponding injured muscles. These data indicate that shrimp hydrolysate can facilitate resolution of inflammation after muscle injury mainly through modulating proinflammatory macrophage accumulation but have less effect on optimal recovery in terms of muscle mass and fiber size.

  9. Risk factors for trauma-induced coagulopathy- and transfusion-associated multiple organ failure in severely injured trauma patients

    Directory of Open Access Journals (Sweden)

    Kirsten eBalvers

    2015-04-01

    Full Text Available Background: Both trauma-induced coagulopathy (TIC as well as transfusion strategies influence early outcome in hemorrhagic trauma patients. Their impact on late outcome is less well characterized. This study systematically reviews risk factors for TIC- and transfusion-associated multiple organ failure (MOF in severely injured trauma patients.Material and methods: A systematic search was conducted in PubMed and Embase. Studies published from 1986 to 2013 on adult trauma patients with an Injury Severity Score (ISS ≥16, investigating TIC or transfusion strategies with MOF as primary or secondary outcome, were eligible for inclusion. Results of the included studies were evaluated with meta-analyses of pooled data. Results: In total 50 studies were included with a total sample size of 63,586 patients. Due to heterogeneity of the study populations and outcome measures, results from 7 studies allowed for pooling of data. Risk factors for TIC-associated MOF were hypocoagulopathy, hemorrhagic shock, activated protein C, increased histone levels and increased levels of markers of fibrinolysis on admission. After at least 24 hours after admission, the occurrence of thromboembolic events was associated with MOF. Risk factors for transfusion-associated MOF were the administration of fluids and red blood cell units within 24 hours post-injury, the age of red blood cells (>14 days and a ratio of FFP:RBC ≥1:1 (OR 1.11, 95% CI 1.04-1.19.Conclusion: Risk factors for TIC-associated MOF in severely injured trauma patients are early hypocoagulopathy and hemorrhagic shock, while a hypercoagulable state with the occurrence of thromboembolic events later in the course of trauma predisposes to MOF. Risk factors for transfusion-associated MOF include administration of crystalloids and red blood cells and a prolonged storage time of red blood cells. Future prospective studies investigating TIC- and transfusion- associated risk factors on late outcome are required.

  10. The impact of playing in matches while injured on injury surveillance findings in professional football.

    Science.gov (United States)

    Hammond, L E; Lilley, J M; Pope, G D; Ribbans, W J

    2014-06-01

    This study aimed to analyze the frequency, nature, and consequences of footballers playing matches while injured, and to examine the impact on injury surveillance findings. High levels of inter-rater reliability and content validity were established for a tool designed to document players who were already injured at the start of a match. The tool was implemented in three English football teams (a Championship, League 1, and League 2 team) for one season, using a "time loss" definition of injury. One hundred forty-three matches were surveyed, revealing 102 match appearances by players who were already injured. Almost half of all games featured at least one injured player, with episodes of playing with injury occurring more frequently and lasting longer in League 2 players compared with higher level players. No association was observed between the number of injured players starting matches and match outcome [χ(2) (4, N = 143) = 3.27, P = 0.514]. Fifteen percent of all injury episodes captured were only through prospective documentation of playing while injured. The findings show that both traumatic and overuse injuries are managed by footballers through competitive matches, and have important implications for aiding understanding of the epidemiology of injury in professional football. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Transplantation of hypoxic preconditioned neural stem cells benefits functional recovery via enhancing neurotrophic secretion after spinal cord injury in rats.

    Science.gov (United States)

    Fan, Wei-Li; Liu, Peng; Wang, Guan; Pu, Jung-Ang; Xue, Xin; Zhao, Jian-Hua

    2017-09-08

    Spinal cord injury (SCI) is a debilitating, costly, and common pathological condition that affects the function of central nervous system (CNS). To date, there are few promising therapeutic strategies available for SCI. To look for a suitable therapeutic strategy, we have developed a sublethal hypoxic preconditioning procedure using Fluorescence-activated cell sorting (FACS) analysis, LDH releasing and cell viability assays in vitro. Meanwhile, we have examined the benefits of neural stem cells (NSCs) transplantation prior to hypoxic preconditioning on functional recovery and potential mechanism via MRI screening, H&E and Nissl staining, immunofluorescence staining and Elisa assays. Our data showed that transplantation of hypoxic prconditioned NSCs could enhance neuronal survival, especially 5-TH(+) and ChAT(+) neurons, in the injured spinal cord to reinforce functional benefits. The hypoxia exposure upregulated HIF-1α, neurotrophic and growth factors including neurotrophin-3 (NT-3), glial cell-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) in vitro and in vivo. Furthermore, functional recovery, including locomotor and hypersensitivities to mechanical and thermal stimulation assessed via behavioral and sensory tests, improved significantly in rats with engraftment of NSCs after hypoxia exposure from day 14 post-SCI, compared with the control and N-NSCs groups. In short, the approach employed in this study could result in functional recovery via upregulating neurotrophic and growth factors, which implies that hypoxic preconditioning strategy could serve as an effective and feasible strategy for cell-based therapy in the treatment of SCI in rats. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  12. Fipronil promotes motor and behavioral changes in honey bees (Apis mellifera) and affects the development of colonies exposed to sublethal doses.

