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Drug repurposing is used to propose new therapeutic perspectives. Here, we introduce "Drug Upgrade", that is, characterizing the mode of action of an old drug to generate new chemical entities and new therapeutics. We proposed a novel methodology covering target identification to pharmacology validation. As an old drug, we chose hydroxychloroquine (HCQ) for its well-documented clinical efficacy in lupus and its side effect, retinal toxicity. Using the Nematic Protein Organization Technique (NPOT®) followed by liquid chromatography-tandem mass spectrometry analyses, we identified myeloperoxidase (MPO) and alpha-crystallin β chain (CRYAB) as primary and secondary targets to HCQ from lupus patients' peripheral blood mononuclear cells (PBMCs) and isolated human retinas. Surface plasmon resonance (SPR) and enzymatic assays confirmed the interaction of HCQ with MPO and CRYAB. We synthesized INS-072 a novel analog of HCQ that increased affinity for MPO and decreased binding to CRYAB compared to HCQ. INS-072 delayed cutaneous eruption significantly compared to HCQ in the murine MRL/lpr model of spontaneous lupus and prevents immune complex vasculitis in mice. In addition, long-term HCQ treatment caused retinal toxicity in mice, unlike INS-072. Our study illustrates a method of drug development, where new applications or improvements can be explored by fully characterizing the drug's mode of action., (© 2024 Wiley Periodicals LLC.)

Background: The authors evaluated the comparative effectiveness of calabadion 2 to reverse non-depolarizing neuromuscular-blocking agents (NMBAs) by binding and inactivation., Methods: The dose-response relationship of drugs to reverse vecuronium-, rocuronium-, and cisatracurium-induced neuromuscular block (NMB) was evaluated in vitro (competition binding assays and urine analysis), ex vivo (n = 34; phrenic nerve hemidiaphragm preparation), and in vivo (n = 108; quadriceps femoris muscle of the rat). Cumulative dose-response curves of calabadions, neostigmine, or sugammadex were created ex vivo at a steady-state deep NMB. In living rats, the authors studied the dose-response relationship of the test drugs to reverse deep block under physiologic conditions, and they measured the amount of calabadion 2 excreted in the urine., Results: In vitro experiments showed that calabadion 2 binds rocuronium with 89 times the affinity of sugammadex (Ka = 3.4 × 10 M and Ka = 3.8 × 10 M-). The results of urine analysis (proton nuclear magnetic resonance), competition binding assays, and ex vivo study obtained in the absence of metabolic deactivation are in accordance with an 1:1 binding ratio of sugammadex and calabadion 2 toward rocuronium. In living rats, calabadion 2 dose-dependently and rapidly reversed all NMBAs tested. The molar potency of calabadion 2 to reverse vecuronium and rocuronium was higher compared with that of sugammadex. Calabadion 2 was eliminated renally and did not affect blood pressure or heart rate., Conclusions: Calabadion 2 reverses NMB induced by benzylisoquinolines and steroidal NMBAs in rats more effectively, i.e., faster than sugammadex. Calabadion 2 is eliminated in the urine and well tolerated in rats.

Background: Cachexia is a complex syndrome defined by weight loss due to an ongoing loss of skeletal muscle mass with or without loss of body fat. It is often associated with anorexia. Numerous results from experimental studies suggest that blockade of the melanocortin-4 receptor (MC4R) could be an effective treatment for anorexia and cachexia. In a previous study, we reported the basic pharmacological properties of a blocking anti-MC4R mAb 1E8a and its scFv derivative in vitro and in vivo., Methods: In the present study, we further characterized the mode of action of the 1E8a scFv, evaluated its pharmacokinetic properties in mice, and assessed its therapeutic potential in a lipopolysaccharide (LPS)-induced cachexia model in rats., Results: In vitro, scFv enhanced the efficacy of the endogenous inverse agonist Agouti-related protein. After intravenous (i.v.) administration in mice, the scFv penetrated the blood-brain barrier (BBB) and reached its central sites of action: the scFv brain-serum concentration ratios increased up to 15-fold which suggests an active uptake into brain tissue. In telemetry experiments, i.v. administration of the scFv in rats was well tolerated and only induced slight cardiovascular effects consistent with MC4R blockade, i.e., a small decrease in mean arterial pressure and heart rate. In the model of LPS-induced anorexia, i.v. administration of scFv 1E8a prevented anorexia and loss of body weight. Moreover, it stimulated a myogenic response which may contribute to the preservation of muscle mass in cachexia., Conclusion: The pharmacological profile of scFv 1E8a suggests its potential value in the treatment of cachexia or anorexia.

