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  1. Article ; Online: Comparison of hydroxocobalamin with other resuscitative fluids in volume-controlled and uncontrolled hemorrhage models in swine ( Sus-scrofa ).

    Paredes, R Madelaine / Castaneda, Maria / Mireles, Allyson A / Rodriguez, Dylan / Maddry, Joseph

    The journal of trauma and acute care surgery

    2023  Volume 95, Issue 2S Suppl 1, Page(s) S120–S128

    Abstract: Background: Traumatic hemorrhage is the leading cause of preventable death in military environments. Treatment with resuscitative fluids and blood components is based on availability, thus, frequently unavailable in the prehospital setting, due to lack ... ...

    Abstract Background: Traumatic hemorrhage is the leading cause of preventable death in military environments. Treatment with resuscitative fluids and blood components is based on availability, thus, frequently unavailable in the prehospital setting, due to lack of resources and costs. Hydroxocobalamin (HOC), increases blood pressure via nitric oxide scavenging. We evaluated HOC as a resuscitation fluid, in two swine hemorrhage models. Our objectives were to (1) evaluate whether HOC treatment following hemorrhagic shock improves hemodynamic parameters and (2) determine whether those effects are comparable to whole blood (WB) and lactated ringers (LR).
    Methods: Yorkshire swine (S us scrofa ) (n = 72) were used in models of controlled hemorrhage (CH) (n = 36) and uncontrolled hemorrhage (UH) (n = 36). Randomized animals received treatment with 500 mL of either WB, LR, HOC (150 mg/kg), followed by a six-hour observation (n = 6 each group). Survival, hemodynamics, blood gases (ABGs) and chemistries were collected. Data reported as mean ± standard error of the mean and statistical analysis by ANOVA ( p < 0.05).
    Results: Blood loss for CH was 41% ± 0.02 versus 33% ± 0.07 for UH. For CH, HOC treatment maintained higher systolic blood pressure (sBP, mm Hg) compared with WB and LR (72 ± 1.1; 60 ± 0.8; 58 ± 1.6; respectively). Heart rate (HR), cardiac output (CO), Sp o2 and vascular resistance were comparable with WB and LR. The ABG values were comparable between HOC and WB. For UH, HOC treatment maintained sBP levels comparable to WB and higher than LR (70 ± 0.9; 73 ± 0.5; 56 ± 1.2). HR, CO, Sp o2 , and systemic vascular resistance were comparable between HOC and WB. Survival, hemodynamics, blood gases were comparable between HOC and WB. No survival differences were found between cohorts.
    Conclusion: Hydroxocobalamin treatment improved hemodynamic parameters and Ca 2+ levels compared with LR and equivalent to WB, in both models. Hydroxocobalamin may be a viable alternative when WB is not available.
    MeSH term(s) Animals ; Disease Models, Animal ; Gases ; Hemodynamics ; Hemorrhage ; Hydroxocobalamin/pharmacology ; Hydroxocobalamin/therapeutic use ; Isotonic Solutions ; Resuscitation ; Shock, Hemorrhagic/drug therapy ; Swine
    Chemical Substances Gases ; Hydroxocobalamin (Q40X8H422O) ; Isotonic Solutions
    Language English
    Publishing date 2023-05-17
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2651070-4
    ISSN 2163-0763 ; 2163-0755
    ISSN (online) 2163-0763
    ISSN 2163-0755
    DOI 10.1097/TA.0000000000004049
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Efficacy of a single-day task trainer-based extracorporeal membrane oxygenation training course.

    Maddry, Joseph K / Paredes, R Madelaine / Abdurashidov, Timur / Davis, William T / Paciocco, Joni A / Castaneda, Maria / Araña, Allyson A / Perez, Crystal A / Baldwin, Darren S / Rodriguez, Dylan C / Medellin, Kimberly L / Ng, Patrick

    AEM education and training

    2022  Volume 6, Issue 6, Page(s) e10806

    Abstract: Background: Extracorporeal membrane oxygenation (ECMO) is an advanced medical technology used to treat respiratory and heart failure. The coronavirus pandemic has resulted in significantly more ECMO patients worldwide. However, the number of hospitals ... ...

