Manfred Weiss | University of Ulm (original) (raw)

Papers by Manfred Weiss

Research paper thumbnail of Effects of G-CSF on Atp Content and Cytokine Release by Endothelial Cells and PMN with or Without LPS

Research paper thumbnail of Course of inflammation and infection markers differ in ICU patients with severe COVID-19 under casirivimab- and/or tocilizumab application: an observational study

Research Square (Research Square), Apr 5, 2024

Research paper thumbnail of Bioassay for Endothelial Damage Mediators Retrieved by Hemoadsorption

Scientific Reports, 2019

Hemoadsorption devices are used to treat septic shock by adsorbing inflammatory cytokines and as ... more Hemoadsorption devices are used to treat septic shock by adsorbing inflammatory cytokines and as yet incompletely defined danger and pathogen associated molecular patterns. In an ideal case, hemoadsorption results in immediate recovery of microvascular endothelial cells’ (mEC) function and rapid recovery from catecholamine-dependency and septic shock. We here tested a single device, which consists of polystyrene-divinylbenzene core particles of 450 μm diameter with a high affinity for hydrophobic compounds. The current study aimed at the proof of concept that endothelial-specific damage mediators are adsorbed and can be recovered from hemoadsorption devices. Because of excellent clinical experience, we tested protein fractions released from a hemoadsorber in a novel endothelial bioassay. Video-based, long-term imaging of mEC proliferation and cell death were evaluated and combined with apoptosis and ATP measurements. Out of a total of 39 fractions recovered from column fractionation...

Research paper thumbnail of Surviving Sepsis Campaign Bundles Adherence and Their Limits in Surgical Patients with Septic Shock in an ICU

British Journal of Medicine and Medical Research, 2013

Aims: The Surviving Sepsis Campaign (SSC) guidelines aimed to reduce heterogeneity of conventiona... more Aims: The Surviving Sepsis Campaign (SSC) guidelines aimed to reduce heterogeneity of conventional therapy and mortality. The present study was performed in septic shock to describe the adherence to the 2008 SSC guidelines, confounding factors, and limitations.

Research paper thumbnail of 999 93: 425-439Granulocyte colony-stimulating factor to prevent the progression of systemic non-responsiveness in SIRS and sepsis

Research paper thumbnail of Phagocytosis and digestion of pH-sensitive fluorescent dye (Eos-FP) transfected E. coli in whole blood assays from patients with severe sepsis and septic shock

Journal of cell communication and signaling, 2011

The function of phagocytic and antigen presenting cells is of crucial importance to sustain immun... more The function of phagocytic and antigen presenting cells is of crucial importance to sustain immune competence against infectious agents as well as malignancies. We here describe a reproducible procedure for the quantification of phagocytosis by leukocytes in whole blood. For this, a pH-sensitive green-fluorescent protein- (GFP) like dye (Eos-FP) is transfected into infectious microroganisms. After UV-irradiation, the transfected bacteria emit green (≈5160 nm) and red (≈581 nm) fluorescent light at 490 nm excitation. Since the red fluorescent light is sensitive to acidic pH, the phagocytosed bacteria stop emitting red fluorescent light as soon as the phagosomes fuse with lysosomes. The green fluorescence is maintained in the phagolysosome until pathogen degradation is completed. Fluorescence emission can be followed by flow cytometry with filter settings documenting fluorescence 1 (FL 1, FITC) and fluorescence 2 (FL 2, phycoerythrin, PE). Eos-FP transfected bacteria can also be trace...

Research paper thumbnail of Authors's reply

Journal of emergencies, trauma, and shock, 2010

Research paper thumbnail of Decreased monocyte surface expression of HLA-DQ is associated with prolonged duration of septic shock

Purpose: To describe the epidemiology of the acute respiratory distress syndrome (ARDS) in a Braz... more Purpose: To describe the epidemiology of the acute respiratory distress syndrome (ARDS) in a Brazilian ICU. Methods: This prospective observational, non-interventional study, included all consecutive patients with ARDS criteria [1] admitted in the ICU of a Brazilian tertiary hospital, between January 1997 and September 2001. Were collected in a prospective fashion the following variables: age, gender, APACHE II score at ICU admission and at ARDS diagnosis, cause of ARDS, presence of AIDS, cancer and immunosuppression, occurrence of barotrauma, performance of traqueostomy, mortality, duration of mechanical ventilation (MV), length of stay (LOS) in ICU and in hospital. The lung injury score (LIS) [2] was used to quantify the degree of pulmonary injury in the first week of ARDS. Results: There was 2182 patients (P) admitted in ICU during the study period, of whom 141 (6.46%) had ARDS criteria. Seventy-six (54%) were men, the mean age was 46 ± 18 years, APACHE II 18 ± 7 and 19 ± 7 at admission and at ARDS diagnosis, respectively. Septic shock accounted for 42% (60 P) of the ARDS causes, sepsis 22% (31 P), diffuse pulmonary infection 16% (23 P), aspiration pneumonia 11% (15 P), non-septic shock 5% (7 P) and others 4% (5 P). Ten percent (14 P) had AIDS, 30% (43 P) cancer and 25% (36 P) immunosuppression. All patients were mechanically ventilated with Tidal Volume between 4 and 8 ml/kg. Only 3.5% (5 P) had barotrauma and 10% (14 P) performed traqueostomy. Mortality rate was 79% in the ICU. The patients required 12 ± 10 days on MV, ranging from 1 to 55 days. The LOS in ICU and hospital was 14 ± 13 (1-69) days and 28 ± 32 (1-325) days, respectively. There was a time delay of 3.7 ± 4.5 days between admission in ICU and the onset of ARDS. The Murray score (mean ± SD) was 3.2 ± 0.4, 3 ± 0.5, 3 ± 0.5, 2.9 ± 0.6, 2.8 ± 0.7, 2.7 ± 0.7 and 2.6 ± 0.8 in the first 7 days, respectively. Conclusions: ARDS in our hospital has a similar incidence of reports in the USA and Europe. There was a higher mortality, which could be explained by a high incidence of infection causes of ARDS, mainly septic shock, and elevated combined occurrence of AIDS, cancer and immunosuppression, along the degree of LIS. The incidence of barotrauma was low, as a consequence of the current mechanical ventilation strategies.