Introduction: The frequency of adverse events, such as cerebral ischemia, following traumatic brain injury (TBI) is often debated. Point-in-time monitoring modalities provide important information, but have limited temporal resolution., Purpose: This study examines the frequency of an adverse event as a point prevalence at 24 and 72 h post-injury, compared with the cumulative burden measured as a frequency of the event over the full duration of monitoring., Methods: Reduced brain tissue oxygenation (PbtO(2) < 10 mmHg) was the adverse event chosen for examination. Data from 100 consecutive children with severe TBI who received PbtO(2) monitoring were retrospectively examined, with data from 87 children found suitable for analysis. Hourly recordings were used to identify episodes of PbtO(2) less than 10 mmHg, at 24 and 72 h post-injury, and for the full duration of monitoring., Results: Reduced PbtO(2) was more common early than late after injury. The point prevalence of reduced PbtO(2) at the selected time points was relatively low (10 % of patients at 24 h and no patients at the 72-h mark post-injury). The cumulative burden of these events over the full duration of monitoring was relatively high: 50 % of patients had episodes of PbtO(2) less than 10 mmHg and 88 % had PbtO(2) less than 20 mmHg., Conclusion: Point-in-time monitoring in a dynamic condition like TBI may underestimate the overall frequency of adverse events, like reduced PbtO(2), particularly when compared with continuous monitoring, which also has limitations, but provides a dynamic assessment over a longer time period.

Melanocortin receptors (MCR) play an important role in the regulation of energy balance and autonomic function. In the present studies, we used active immunization against peptide sequences from the first and the third extracellular loop (EL1 and EL3) of the MC3R to generate selective antibodies (Abs) against this MCR subtype in rats. Immunization with the EL1 peptide resulted in Abs that enhanced the effects of the endogenous ligand α-melanocyte-stimulating hormone (α-MSH), whereas immunization with the EL3 peptide resulted in Abs acting as non-competitive antagonists. The phenotype of immunized rats chronically instrumented with telemetry transducers was studied under four different conditions: a high-fat diet was followed by standard lab chow, by fasting, and finally by an intraperitoneal injection of lipopolysaccharide (LPS). Under high-fat diet, food intake and body weight were higher in the EL3 than in the EL1 or the control group. Blood pressure was increased in EL3 rats and locomotor activity was reduced. Plasma concentrations of triglycerides, insulin, and leptin tended to rise in the EL3 group. After switching to standard lab chow, the EL1 group showed a small significant increase in blood pressure that was more pronounced and associated with an increase in heart rate during food restriction. No differences between the EL1 or the EL3 group were observed after LPS injection. These results show that immunization against the MC3R resulted in the production of Abs with positive or negative allosteric properties. The presence of such Abs induced small changes in metabolic and cardiovascular parameters.

Background: This study examines the effect of an increase in the inspired fraction of oxygen (FiO2) on brain tissue oxygen (PbO2) in children with severe traumatic brain injury (TBI)., Methods: A prospective observational study of patients who underwent PbO2 monitoring and an oxygen challenge test (temporary increase of FiO2 for 15 min) was undertaken. Pre- and post-test values for arterial partial pressure of oxygen (PaO2), PbO2, and arterial oxygen content (CaO2) were examined while controlling for any changes in arterial carbon dioxide tension and cerebral perfusion pressure during the test. Baseline transcranial Doppler studies were done. Outcome was assessed at 6 months., Results: A total of 43 tests were performed in 28 patients. In 35 tests in 24 patients, the PbO2 monitor was in normal-appearing white matter and in eight tests in four patients, the monitor was in a pericontusional location. When catheters were pericontusional or in normal white matter the baseline PbO2/PaO2 ratio was similar. PaO2 (P < 0.0001) and PbO2 (P < 0.0001) significantly increased when FiO2 was increased. The magnitude of the PbO2 response (PbO2) was correlated with PaO2 (P < 0.0001, R(2) = 0.37) and CaO2 (P = 0.001, R(2) = 0.23). The PbO2/PaO2 ratio (oxygen reactivity) varied between patients, was related to the baseline PbO2 (P = 0.001, r = 0.54) and was inversely related to outcome (P = 0.02, confidence interval 0.03-0.78)., Conclusion: Normobaric hyperoxia increases PbO2 in children with severe TBI, but the response is variable. The magnitude of this response is related to the change in PaO2 and the baseline PbO2. A greater response appears to be associated with worse outcome.

The hypothalamic melanocortin-4 receptor (MC4R) is a constituent of an important pathway regulating food intake and energy expenditure. We produced a monoclonal antibody (mAb) directed against the N-terminal domain of the MC4R and evaluated its potential as a possible therapeutic agent. This mAb (1E8a) showed specific binding to the MC4R in human embryonic kidney 293 cells expressing the human MC4R and blocked the activity of the MC4R under basal conditions and after stimulation with alpha-melanocyte-stimulating hormone (alpha-MSH). The inverse agonist action of Agouti-related protein was significantly enhanced in the presence of mAb 1E8a. After a single intracerebroventricular injection into the third ventricle, mAb 1E8a (1 microg) increased 24-h food intake in rats. After 7 days of continuous intracerebroventricular administration, mAb 1E8a increased food intake, body weight, and fat pad weight and induced hyperglycemia. Because the complete mAb was ineffective after intravenous injection, we produced single-chain variable fragments (scFvs) derived from mAb 1E8a. In pharmacokinetic studies it was demonstrated that these scFvs crossed the blood-brain barrier and reached the hypothalamus. Consequently, the scFv 1E8a increased significantly food intake and body weight in rats after intravenous administration (300 mug/kg). The pharmacological profile of mAb 1E8a and the fact that its scFv was active after peripheral administration suggest that derivatives of anti-MC4R mAbs may be useful in the treatment of patients with anorexia or cachexia.

