Gábor Woth

Dynamic changes of matrix metalloproteinases and their tissue inhibitors in severe sepsis

PURPOSE Little is known about the dynamic changes of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in sepsis. Our aim was therefore to investigate the time course of MMPs and their inhibitors in patients experiencing severe sepsis. METHODS Our prospective controlled analysis included 38 patients with severe sepsis. Plasma levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured daily at a 5-day-long period with enzyme-linked immunosorbent assay. Seventeen healthy volunteers were invited as controls. RESULTS MMP-2 showed no difference compared to controls, whereas significantly elevated MMP-9 levels were detected on admission (P < .005). Significantly elevated but declining TIMP-1 levels were measured during the whole trial (P < .002-.004). Except for the second day, TIMP-2 levels were significantly lower than controls (P < .05-.009). MMP2/TIMP-1 ratios were significantly lower in septic patients (P < .03-.006), whereas MMP-2/TIMP-2 ratios were elevated throughout our study (P < .03-.006). MMP-9/TIMP-1 ratios were significantly lower at the first 3 days (P < .05-.008). MMP-9/TIMP-2 was significantly elevated on admission (P < .006). CONCLUSIONS Our research is the first follow-up study dealing with MMPs, TIMPs, and their ratios in severe sepsis. Our results indicate that MMPs and TIMPs may play a crucial role in severe sepsis, especially TIMP-1, MMP-9, and possibly TIMP-2, after an extensive study.

Microparticles and acute renal dysfunction in septic patients

PURPOSE The role of microparticles (MPs) in the pathogenesis of sepsis is not completely elucidated. We aimed to assess changes in the number of MPs during severe sepsis to follow the effect of sepsis-related organ failures, particularly renal impairment, an independent mortality factor of sepsis. MATERIALS AND METHODS Thirty-seven severe septic patients and 20 controls were enrolled. Patient status as well as organ failure-related laboratory markers was followed up to 5 consecutive days. Microparticles (annexin V+ events in MP size gate) of platelet (CD41, CD42a, and PAC1), monocyte (CD14), and myeloid cell line (CD13) origin were measured using flow cytometry. RESULTS Significantly increased total MP and CD41-, CD42a-, and PAC1-positive particle numbers were found in septic patients compared with controls. Actual number of organ dysfunctions on sample collection showed no correlation with MP numbers. Septic patients with renal dysfunction showed an increase in total MP, CD41(+), and CD13(+) particle numbers on admission. Amounts of platelet-derived CD42a(+) particles from patients with sepsis-related renal injury correlated negatively with actual blood urea nitrogen and creatinine concentrations. CONCLUSION The increased numbers of platelet-derived MPs in severe septic patients emphasize the possible contribution of the hemostasis system in the development of sepsis-related renal impairments.

Activated platelet-derived microparticle numbers are elevated in patients with severe fungal (Candida albicans) sepsis

Background The treatment of severe sepsis highly depends on the identification of bacteria or fungi from blood and/or other body materials. Although widely available blood culturing and risk assessment scores are not completely reliable, current guidelines do not recommend the wide empirical use of antifungal medications based on questionable benefit or possible side-effects. We aimed to test whether platelet-derived microparticle (MP) measurements can improve the early detection of the infective agent behind sepsis. Methods Thirty-three consecutive severe septic patients from our university intensive care unit were included in our prospective study. MP number and surface antigen characteristics were followed by flow cytometry on days 1 (admission), 3 and 5. For microbiological identification, various specimens were collected on admission and in case of overall status deterioration. Results On admission, septic patients showed elevated annexin V and constitutive platelet marker (CD41)-positive MP numbers compared with volunteers. Mixed fungal septic patients showed significantly elevated annexin V and CD41-positive particle numbers on day 1 (P < 0.05) compared with the non-fungal septic group. Adhesive platelet marker (CD42a) harbouring vesicles were negligible in the non-fungal group, while fungal septic patients showed significantly elevated numbers in all measurements (P < 0.01). Particles from activated platelets (PAC1) had elevated numbers in the first and fifth study days compared with non-fungal septic patients (P < 0.05). Conclusions The measurement of CD42a- and PAC1-positive microparticles may provide important additional information which can help to improve the early instalment of antifungal therapy of severe septic patients.

The effect of Na-selenite treatment on the oxidative stress–antioxidants balance of multiple organ failure

PURPOSE Our study tested the hypothesis that sodium (Na)-selenite expression treatment can reduce oxidative stress and increase plasma antioxidants, whereas modulating white blood cell antigen expression in severe sepsis. Selenite is a well known cofactor of glutathione peroxidases and other antioxidant enzymes; therefore, one may expect an antioxidant effect of treatment. MATERIALS We randomized 40 severe septic patients into treatment and control groups. Treatment group (n = 21) received 1000-μg/2 hours Na-selenite load, followed by a 1000-μg/die medication. Oxidative stress markers, including malondialdehyde, maximal free radical production, and plasma antioxidants: free sulfhydryl groups, glutathione levels, and superoxide dismutase and catalase enzyme activity were measured. RESULTS According to our results, the treatment regime successfully restored serum selenium levels. Treatment group developed a significant malondialdehyde increase by the fifth study day, whereas reactive oxygen species production decreased significantly. Reduced glutathione and plasma sulfhydryl groups showed no significant difference. Treatment group showed deteriorated expression of CD11a and slight increase of CD49d expression on monocytes throughout our study. CONCLUSIONS Although our Na-selenite treatment regime successfully restored the selenium deficiency of severe septic patients, antioxidant and white blood cell antigen expression modulating effect of the therapy was not observed in our patient group.