Research paper thumbnail of Complement Activation During Sepsis in Humans

Advances in Experimental Medicine and Biology

In critically ill patients, sepsis is currently one of the leading causes of death in both the Un... more In critically ill patients, sepsis is currently one of the leading causes of death in both the United States and Europe1, 2. Despite intensive scientific and clinical efforts, the deleterious pathophysiological consequences of sepsis remain enigmatic. During experimental ...

Research paper thumbnail of EARLY EXPRESSION CHANGES OF COMPLEMENT REGULATORY PROTEINS AND C5a RECEPTOR (CD88) ON LEUKOCYTES AFTER MULTIPLE INJURY IN HUMANS

Shock, 2010

As a crucial element of innate immunity, the complement cascade becomes activated after severe tr... more As a crucial element of innate immunity, the complement cascade becomes activated after severe trauma. Regulation of the complement cascade and protection against complement-mediated tissue destruction is provided by a selection of soluble and membrane-bound complement regulatory proteins (CRegs). To date, the leukocyte expression profile of CRegs in multiple injured patients is unknown. In the present study, expression of CRegs and the C5a receptor (CD88) was analyzed on neutrophils, monocytes, and lymphocytes by flow cytometry. Whole blood samples were obtained from healthy volunteers (n = 16) or multiple injured patients (n = 12) on admission in the emergency department and 4, 12, 24, 120, and 240 h after trauma. The content of CRegs and CD88 on leukocytes was significantly altered posttrauma: CD55 (decay accelerating factor) displayed a time-dependent, elevated expression pattern on neutrophils and monocytes, but not on lymphocytes. CD59 (membrane attack complex inhibitor) expression was significantly increased on neutrophils and monocytes at the time of admission and after 5 to 10 days in lymphocytes. CD46 (membrane cofactor protein) was significantly down-regulated in all three cell types posttrauma. CD35 (complement receptor 1) expression on neutrophils was initially decreased, whereas monocytes presented a significant increase in CD35 expression. CD35 on lymphocyte remained unchanged throughout the observation period. CD88 expression was considerably reduced on leukocytes between 0 and 240 h after injury. CD59, CD46, and CD88 expression values on neutrophils reversely correlated with severity of injury. In summary, expression profiles of CRegs and CD88 on leukocytes are specifically altered after polytrauma in humans, indicating a trauma-induced "complementopathy."

Research paper thumbnail of The Q460R Mutation of the Atp Sensitive P2X7 Ion Channel is Likely to Influence Trauma-Associated Hypercytokinemia