    Abstract Background: Extracorporeal membrane oxygenation (ECMO) is an advanced medical technology used to treat respiratory and heart failure. The coronavirus pandemic has resulted in significantly more ECMO patients worldwide. However, the number of hospitals with ECMO capabilities and ECMO-trained staff are limited. Training of personnel in ECMO could supplement this demand.
    Objective: To evaluate our previously developed ECMO course using a task trainer-based training, as opposed to an existing live tissue-training model, and determine if such a program was adequate and could be expanded to other facilities.
    Methods: Seventeen teams, each consisting of a physician and nurse, underwent a 5 hour accelerated ECMO course in which they watched prerecorded ECMO training lectures, primed circuit, cannulated, initiated ECMO, and corrected common complications. Training success was evaluated via knowledge and confidence assessments and observation of each team attempting to initiate ECMO while troubleshooting complications on a Yorkshire swine.
    Results: Seventeen teams successfully completed the course. Sixteen teams (94%, 95% CI = 71%-100%) successfully placed the swine on veno-arterial ECMO. Of those 16 teams, 15 successfully transitioned to veno-arterial-venous ECMO. The knowledge assessments and confidence levels of physicians and nurses increased by 24.3% from pretest (mean of 65.3%, SD 14.4%) to posttest (mean of 89.6%, SD 10.3%),
    Conclusions: An abbreviated one day lecture and hands-on task trainer-based ECMO course resulted in a high rate of successful skill demonstration and improvement of physicians' and nurses' knowledge assessments and confidence levels, similar to our previous live tissue training program.
    Language English
    Publishing date 2022-12-05
    Publishing country United States
    Document type Journal Article
    ISSN 2472-5390
    ISSN (online) 2472-5390
    DOI 10.1002/aet2.10806
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Cryopreservation of human whole blood allows immunophenotyping by flow cytometry up to 30days after cell isolation

    Madelaine Paredes, R / Douglas K. Tadaki / Amanda Sooter / Fabia Gamboni / C.D.R. Forest Sheppard

    Journal of Immunological Methods. 2017,

    2017  

    Abstract: Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed ... ...

    Abstract Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed within seven days. This data acquisition window can be extended to up to one year post-labeling, but this requires cryopreservation of the samples at ultra-low temperatures (≤−80°C or in liquid nitrogen). In this study we optimized a standardized cryopreservation protocol to enable preservation of immunolabeled, human WB samples at −20°C for FC and tested its effectiveness after 0, 5, 15 or 30days. Analysis of stored samples shows that this protocol effectively preserves immunolabeled WB samples and that the duration of storage has no effect on morphology, viability or frequency of WB cell subpopulations, and that the intensity of fluorescent signal from labeled extracellular markers is fully preserved for at least 15days, and up to 30days for some markers. We demonstrate that using this protocol, we are able to differentiate resting versus activated WB cells as demonstrated by detection of significantly increased expression of CD11b by myeloid cells in WB samples pretreated with LPS (100μg/mL for 12h). Finally, we show that this method allows for labeling and detection of the intracellular cytokine (IL-8) up to 30days following cryopreservation from myeloid cells, in previously labeled and cryopreserved WB samples.
    Keywords biomedical research ; blood ; cryopreservation ; flow cytometry ; fluorescence ; humans ; interleukin-8 ; liquids ; nitrogen ; patients ; temperature ; viability
    Language English
    Size p. .
    Publishing place Elsevier B.V.
    Document type Article
    Note Pre-press version
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2017.08.013
    Database NAL-Catalogue (AGRICOLA)

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  4. Article ; Online: Efficacy of Intravenous Hydroxocobalamin for Treatment of Sodium Methanethiolate Exposure in a Swine Model (Sus scrofa) of Severe Methanethiol Toxicity.

    Maddry, Joseph K / Paredes, R Madelaine / Rebeles, Jennifer / Olson, Glen / Castaneda, Maria / Canellis, Kaysie / Ng, Patrick C / Bebarta, Vikhyat S

    Journal of medical toxicology : official journal of the American College of Medical Toxicology

    2020  Volume 16, Issue 4, Page(s) 388–397

    Abstract: Introduction: Methanethiol is a highly toxic chemical present in crude oil and natural gas. At high concentrations, methanethiol causes metabolic acidosis, seizures, myocardial infarction, coma, and death. Occupational Health and Safety Administration ... ...