Intracranial volume inversely correlates with serum 25(OH)D level in healthy young women

Abstract Objectives Vitamin D is important in normal brain development. In animals low vitamin D level is associated with brain morphological alterations including enlargement of the brain. Whether a similar association exists in humans is unknown. Here we investigated the relationship between vitamin D and total intracranial volume as well as total volume of the cortical grey and cerebral white matter and that of the ventricles in young healthy women. Methods To assess volumes we applied semi-automatic user-independent MR volumetry. For the vitamin D measurements automated electrochemiluminescence immunoassay was used. Results We found a significant negative correlation between vitamin D and total intracranial volume as well as total cortical grey and cerebral white matter volumes. Discussion This association may reflect a trait-like relationship between vitamin D and brain size possibly determined in early development.

Comparison of VividTrac®, Airtraq®, King Vision®, Macintosh Laryngoscope and a Custom-Made Videolaryngoscope for difficult and normal airways in mannequins by novices

BackgroundDirect laryngoscopy remains the gold standard for endotracheal intubation and is preferred by experienced operators. However, an increasing number of reports currently support videolaryngoscopy, particularly for novice users. The widespread use of videolaryngoscopy may be limited due to financial limitations, especially in low-income countries. Therefore, affordable single-use scopes are now becoming increasingly popular. We sought to compare these new scopes with direct laryngoscopes and the previously tested videolaryngoscopes in mannequins by novices.MethodsFifty medical students were recruited to serve as novice users. Following brief, standardized training, students were asked to execute endotracheal intubation with each of the devices, including the Airtraq®, a custom-made videolaryngoscope, the King Vision®, the Macintosh laryngoscope and the VividTrac®, on an airway trainer (Laerdal Airway Management Trainer®) in normal and difficult airway scenarios. We evaluated the time to and the proportion of successful intubation, the best view of the glottis, esophageal intubation, dental trauma and user satisfaction.ResultsWe observed no differences in esophageal intubation. However, intubation-related times, the view of the glottis and operator satisfaction were significantly better throughout the study with the commercial videolaryngoscopes. In comparison, the custom-made videolaryngoscope performance proved to be similar to that of the Macintosh laryngoscope. The VividTrac® performance was similar (P > 0.05) or significantly better than that of the King Vision® in both scenarios.ConclusionsBased upon our results, the Airtraq®, King Vision® and VividTrac® were superior to the Macintosh laryngscope in both normal and difficult airway scencarios for novice users. In particular, our study is the first to report that the VividTrac® shows promise for further clinical evaluation.

Perioperative time course of matrix metalloproteinase-9 (MMP-9), its tissue inhibitor TIMP-1 & S100B protein in carotid surgery

Background & objectives: Ischaemic stroke is a life burdening disease for which carotid endarterectomy (CEA) is considered a gold standard intervention. Pro-inflammatory markers like matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) and S-100 Beta (S100B) may have a role in the early inflammation and cognitive decline following CEA. This study was aimed to describe the perioperative time courses and correlations between of MMP-9, TIMP-1 and S100B following CEA. Methods: Fifty four patients scheduled for CEA were enrolled. Blood samples were collected at four time points, T1: preoperative, T2: 60 min after cross-clamp release, T3: first postoperative morning, T4: third postoperative morning. Twenty atherosclerotic patients were included as controls. Plasma MMP-9, TIMP-1 and S100B levels were estimated by ELISA. Results: TIMP-1 was decreased significantly in the CEA group (P<0.01). Plasma MMP-9 was elevated and remained elevated from T1-4 in the CEA group (P<0.05) with a marked elevation in T3 compared to T1 (P<0.05). MMP-9/TIMP-1 was elevated in the CEA group and increased further by T2 and T3 (P<0.05). S100B was elevated on T2 and decreased on T3-4 compared to T1. Interpretation & conclusions: Our study provides information on the dynamic changes of MMP-9-TIMP-1 system and S100B in the perioperative period. Preoperative reduction of TIMP-1 might be predictive for shunt requirement but future studies are required for verification.

Comparison and evaluation of seven different bench-top flow cytometers with a modified six-plexed mycotoxin kit.