    Science.gov (United States)

    Zaluski, Rodrigo; Kadri, Samir Moura; Alonso, Diego Peres; Martins Ribolla, Paulo Eduardo; de Oliveira Orsi, Ricardo

    2015-05-01

    Bees play a crucial role in pollination and generate honey and other hive products; therefore, their worldwide decline is cause for concern. New broad-spectrum systemic insecticides such as fipronil can harm bees and their use has been discussed as a potential threat to bees' survival. In the present study, the authors evaluate the in vitro toxicity of fipronil and note behavioral and motor activity changes in Africanized adult Apis mellifera that ingest or come into contact with lethal or sublethal doses of fipronil. The effects of sublethal doses on brood viability, population growth, behavior, and the expression of the defensin 1 gene in adult bees were studied in colonies fed with contaminated sugar syrup (8 µg fipronil L(-1) ). Fipronil is highly toxic to bees triggering agitation, seizures, tremors, and paralysis. Bees that are exposed to a lethal or sublethal doses showed reduced motor activity. The number of eggs that hatched, the area occupied by worker eggs, and the number of larvae and pupae that developed were reduced, adult bees showed lethargy, and colonies were abandoned when they were exposed to sublethal doses of fipronil. No change was seen in the bees' expression of defensin 1. The authors conclude that fipronil is highly toxic to honey bees and even sublethal doses may negatively affect the development and maintenance of colonies. © 2015 SETAC.

  13. Assessment of the lethal and sublethal effects of 20 environmental chemicals in zebrafish embryos and larvae by using OECD TG 212.

    Science.gov (United States)

    Horie, Yoshifumi; Yamagishi, Takahiro; Takahashi, Hiroko; Shintaku, Youko; Iguchi, Taisen; Tatarazako, Norihisa

    2017-10-01

    Fish embryo toxicity tests are used to assess the lethal and sublethal effects of environmental chemicals in aquatic organisms. Previously, we used a short-term toxicity test published by the Organization for Economic Co-operation and Development (test no. 212: Fish, Short-term Toxicity Test on Embryo and Sac-Fry Stages [OECD TG 212]) to assess the lethal and sublethal effects of aniline and several chlorinated anilines in zebrafish embryos and larvae. To expand upon this previous study, we used OECD TG 212 in zebrafish embryos and larvae to assess the lethal and sublethal effects of 20 additional environmental chemicals that included active pharmaceutical ingredients, pesticides, metals, aromatic compounds or chlorinated anilines. Zebrafish embryos (Danio rerio) were exposed to the test chemicals until 8 days post-fertilization. A delayed lethal effect was induced by 16 of the 20 test chemicals, and a positive correlation was found between heart rate turbulence and mortality. We also found that exposure to the test chemicals at concentrations lower than the lethal concentration induced the sublethal effects of edema, body curvature and absence of swim-bladder inflation. In conclusion, the environmental chemicals assessed in the present study induced both lethal and sublethal effects in zebrafish embryos and larvae, as assessed by using OECD TG 212. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  14. Effects of Sublethal Cadmium Exposure on Antipredator Behavioural and Antitoxic Responses in the Invasive Amphipod Dikerogammarus villosus

    Science.gov (United States)

    Sornom, Pascal; Gismondi, Eric; Vellinger, Céline; Devin, Simon; Férard, Jean-François; Beisel, Jean-Nicolas