Shock, 2006

A separation method is presented that yields liquid fractions with sample constituents conserved ... more A separation method is presented that yields liquid fractions with sample constituents conserved in their native states. Size exclusion, anion exchange, and lectin chromatography are used for the first, second, and third separation dimension, respectively. 96 parallel chromatographic micro-columns are used with micro-plates and the automated CyBio AG analysis platform so that each Separation step may be applied to all fractions of the foregoing one. Up to 100 mg of sample protein can be separated into up to 6000 fractions. Routinely, two samples are separated within 14 h simultaneously into 3360 2D-fractions each. 1D-and 2D recovery is 96.2 T 7.5% (n = 178) and 103.2 T 16.1% (n = 26), respectively. The protein concentrations obtained are highly reproducible. Within 2-D fractions of six runs, CV was 12.9% T 7.7% (mean + SD), 9.5% T 3.3%, and 6.1% T 2.0% in the range 0.35 to 0.5, 0.7 to 1.0, and 1.4 to 2 mg protein/ml, respectively. Spectrophotometric, enzymatic, and immunological analyses as well as MALDI-MS and ESI-MS/MS can be applied to the chromatographic fractions. More than 1000 proteins were identified with this method in healthy human serum (www.db-thuerinqen.de/servlets/ DocumentServlet?id=4054). Along with protein entities, various functional protein complexes (interactomes), proteolytic fragments, glycosylated and glycated proteins, and non-protein constituents were evidenced. Firstly, 2D fraction loci are considered to contain biomarkers, if their UV-absorbance correlate clearly and reproducibly with the clinical findings. Secondly, Proteins exhibiting altered concentrations in such patient-serum fractions are identified by a robust semi-quantitative mass spectrometric approach. The method was evaluated by confirming known protein changes in sera of patients with renal failure and severe inflammatory diseases. Lists of new biomarker candidates obtained are still under proof. 4 COMBINED IN SILICO AND IN VIVO STUDIES ON ACUTE INFLAMMATION. Yoram Vodovotz Trauma and hemorrhagic shock elicit an acute inflammatory response, predisposing patients to sepsis, organ dysfunction, and death. Few approved therapies exist for these acute inflammatory states, mainly due to the complex interplay of interacting inflammatory and physiologic elements working at multiple levels. Mathematical modeling of complex systems is an approach for understanding the interplay among the plethora of biological interactions. We constructed a mathematical model using ordinary differential equations that encompass the dynamics of cells and cytokines of the acute inflammatory response, as well as global tissue dysfunction. More recently, the model was augmented with elements of adaptive immunity. The model was calibrated separately in various strains of mice, rats, swine, and humans subjected to various acute inflammatory insults. Software was created that facilitated fitting of the mathematical model to experimental data, as well as for simulation of experiments with various inflammatory challenges and associated variations (gene knockouts, inhibition of specific cytokines, etc.). Using this software, the C57Bl/6 mousespecific model was recalibrated for inflammatory analyte data in CD14 j/j mice, and was used to elucidate altered features of inflammation in these animals. Similar methodology was used to elucidate mechanisms of inflammatory changes associated with Abstracts 1 Copyright @ 2006 by the Shock Society. Unauthorized reproduction of this article is prohibited. aging and novel drug therapy in mice. A model-derived hypothesis that underlying trauma is the central driver of inflammation and organ damage following hemorrhagic shock was validated with data on circulating markers and on the liver transcriptome in mice. Simulations of reversible and irreversible hemorrhagic shock in mice were also carried out. Studies in other pre-clinical paradigms of inflammation and wound healing, as well as other experimental animal species, are ongoing. Mathematical modeling may provide insights into the complex dynamics of acute inflammation in a manner that can be tested in vivo.

Research paper thumbnail of Changes of Complement-Regulatory Proteins During Sepsis in Humans

Shock, 2006

A separation method is presented that yields liquid fractions with sample constituents conserved ... more A separation method is presented that yields liquid fractions with sample constituents conserved in their native states. Size exclusion, anion exchange, and lectin chromatography are used for the first, second, and third separation dimension, respectively. 96 parallel chromatographic micro-columns are used with micro-plates and the automated CyBio AG analysis platform so that each Separation step may be applied to all fractions of the foregoing one. Up to 100 mg of sample protein can be separated into up to 6000 fractions. Routinely, two samples are separated within 14 h simultaneously into 3360 2D-fractions each. 1D-and 2D recovery is 96.2 T 7.5% (n = 178) and 103.2 T 16.1% (n = 26), respectively. The protein concentrations obtained are highly reproducible. Within 2-D fractions of six runs, CV was 12.9% T 7.7% (mean + SD), 9.5% T 3.3%, and 6.1% T 2.0% in the range 0.35 to 0.5, 0.7 to 1.0, and 1.4 to 2 mg protein/ml, respectively. Spectrophotometric, enzymatic, and immunological analyses as well as MALDI-MS and ESI-MS/MS can be applied to the chromatographic fractions. More than 1000 proteins were identified with this method in healthy human serum (www.db-thuerinqen.de/servlets/ DocumentServlet?id=4054). Along with protein entities, various functional protein complexes (interactomes), proteolytic fragments, glycosylated and glycated proteins, and non-protein constituents were evidenced. Firstly, 2D fraction loci are considered to contain biomarkers, if their UV-absorbance correlate clearly and reproducibly with the clinical findings. Secondly, Proteins exhibiting altered concentrations in such patient-serum fractions are identified by a robust semi-quantitative mass spectrometric approach. The method was evaluated by confirming known protein changes in sera of patients with renal failure and severe inflammatory diseases. Lists of new biomarker candidates obtained are still under proof. 4 COMBINED IN SILICO AND IN VIVO STUDIES ON ACUTE INFLAMMATION. Yoram Vodovotz Trauma and hemorrhagic shock elicit an acute inflammatory response, predisposing patients to sepsis, organ dysfunction, and death. Few approved therapies exist for these acute inflammatory states, mainly due to the complex interplay of interacting inflammatory and physiologic elements working at multiple levels. Mathematical modeling of complex systems is an approach for understanding the interplay among the plethora of biological interactions. We constructed a mathematical model using ordinary differential equations that encompass the dynamics of cells and cytokines of the acute inflammatory response, as well as global tissue dysfunction. More recently, the model was augmented with elements of adaptive immunity. The model was calibrated separately in various strains of mice, rats, swine, and humans subjected to various acute inflammatory insults. Software was created that facilitated fitting of the mathematical model to experimental data, as well as for simulation of experiments with various inflammatory challenges and associated variations (gene knockouts, inhibition of specific cytokines, etc.). Using this software, the C57Bl/6 mousespecific model was recalibrated for inflammatory analyte data in CD14 j/j mice, and was used to elucidate altered features of inflammation in these animals. Similar methodology was used to elucidate mechanisms of inflammatory changes associated with Abstracts 1 Copyright @ 2006 by the Shock Society. Unauthorized reproduction of this article is prohibited. aging and novel drug therapy in mice. A model-derived hypothesis that underlying trauma is the central driver of inflammation and organ damage following hemorrhagic shock was validated with data on circulating markers and on the liver transcriptome in mice. Simulations of reversible and irreversible hemorrhagic shock in mice were also carried out. Studies in other pre-clinical paradigms of inflammation and wound healing, as well as other experimental animal species, are ongoing. Mathematical modeling may provide insights into the complex dynamics of acute inflammation in a manner that can be tested in vivo.

