The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models.

A better understanding of the function of neutrophil extracellular traps (NETs) may facilitate the development of interventions for sepsis. The study aims to investigate the formation and degradation of NETs in three murine sepsis models and to analyze the production of reactive oxygen species (ROS) during NET formation. Murine sepsis was induced by midgut volvulus (720° for 15 min), cecal ligation and puncture (CLP), or the application of lipopolysaccharide (LPS) (10 mg/kg body weight i.p.). NET formation and degradation was modulated using mice that were genetically deficient for peptidyl arginine deiminase-4 (PAD4-KO) or DNase1 and 1L3 (DNase1/1L3-DKO). After 48 h, mice were killed. Plasma levels of circulating free DNA (cfDNA) and neutrophil elastase (NE) were quantified to assess NET formation and degradation. Plasma deoxyribonuclease1 (DNase1) protein levels, as well as tissue malondialdehyde (MDA) activity and glutathione peroxidase (GPx) activity, were quantified. DNase1 and DNase1L3 in liver, intestine, spleen, and lung tissues were assessed. The applied sepsis models resulted in a simultaneous increase in NET formation and oxidative stress. NET formation and survival differed in the three models. In contrast to LPS and Volvulus, CLP-induced sepsis showed a decreased and increased 48 h survival in PAD4-KO and DNase1/1L3-DKO mice, when compared to WT mice, respectively. PAD4-KO mice showed decreased formation of NETs and ROS, while DNase1/1L3-DKO mice with impaired NET degradation accumulated ROS and chronicled the septic state. The findings indicate a dual role for NET formation and degradation in sepsis and ischemia-reperfusion (I/R) injury: NETs seem to exhibit a protective capacity in certain sepsis paradigms (CLP model), whereas, collectively, they seem to contribute adversely to scenarios where sepsis is combined with ischemia-reperfusion (volvulus).

Barriers and prospects for skin grafting in burn treatment across African countries.

INTRODUCTION: The current standard management of full-thickness or deep dermal burns is early tangential excision and skin grafting. A conservative approach to deep burns without the option of skin grafting results in delayed wound healing, possibly leading to wound infection and is associated with hypertrophic scarring and increased morbidity and mortality. The aim of this study was to improve the understanding of the management and availability to perform skin grafting for burns on the African continent. It also sought to identify challenges and perceived improvements. METHODS: A web-based, structured, closed-formatted, multinational survey was designed to gather information on the current state and availability of skin grafting of burn wounds on the African continent. The questionnaire consisted of 27 questions, available in English and French. It was reviewed within the GAP-Burn collaboration network and sent to 271 health care professionals who had participated in a previous study and had initially been recruited by means of the snowball system. RESULTS: The questionnaire was completed 84 times (response rate: 31.0%), of which 3 were excluded. Responses originated from 22 African countries. The majority 71 (87.7%) resulted from countries with a low Human Development Index (HDI), 7 (8.6%) from medium HDI countries. Split thickness skin grafting (STSG) is performed in 51 (63.0%) centers. The majority considers STSG to reduce length of stay (72.8%) and improve scarring (54.3%), yet some indicated that STSG is associated with increased risk of donor site infection (8.6%) and severe bleeding (7.4%). Factors preventing increased grafting included lack of equipment and training. CONCLUSION: Skin grafting is not performed in a significant number of hospitals treating burns. The majority of the staff believe that more skin grafting would lead to a better outcome. Advocacy and improved infrastructure, human resources coupled with introduction to well-structured health coverage for all in African countries could help to better access and affordability in burn care.

As time goes by - Overlooking 40 years of inpatient burn treatment at a national pediatric burn center in Switzerland.