    2012-01-01

    Amphipods are recognised as an important component of freshwater ecosystems and are frequently used as an ecotoxicological test species. Despite this double interest, there is still a lack of information concerning toxic impacts on ecologically relevant behaviours. The present study investigated the influence of cadmium (Cd), a non-essential heavy metal, on both antipredator behaviours and antitoxic responses in the invasive amphipod Dikerogammarus villosus under laboratory conditions. Amphipod behaviour (i.e. refuge use, aggregation with conspecifics, exploration and mobility) was recorded following a 4-min test-exposure to 500 µg Cd/L with or without a 24-h Cd pre-exposure and in the presence or absence of a high perceived risk of predation (i.e. water scented by fish predators and injured conspecifics). Following behavioural tests, malondialdehyde (MDA) levels, a biomarker for toxic effect, and energy reserves (i.e. lipid and glycogen contents) were assessed. Cd exposures induced (1) cell damage reflected by high MDA levels, (2) erratic behaviour quantified by decreasing refuge use and exploration, and increasing mobility, and (3) a depletion in energy reserves. No significant differences were observed between 4-min test-exposed and 24-h pre-exposed individuals. Gammarids exposed to Cd had a disturbed perception of the alarm stimuli, reflected by increased time spent outside of refuges and higher mobility compared to gammarids exposed to unpolluted water. Our results suggest that Cd exposure rapidly disrupts the normal behavioural responses of gammarids to alarm substances and alters predator-avoidance strategies, which could have potential impacts on aquatic communities. PMID:22879985

  15. Effects of sublethal cadmium exposure on antipredator behavioural and antitoxic responses in the invasive amphipod Dikerogammarus villosus.

    Directory of Open Access Journals (Sweden)

    Pascal Sornom

    Full Text Available Amphipods are recognised as an important component of freshwater ecosystems and are frequently used as an ecotoxicological test species. Despite this double interest, there is still a lack of information concerning toxic impacts on ecologically relevant behaviours. The present study investigated the influence of cadmium (Cd, a non-essential heavy metal, on both antipredator behaviours and antitoxic responses in the invasive amphipod Dikerogammarus villosus under laboratory conditions. Amphipod behaviour (i.e. refuge use, aggregation with conspecifics, exploration and mobility was recorded following a 4-min test-exposure to 500 µg Cd/L with or without a 24-h Cd pre-exposure and in the presence or absence of a high perceived risk of predation (i.e. water scented by fish predators and injured conspecifics. Following behavioural tests, malondialdehyde (MDA levels, a biomarker for toxic effect, and energy reserves (i.e. lipid and glycogen contents were assessed. Cd exposures induced (1 cell damage reflected by high MDA levels, (2 erratic behaviour quantified by decreasing refuge use and exploration, and increasing mobility, and (3 a depletion in energy reserves. No significant differences were observed between 4-min test-exposed and 24-h pre-exposed individuals. Gammarids exposed to Cd had a disturbed perception of the alarm stimuli, reflected by increased time spent outside of refuges and higher mobility compared to gammarids exposed to unpolluted water. Our results suggest that Cd exposure rapidly disrupts the normal behavioural responses of gammarids to alarm substances and alters predator-avoidance strategies, which could have potential impacts on aquatic communities.

  16. Lethal and Sublethal Effects of Essential Oils From Artemisia khorassanica and Vitex pseudo-negundo Against Plodia interpunctella (Lepidoptera: Pyralidae).

    Science.gov (United States)

    Borzoui, Ehsan; Naseri, Bahram; Abedi, Zahra; Karimi-Pormehr, Mohammad Sadegh

    2016-10-01

    Plodia interpunctella (Hübner, 1813) is a polyphagous and key pest of different stored products worldwide. The lethal and sublethal effects of essential oils of Artemisia khorassanica Podl. and Vitex pseudo-negundo (Hausskn) were studied on P. interpunctella The chemical constituents of the essential oils were also assessed using gas chromatography-mass spectrometry. Assays showed that the fumigant toxicity of A. khorassanica (LC50: 9.60 µl/liter air) was higher than V. pseudo-negundo (LC50: 23.05 µl/liter air). Moreover, the speed of mortality caused by A. khorassanica oil (LT50: 2.07 h) was higher than V. pseudo-negundo (LT50: 3.11 h). To assess the sublethal effects of the essential oils, adult moths were exposed to the LC30 of each essential oil, and life table parameters and energy contents of the surviving P. interpunctella were studied. Exposure to sublethal concentration of A. khorassanica negatively affected the life table of P. interpunctella, and also the protein, lipid, and glycogen contents of the larvae that came from treated adults. Vitex pseudo-negundo also affected lipid, protein, and glycogen contents of P. interpunctella The intrinsic rate of increase (rm), finite rate of increase ([Formula: see text]), and doubling time (DT) were not significantly different between control and V. pseudo-negundo treatment. According with these results, both tested essential oils, especially one extracted from A. khorassanica, have potential applications for the integrated management of P. interpunctella. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. Exposure to Sublethal Doses of Fipronil and Thiacloprid Highly Increases Mortality of Honeybees Previously Infected by Nosema ceranae