Research paper thumbnail of Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases

Medical Engineering & Physics, 2008

Miniaturized and parallelized sandwich immunoassays allow the simultaneous analysis of a variety ... more Miniaturized and parallelized sandwich immunoassays allow the simultaneous analysis of a variety of parameters in a single experiment. Bead-based protein array systems or suspension microarrays are well-established multiplex sandwich immunoassay formats. To study inflammatory diseases, protein arrays can be used to analyze changes in plasma protein levels, such as cytokines, chemokines, soluble receptors, and matrix metalloproteinases. Using the bead-based Luminex system, multiplexed sandwich immunoassays have been developed to analyze the plasma concentrations of soluble receptors: sTNF-RI, sTNF-RII, sIL-2R, sgp130, sFas, sRAGE, sE-selectin, sICAM-1, sVCAM-1, sMIF-1 and sFasL. This newly established 11-plex soluble receptors assay demonstrated acceptable intra-assay and inter-assay precision, appropriate accuracy, and no crossreactivity between analytes. Using this assay, 100 plasma samples derived from 36 critically ill intensive care unit (ICU) patients with trauma or sepsis were analyzed for their soluble receptor plasma concentrations. Results obtained allowed grouping of patients' samples into a trauma and a sepsis group. Four candidate molecules: sFas, sICAM-1, sTNF-RI, and sTNF-RII had higher concentrations in patients with sepsis than in those with trauma, contributing the highest discriminatory values to define the nature of the inflammatory disease originating from pathogen-involved (sepsis) or pathogen-independent inflammation.

Research paper thumbnail of Changes in the Novel Orphan, C5a Receptor (C5L2), during Experimental Sepsis and Sepsis in Humans

The Journal of Immunology, 2005

Sepsis is associated with extensive complement activation, compromising innate immune defenses, e... more Sepsis is associated with extensive complement activation, compromising innate immune defenses, especially in neutrophils (PMN). Recently, a second C5a receptor (C5L2) was detected on PMN without evidence of intracellular signaling. The current study was designed to determine changes in C5L2 in blood PMN during sepsis. In vitro exposure of PMN to C5a, but not to fMLP, led to reduced content of C5L2. Following cecal ligation and puncture-induced sepsis in rats, PMN demonstrated a time-dependent decrease in C5L2. In vivo blockade of C5a during experimental sepsis resulted in preservation of C5L2. Similarly, PMN from patients with progressive sepsis showed significantly reduced C5L2 expression (n ‫؍‬ 26), which was virtually abolished in patients who developed multiorgan failure (n ‫؍‬ 10). In contrast, sepsis survivors exhibited retention of C5L2 (n ‫؍‬ 12/13). The data suggest that C5L2 on PMN diminishes during sepsis due to systemic generation of C5a, which is associated with a poor prognosis.

Research paper thumbnail of Enhancement of neutrophil function by in vivo filgrastim treatment for prophylaxis of sepsis in surgical intensive care patients

Journal of Critical Care, 1995

Research paper thumbnail of P1530 CMV reactivation and antiviral immune response in patients with septic shock

International Journal of Antimicrobial Agents, 2007

Research paper thumbnail of Hepato-splanchnic metabolic effects of the stable prostacyclin analogue iloprost in patients with septic shock

Intensive Care Medicine, 2001

Research paper thumbnail of CD64 surface expression on neutrophils is transiently upregulated in patients with septic shock

Intensive Care Medicine, 2001

Research paper thumbnail of Stereoselective Suppression of Neutrophil Function by Ketamine?

Immunopharmacology and Immunotoxicology, 1995

The effects of the commercially available ketamine preparation (Ketanest), the ketamine racemate ... more The effects of the commercially available ketamine preparation (Ketanest), the ketamine racemate and of the two enantiomers, the R(-)-racemate and the S(+)-racemate, as well as its drug-free solvent were examined by N-formyl-methionyl-leucyl-phenylalanine-(FMLP)- and zymosan-induced oxygen radical production of polymorphonuclear cells (PMN). The racemate and the two enantiomers of ketamine suppressed FMLP- and zymosan-induced chemiluminescence of PMN in a dose-dependent fashion to the same extent. Therefore suppression of chemiluminescence of PMN by ketamine does not result from a specific receptor interaction.

Research paper thumbnail of Effects of sedatives and their solvents on the neutrophil granulocyte function

Journal of Huazhong University of Science and Technology [Medical Sciences], 1994

We have examined the effects of commercially available preparations and drug-free solvents of Mid... more We have examined the effects of commercially available preparations and drug-free solvents of Midazolam (Dormicum) and Propofol (Disoprivan) by N-formyl-methionyl-leucyl-phenylalanine (FMLP) -induced polymorphonuclear cell (PMN) chemiluminescence system. In the case of propofol and drug-free solvent, PMN chemiluminescence was suppressed. With Midazolam, only the active drug depressed PMN chemiluminescence.