BACKGROUND: Despite extensive prevention programs, burns remain a frequent cause of injury in Switzerland with a known age peak in children. Pediatric burns may cause substantial morbidity, a psyochological burden and therapy related high economic costs. To improve preventive measures, precise knowledge of etiology and treatment of pediatric burns in Switzerland as well as their temporal evolution is indispensable. METHODS: The present retrospective analysis included pediatric burn patients admitted for acute treatment to the Pediatric Burn Center of the University Children`s Hospital Zurich over the last four decades. Sociodemographic, injury related, and treatment related data were extracted from medical records. Linear regression analysis was applied to determine temporal changes during the past four decades and chi-square and t-tests were applied wherever applicable. RESULTS: A total of 3425 acute burn patients were included in the study between 1977 and 2020, corresponding to a mean of 89 patients/year. Mean age was 3.60 ± 4.12 years, three quarters of all patients were preschool children (0-5 years) and mean total body surface area (TBSA) burned was 8.01% ± 9.57%, however only around one fifth had severe burns (>10% TBSA). Scald burns (65.31%) and flame burns (32.99%) were most commonly seen. Linear regression analysis showed the total number of thermal injuries treated at our center to have increased significantly as of 2004 (p < 0.001). Separate analysis showed the same for small and medium (<10% TBSA) burns (p < 0.001), whereas the number of severe burns did not increase significantly. Length of stay (LOS) was highly associated with %TBSA burned. The percentage of female patients amongst all patients increased over time (p = 0.012). LOS per TBSA burned decreased significantly (p < 0.001). CONCLUSION: The present data show pediatric burns to remain a major health burden in Switzerland, especially small and medium burns in preschool children. Prevention programs should focus on this age population as well as on scald and flame burns as most common etiologies. The observed decrease in length of stay suggests a major improvement in overall quality of care in pediatric burns and supports centralization of care.

Murine scald models characterize the role of neutrophils and neutrophil extracellular traps in severe burns.

Introduction: Severe burns cause unique pathophysiological alterations especially on the immune system. A murine scald model was optimized as a basis for the understanding of immunological reactions in response to heat induced injury. The understanding of the roles of neutrophil extracellular traps (NETs) and DNases will support the development of new surgical or pharmacological strategies for the therapy of severe burns. Methods: We studied C57BL/6 mice (n=30) and employed four scalding protocols with varying exposure times to hot water. An additional scald group with a shorter observational time was generated to reduce mortality and study the very early phase of pathophysiology. At 24h or 72h, blood was drawn and tissue (wound, liver, lung, spleen) was analyzed for the presence of NETs, oxidative stress, apoptosis, bacterial translocation, and extracellular matrix re-organization. In addition, we analyzed the transcriptome from lung and liver tissues. Results: Exposure to hot water for 7s led to significant systemic and local effects and caused considerable late mortality. Therefore, we used an observation time of 24h in this groups. To study later phases of burns (72h) an exposure time of 6s is optimal. Both conditions led to significant disorganization of collagen, increased oxidative stress, NET formation (by immunodetection of H3cit, NE, MPO), apoptosis (cC3) and alterations of the levels of DNase1 and DNase1L3. Transcriptome analysis revealed remarkable alterations in genes involved in acute phase signaling, cell cohesion, extracellular matrix organization, and immune response. Conclusion: We identified two scald models that allow the analysis of early (24h) or late (72h) severe burn effects, thereby generating reproducible and standardized scald injuries. The study elucidated the important involvement of neutrophil activity and the role of NETs in burns. Extensive transcriptome analysis characterized the acute phase and tissue remodeling pathways involved in the process of healing and may serve as crucial basis for future in-depth studies.

State of burns management in Africa: Challenges and solutions.

INTRODUCTION: Understand the availability of human resources, infrastructure and medical equipment and perceived improvement helps to address interventions to improve burn care. METHODS: Online survey covering human resources, infrastructure, and medical equipment of burn centers as well as perceived challenges and points for improvement. The survey was distributed in English and French via snowball method. Descriptive statistics and AI-based technique random forest analysis was applied to identify determinants for a reduction of the reported mortality rate. RESULTS: 271 questionnaires from 237 cities in 40 African countries were analyzed. 222 (81.9 %) from countries with a very low Human Development Index (HDI) (4th quartile). The majority (154, 56.8 %) of all responses were from tertiary health care facilities. In only 18.8 % (n = 51) therapy was free of charge for the patients. The majority (n = 131, 48.3 %) had between 1 and 3 specialist doctors (n = 131, 48.3 %), 1 to 3 general doctors (n = 138, 50.9 %) and more than 4 nurses (n = 175, 64.6 %). A separate burn ward was available in 94 (34.7 %) centers. Regular skin grafting was performed in 165 (39.1 %) centers. Random forest-based analysis revealed a significant association between HDI (feature importance: 0.38) and mortality. The most important reason for poor outcome was perceived late presentation (212 institutions, 78.2 %). The greatest perceived potential for improvement was introduction of intensive care units (229 institutions, 84.5 %), and prevention or education (227 institutions, 83.7 %). INTERPRETATION: A variety of factors, including a low HDI, delayed hospital presentation e.g. due to prior care by non-physicians and lack of equipment seem to worsen the outcome. Introduction of an intensive care unit and communal education are perceived to be important steps in improving health care in burns.