    Science.gov (United States)

    Vidau, Cyril; Diogon, Marie; Aufauvre, Julie; Fontbonne, Régis; Viguès, Bernard; Brunet, Jean-Luc; Texier, Catherine; Biron, David G.; Blot, Nicolas; El Alaoui, Hicham; Belzunces, Luc P.; Delbac, Frédéric

    2011-01-01

    Background The honeybee, Apis mellifera, is undergoing a worldwide decline whose origin is still in debate. Studies performed for twenty years suggest that this decline may involve both infectious diseases and exposure to pesticides. Joint action of pathogens and chemicals are known to threaten several organisms but the combined effects of these stressors were poorly investigated in honeybees. Our study was designed to explore the effect of Nosema ceranae infection on honeybee sensitivity to sublethal doses of the insecticides fipronil and thiacloprid. Methodology/Finding Five days after their emergence, honeybees were divided in 6 experimental groups: (i) uninfected controls, (ii) infected with N. ceranae, (iii) uninfected and exposed to fipronil, (iv) uninfected and exposed to thiacloprid, (v) infected with N. ceranae and exposed 10 days post-infection (p.i.) to fipronil, and (vi) infected with N. ceranae and exposed 10 days p.i. to thiacloprid. Honeybee mortality and insecticide consumption were analyzed daily and the intestinal spore content was evaluated 20 days after infection. A significant increase in honeybee mortality was observed when N. ceranae-infected honeybees were exposed to sublethal doses of insecticides. Surprisingly, exposures to fipronil and thiacloprid had opposite effects on microsporidian spore production. Analysis of the honeybee detoxification system 10 days p.i. showed that N. ceranae infection induced an increase in glutathione-S-transferase activity in midgut and fat body but not in 7-ethoxycoumarin-O-deethylase activity. Conclusions/Significance After exposure to sublethal doses of fipronil or thiacloprid a higher mortality was observed in N. ceranae-infected honeybees than in uninfected ones. The synergistic effect of N. ceranae and insecticide on honeybee mortality, however, did not appear strongly linked to a decrease of the insect detoxification system. These data support the hypothesis that the combination of the increasing

  18. Sublethal Exposure to Diatomaceous Earth Increases Net Fecundity of Flour Beetles (Tribolium confusum) by Inhibiting Egg Cannibalism

    Science.gov (United States)

    Shostak, Allen W.

    2014-01-01

    Population regulation results from an interplay of numerous intrinsic and external factors, and for many insects cannibalism is such a factor. This study confirms a previously-reported observation that sublethal exposure to the fossilized remains of diatoms (i.e. diatomaceous earth) increases net fecundity (eggs produced minus eggs destroyed/day) of flour beetles, Tribolium confusum. The aim was to experimentally test two non-mutually-exclusive ecological mechanisms potentially responsible for the increased net fecundity: higher egg production and lower egg cannibalism. Adult T. confusum were maintained at low or high density in medium containing sublethal (0–4%) diatomaceous earth. Net fecundity increased up to 2.1× control values during diatomaceous earth exposure, and returned to control levels following removal from diatomaceous earth. Cannibalism assays on adults showed that diatomaceous earth reduced the number of eggs produced to 0.7× control values at low density and to 0.8× controls at high density, and also reduced egg cannibalism rates of adults to as little as 0.4× control values, but at high density only. Diatomaceous earth also reduced cannibalism by larvae on eggs to 0.3× control values. So, while the presence of diatomaceous earth reduced egg production, net fecundity increased as a result of strong suppression of the normal egg cannibalism by adults and larvae that occurs at high beetle density. Undisturbed cultures containing sublethal diatomaceous earth concentrations reached higher population densities than diatomaceous earth-free controls. Cohort studies on survival from egg to adult indicated that this population increase was due largely to decreased egg cannibalism by adult females. This is the first report of inhibition of egg cannibalism by diatomaceous earth on larval or adult insects. The ability of diatomaceous earth to alter cannibalism behavior without causing mortality makes it an ideal investigative tool for cannibalism

  19. Sublethal exposure to diatomaceous earth increases net fecundity of flour beetles (Tribolium confusum by inhibiting egg cannibalism.