    Abstract Introduction: Methanethiol is a highly toxic chemical present in crude oil and natural gas. At high concentrations, methanethiol causes metabolic acidosis, seizures, myocardial infarction, coma, and death. Occupational Health and Safety Administration lists methanethiol as a potential terrorist weapon. Methanethiol blocks the electron transport chain, resulting in lactic acidosis and acidemia. There is no specific treatment for methanethiol. Our objective was to measure the efficacy of intravenous (IV) hydroxocobalamin (HOC) versus no treatment (control) in methanethiol-induced apnea in a swine model.
    Methods: Sixteen anesthetized swine received IV sodium methanethiolate to apnea and were randomized to receive either IV HOC or no treatment. Physiologic and laboratory parameters were monitored throughout the study. Power analysis indicated that 8 animals per group would be sufficient to find a moderate effect (f = 0.24) with 2 groups, α = 0.05, and 80% power.
    Results: Both groups were similar in baseline characteristics. Following treatment, the HOC group had significantly higher heart rate and blood pressure at 5-10 minutes post-apnea, higher systemic vascular resistance at 5 minutes post-apnea, higher tidal volume, higher end-tidal carbon dioxide, and lower end-tidal oxygen 10-15 minutes post-apnea compared with controls. None of the animals survived to the end of the study (60 minutes). The Kaplan-Meier survival curves were significantly different between cohorts (log-rank p = 0.0321), with the HOC group surviving longer than controls (32.4 ± 7.3 vs. 25.8 ± 1.0 minutes).
    Conclusions: In our model of intravenous methanethiolate poisoning, IV HOC administration resulted in a transient improvement in vital signs and prolonged time to death; however, it did not improve survival.
    MeSH term(s) Animals ; Antidotes/administration & dosage ; Apnea/chemically induced ; Apnea/drug therapy ; Apnea/physiopathology ; Disease Models, Animal ; Hydroxocobalamin/administration & dosage ; Infusions, Intravenous ; Lung/drug effects ; Lung/physiopathology ; Sulfhydryl Compounds ; Sus scrofa ; Time Factors
    Chemical Substances Antidotes ; Sulfhydryl Compounds ; methylmercaptan (2X8406WW9I) ; Hydroxocobalamin (Q40X8H422O)
    Language English
    Publishing date 2020-04-01
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2435016-3
    ISSN 1937-6995 ; 1556-9039
    ISSN (online) 1937-6995
    ISSN 1556-9039
    DOI 10.1007/s13181-020-00767-7
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Generation of complement molecular complex C5b-9 (C5b-9) in response to poly-traumatic hemorrhagic shock and evaluation of C5 cleavage inhibitors in non-human primates.

    Paredes, R Madelaine / Reyna, Sarah / Vernon, Philip / Tadaki, Douglas K / Dallelucca, Jurandir J / Sheppard, Forest

    International immunopharmacology

    2018  Volume 54, Page(s) 221–225

    Abstract: Severe trauma initiates a systemic inflammatory cascade and that involves early activation of complement and cleavage of C5 into C5a (anaphylatoxin) and C5b (C5b-9 membrane attack complex). We examined activation of C5 in non-human primate (NHP) models ... ...

    Abstract Severe trauma initiates a systemic inflammatory cascade and that involves early activation of complement and cleavage of C5 into C5a (anaphylatoxin) and C5b (C5b-9 membrane attack complex). We examined activation of C5 in non-human primate (NHP) models of hemorrhagic shock. Blood plasma concentrations of C5b-9 were significantly increased in NHPs in response to hemorrhage alone and were further increased with the addition of tissue trauma. The onset of increased C5 cleavage was accelerated in NHPs that experienced decompensated poly-traumatic hemorrhagic shock. Next, to identify an effective inhibitor of NHP C5 cleavage in vitro, as a first step in the development of a potential therapy, three inhibitors of human C5 cleavage and hemolysis were tested in vitro. NHP C5 cleavage and complement-mediated hemolysis were successfully inhibited by pre-treatment of serum samples with a small, inhibitory peptide RA101348. Commercially-available C5 inhibitory antibodies were found to exhibit species-specific efficacy in vitro. Quidel's A217 antibody demonstrated dose-dependent inhibition of C5 cleavage and hemolysis in NHP samples, whereas LGM-Eculizumab only inhibited complement-mediated hemolysis in human samples. This study shows that complement activation in NHPs following experimental poly-traumatic hemorrhagic shock is consistent with clinical reports, and that cleavage of C5 and complement-mediated hemolysis can be effectively inhibited in vitro using a small peptide inhibitor. Taken together, these findings offer a clinically-relevant vehicle and a potential strategy for treatment of hemorrhagic shock with poly-traumatic injury.
    MeSH term(s) Animals ; Antibodies, Blocking/therapeutic use ; Antibodies, Monoclonal, Humanized/therapeutic use ; Complement Activation ; Complement C5/metabolism ; Complement Membrane Attack Complex/metabolism ; Enzyme Inhibitors/therapeutic use ; Hemolysis ; Humans ; Multiple Trauma/immunology ; Peptides/therapeutic use ; Primates ; Proteolysis ; Shock, Hemorrhagic/immunology
    Chemical Substances Antibodies, Blocking ; Antibodies, Monoclonal, Humanized ; Complement C5 ; Complement Membrane Attack Complex ; Enzyme Inhibitors ; Peptides ; eculizumab (A3ULP0F556)
    Language English
    Publishing date 2018-01
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2043785-7
    ISSN 1878-1705 ; 1567-5769
    ISSN (online) 1878-1705
    ISSN 1567-5769
    DOI 10.1016/j.intimp.2017.10.033
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Cryopreservation of human whole blood allows immunophenotyping by flow cytometry up to 30days after cell isolation.