Research paper thumbnail of Effects of G-CSF on Atp Content and Cytokine Release by Endothelial Cells and PMN with or Without LPS

Research paper thumbnail of Course of inflammation and infection markers differ in ICU patients with severe COVID-19 under casirivimab- and/or tocilizumab application: an observational study

Research Square (Research Square), Apr 5, 2024

Research paper thumbnail of Bioassay for Endothelial Damage Mediators Retrieved by Hemoadsorption

Scientific Reports, 2019

Hemoadsorption devices are used to treat septic shock by adsorbing inflammatory cytokines and as ... more Hemoadsorption devices are used to treat septic shock by adsorbing inflammatory cytokines and as yet incompletely defined danger and pathogen associated molecular patterns. In an ideal case, hemoadsorption results in immediate recovery of microvascular endothelial cells’ (mEC) function and rapid recovery from catecholamine-dependency and septic shock. We here tested a single device, which consists of polystyrene-divinylbenzene core particles of 450 μm diameter with a high affinity for hydrophobic compounds. The current study aimed at the proof of concept that endothelial-specific damage mediators are adsorbed and can be recovered from hemoadsorption devices. Because of excellent clinical experience, we tested protein fractions released from a hemoadsorber in a novel endothelial bioassay. Video-based, long-term imaging of mEC proliferation and cell death were evaluated and combined with apoptosis and ATP measurements. Out of a total of 39 fractions recovered from column fractionation...

Research paper thumbnail of Surviving Sepsis Campaign Bundles Adherence and Their Limits in Surgical Patients with Septic Shock in an ICU

British Journal of Medicine and Medical Research, 2013

Aims: The Surviving Sepsis Campaign (SSC) guidelines aimed to reduce heterogeneity of conventiona... more Aims: The Surviving Sepsis Campaign (SSC) guidelines aimed to reduce heterogeneity of conventional therapy and mortality. The present study was performed in septic shock to describe the adherence to the 2008 SSC guidelines, confounding factors, and limitations.

Research paper thumbnail of 999 93: 425-439Granulocyte colony-stimulating factor to prevent the progression of systemic non-responsiveness in SIRS and sepsis

Research paper thumbnail of Phagocytosis and digestion of pH-sensitive fluorescent dye (Eos-FP) transfected E. coli in whole blood assays from patients with severe sepsis and septic shock

Journal of cell communication and signaling, 2011

The function of phagocytic and antigen presenting cells is of crucial importance to sustain immun... more The function of phagocytic and antigen presenting cells is of crucial importance to sustain immune competence against infectious agents as well as malignancies. We here describe a reproducible procedure for the quantification of phagocytosis by leukocytes in whole blood. For this, a pH-sensitive green-fluorescent protein- (GFP) like dye (Eos-FP) is transfected into infectious microroganisms. After UV-irradiation, the transfected bacteria emit green (≈5160 nm) and red (≈581 nm) fluorescent light at 490 nm excitation. Since the red fluorescent light is sensitive to acidic pH, the phagocytosed bacteria stop emitting red fluorescent light as soon as the phagosomes fuse with lysosomes. The green fluorescence is maintained in the phagolysosome until pathogen degradation is completed. Fluorescence emission can be followed by flow cytometry with filter settings documenting fluorescence 1 (FL 1, FITC) and fluorescence 2 (FL 2, phycoerythrin, PE). Eos-FP transfected bacteria can also be trace...

Research paper thumbnail of Authors's reply

Journal of emergencies, trauma, and shock, 2010

Research paper thumbnail of Decreased monocyte surface expression of HLA-DQ is associated with prolonged duration of septic shock

Purpose: To describe the epidemiology of the acute respiratory distress syndrome (ARDS) in a Braz... more Purpose: To describe the epidemiology of the acute respiratory distress syndrome (ARDS) in a Brazilian ICU. Methods: This prospective observational, non-interventional study, included all consecutive patients with ARDS criteria [1] admitted in the ICU of a Brazilian tertiary hospital, between January 1997 and September 2001. Were collected in a prospective fashion the following variables: age, gender, APACHE II score at ICU admission and at ARDS diagnosis, cause of ARDS, presence of AIDS, cancer and immunosuppression, occurrence of barotrauma, performance of traqueostomy, mortality, duration of mechanical ventilation (MV), length of stay (LOS) in ICU and in hospital. The lung injury score (LIS) [2] was used to quantify the degree of pulmonary injury in the first week of ARDS. Results: There was 2182 patients (P) admitted in ICU during the study period, of whom 141 (6.46%) had ARDS criteria. Seventy-six (54%) were men, the mean age was 46 ± 18 years, APACHE II 18 ± 7 and 19 ± 7 at admission and at ARDS diagnosis, respectively. Septic shock accounted for 42% (60 P) of the ARDS causes, sepsis 22% (31 P), diffuse pulmonary infection 16% (23 P), aspiration pneumonia 11% (15 P), non-septic shock 5% (7 P) and others 4% (5 P). Ten percent (14 P) had AIDS, 30% (43 P) cancer and 25% (36 P) immunosuppression. All patients were mechanically ventilated with Tidal Volume between 4 and 8 ml/kg. Only 3.5% (5 P) had barotrauma and 10% (14 P) performed traqueostomy. Mortality rate was 79% in the ICU. The patients required 12 ± 10 days on MV, ranging from 1 to 55 days. The LOS in ICU and hospital was 14 ± 13 (1-69) days and 28 ± 32 (1-325) days, respectively. There was a time delay of 3.7 ± 4.5 days between admission in ICU and the onset of ARDS. The Murray score (mean ± SD) was 3.2 ± 0.4, 3 ± 0.5, 3 ± 0.5, 2.9 ± 0.6, 2.8 ± 0.7, 2.7 ± 0.7 and 2.6 ± 0.8 in the first 7 days, respectively. Conclusions: ARDS in our hospital has a similar incidence of reports in the USA and Europe. There was a higher mortality, which could be explained by a high incidence of infection causes of ARDS, mainly septic shock, and elevated combined occurrence of AIDS, cancer and immunosuppression, along the degree of LIS. The incidence of barotrauma was low, as a consequence of the current mechanical ventilation strategies.