    Directory of Open Access Journals (Sweden)

    Allen W Shostak

    Full Text Available Population regulation results from an interplay of numerous intrinsic and external factors, and for many insects cannibalism is such a factor. This study confirms a previously-reported observation that sublethal exposure to the fossilized remains of diatoms (i.e. diatomaceous earth increases net fecundity (eggs produced minus eggs destroyed/day of flour beetles, Tribolium confusum. The aim was to experimentally test two non-mutually-exclusive ecological mechanisms potentially responsible for the increased net fecundity: higher egg production and lower egg cannibalism. Adult T. confusum were maintained at low or high density in medium containing sublethal (0-4% diatomaceous earth. Net fecundity increased up to 2.1× control values during diatomaceous earth exposure, and returned to control levels following removal from diatomaceous earth. Cannibalism assays on adults showed that diatomaceous earth reduced the number of eggs produced to 0.7× control values at low density and to 0.8× controls at high density, and also reduced egg cannibalism rates of adults to as little as 0.4× control values, but at high density only. Diatomaceous earth also reduced cannibalism by larvae on eggs to 0.3× control values. So, while the presence of diatomaceous earth reduced egg production, net fecundity increased as a result of strong suppression of the normal egg cannibalism by adults and larvae that occurs at high beetle density. Undisturbed cultures containing sublethal diatomaceous earth concentrations reached higher population densities than diatomaceous earth-free controls. Cohort studies on survival from egg to adult indicated that this population increase was due largely to decreased egg cannibalism by adult females. This is the first report of inhibition of egg cannibalism by diatomaceous earth on larval or adult insects. The ability of diatomaceous earth to alter cannibalism behavior without causing mortality makes it an ideal investigative tool for

  20. Sublethal doses of neonicotinoid imidacloprid can interact with honey bee chemosensory protein 1 (CSP1) and inhibit its function.

    Science.gov (United States)

    Li, Hongliang; Tan, Jing; Song, Xinmi; Wu, Fan; Tang, Mingzhu; Hua, Qiyun; Zheng, Huoqing; Hu, Fuliang

    2017-04-29

    As a frequently used neonicotinoid insecticide, imidacloprid can impair the chemoreceptive behavior of honey bees even at sublethal doses, while the physiochemical mechanism has not been further revealed. Here, multiple fluorescence spectra, thermodynamic method, and molecular docking were used to study the interaction and the functional inhibition of imidacloprid to the recombinant CSP1 protein in Asian honey bee, Apis cerana. The results showed that the fluorescence intensity (λ em  = 332 nm) of CSP1 could be significantly quenched by imidacloprid in a dynamic mode. During the quenching process, ΔH > 0, ΔS > 0, indicating that the acting forces of imidacloprid with CSP1 are mainly hydrophobic interactions. Synchronous fluorescence showed that the fluorescence of CSP1 was mainly derived from tryptophan, and the hydrophobicity of tryptophan decreased with the increase of imidacloprid concentration. Molecular docking predicted the optimal pose and the amino acid composition of the binding process. Circular dichroism (CD) spectra showed that imidacloprid reduced the α-helix of CSP1 and caused the extension of the CSP1 peptide chain. In addition, the binding of CSP1 to floral scent β-ionone was inhibited by nearly 50% of the apparent association constant (K A ) in the presence of 0.28-2.53 ng/bee of imidacloprid, and the inhibition rate of nearly 95% at 3.75 ng/bee of imidacloprid at sublethal dose level. This study initially revealed the molecular physiochemical mechanism that sublethal doses of neonicotinoid still interact and inhibit the physiological function of the honey bees' chemoreceptive system. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Lethal and Sub-lethal Effects of Four Insecticides on the Aphidophagous Coccinellid Adalia bipunctata (Coleoptera: Coccinellidae).

    Science.gov (United States)

    Depalo, Laura; Lanzoni, Alberto; Masetti, Antonio; Pasqualini, Edison; Burgio, Giovanni