    Paredes, R Madelaine / Tadaki, Douglas K / Sooter, Amanda / Gamboni, Fabia / Sheppard, Forest

    Journal of immunological methods

    2017  

    Abstract: Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed ... ...

    Abstract Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed within seven days. This data acquisition window can be extended to up to one year post-labeling, but this requires cryopreservation of the samples at ultra-low temperatures (≤-80°C or in liquid nitrogen). In this study we optimized a standardized cryopreservation protocol to enable preservation of immunolabeled, human WB samples at -20°C for FC and tested its effectiveness after 0, 5, 15 or 30days. Analysis of stored samples shows that this protocol effectively preserves immunolabeled WB samples and that the duration of storage has no effect on morphology, viability or frequency of WB cell subpopulations, and that the intensity of fluorescent signal from labeled extracellular markers is fully preserved for at least 15days, and up to 30days for some markers. We demonstrate that using this protocol, we are able to differentiate resting versus activated WB cells as demonstrated by detection of significantly increased expression of CD11b by myeloid cells in WB samples pretreated with LPS (100μg/mL for 12h). Finally, we show that this method allows for labeling and detection of the intracellular cytokine (IL-8) up to 30days following cryopreservation from myeloid cells, in previously labeled and cryopreserved WB samples.
    Language English
    Publishing date 2017-09-18
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 120142-6
    ISSN 1872-7905 ; 0022-1759
    ISSN (online) 1872-7905
    ISSN 0022-1759
    DOI 10.1016/j.jim.2017.08.013
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Development and Evaluation of An Abbreviated Extracorporeal Membrane Oxygenation (ECMO) Course for Nonsurgical Physicians and Nurses.

    Maddry, Joseph K / Paredes, R Madelaine / Paciocco, Joni A / Castaneda, Maria / Araña, Allyson A / Perez, Crystal A / Reeves, Lauren K / Newberry, Ryan K / Bebarta, Vikhyat S / Kester, Nurani / Mason, Phillip E

    AEM education and training

    2020  Volume 4, Issue 4, Page(s) 347–358

    Abstract: Background: Extracorporeal membrane oxygenation (ECMO) is a modification of cardiopulmonary bypass that allows prolonged support of patients with severe respiratory or cardiac failure. ECMO indications arse rapidly evolving and there is growing interest ...

    Abstract Background: Extracorporeal membrane oxygenation (ECMO) is a modification of cardiopulmonary bypass that allows prolonged support of patients with severe respiratory or cardiac failure. ECMO indications arse rapidly evolving and there is growing interest in its use for cardiac arrest and cardiogenic shock. However, ECMO training programs are limited. Training of emergency medicine and critical care clinicians could expand the use of this lifesaving intervention. Our objective was to develop and evaluate an abbreviated ECMO course that can be taught to emergency and critical care physicians and nurses.
    Methods: We developed a training model using Yorkshire swine (
    Results: Seventeen teams (34 clinicians) completed the abbreviated ECMO course. None had previously completed an ECMO certification course. Immediately following the course, all teams successfully primed and prepared the ECMO circuit. Fifteen teams (88%, 95% confidence interval [CI] = 64% to 99%) successfully initiated ECMO. Participants improved their knowledge (difference 21.2, 95% CI = 16.5 to 25.8) and confidence (difference 40.3, 95% CI = 35.6 to 45.0) scores after completing the course.
    Conclusions: We developed an accelerated 1-day ECMO course. Clinicians' confidence and knowledge assessments improved and 88% of teams could successfully initiate venoarterial ECMO after the course.
    Language English
    Publishing date 2020-04-16
    Publishing country United States
    Document type Journal Article
    ISSN 2472-5390
    ISSN (online) 2472-5390
    DOI 10.1002/aet2.10447
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Luminal Ca2+ depletion during the unfolded protein response in Xenopus oocytes: cause and consequence.