Midgut Volvulus Adds a Murine, Neutrophil-Driven Model of Septic Condition to the Experimental Toolbox.

BACKGROUND: Severe infections that culminate in sepsis are associated with high morbidity and mortality. Despite continuous efforts in basis science and clinical research, evidence based-therapy is mostly limited to basic causal and supportive measures. Adjuvant therapies often remain without clear evidence. The objective of this study was to evaluate the septic volvulus ischemia-reperfusion model in comparison to two already established models and the role of neutrophil extacellular traps (NETs) in this model. METHODS: The technique of the murine model of midgut volvulus was optimized and was compared to two established models of murine sepsis, namely cecal ligation and puncture (CLP) and intra-peritoneal (i.p.) injection of lipopolysaccharide (LPS). RESULTS: Midgut volvulus for 15 min caused a comparable mortality (38%) as CLP (55%) and peritoneal LPS injection (25%) at 48 h. While oxidative stress was comparable, levels of circulating free DNA (cfDNA), and splenic/hepatic and pulmonary translocation of bacteria were decreased and increased, respectively at 48 h. DNases were increased compared to the established models. Proteomic analysis revealed an upregulation of systemic Epo, IL-1b, Prdx5, Parp1, Ccl2 and IL-6 at 48 h in comparison to the healthy controls. DISCUSSION AND CONCLUSION: Midgut volvulus is a stable and physiological model for sepsis. Depending on the duration and subsequent tissue damage, it represents a combination of ischemia-reperfusion injury and hyperinflammation.

Are We Hitting the Target?: The Effectiveness of Distinct Antibiotic Regimens in Pediatric Appendectomies.

BACKGROUND: Currently, a range of different antibiotics are administered perioperatively during appendectomy surgery in the pediatric population. One reason for the lack of a uniform treatment regimen is the paucity of large studies on microbiologic data and susceptibility patterns. METHODS: A retrospective, monocentric study on antibiotic susceptibility, including 1258 pediatric patients undergoing appendectomy from 2012 to 2020 was performed. Predictors for prolonged length of stay (LOS) and complications were determined using regression analysis, and objective criteria for broad-spectrum antibiotic regimens were identified. Simulations of the effectiveness of different antibiotic regimens against identified bacterial susceptibility results were conducted. RESULTS: The most important predictors for prolonged LOS and an increased incidence of infectious complications are elevated preoperative C-reactive protein levels, high intraoperative severity gradings and insufficient primary antibiotic treatment. Based on our results, a simple antibiotic treatment, consisting of cefuroxime-metronidazole (C-M) or ampicillin-sulbactam (AS), is sufficient in around 85% of all patients; both options being equally efficient. In severe appendicitis, the likelihood of resistance is 31% and 32% for C-M and AS, respectively. For piperacillin-tazobactam and C-M-tobramycin, the incidence of bacterial resistance decreases to 12%. Resistance is reduced to 8% for AS-tobramycin treatment. The patient populations benefiting most from an extended antibiotic regimen are those with a high intraoperative severity grading. CONCLUSIONS: Evidence suggests that an insufficient antibiotic regimen is associated with a prolonged LOS and a higher rate of infectious complications, justifying the expansion of a simple antibiotic regimen in cases of severe intraabdominal findings. In all other cases, a simple antibiotic treatment with AS is recommended.

Fetal-Maternal Surgery for Spina Bifida in a HIV-Infected Mother.