    2017-12-05

    Conventional insecticide assays, which measure the effects of insecticide exposure on short-term mortality, overlook important traits, including persistence of toxicity or sub-lethal effects. Therefore, such approaches are especially inadequate for prediction of the overall impact of insecticides on beneficial arthropods. In this study, the side effects of four modern insecticides (chlorantraniliprole, emamectin benzoate, spinosad, and spirotetramat) on Adalia bipunctata (L.) (Coleoptera: Coccinellidae) were evaluated under laboratory conditions by exposition on treated potted plants. In addition to investigation of acute toxicity and persistence of harmful activity in both larvae and adults of A. bipunctata, demographic parameters were evaluated, to provide a comprehensive picture of the nontarget effects of these products. Field doses of the four insecticides caused detrimental effects to A. bipunctata; but in different ways. Overall, spinosad showed the best toxicological profile among the products tested. Emamectin benzoate could be considered a low-risk insecticide, but had high persistence. Chlorantraniliprole exhibited lethal effects on early instar larvae and adults, along with a long-lasting activity, instead spirotetramat showed a low impact on larval and adult mortality and can be considered a short-lived insecticide. However, demographic analysis demonstrated that chlorantraniliprole and spirotetramat caused sub-lethal effects. Our findings highlight that sole assessment of mortality can lead to underestimation of the full impact of pesticides on nontarget insects. Demographic analysis was demonstrated to be a sensitive method for detection of the sub-lethal effects of insecticides on A. bipunctata, and this approach should be considered for evaluation of insecticide selectivity. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Effect of sub-lethal exposure to ultraviolet radiation on the escape performance of Atlantic cod larvae (Gadus morhua.

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    Yuichi Fukunishi

    Full Text Available The amount of ultraviolet (UV radiation reaching the earth's surface has increased due to depletion of the ozone layer. Several studies have reported that UV radiation reduces survival of fish larvae. However, indirect and sub-lethal impacts of UV radiation on fish behavior have been given little consideration. We observed the escape performance of larval cod (24 dph, SL: 7.6±0.2 mm; 29 dph, SL: 8.2±0.3 mm that had been exposed to sub-lethal levels of UV radiation vs. unexposed controls. Two predators were used (in separate experiments: two-spotted goby (Gobiusculus flavescens; a suction predator and lion's mane jellyfish (Cyanea capillata; a "passive" ambush predator. Ten cod larvae were observed in the presence of a predator for 20 minutes using a digital video camera. Trials were replicated 4 times for goby and 5 times for jellyfish. Escape rate (total number of escapes/total number of attacks ×100, escape distance and the number of larvae remaining at the end of the experiment were measured. In the experiment with gobies, in the UV-treated larvae, both escape rate and escape distance (36%, 38±7.5 mm respectively were significantly lower than those of control larvae (75%, 69±4.7 mm respectively. There was a significant difference in survival as well (UV: 35%,63%. No apparent escape response was observed, and survival rate was not significantly different, between treatments (UV: 66%,74% in the experiment with jellyfish. We conclude that the effect and impact of exposure to sub-lethal levels of UV radiation on the escape performance of cod larvae depends on the type of predator. Our results also suggest that prediction of UV impacts on fish larvae based only on direct effects are underestimations.

  3. Low salinity enhances NI-mediated oxidative stress and sub-lethal toxicity to the green shore crab (Carcinus maenas).

    Science.gov (United States)

    Blewett, Tamzin A; Wood, Chris M

    2015-12-01

    Nickel (Ni) is a metal of environmental concern, known to cause toxicity to freshwater organisms by impairing ionoregulation and/or respiratory gas exchange, and by inducing oxidative stress. However, little is known regarding how nickel toxicity is influenced by salinity. In the current study we investigated the salinity-dependence and mechanisms of sub-lethal Ni toxicity in a euryhaline crab (Carcinus maenas). Crabs were acclimated to three experimental salinities--20, 60 and 100% seawater (SW)--and exposed to 3mg/L Ni for 24h or 96 h. Tissues were dissected for analysis of Ni accumulation, gills were taken for oxidative stress analysis (catalase activity and protein carbonyl content), haemolymph ions were analysed for ionoregulatory disturbance, and oxygen consumption was determined in exercised crabs after 96 h of Ni exposure. Total Ni accumulation was strongly dependant on salinity, with crabs from 20% SW displaying the highest tissue Ni burdens after both 24 and 96-h exposures. After 96 h of exposure, the highest accumulation of Ni occurred in the posterior (ionoregulatory) gills at the lowest salinity, 20% SW. Posterior gill 8 exhibited elevated protein carbonyl levels and decreased catalase activity after Ni exposure, but only in 20% SW. Similarly, decreased levels of haemolymph Mg and K and an increased level of Ca were recorded but only in crabs exposed to Ni for 96 h in 20% SW. Oxygen consumption after exercise was also inhibited in crabs exposed to Ni in 20% SW. These data show for the first time the simultaneous presence of all three modes of sub-lethal Ni toxicity in exposed animals, and indi