    Paredes, R Madelaine / Bollo, Mariana / Holstein, Deborah / Lechleiter, James D

    Cell calcium

    2013  Volume 53, Issue 4, Page(s) 286–296

    Abstract: The endoplasmic reticulum (ER) is a Ca(2+) storing organelle that plays a critical role in the synthesis, folding and post-translational modifications of many proteins. The ER enters into a condition of stress when the load of newly synthesized proteins ... ...

    Abstract The endoplasmic reticulum (ER) is a Ca(2+) storing organelle that plays a critical role in the synthesis, folding and post-translational modifications of many proteins. The ER enters into a condition of stress when the load of newly synthesized proteins exceeds its folding and processing capacity. This activates a signal transduction pathway called the unfolded protein response (UPR) that attempts to restore homeostasis. The precise role of ER Ca(2+) in the initiation of the UPR has not been defined. Specifically, it has not been established whether ER Ca(2+) dysregulation is a cause or consequence of ER stress. Here, we report that partial depletion of ER Ca(2+) stores induces a significant induction of the UPR, and leads to the retention of a normally secreted protein Carboxypeptidase Y. Moreover, inhibition of protein glycosylation by tunicamycin rapidly induced an ER Ca(2+) leak into the cytosol. However, blockade of the translocon with emetine inhibited the tunicamycin-induced Ca(2+) release. Furthermore, emetine treatment blocked elF2α phosphorylation and reduced expression of the chaperone BiP. These findings suggest that Ca(2+) may be both a cause and a consequence of ER protein misfolding. Thus, it appears that ER Ca(2+) leak is a significant co-factor for the initiation of the UPR.
    MeSH term(s) Animals ; Calcium/metabolism ; Cathepsin A/antagonists & inhibitors ; Cathepsin A/metabolism ; Cytosol/drug effects ; Cytosol/metabolism ; Endoplasmic Reticulum/drug effects ; Endoplasmic Reticulum/metabolism ; Endoplasmic Reticulum Stress/drug effects ; Glycosylation/drug effects ; Oocytes/cytology ; Oocytes/drug effects ; Oocytes/metabolism ; Protein Unfolding ; Tunicamycin/pharmacology ; Unfolded Protein Response ; Xenopus laevis
    Chemical Substances Tunicamycin (11089-65-9) ; Cathepsin A (EC 3.4.16.5) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2013-02-12
    Publishing country Netherlands
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 757687-0
    ISSN 1532-1991 ; 0143-4160
    ISSN (online) 1532-1991
    ISSN 0143-4160
    DOI 10.1016/j.ceca.2013.01.002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Inflammatory Profile in Response to Uncontrolled Hemorrhage in a Non-Human Primate (Rhesus Macaque) Model.

    Burdette, Alexander J / Paredes, Ruth Madelaine / Crossland, Randy F / Macko, Antoni R / Aden, James / Sheppard, Forest R

    Shock (Augusta, Ga.)

    2016  Volume 46, Issue 3 Suppl 1, Page(s) 115–122

    Abstract: Background: Uncontrolled hemorrhage (UH), the leading cause of potentially survivable combat-related death, elicits a deleterious inflammatory response. Our group previously reported an increased secretion of pro-inflammatory cytokines in a novel non- ... ...