Research paper thumbnail of Complement Activation During Sepsis in Humans

Advances in Experimental Medicine and Biology

In critically ill patients, sepsis is currently one of the leading causes of death in both the Un... more In critically ill patients, sepsis is currently one of the leading causes of death in both the United States and Europe1, 2. Despite intensive scientific and clinical efforts, the deleterious pathophysiological consequences of sepsis remain enigmatic. During experimental ...

Research paper thumbnail of EARLY EXPRESSION CHANGES OF COMPLEMENT REGULATORY PROTEINS AND C5a RECEPTOR (CD88) ON LEUKOCYTES AFTER MULTIPLE INJURY IN HUMANS

Shock, 2010

As a crucial element of innate immunity, the complement cascade becomes activated after severe tr... more As a crucial element of innate immunity, the complement cascade becomes activated after severe trauma. Regulation of the complement cascade and protection against complement-mediated tissue destruction is provided by a selection of soluble and membrane-bound complement regulatory proteins (CRegs). To date, the leukocyte expression profile of CRegs in multiple injured patients is unknown. In the present study, expression of CRegs and the C5a receptor (CD88) was analyzed on neutrophils, monocytes, and lymphocytes by flow cytometry. Whole blood samples were obtained from healthy volunteers (n = 16) or multiple injured patients (n = 12) on admission in the emergency department and 4, 12, 24, 120, and 240 h after trauma. The content of CRegs and CD88 on leukocytes was significantly altered posttrauma: CD55 (decay accelerating factor) displayed a time-dependent, elevated expression pattern on neutrophils and monocytes, but not on lymphocytes. CD59 (membrane attack complex inhibitor) expression was significantly increased on neutrophils and monocytes at the time of admission and after 5 to 10 days in lymphocytes. CD46 (membrane cofactor protein) was significantly down-regulated in all three cell types posttrauma. CD35 (complement receptor 1) expression on neutrophils was initially decreased, whereas monocytes presented a significant increase in CD35 expression. CD35 on lymphocyte remained unchanged throughout the observation period. CD88 expression was considerably reduced on leukocytes between 0 and 240 h after injury. CD59, CD46, and CD88 expression values on neutrophils reversely correlated with severity of injury. In summary, expression profiles of CRegs and CD88 on leukocytes are specifically altered after polytrauma in humans, indicating a trauma-induced "complementopathy."

Research paper thumbnail of The Q460R Mutation of the Atp Sensitive P2X7 Ion Channel is Likely to Influence Trauma-Associated Hypercytokinemia