Many bench‐top flow cytometers (b‐FCs) are compatible with microsphere‐based multiplexed assays. Disciplines implementing b‐FCs–based assays are expanding; they include monitoring and validating food quality. A multiplexed platform protocol was evaluated for poly‐mycotoxin assays, which is compatible with a variety of b‐FC models. The seven instruments included: BD FACSCalibur™, BD FACSArray™ Bioanalyzer, Accuri C6, Partec CyFlow® Space, Beckman Coulter FC 500, Guava EasyCyte Mini, and Luminex 100 ™. Current reports related to the food industry describe fungal co‐infections leading to poly‐mycotoxin contamination in grain (Sulyok M, Berthiller F, Krska R, Schuhmacher R, Rapid Commun Mass Spectrom 2006;20:2649–2659). It is imperative to determine whether b‐FC–based assays can replace traditional single‐mycotoxin enzyme‐linked immunosorbent assay (ELISA). A six‐plexed poly‐mycotoxin kit was tested on seven different b‐FCs. The modified kit was initially developed for the BD FACSArray™ Bioanalyzer (BD Biosciences) (Czeh A, Mandy F, Feher‐Toth S, Torok L, Mike Z, Koszegi B, Lustyik G, J Immunol Methods 2012;384:71–80). With the multiplexed platform, it is possible to identify up to six mycotoxin contaminants simultaneously at regional grain collection/transfer/inspection facilities. In the future, elimination of contaminated food threat may be better achieved with the inclusion of b‐FCs in the food protection arsenal. A universal protocol, matched with postacquisition software, offers an effective alternative platform compared to using a series of ELISA kits. To support side‐by‐side evaluation of seven flow cytometers, an instrument‐independent fluorescence emission calibration was added to the protocol. All instrument performances were evaluated for strength of agreement based on paired sets of evaluation to predicate method. The results suggest that all b‐FCs were acceptable of performing with the multiplexed kit for five of six mycotoxins. For OTA, the detection sensitivity was consistent only for five of the seven instruments.

Time courses of changes of para-, meta-, and ortho-tyrosine in septic patients: A pilot study

Objectives: Sepsis is associated with oxidative stress. Due to oxidative stress, three tyrosine isoforms, para-, meta-, and ortho-tyrosine (p-, m-, and o-Tyr), can be formed non-enzymatically in smaller amounts. p-Tyr is mainly formed physiologically in the kidneys through the activity of the phenylalanine hydroxylase enzyme. The three tyrosine isoforms may undergo different renal handling. Methods: Twenty septic patients were involved in the study and 25 healthy individuals served as controls. Blood and urine levels of p-, m-, and o-Tyr were measured on admission and four consecutive days. Results: Serum m-Tyr levels were higher in septic patients than in controls on days 2 (P = 0.031) and 3 (P = 0.035). Serum p-Tyr levels were lower in the cases than in controls on days 1 (P = 0.005) and 2 (P = 0.040), and subsequently normalized due to a day-by-day elevation (P = 0.002). The tendency of urinary m-Tyr concentration was decreasing (P = 0.041), while that of urinary p-Tyr concentration was increasing (P = 0.001). Fractional excretion of m-Tyr (FEm-Tyr) showed a decreasing tendency (P = 0.009), and was, on all days, higher than FEp-Tyr, which remained near-normal, less than 4%. Procalcitonin showed significant correlation with FEm-Tyr (r = 0.454; P < 0.001). Discussion: Our data suggest that the oxidative stress marker m-Tyr and physiologic p-Tyr may be handled differently in septic patients. The excretion of m-Tyr correlates with inflammation. m-Tyr may be actively secreted or produced in the kidney in some patients, whereas the decreased serum level of p-Tyr is a consequence of diminished renal production and not of renal loss.

Effects of therapeutic hypothermia and kinetics of serum protein S100B after cardiopulmonary resuscitation

Introduction. Post-resuscitation care is regulated by international guidelines. A milestone of these is the application of therapeutic hypothermia (TH). The aims of our study were: to determine the 30-day-mortality for our patients, to monitor the efficacy and effects of TH, and to investigate serum protein S100B – as an early prognostic marker.Materials and Methods. In our study, 57 patients, treated after cardiopulmonary resuscitation (CPR) on a multidisciplinary intensive care unit, were included. Patients were divided into groups who received and who didn’t receive TH. 30-day-mortality was determined as an end-point. Effects of TH were monitored using statistical analysis according to clinical parameters and laboratory tests. Serum protein S100B levels were measured with ELISA technique on 20 randomised patients atadmission and the 1st, 3rd and 5th day after CPR. Results. Total 30-day-mortality was 74%. TH did not reduced the 30-day-mortality (73% vs. 74%, p>0.05). We found a significant correlation between TH and serum lactate concentration after admission (0h,p=0.006) and at 12 (p=0.045) and 36 (p=0.049) hours after CPR. On the3rd (p=0.005) and 4th (p=0.043) day after CPR, as a result of TH, platelet count was significantly higher compared to normothermic samples. There was no significant difference in protein S100B levels between the normothermic and TH group and protein S100B levels did not correlate with 30-day-mortality.Conclusion. Despite recommendations of international guidelines, we cannot prove the beneficial effect of TH, or a correlation of protein S100B levels with a positive outcome.