Introduction: Neurotrauma remains a major global burden of injury, especially for young patients, and will consequently always be a condition that pediatric neurosurgeons are called upon to treat. However, the face of modern neurotrauma management is changing, presenting important challenges to today's pediatric neurosurgeons., Objective: This article summarizes some of the issues in neurotrauma facing clinicians whose responsibility it is to treat these children., Conclusion: It is up to the individual neurosurgeon to familiarize him- or herself with the emerging literature on the modern management of pediatric neurotrauma.

Introduction: The aim of this study was to evaluate the mechanical status of the spine in patients with spastic diplegia 17-26 years after selective dorsal rhizotomy (SDR)., Methods: We compared original radiographic reports from our earlier short-term follow-up study with current X-rays. In addition, we obtained magnetic resonance images (MRI) of the spine and additional information regarding back pain and clinical assessments., Results: Thirty patients (17 males and 13 females; median age 26.8 years) participated in the current study, with median follow-up times of 4.0 and 21.4 years. Comparison of the X-ray results showed respectively: scoliosis 0% and 57%; kyphosis 0% and 7%; lordosis 21% and 40%; spondylolysis 18% and 37%; and spondylolisthesis grade I occurred in one patient. The only statistically significant difference was found for scoliosis (p < 0.01). The majority had Cobb angles <30 degrees with only two patients with curves of 35 degrees. MRI scans showed spinal stenosis in 27%, black discs in 10%, and disc protrusion in 3%. Daily back pain was reported in 17%, while 23% reported "moderate disability" as a result of back and leg pain. No patient to date has required any surgical intervention on the spine., Conclusions: Except for spondylolisthesis, spinal deformities did appear to progress with time. However, this increase was not marked, and the development of relatively mild scoliosis was the only statistically significant increase. This group of patients requires continued follow-up. Further studies are required to ascertain the natural history of spinal deformity in adults with spastic diplegia who have not had SDR.

Object: Cerebral pressure autoregulation is an important neuroprotective mechanism that stabilizes cerebral blood flow when blood pressure (BP) changes. In this study the authors examined the association between autoregulation and clinical factors, BP, intracranial pressure (ICP), brain tissue oxygen tension (PbtO(2)), and outcome after pediatric severe traumatic brain injury (TBI). In particular we examined how the status of autoregulation influenced the effect of BP changes on ICP and PbtO(2)., Methods: In this prospective observational study, 52 autoregulation tests were performed in 24 patients with severe TBI. The patients had a mean age of 6.3 +/- 3.2 years, and a postresuscitation Glasgow Coma Scale score of 6 (range 3-8). All patients underwent continuous ICP and PbtO(2) monitoring, and transcranial Doppler ultrasonography was used to examine the autoregulatory index (ARI) based on blood flow velocity of the middle cerebral artery after increasing mean arterial pressure by 20% of the baseline value. Impaired autoregulation was defined as an ARI < 0.4 and intact autoregulation as an ARI >or= 0.4. The relationships between autoregulation (measured as both a continuous and dichotomous variable), outcome, and clinical and physiological variables were examined using multiple logistic regression analysis., Results: Autoregulation was impaired (ARI < 0.4) in 29% of patients (7 patients). The initial Glasgow Coma Scale score was significantly associated with the ARI (p = 0.02, r = 0.32) but no other clinical factors were associated with autoregulation status. Baseline values at the time of testing for ICP, PbtO(2), the ratio of PbtO(2)/PaO(2), mean arterial pressure, and middle cerebral artery blood flow velocity were similar in the patients with impaired or intact autoregulation. There was an inverse relationship between ARI (continuous and dichotomous) with a change in ICP (continuous ARI, p = 0.005; dichotomous ARI, p = 0.02); that is, ICP increased with the BP increase when ARI was low (weak autoregulation). The ARI (continuous and dichotomous) was also inversely associated with a change in PbtO(2) (continuous ARI, p = 0.002; dichotomous ARI, p = 0.02). The PbtO(2) increased when BP was increased in most patients, even when the ARI was relatively high (stronger autoregulation), but the magnitude of this response was still associated with the ARI. There was no relationship between the ARI and outcome., Conclusions: These data demonstrate the influence of the strength of autoregulation on the response of ICP and PbtO(2) to BP changes and the variability of this response between individuals. The findings suggest that autoregulation testing may assist clinical decision-making in pediatric severe TBI and help better define optimal BP or cerebral perfusion pressure targets for individual patients.