Shock, 2006

A separation method is presented that yields liquid fractions with sample constituents conserved ... more A separation method is presented that yields liquid fractions with sample constituents conserved in their native states. Size exclusion, anion exchange, and lectin chromatography are used for the first, second, and third separation dimension, respectively. 96 parallel chromatographic micro-columns are used with micro-plates and the automated CyBio AG analysis platform so that each Separation step may be applied to all fractions of the foregoing one. Up to 100 mg of sample protein can be separated into up to 6000 fractions. Routinely, two samples are separated within 14 h simultaneously into 3360 2D-fractions each. 1D-and 2D recovery is 96.2 T 7.5% (n = 178) and 103.2 T 16.1% (n = 26), respectively. The protein concentrations obtained are highly reproducible. Within 2-D fractions of six runs, CV was 12.9% T 7.7% (mean + SD), 9.5% T 3.3%, and 6.1% T 2.0% in the range 0.35 to 0.5, 0.7 to 1.0, and 1.4 to 2 mg protein/ml, respectively. Spectrophotometric, enzymatic, and immunological analyses as well as MALDI-MS and ESI-MS/MS can be applied to the chromatographic fractions. More than 1000 proteins were identified with this method in healthy human serum (www.db-thuerinqen.de/servlets/ DocumentServlet?id=4054). Along with protein entities, various functional protein complexes (interactomes), proteolytic fragments, glycosylated and glycated proteins, and non-protein constituents were evidenced. Firstly, 2D fraction loci are considered to contain biomarkers, if their UV-absorbance correlate clearly and reproducibly with the clinical findings. Secondly, Proteins exhibiting altered concentrations in such patient-serum fractions are identified by a robust semi-quantitative mass spectrometric approach. The method was evaluated by confirming known protein changes in sera of patients with renal failure and severe inflammatory diseases. Lists of new biomarker candidates obtained are still under proof. 4 COMBINED IN SILICO AND IN VIVO STUDIES ON ACUTE INFLAMMATION. Yoram Vodovotz Trauma and hemorrhagic shock elicit an acute inflammatory response, predisposing patients to sepsis, organ dysfunction, and death. Few approved therapies exist for these acute inflammatory states, mainly due to the complex interplay of interacting inflammatory and physiologic elements working at multiple levels. Mathematical modeling of complex systems is an approach for understanding the interplay among the plethora of biological interactions. We constructed a mathematical model using ordinary differential equations that encompass the dynamics of cells and cytokines of the acute inflammatory response, as well as global tissue dysfunction. More recently, the model was augmented with elements of adaptive immunity. The model was calibrated separately in various strains of mice, rats, swine, and humans subjected to various acute inflammatory insults. Software was created that facilitated fitting of the mathematical model to experimental data, as well as for simulation of experiments with various inflammatory challenges and associated variations (gene knockouts, inhibition of specific cytokines, etc.). Using this software, the C57Bl/6 mousespecific model was recalibrated for inflammatory analyte data in CD14 j/j mice, and was used to elucidate altered features of inflammation in these animals. Similar methodology was used to elucidate mechanisms of inflammatory changes associated with Abstracts 1 Copyright @ 2006 by the Shock Society. Unauthorized reproduction of this article is prohibited. aging and novel drug therapy in mice. A model-derived hypothesis that underlying trauma is the central driver of inflammation and organ damage following hemorrhagic shock was validated with data on circulating markers and on the liver transcriptome in mice. Simulations of reversible and irreversible hemorrhagic shock in mice were also carried out. Studies in other pre-clinical paradigms of inflammation and wound healing, as well as other experimental animal species, are ongoing. Mathematical modeling may provide insights into the complex dynamics of acute inflammation in a manner that can be tested in vivo.

Research paper thumbnail of Changes of Complement-Regulatory Proteins During Sepsis in Humans

Shock, 2006

A separation method is presented that yields liquid fractions with sample constituents conserved ... more A separation method is presented that yields liquid fractions with sample constituents conserved in their native states. Size exclusion, anion exchange, and lectin chromatography are used for the first, second, and third separation dimension, respectively. 96 parallel chromatographic micro-columns are used with micro-plates and the automated CyBio AG analysis platform so that each Separation step may be applied to all fractions of the foregoing one. Up to 100 mg of sample protein can be separated into up to 6000 fractions. Routinely, two samples are separated within 14 h simultaneously into 3360 2D-fractions each. 1D-and 2D recovery is 96.2 T 7.5% (n = 178) and 103.2 T 16.1% (n = 26), respectively. The protein concentrations obtained are highly reproducible. Within 2-D fractions of six runs, CV was 12.9% T 7.7% (mean + SD), 9.5% T 3.3%, and 6.1% T 2.0% in the range 0.35 to 0.5, 0.7 to 1.0, and 1.4 to 2 mg protein/ml, respectively. Spectrophotometric, enzymatic, and immunological analyses as well as MALDI-MS and ESI-MS/MS can be applied to the chromatographic fractions. More than 1000 proteins were identified with this method in healthy human serum (www.db-thuerinqen.de/servlets/ DocumentServlet?id=4054). Along with protein entities, various functional protein complexes (interactomes), proteolytic fragments, glycosylated and glycated proteins, and non-protein constituents were evidenced. Firstly, 2D fraction loci are considered to contain biomarkers, if their UV-absorbance correlate clearly and reproducibly with the clinical findings. Secondly, Proteins exhibiting altered concentrations in such patient-serum fractions are identified by a robust semi-quantitative mass spectrometric approach. The method was evaluated by confirming known protein changes in sera of patients with renal failure and severe inflammatory diseases. Lists of new biomarker candidates obtained are still under proof. 4 COMBINED IN SILICO AND IN VIVO STUDIES ON ACUTE INFLAMMATION. Yoram Vodovotz Trauma and hemorrhagic shock elicit an acute inflammatory response, predisposing patients to sepsis, organ dysfunction, and death. Few approved therapies exist for these acute inflammatory states, mainly due to the complex interplay of interacting inflammatory and physiologic elements working at multiple levels. Mathematical modeling of complex systems is an approach for understanding the interplay among the plethora of biological interactions. We constructed a mathematical model using ordinary differential equations that encompass the dynamics of cells and cytokines of the acute inflammatory response, as well as global tissue dysfunction. More recently, the model was augmented with elements of adaptive immunity. The model was calibrated separately in various strains of mice, rats, swine, and humans subjected to various acute inflammatory insults. Software was created that facilitated fitting of the mathematical model to experimental data, as well as for simulation of experiments with various inflammatory challenges and associated variations (gene knockouts, inhibition of specific cytokines, etc.). Using this software, the C57Bl/6 mousespecific model was recalibrated for inflammatory analyte data in CD14 j/j mice, and was used to elucidate altered features of inflammation in these animals. Similar methodology was used to elucidate mechanisms of inflammatory changes associated with Abstracts 1 Copyright @ 2006 by the Shock Society. Unauthorized reproduction of this article is prohibited. aging and novel drug therapy in mice. A model-derived hypothesis that underlying trauma is the central driver of inflammation and organ damage following hemorrhagic shock was validated with data on circulating markers and on the liver transcriptome in mice. Simulations of reversible and irreversible hemorrhagic shock in mice were also carried out. Studies in other pre-clinical paradigms of inflammation and wound healing, as well as other experimental animal species, are ongoing. Mathematical modeling may provide insights into the complex dynamics of acute inflammation in a manner that can be tested in vivo.