INTRODUCTION: In select cases, in utero surgery for myelomeningocele (MMC) leads to better outcomes than postnatal repair. However, maternal HIV infection constitutes a formal exclusion criterion due to the potential of vertical HIV transmission. Encouraged by a previous case of a successful fetal spina bifida repair in a Hepatitis Bs antigen-positive woman, a plan was devised allowing for fetal surgery. CASE REPORT: In utero MMC repair was performed although the mother was HIV-infected. To minimize the risk of in utero HIV transmission, the mother was treated by highly active antiretroviral therapy throughout gestation as well as intravenous zi-dovudine administration during maternal-fetal surgery. The mother tolerated all procedures very well without any sequelae. The currently 20 month-old toddler is HIV negative and has significantly benefitted from fetal surgery. DISCUSSION/CONCLUSION: This case shows that maternal HIV is not a priori a diagnosis that excludes fetal surgery. Rather, it might be a surrogate for moving towards personalized medicine and away from applying too rigorous exclusion criteria in the selection of candidates for maternal-fetal surgery.

[Impact and Temporal Evolution of the Corona Pandemic on Pediatric Emergency Care]. / Auswirkungen der Corona Pandemie auf die Notfallversorgung im Kindesalter und die zeitliche Entwicklung im Verlauf der Pandemie.

HINTERGRUND: Die COVID-19 Pandemie hat zu massiven gesellschaftlichen und wirtschaftlichen Einschränkungen geführt. Im Kindesalter wurden elektive Vorsorgeuntersuchung oftmals nicht wahrgenommen und es zeigte sich eine Reduktion der Zahl der Notfallvorstellungen. MATERIAL UND METHODEN: In einer retrospektiven Studie erfolgte eine quantitative Auswertung aller Notfallpatienten der 5 Kindernotaufnahmen Hamburgs und der Kinder- und Jugendarztpraxen der Jahre 2019 und 2020. Zusätzlich erfolgte die detaillierte Analyse der Notfallbehandlungen des Altonaer Kinderkrankenhauses während der ersten Phase der Pandemie im Vergleich zum Vorjahr. Zusammenhänge zu den Eckpunkten der Pandemieentwicklung wurden analysiert. Die Berechnung signifikanter Unterschiede in Bezug auf die demographischen Daten und Krankheitsbilder erfolgte mittels Chi- Quadrat und t-Test. ERGEBNISSE: In allen 5 pädiatrischen Notaufnahmen Hamburgs zeigte sich eine nahezu gleichförmige Abnahme der Patientenvorstellungen während der Pandemie, ähnlich in den Kinder- und Jugendarztpraxen. Die Zahl der Behandlungen verhielt sich dabei nicht streng gegenläufig zu den Neuinfektionen, sondern korreliert eher mit den Mobilitätsdaten und entsprach somit den gesamtgesellschaftlichen Veränderungen. Während der Pandemie stieg der Anteil häuslicher Unfälle an der Gesamtzahl der Vorstellungen signifikant an. Die Analyse der Diagnosen zeigte unter anderem eine relative Abnahme von (viralen) Infektionskrankheiten. Nicht behandlungspflichtige Erkrankungen führten in der Pandemie seltener zur Vorstellung. Diese Phänomene entlasteten einerseits die Notaufnahmen, bergen aber auch die Gefahr, dass Erkrankungen durch Eltern falsch eingeschätzt werden und eine notwendige ärztliche Vorstellung somit zu spät erfolgt.

Patient- and Physician-Reported Outcome of Combined Fractional CO2 and Pulse Dye Laser Treatment for Hypertrophic Scars in Children.

BACKGROUND: Hypertrophic scars are commonly seen in children and associated with pruritus, pain, functional impairment, and aesthetic disfigurement. Ablative fractional CO2 and pulse dye laser are emerging techniques to improve scar quality. Only limited data are available on children, nonburn scars, and patient-reported outcome. We aimed to investigate safety and outcome of repeated laser therapy for hypertrophic scars originating from burns and other conditions by means of patient- and physician-reported outcome measures. METHODS: This was a retrospective before-after analysis of laser treatments in children with hypertrophic scars. Outcome was measured using Patient and Observer Scar Assessment Scale, Vancouver Scar Scale and Itch Man Scale. With respect to safety, laser- and anesthesia-related complications were analyzed. RESULTS: Seventeen patients, aged 11.37 ± 4.82 years with 27 scars, underwent 102 distinct laser treatments, mainly combined CO2 and pulse dye laser (94%), with few CO2 only (6%). Vancouver Scar Scale total score before the first and after the first session decreased significantly from 7.65 ± 2.12 to 4.88 ± 1.73; Patient and Observer Scar Assessment Scale observer overall opinion also dropped from 5.88 ± 1.57 to 4.25 ± 1.70. Pruritus improved significantly. Patient age and timing of laser intervention did not have an impact on treatment response. Complications related to laser treatment were seen in 2% (wound infection, n = 2) and to anesthesia in 4% (insignificant n = 2, minor n = 1). CONCLUSIONS: Combined laser therapy significantly improves quality, pain, and pruritus of hypertrophic scars in children. When provided by experienced laser and anesthesia teams, it is safe with a low rate of complications.