Introduction: Brain tissue oxygen tension (PbtO(2)) monitoring is used increasingly in adult severe traumatic brain injury (TBI) management. Several factors are known to influence PbtO(2) in adults, but the variables that affect PbtO(2) in pediatric TBI are not well described. This study examines the relationships between PbtO(2) and (1) physiological markers of potential secondary insults commonly used in pediatric TBI, in particular intracranial pressure (ICP), cerebral perfusion pressure (CPP), and systemic hypoxia, and (2) other clinical factors and treatment received that may influence PbtO(2)., Materials and Methods: In this prospective observational study, 52 children (mean age, 6.5 +/- 3.4 years; range, 9 months to 14 years old) with severe TBI and a median post-resuscitation Glasgow Coma Score (GCS) of 5 were managed with continuous PbtO(2) monitoring. The relationships between PbtO(2) parameters (Pbt)(2)(low), PbtO(2) < 5, PbtO(2) < 10, and mPbtAO(2)(24)) and clinical, physiological, and treatment factors were explored using time-linked data and Spearman's correlation coefficients., Results: No clinical, physiological, or treatment variable was significantly associated with all PbtO(2) parameters, but individual associations were found with initial GCS (PbtO(2) < 5, p = 0.0113), admission Pediatric Trauma Score (PbtO(2) < 10, 0.0175), mICP > 20 (mPbtO(2)(24), p = 0.0377), CPP(low) (PbtO(2)(low), p = 0.0065), CPP < 40 (PbtO(2)(low), p = 0.0269; PbtO(2) < 5, p = 0.0212), P(a)O(2) < 60 (mPbtO(2)(24), p = 0.0037), S(a)O(2) < 90 (PbtO(2)(low), p = 0.0438), and use of inotropes during ICU care (PbtO(2)(low), p = 0.0276; PbtO(2) < 10, p = 0.0277; p = mPbtO(2)(24))., Conclusion: Delivery of oxygen to the brain is important to limit secondary neuronal injury after severe TBI. Our data show that PbtO(2) is poorly predicted by clinical and physiological factors commonly measured in the pediatric ICU. Multimodality monitoring may be needed to detect all secondary cerebral insults in pediatric TBI.

Introduction: Intracranial pressure (ICP) monitoring and cerebral perfusion pressure (CPP) management are the current standards to guide care of severe traumatic brain injury (TBI). However, brain hypoxia and secondary brain injury can occur despite optimal ICP and CPP. In this study, we used brain tissue oxygen tension (PbtO(2)) monitoring to examine the association between multiple patient factors, including PbtO(2), and outcome in pediatric severe TBI., Materials and Methods: In this prospective observational study, 52 children (less than 15 years) with severe TBI were managed with continuous PbtO(2) and ICP monitoring. The relationships between outcome [Glasgow Outcome Score (GOS) and Pediatric Cerebral Performance Category Scale] and clinical, radiologic, treatment, and physiological variables, including PbtO(2), were examined using multiple logistic regression analysis., Results: Outcome was favorable in 40 patients (77%) and unfavorable (mortality, 9.6%; n = 5) in 12 (23%). In univariate analysis, the following variables had a significant association with unfavorable outcome: initial GCS, computed tomography classification, ICP(peak), mICP(24), mICP, CPP(low), CPP(<40), pupil reactivity, PbtO(2)(low), PbtO(2) < 5 mmHg, PbtO(2) < 10 mmHg, mPbtO(2)(24), and time-severity product. PbtO(2) parameters had the strongest independent association with poor outcome in multiple regression analysis. In particular, when PbtO(2) was <5 mmHg for >1 h, the adjusted OR for poor outcome was 27.4 (95% confidence interval, 1.9-391). No variables apart from PbtO(2) were independently associated with mortality when controlled for PbtO(2)., Conclusion: Reduced PbtO(2) is shown to be an independent factor associated with poor outcome in pediatric severe TBI in the largest study to date. It appears to have a stronger association with outcome than conventionally evaluated measures.

Background: The TCD-derived PI has been associated with ICP in adult studies but has not been well investigated in children. We examined the relationship between PI and ICP and CPP in children with severe TBI., Methods: Data were prospectively collected from consecutive TCD studies in children with severe TBI undergoing ICP monitoring. Ipsilateral ICP and CPP values were examined with Spearman correlation coefficient (mean values and raw observations), with a GEE, and as binary values (1 and 20 mm Hg, respectively)., Results: Thirty-four children underwent 275 TCD studies. There was a weak relationship between mean values of ICP and PI (P = .04, r = 0.36), but not when raw observations (P = .54) or GEE (P = .23) were used. Pulsatility index was 0.76 when ICP was lower than 20 mm Hg and 0.86 when ICP was 20 mm Hg or higher. When PI was 1 or higher, ICP was lower than 20 mm Hg in 62.5% (25 of 40 studies), and when ICP was 20 mm Hg or higher, PI was lower than 1 in 75% (46 of 61 studies). The sensitivity and specificity of a PI threshold of 1 for examining the ICP threshold of 20 mm Hg were 25% and 88%, respectively. The relationship between CPP and PI was stronger (P = .001, r = -0.41), but there were too few observations below 50 mm Hg to examine PI at this threshold., Conclusion: The absolute value of the PI is not a reliable noninvasive indicator of ICP in children with severe TBI. Further study is required to examine the relationship between PI and a CPP threshold of 50 mm Hg.