    Abstract Background: Uncontrolled hemorrhage (UH), the leading cause of potentially survivable combat-related death, elicits a deleterious inflammatory response. Our group previously reported an increased secretion of pro-inflammatory cytokines in a novel non-human primate model of UH; however, to better understand the molecular profile of the inflammatory response to UH, we performed a comprehensive evaluation of inflammation at the proteomic and transcriptomic level.
    Methods: Anesthetized rhesus macaques (n = 8) underwent UH by 60% left lobe hepatectomy T = 0 min. At T = 5 min, animals received 11 mL of 5% albumin followed by normal saline infusion to a total resuscitation volume of 20 mL/kg by T = 120 min. Blood (T = 0, 5, 20, 120, 480 min) was collected for qPCR and multiplex cytokine quantification. Results from each non-human primate (NHP) per time-point are shown. Statistical analysis by one-way ANOVA with repeated measures, P <0.05 was considered significant.
    Results: Luminex analysis in serum revealed significant up-regulation compared with baseline of 8 cytokines/chemokines starting T = 120 min postinjury and significant down-regulation of 4 cytokines/chemokines as early as T = 20 min postinjury. Gene expression analysis in white blood cells uncovered 10 genes that were up-regulated greater than 3-fold compared with baseline and 29 genes that were down-regulated greater than 3-fold.
    Conclusion: The present study confirms the presence of systemic inflammation after UH at the proteomic and transcriptomic level providing insight into the inflammatory mediators that are involved as well as their kinetics following UH. The data demonstrates that NHP hemorrhage models may be suitable for evaluating therapeutics to control inflammation following hemorrhage.
    Language English
    Publishing date 2016-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1185432-7
    ISSN 1540-0514 ; 1073-2322
    ISSN (online) 1540-0514
    ISSN 1073-2322
    DOI 10.1097/SHK.0000000000000638
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Severe Hemorrhagic Shock Induces Acute Activation and Expansion of IL-8+/IL-10+ Neutrophils with Enhanced Oxidative Reactivity in Non-Human Primates.

    Vernon, Philip J / Paredes, Ruth Madelaine / Sooter, Amanda J / Schaub, Leasha J / Grossman, Heather M / Pusateri, Anthony E / Glaser, Jacob J / Sheppard, Forest R

    Shock (Augusta, Ga.)

    2016  Volume 46, Issue 3 Suppl 1, Page(s) 129–136

    Abstract: Background: Neutrophilic inflammation is a mediator of morbidity and mortality in response to hemorrhagic shock. Although injury-induced neutrophil margination has long been observed, the nature of neutrophils' role in the "second hit" paradigm remains ... ...

    Abstract Background: Neutrophilic inflammation is a mediator of morbidity and mortality in response to hemorrhagic shock. Although injury-induced neutrophil margination has long been observed, the nature of neutrophils' role in the "second hit" paradigm remains to be fully elucidated. We sought to extensively characterize neutrophil phenotype and functionality in response to severe hemorrhage in non-human primates (NHPs).
    Methods: NHPs (n = 8) were subjected to severe hemorrhagic shock and resuscitation. Blood was obtained at baseline (T = 0 min), end of shock (T = 60 min), end of resuscitation (T = 180 min), T = 360 min, and 24 h (T = 1440 min). Neutrophils were quantified by complete blood count and flow cytometry. IL-8 and IL-10 production was determined by intracellular flow cytometry. Oxidation of dihydrorhodamine-123 (DHR-123) was used to determine neutrophil oxidative bursts (untreated), priming (+fMLP), and burst capacity (+PMA/ionomycin) via microplate reader ex vivo. Data are reported as mean ± SEM; statistical significance was measured using repeated measures ANOVA with Bonferroni adjustment. P < 0.05 is considered significant.
    Results: CD45CD11bCD16 neutrophils doubled postinjury (P < 0.0001); this was due to activated IL-8/IL-10 neutrophils that increased in frequency in relation to resting IL-8IL-10 cells. At 24 h, the proportions of activated to resting neutrophils returned to baseline levels. Resuscitative measures initially decreased neutrophil oxidative output; however, oxidative bursts, priming, and burst capacity were significantly increased at 24 h (P < 0.0025, 0.0124, and 0.0118, respectively).
    Conclusion: These results demonstrate an acute expansion and phenotypic activation of circulating neutrophils postinjury followed by a return to homeostatic proportions within 24 h; paradoxically, phenotypically "resting" neutrophils at 24 h have significantly higher oxidative potential, predisposing for exaggerated inflammatory responses. These data are consistent with clinical literature and provide important functional insight into neutrophil-mediated shock pathology.
    Language English
    Publishing date 2016-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1185432-7
    ISSN 1540-0514 ; 1073-2322
    ISSN (online) 1540-0514
    ISSN 1073-2322
    DOI 10.1097/SHK.0000000000000643
    Database MEDical Literature Analysis and Retrieval System OnLINE

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