Comparative analysis of functional and aesthetic outcomes of retroauricular full thickness versus plantar glabrous split thickness skin grafts in pediatric palmar hand burns.

BACKGROUND: Optimal management of palmar hand burns in children is controversial. We aimed to compare function and aesthetics of retroauricular full thickness skin grafts (FTSG) to plantar glabrous split thickness skin grafts (STSG). METHODS: 32 palmar grafts in paediatric burn patients were analysed: 19 retroauricular FTSG (group 1) and 13 thick plantar glabrous STSG (group 2). The latter were harvested at a thickness of 0.5 mm. The resulting plantar donor defects were covered with a STSG from the scalp, a sequential surgical technique we termed the "Zurich move". Clinical examination, Cutometer and Colorimeter assessment and validated patient and observer questionnaires were used. Donor site complications and subjective complaints were recorded. RESULTS: Colorimeter results were superior in group 2 with an erythema score of 5.73 ± 2.64 (group 1) versus 2.33 ± 1.97 (group 2, p < 0.001) and a pigmentation score of 9.82 ± 5.42 (group 1) and 1.89 ± 1.92 (group 2, p < 0.001). Observers` scar evaluation using VSS and POSAS showed significantly superior results in group 2 for almost all items. Conversely, group 1 grafts were less stiff with mean normalized tissue extension R0 of 0.80 ± 0.21 versus 0.57 ± 0.24 in group 2 grafts (p < 0.05). In both groups donor sites complications were rare. CONCLUSION: Plantar glabrous STSG showed superior functional and aesthetic results when compared to FTSG in pediatric palmar hand burns. In addition, the "Zurich Move" is safe and provides uncomplicated donor site healing on the scalp and the foot allowing rapid restoration of full function.

Using Artificial Intelligence to Obtain More Evidence? Prediction of Length of Hospitalization in Pediatric Burn Patients.

Background: It is not only important for counseling purposes and for healthcare management. This study investigates the prediction accuracy of an artificial intelligence (AI)-based approach and a linear model. The heuristic expecting 1 day of stay per percentage of total body surface area (TBSA) serves as the performance benchmark. Methods: The study is based on pediatric burn patient's data sets from an international burn registry (N = 8,542). Mean absolute error and standard error are calculated for each prediction model (rule of thumb, linear regression, and random forest). Factors contributing to a prolonged stay and the relationship between TBSA and the residual error are analyzed. Results: The random forest-based approach and the linear model are statistically superior to the rule of thumb (p < 0.001, resp. p = 0.009). The residual error rises as TBSA increases for all methods. Factors associated with a prolonged LOS are particularly TBSA, depth of burn, and inhalation trauma. Conclusion: Applying AI-based algorithms to data from large international registries constitutes a promising tool for the purpose of prediction in medicine in the future; however, certain prerequisites concerning the underlying data sets and certain shortcomings must be considered.

A Prospective Controlled Study on Long-Term Outcomes of Facial Lacerations in Children.