Objective: To determine functional status of patients with cerebral palsy 20 years after they received selective dorsal rhizotomy (SDR)., Design: A prospective 20-year follow-up study., Setting: Red Cross Children's Hospital (SDR operation and 1-year follow-up assessment) and at institutional or private locations nearby patients' homes (20-year follow-up assessment)., Participants: Referred sample of 14 patients with spastic diplegia (6 women, 8 men; mean age, 27y; range, 22-33y) who were preoperatively ambulant and fulfilled strict selection criteria for SDR operation in 1985., Interventions: Patients were assessed before and 1 and 20 years after SDR., Main Outcome Measures: Standardized assessments of function according to 2 dimensions of the International Classification of Functioning, Disability and Health (ICF) model: (1) body structure and function (muscle tone, joint stiffness, voluntary movement) and (2) activity (rolling, sitting, kneeling, crawling, standing, walking, transitions) were obtained. In addition, based on assessments and questionnaires, Gross Motor Function Classification System (GMFCS) levels were determined before and at 1 year after SDR retrospectively and currently at 20 years after SDR., Results: One year after SDR, functional outcomes based on the 2 dimensions of the ICF model improved significantly, and these improvements were maintained at 20 years after surgery. Patients showed a shift in their GMFCS levels 1 and 20 years after SDR., Conclusions: In line with our 20-year follow-up study with gait parameters as outcome measures, patients with spastic diplegia still show improvements in their functional status 20 years after SDR. We acknowledge the presence of possible confounding factors and a small sample size, but we argue that the improvements found in this study were caused mainly by SDR. Finally, changes in GMFCS levels suggest a possible role for this tool to detect changes after an intervention.

Background: The melanocortin-4 receptor (MC4R) is part of an important pathway regulating energy balance. Here we report the existence of autoantibodies (autoAbs) against the MC4R in sera of obese patients., Methods: The autoAbs were detected after screening of 216 patients' sera by using direct and inhibition ELISA with an N-terminal sequence of the MC4R. Binding to the native MC4R was evaluated by flow cytometry, and pharmacological effects were evaluated by measuring adenylyl cyclase activity., Results: Positive results in all tests were obtained in patients with overweight or obesity (prevalence, 3.6%) but not in normal weight patients. The selective binding properties of anti-MC4R autoAbs were confirmed by surface plasmon resonance and by immunoprecipitation with the native MC4R. Finally, it was demonstrated that these autoAbs increased food intake in rats after passive transfer via intracerebroventricular injection., Conclusion: These observations suggest that inhibitory anti-MC4R autoAbs might contribute to the development of obesity in a small subpopulation of patients.

Object: The goal of this paper was to examine the relationship between methods of acute clinical assessment and measures of secondary cerebral insults in severe traumatic brain injury in children., Methods: Patients who underwent intracranial pressure (ICP), cerebral perfusion pressure (CPP), and brain oxygenation (PbtO(2)) monitoring and who had an initial Glasgow Coma Scale score, Pediatric Trauma Score, Pediatric Index of Mortality 2 score, and CT classification were evaluated. The relationship between these acute clinical scores and secondary cerebral insult measures, including ICP, CPP, PbtO(2), and systemic hypoxia were evaluated using univariate and multivariate analysis., Results: The authors found significant associations between individual acute clinical scores and select physiological markers of secondary injury. However, there was a large amount of variability in these results, and none of the scores evaluated predicted each and every insult. Furthermore, a number of physiological measures were not predicted by any of the scores., Conclusions: Although they may guide initial treatment, grading systems used to classify initial injury severity appear to have a limited value in predicting who is at risk for secondary cerebral insults.

Objective: Active immunization in rats may serve several purposes: the production of a disease-like phenotype, the generation of pharmacologic tools, and the development of clinically useful therapies. We selected the melanocortin-4 receptor (MC4R) as a target because its blockade could provide a treatment for anorexia and cachexia., Methods: We used a sequence of the N-terminal (NT) domain of the MC4R as an antigen. Rats immunized against the NT peptide produced specific MC4R antibodies (Abs) that were purified and characterized in vitro and in vivo., Results: The Abs acted as inverse agonists and reduced under basal conditions the production of cyclic adenosine monophosphate in HEK-293 cells expressing the human MC4R. Rats immunized against the NT peptide developed a phenotype consistent with hypothalamic MC4R blockade, i.e., increased food intake and body weight, liver and fat-pad weights, hepatic steatosis, and increased plasma triacylglycerols. With a high-fat diet, plasma insulin levels were significantly increased. In separate experiments an increase in food intake was observed after injection of purified MC4R Abs into the third ventricle. When lipopolysaccharide was administered in NT-immunized rats the reduction of food intake was partly prevented in this model of cytokine-induced anorexia., Conclusion: Our results show that active immunization of rats against the MC4R resulted in the generation of specific Abs that stimulated food intake by acting as inverse agonists of the hypothalamic MC4R. Pharmacologically active monoclonal MC4R Abs could be the starting point for the development of novel treatments for patients with anorexia or cachexia.