Research paper thumbnail of Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases

Medical Engineering & Physics, 2008

Miniaturized and parallelized sandwich immunoassays allow the simultaneous analysis of a variety ... more Miniaturized and parallelized sandwich immunoassays allow the simultaneous analysis of a variety of parameters in a single experiment. Bead-based protein array systems or suspension microarrays are well-established multiplex sandwich immunoassay formats. To study inflammatory diseases, protein arrays can be used to analyze changes in plasma protein levels, such as cytokines, chemokines, soluble receptors, and matrix metalloproteinases. Using the bead-based Luminex system, multiplexed sandwich immunoassays have been developed to analyze the plasma concentrations of soluble receptors: sTNF-RI, sTNF-RII, sIL-2R, sgp130, sFas, sRAGE, sE-selectin, sICAM-1, sVCAM-1, sMIF-1 and sFasL. This newly established 11-plex soluble receptors assay demonstrated acceptable intra-assay and inter-assay precision, appropriate accuracy, and no crossreactivity between analytes. Using this assay, 100 plasma samples derived from 36 critically ill intensive care unit (ICU) patients with trauma or sepsis were analyzed for their soluble receptor plasma concentrations. Results obtained allowed grouping of patients' samples into a trauma and a sepsis group. Four candidate molecules: sFas, sICAM-1, sTNF-RI, and sTNF-RII had higher concentrations in patients with sepsis than in those with trauma, contributing the highest discriminatory values to define the nature of the inflammatory disease originating from pathogen-involved (sepsis) or pathogen-independent inflammation.

Research paper thumbnail of Changes in the Novel Orphan, C5a Receptor (C5L2), during Experimental Sepsis and Sepsis in Humans

The Journal of Immunology, 2005

Sepsis is associated with extensive complement activation, compromising innate immune defenses, e... more Sepsis is associated with extensive complement activation, compromising innate immune defenses, especially in neutrophils (PMN). Recently, a second C5a receptor (C5L2) was detected on PMN without evidence of intracellular signaling. The current study was designed to determine changes in C5L2 in blood PMN during sepsis. In vitro exposure of PMN to C5a, but not to fMLP, led to reduced content of C5L2. Following cecal ligation and puncture-induced sepsis in rats, PMN demonstrated a time-dependent decrease in C5L2. In vivo blockade of C5a during experimental sepsis resulted in preservation of C5L2. Similarly, PMN from patients with progressive sepsis showed significantly reduced C5L2 expression (n ‫؍‬ 26), which was virtually abolished in patients who developed multiorgan failure (n ‫؍‬ 10). In contrast, sepsis survivors exhibited retention of C5L2 (n ‫؍‬ 12/13). The data suggest that C5L2 on PMN diminishes during sepsis due to systemic generation of C5a, which is associated with a poor prognosis.

Research paper thumbnail of Enhancement of neutrophil function by in vivo filgrastim treatment for prophylaxis of sepsis in surgical intensive care patients

Journal of Critical Care, 1995

Research paper thumbnail of P1530 CMV reactivation and antiviral immune response in patients with septic shock

International Journal of Antimicrobial Agents, 2007

Research paper thumbnail of Hepato-splanchnic metabolic effects of the stable prostacyclin analogue iloprost in patients with septic shock

Intensive Care Medicine, 2001

Research paper thumbnail of CD64 surface expression on neutrophils is transiently upregulated in patients with septic shock

Intensive Care Medicine, 2001

Research paper thumbnail of Stereoselective Suppression of Neutrophil Function by Ketamine?

Immunopharmacology and Immunotoxicology, 1995

The effects of the commercially available ketamine preparation (Ketanest), the ketamine racemate ... more The effects of the commercially available ketamine preparation (Ketanest), the ketamine racemate and of the two enantiomers, the R(-)-racemate and the S(+)-racemate, as well as its drug-free solvent were examined by N-formyl-methionyl-leucyl-phenylalanine-(FMLP)- and zymosan-induced oxygen radical production of polymorphonuclear cells (PMN). The racemate and the two enantiomers of ketamine suppressed FMLP- and zymosan-induced chemiluminescence of PMN in a dose-dependent fashion to the same extent. Therefore suppression of chemiluminescence of PMN by ketamine does not result from a specific receptor interaction.

Research paper thumbnail of Effects of sedatives and their solvents on the neutrophil granulocyte function

Journal of Huazhong University of Science and Technology [Medical Sciences], 1994

We have examined the effects of commercially available preparations and drug-free solvents of Mid... more We have examined the effects of commercially available preparations and drug-free solvents of Midazolam (Dormicum) and Propofol (Disoprivan) by N-formyl-methionyl-leucyl-phenylalanine (FMLP) -induced polymorphonuclear cell (PMN) chemiluminescence system. In the case of propofol and drug-free solvent, PMN chemiluminescence was suppressed. With Midazolam, only the active drug depressed PMN chemiluminescence.