Background: Although skin adhesives have been used for decades to treat skin lacerations, uncertainty remains about long-term results, and complications. Methods: In this prospective, controlled, single-blinded, observational cohort study, outcomes were assessed by five plastic surgeons with standardized photographs at 6-12 months using a modified Patient and Observer Scar Assessment Scale (POSAS) and Vancouver Scar Scale (VSS); additionally, the POSAS was performed by the patients/caregivers and the physician; pain, requirement of anesthesia, treatment time, costs, complications, and quality of live (QoL) were assessed. Results: A total of 367 patients were enrolled; 230 were included in the main analysis; 96 wounds were closed using tissue adhesives (group 1); 134 were sutured (group 2). Assessment by the independent observers revealed an improved mean modified overall POSAS score in group 1 in comparison with group 2 [2.1, 95% CI [1.97-2.25] vs. 2.5, 95% CI [2.39-2.63]; p < 0.001, d = 0.58] and mean VSS score [1.2, 95% CI [0.981-1.34] vs. 1.6, 95% CI [1.49-1.79], p < 0.001, d = 0.53]. At the early follow-up, dehiscence rate was 12.5% in group 1 and 3.7% in group 2 (p < 0.001); later on, one dehiscence remained per group. Mild impairment of QoL was found at the early follow-up in both groups, with no impairment remaining later on. Duration of treatment and treatment costs were lower in group 1. Conclusion: Both modalities of wound closure yield favorable esthetic results, and complications are rare. Adhesives are more cost-effective, and its application is less time-consuming; therefore, tissue adhesives offer considerable advantages when used appropriately. Trial Registration: Public trial registration was performed at www.ClinicalTrials.gov (Identifier: NCT03080467).

Incidence, severity and pattern of burns in children and adolescents: An epidemiological study among immigrant and Swiss patients in Switzerland.

BACKGROUND: Despite advances in surgical management and critical care for burn surgery, morbidity and mortality of patients with severe burns remains high. Especially in the pediatric population, burns often lead to devastating consequences such as the necessity of corrective surgery until adulthood. Worldwide, 80%-90% of all severe burns occur in low to middle income countries. But also in high income countries, burns are distributed inequitably. Risk factors include age, sex, socioeconomic status, and ethnicity. AIM: The objective of this study was to determine the typical demographics and injury-related data of pediatric burn patients in order to contribute to preventive measures. Special interest was paid to the question of whether the incidence of severe burns is higher among patients with an immigration background. PATIENTS AND METHODS: Patient records of the 4373 patients admitted to the Pediatric Burn Unit of the University Children's Hospital of Zurich from 2006 to 2018 were analyzed. Demographic data and injury patterns are presented descriptively. Temporal trends concerning duration of hospitalization and setting (inpatient versus outpatient), differences in relative incidence and in burn mechanism in distinct cohorts (by nation and Human Development Index (HDI)) and seasonal trends were analyzed. Furthermore, risk factors for large burns and for (prolonged) inpatient treatment were examined using a multivariate approach. RESULTS: Temporal resolution shows considerable variation between inpatient and outpatient treatment (p>0.001) and with shorter hospital stays (p=0.004). Swiss citizens and patients with an immigration background from very highly developed countries sustain a significantly lower incidence of heat-related injuries than all others (p<0.001). The most common burn causes among all children, independent of their country of citizenship, are related to kitchen items or hot tea and coffee (35.57%±4.01% resp. 32.39%±5.95%). Logistic regression revealed that migration background from a low HDI country is significantly associated with larger burns (>5% TBSA) and with a need for inpatient treatment. CONCLUSION: The study emphasizes the need for highly specific measures of burn prevention and indicates the necessity of focusing on certain target groups who are especially vulnerable to burns, such as immigrants from less developed countries.

An effective procedure for skin stiffness measurement to improve Paediatric Burn Care.

BACKGROUND: The objective evaluation of scar quality plays a crucial role in improving burn surgery and scar rehabilitation. Suction-based skin measurements were proposed as a method to objectively determine the mechanical properties of scars, yet their use is limited, in particular for paediatric burn care. A new device was developed which provides essential advantages for scar assessment. The aim of this study was to assess its reliability, intra- and interobserver variability. METHODS: The new device, "Nimble", consists of a lightweight probe that measures the negative pressure needed to obtain a predefined tissue elevation, revealing information on the stiffness of the tissue. 29 former paediatric burn patients were included. Three observers measured the tissue stiffness of a predefined location on the scar and on healthy skin using the Nimble, and the established suction device, the Cutometer®. The reliability of both instruments in distinguishing between healthy skin and scar was assessed by means of the intraclass correlation coefficient. RESULTS: The Nimble successfully differentiated between scar tissue and healthy skin in 92%, the Cutometer in 80% of the patients (p<0.05). Inter- and intraobserver variability of the Nimble (ICCs) were excellent. For the majority of the calculated ICC values the Nimble exceeded the Cutometer®. CONCLUSION: The new device enables reliable and safe measurement of the stiffness of scars. Measurements are less susceptible to patient non-compliance and observer dependency. The Nimble might therefore constitute an easy to use tool for the systematic assessment of scars, thus supporting decision-making in paediatric burn care.