Objective: Tuberculous meningitis (TBM) is a massive global problem. The mortality and morbidity associated with the severe form of the disease are exceptionally high. Even when increased intracranial pressure is treated and full conventional therapy is commenced, cerebral ischemia can develop and is associated with a particularly poor prognosis. We sought to evaluate our experience with two patients with severe TBM and cerebral oxygenation monitoring., Design: Case report., Setting: Red Cross Children's Hospital, Cape Town., Patients: Two comatose patients with TBM., Interventions: Targeted interventions against low cerebral oxygenation in one patient., Measurements and Main Results: Cerebral tissue oxygenation (Ptio2) was measured. In both patients, Ptio2 monitoring demonstrated delayed cerebral ischemia despite the institution of full conventional therapy and the control of intracranial pressure. These data confirm that the vascular involvement in TBM is potentially progressive and that failure to diagnose infarction initially is not merely due to a delay in the radiologic appearance. The first patient developed extensive infarction, consistent with Ptio2 readings, and subsequently died after treatment withdrawal. Intervention in the second patient successfully reversed a precipitous decline of the Ptio2 readings and may have prevented infarction in this patient., Conclusions: The development of delayed cerebral ischemia in TBM despite treatment is confirmed in these two patients. The reversal of a decline in Ptio2 readings suggests a possible benefit for cerebral oxygenation monitoring in selected patients with severe TBM.

Objective: Most physicians rely on conventional treatment targets for intracranial pressure, cerebral perfusion pressure, systemic oxygenation, and hemoglobin to direct management of traumatic brain injury (TBI) in children. In this study, we used brain tissue oxygen tension (PbtO2) monitoring to examine the association between PbtO2 values and outcome in pediatric severe TBI and to determine the incidence of compromised PbtO2 in patients for whom acceptable treatment targets had been achieved., Methods: In this prospective observational study, 26 children with severe TBI and a median postresuscitation Glasgow Coma Scale score of 5 were managed with continuous PbtO2 monitoring. The relationships between outcome and the 6-hour period of lowest PbtO2 values and the length of time that PbtO2 was less than 20, 15, 10, and 5 mmHg were examined. The incidence of reduced PbtO2 for each threshold was evaluated where the following targets were met: intracranial pressure less than 20 mmHg, cerebral perfusion pressure greater than 50 mmHg, arterial oxygen tension greater than 60 mmHg (and peripheral oxygen saturation > 90%), and hemoglobin greater than 8 g/dl., Results: There was a significant association between poor outcome and the 6-hour period of lowest PbtO2 and length of time that PbtO2 was less than 15 and 10 mmHg. Multiple logistic regression analysis showed that low PbtO2 had an independent association with poor outcome. Despite achieving the management targets described above, 80% of patients experienced one or more episodes of compromised PbtO2 (< 20 mmHg), and almost one-third experienced episodes of brain hypoxia (PbtO2 < 10 mmHg)., Conclusion: Reduced PbtO2 is associated with poor outcome in pediatric severe TBI. In addition, many patients experience episodes of compromised PbtO2 despite achieving acceptable treatment targets.

Objectives: Selective dorsal rhizotomy (SDR) has been widely performed for the reduction of spasticity in patients with cerebral palsy during the past 2 decades. The objective of this study was to determine whether the surgery has yielded long-term functional benefits for these patients., Methods: The authors present results from a prospective 20-year follow-up study of locomotor function in 13 patients who underwent an SDR in 1985. For comparison, we also present gait data for 48 age-matched healthy controls (12 at each of 4 time points). Patients were studied preoperatively and then at 1, 3, 10, and 20 years after surgery. Study participants were recorded in the sagittal plane while walking using a digital video camera, and 6 standard gait parameters were measured., Results: In this group of patients 20 years after surgery, knee range of motion (ROM) was on average 12 degrees greater than preoperative values (p < 0.001). Hip ROM before surgery was no different from that in the healthy control group. This parameter increased markedly immediately after surgery (p < 0.001) but had returned to normal after 20 years. The knee and hip midrange values-a measure of the degree of "collapse" due to muscle weakness after surgery-had returned to preoperative levels after 20 years, although they were respectively 11 and 8 degrees greater than those in healthy controls. Both temporal-distance parameters (dimensionless cadence and dimensionless step length) were significantly greater at 20 years than preoperative values (cadence, p = 0.003; step length, p = 0.02), leading to improved walking speed., Conclusions: Twenty years after undergoing SDR, our patients showed improved locomotor function compared with their preoperative status.

Introduction: There has been a resurgence of interest in decompressive craniectomy for traumatic brain injury (TBI), but the impact of craniectomy on intracranial pressure (ICP) and cerebral oxygenation has not been well described for diffuse injury in children., Methods: ICP and brain tissue oxygenation (PbtO2) changes after decompressive craniectomy for diffuse brain swelling after TBI in children were analysed., Findings: Decompressive craniectomy was performed for diffuse brain swelling in 18 children under 15 years old. For 8 patients, craniectomy was performed as an emergency for malignant brain swelling, and in 10, for sustained ICP > 25 mmHg refractory to conventional medical treatment. In 6 of these patients, PbtO2 was also monitored. Median ICP was reduced from 40 mmHg before craniectomy to 16 mmHg for 24 hours thereafter, and PbtO2 improved from a median of 17.4 to 43.4 mmHg. Clinical outcome was favourable in 78%., Conclusions: In selected pediatric patients with TBI, craniectomy for diffuse brain swelling can significantly improve ICP and cerebral oxygenation control. The use of the procedure in appropriate settings does not appear to increase the proportion of disabled survivors.

In the present study, we aimed to investigate the neuromodulatory role played by hypothalamic brain-derived neurotrophic factor (BDNF) in the regulation of acute cardiovascular and feeding responses to melanocortin-4 receptor (MC4R) activation. In vitro, a selective MC4R agonist, MK1, stimulated BDNF release from isolated rat hypothalami and this effect was blocked by preincubation with the MC3/4R antagonist SHU-9119. In vivo, peripheral administration of MK1 decreased food intake in rats and this effect was blocked by pretreatment with an anti-BDNF antibody administered into the third ventricle. When anorexia was induced with the cannabinoid-1 receptor (CB1R) antagonist AM251, the anti-BDNF antibody did not prevent the reduction in food intake. Peripheral administration of MK1 also increased mean arterial pressure, heart rate and body temperature. These effects were prevented by pretreatment with the anti-BDNF antibody whereas the intracerebroventricular administration of BDNF caused changes similar to those of MK1. These findings demonstrate for the first time that activation of MC4R leads to an acute release of BDNF in the hypothalamus. This release is a prerequisite for MC4R-induced effects on appetite, body temperature and cardiovascular function. By contrast, CB1R antagonist-mediated anorexia is independent of the MC4R/BDNF pathway. Overall, these results show that BDNF is an important downstream mediator of the MC4R pathway.

Case Report: The authors present the case of a 5-year-old child with severe traumatic brain injury in whom decompressive hemicraniectomy was performed for progressive increased intracranial pressure (ICP) unresponsive to medical treatment. Data from ICP and cerebral tissue oxygenation monitoring in the contralateral hemisphere were recorded, which demonstrated the immediate and delayed mechanical and physiological changes occurring after bony and dural decompression., Discussion: The role of the procedure and that of the monitoring approach are discussed.

Introduction: Given the large number of cerebral palsy patients who have undergone selective dorsal rhizotomy in the past two decades, it is clearly imperative that the clinical community be provided with objective and compelling evidence of the long-term sequelae of the procedure., Materials and Methods: In the early 1980s, Peacock in Cape Town shifted the site of the rhizotomy from the conus medullaris to the cauda equina, and in the past 25 years, more than 200 children have been operated on. We have studied the incidence of spinal deformities after multiple-level laminectomy and recorded a 20% incidence of isthmic spondylolysis or grade-I spondylolisthesis. We have also conducted a long-term prospective gait analysis study on a cohort of 14 ambulatory patients who were operated on in 1985., Results: Ten years after surgery, our patients had increased ranges of motion that were within normal limits. Step length was significantly improved, although cadence was unchanged postoperatively and was significantly less than normal age-matched control subjects., Discussion: We have recently tracked down all 14 patients from the original cohort and are currently completing a 20-year prospective follow-up analysis of their neuromuscular function and gait. Our preliminary data suggest that selective dorsal rhizotomy is not only an effective method for alleviating spasticity but it also leads to long-term functional benefits.

Functionally active antibodies (Abs) against central G-protein-coupled receptors have not yet been reported. We selected the hypothalamic melanocortin-4 receptor (MC4-R) as a target because of its crucial role in the regulation of energy homeostasis. A 15 amino acid sequence of the N-terminal (NT) domain was used as an antigen. This peptide showed functional activity in surface plasmon resonance experiments and in studies on HEK-293 cells overexpressing the human MC4-R (hMC4-R). Rats immunized against the NT peptide produced specific antibodies, which were purified and characterized in vitro. In HEK-293 cells, rat anti-NT Abs showed specific immunofluorescence labeling of hMC4-R. They reduced the production of cAMP under basal conditions and after stimulation with a synthetic MC4-R agonist. Rats immunized against the NT peptide developed a phenotype consistent with MC4-R blockade, that is, increased food intake and body weight, increased liver and fat pad weight, and elevated plasma triglycerides. In a separate experiment in rats, an increase in food intake could be produced after injection of purified Abs into the third ventricle. Similar results were obtained in rats injected with anti-NT Abs raised in rabbits. Our data show for the first time that active immunization of rats against the NT sequence of the MC4-R results in specific Abs, which appear to stimulate food intake by acting as inverse agonists in the hypothalamus.

Introduction: The role of endoscopy in hydrocephalus due to infectious aetiology is unclear. Tuberculous hydrocephalus is a useful model to study because it presents particular challenges and the pathophysiology of the cerebrospinal fluid disturbance is well known., Materials and Methods: We present the results of 24 endoscopic operations in tuberculous meningitis., Result: Endoscopic third ventriculostomy (ETV) was attempted in 17 patients: seven were successful, five failed, and five were not completed due to abnormal anatomy. There were five fenestration procedures, three of which were successful. Endoscopic biopsy of two tuberculomas failed to yield a bacteriological result. These operations were more difficult to perform than for hydrocephalus due to other aetiologies., Conclusion: Although ETV is technically possible in this situation, it is imperative that the patients are adequately selected for the procedure to ensure optimal treatment and that the surgeon has experience with difficult cases.