Plastic and Orthopaedic Interventions and Long-Term Sequelae in Children with Meningococcal Septicemia-40 Years of Experience at the University Children's Hospital Zurich.

INTRODUCTION: Meningococcal septicemia is not merely an acute disease with a high lethality, but patients surviving the fulminant phase can suffer from serious long-term sequelae. MATERIALS AND METHODS: The records of 165 patients admitted to the hospital from 1968 to 2008 with the diagnosis of meningococcal septicemia were retrospectively reviewed for early signs at presentation, intensive care management, acute symptoms, the necessity of plastic and orthopaedic surgical management, and long-term orthopaedic sequelae. Possible predictors of these conditions were determined. RESULTS: Overall lethality was 17.5%, and mean time of hospitalization time was 28 days. Integument involvement occurred in 45%, often resulting in the necessity of plastic surgical procedures. Young age is a significant indicator or skin involvement. Amputations were necessary in 9% of all cases, affecting the lower extremities more often than the upper extremities. Six percent of all children suffered from long-term orthopaedic sequelae, such as growth retardation and angular deformities, appearing up to 11 years (mean 4.9 years) after onset of the acute disease. The incidence of amputations and long-term orthopaedic sequelae correlated significantly with severity of the disease. CONCLUSION: Meningococcal septicemia can entail devastating long-term consequences in children surviving the acute phase of the disease. Sequelae may become apparent only years later and cause further damage. To prevent this, a systematic follow-up till adulthood is necessary.

A MALDI MS investigation of the lysophosphatidylcholine/phosphatidylcholine ratio in human spermatozoa and erythrocytes as a useful fertility marker.

The human spermatozoa membrane is characterized by a unique fatty acyl composition with significant amounts of highly unsaturated fatty acids, particularly docosahexaenoic acid (22:6), whereby phosphatidylcholine (PtdCho) (16:0/22:6) is the most abundant glycerophospholipid. The large amount of highly unsaturated fatty acyl residues is crucial for the fluidity of the membrane and, therefore, the successful fertilization process. Consequently, however, the spermatozoa are very sensitive to reactive oxygen species (ROS) that are generated under conditions of "oxidative stress" and key players in many pathological conditions. Lipid oxidation of the sperm membrane is accompanied by the loss of the oxidatively modified unsaturated residue (normally in the sn-2 position) and the generation of saturated lysophosphatidylcholine (LysoPtdCho). Although other lysolipids are also generated, LysoPtdCho is the "marker" lipid of choice due to the high abundance of PtdCho. In particular, obesity (body mass index >30 kg/m(2)) is characterized by increased ROS generation and negatively affects the reproductive potential. We will show here that the LysoPtdCho/PtdCho ratio can be easily determined by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The data found do correlate with clinical markers of sperm quality. A very interesting aspect is that the LysoPtdCho/PtdCho ratios determined in the spermatozoa extracts correlate with the LysoPtdCho/PtdCho values determined in the organic extracts of erythrocytes. Thus, there is no absolute need for a sperm investigation, but an estimation of the fertilizing ability of the corresponding male could be also made directly from the blood which is more readily available than the spermatozoa.

ADME related profiling in 96 and 384 well plate format--a novel and robust HT-assay for the determination of lipophilicity and serum albumin binding.

The failure of about half of the drug candidates is associated with poor pharmacokinetic properties leading to a huge loss of time and money [1]. Early profiling of drug like properties provides important information in order to screen out insoluble, poorly absorbed and toxic compounds. Today, large compound libraries have to be screened, and of course the total number of compounds will rise over the next years leading to a growing demand for fully automated assays. A balance between quality, speed, throughput, cost and information content can be accomplished by the careful selection of assays and experimental conditions. Here we describe a novel 384 well format assay for two important ADME related descriptors (lipophilicity and serum protein binding) as input parameters for a precise prediction of fraction absorbed, blood/organ distribution coefficients and permeability, in order to maximize the information about a compound at an early stage of discovery.