Conjoined twins represent a rare but fascinating congenital condition, the aetiology of which remains obscure. Over the past four decades, the paediatric surgeons at Red Cross Children's Hospital have been involved in the management of 46 pairs of conjoined twins, of which 33 have been symmetrical and 12 asymmetrical. Seventeen symmetrical twins have undergone separation with 22 children (65%) surviving; all of the live asymmetrical twins survived separation. We describe the important features of this unique cohort, outline our approach to management and present the results of this approach. We consider some of the ethical and moral dilemmas we have confronted, and discuss the prenatal diagnosis, obstetric implications and postnatal care of these children, including the relevant investigations and anaesthetic and surgical management. Specific aspects related to the cardiovascular system, hepatobiliary and gastrointestinal tracts, urogenital tract, central nervous system and musculoskeletal system are highlighted.

Objectives: Traumatic brain injury accounts for a high percentage of deaths in children. Raised intracranial pressure (ICP) due to brain swelling within the closed compartment of the skull leads to death or severe neurological disability if not effectively treated. We report our experience with 12 children who presented with cerebral herniation due to traumatic brain swelling in whom decompressive craniectomy was used as an emergency., Design: Prospective, observational., Setting: Red Cross Children's Hospital., Subjects: Children with severe traumatic brain injury and cerebral swelling., Outcome Measures: Computed tomography (CT) scanning, ICP control, clinical outcome., Results: Despite the very poor clinical condition of these children preoperatively, aggressive management of the raised pressure resulted in unexpectedly good outcomes., Conclusion: Aggressive surgical measures to decrease ICP in the emergency situation can be of considerable benefit; the key concepts are selection of appropriate patients and early intervention.

Autoantibodies against beta-adrenoceptors might be involved in different cardiomyopathic diseases such as idopathic dilated cardiomyopathy, Chagas' disease and ventricular arrhythmias. To study the effects of such antibodies on the whole heart, we made use of a new technique allowing the measurement of Ca++ transients as well as action potentials in Langendorff preparations of mouse hearts. Mouse antibodies directed against the second extracellular loop of the beta2-adrenoceptor induced conduction blocks which could be washed away by the beta2-adrenoceptor inverse agonist ICI118,551, confirming the specificity and non-toxicity of these events. These results were confirmed by the use of a monoclonal antibody, monospecific for the beta2-adrenoceptor and the beta2-specific full agonist, clenbuterol. Both increased slightly, but significantly, the beating frequency but their main effect was the production of conduction blocks. In contrast, a monoclonal antibody, monospecific for the beta1-adrenoceptor, highly increased the beating frequency without interfering with the conduction. Our results suggest that stimulation of the beta2-adrenoceptor by anti-receptor antibodies in the conduction tissues leads to conduction disturbances, probably mediated by coupling to a different pathway than the classical Gs pathway. They confirm that anti-beta2 adrenoceptor antibodies could be responsible for ventricular arrhythmias.

Interaction between CD40, a member of the tumor necrosis factor receptor (TNFR) superfamily, and its ligand CD40L, a 39-kDa glycoprotein, is essential for the development of humoral and cellular immune responses. Selective blockade or activation of this pathway provides the ground for the development of new treatments against immunologically based diseases and malignancies. Like other members of the TNF superfamily, CD40L monomers self-assemble around a threefold symmetry axis to form noncovalent homotrimers that can each bind three receptor molecules. Here, we report on the structure-based design of small synthetic molecules with C3 symmetry that can mimic CD40L homotrimers. These molecules interact with CD40, compete with the binding of CD40L to CD40, and reproduce, to a certain extent, the functional properties of the much larger homotrimeric soluble CD40L. Architectures based on rigid C3-symmetric cores may thus represent a general approach to mimicking homotrimers of the TNF superfamily.

Splenocytes from a BALB/c mouse immunised with a synthetic peptide corresponding to the second extracellular loop of the 5-HT4 receptor were fused with SP2/O myeloma cells to produce a monoclonal antibody. The monoclonal antibody was of the IgG2b isotype. The antibody recognised the human 5-HT4(g) (h5-HT4(g)) receptor by immunoblots and by immunofluorescence on chinese hamster ovary (CHO) cells expressing this 5-HT4 receptor isoform. Epitope mapping of the antibody suggested the recognition of a conformational epitope, encompassing the N- and C-terminal fragments of the second extracellular loop. Kinetic experiments using surface plasmon resonance showed that the antibody had a picomolar affinity for its cognate peptide. Inhibition experiments using the same methodology confirmed the specificity of the interaction. The antibody at a concentration of 500 pM competitively inhibited inverse agonist GR113808 binding and showed an inverse agonist effect on the basal activity of CHO cells expressing the 5-HT4(g) receptor. The antibody decreased the effect of 5-HT at 500 and 50 pM concentrations but it increased 5-HT-induced cAMP levels at 5 pM. The dual effect of the monoclonal antibody could be ascribed to mono- or bivalent recognition of the receptor. The antibody described here is the first example of a high-affinity modulator of the 5-HT4 receptor.