The effect of air-free administration of intravenous drugs on microemboli during cardiopulmonary bypass.

OBJECTIVE: During cardiopulmonary bypass (CPB), gaseous microemboli (GME) that originate from the extracorporeal circuit are released into the arterial blood stream of the patient. Gaseous microemboli may contribute to adverse outcome after cardiac surgery with CPB. Possibly, air may be collected in the right atrium during induction of anesthesia and released during CPB start. The aim of this study was to assess if the GME load entering the venous line of the CPB circuit could be reduced by training of anesthesia personal in avoiding air introduction during administration of intravenous medication. METHODS: In 94 patients undergoing coronary artery bypass grafting with CPB, GME number and volume were measured intraoperatively with a bubble counter (BCC300). The quantity and the relationship between GME number and volume in the venous and arterial line were determined in 2 periods before and after education of the anesthesiologists and nurses. RESULTS: In the venous line no significant differences were observed between numbers and volumes of GME between groups. Comparing patients with low versus high GME load, showed significantly more patients from the intervention group in the low GME-load group, namely 29 versus 18. Administration of medication by anesthesia was confirmed as a clear cause of GME/air-introduction into the venous circulation. Scavenging properties of the CPB circuit including the oxygenator showed a 99.9% reduction of GME. CONCLUSIONS: A wide spread of GME generation during perfusion was present with no difference in generation of GME between groups. Lower GME load observed in patients (intervention group) and examples of air introduction during drug administration suggest that air introduced by anesthesia contributes to the GME load during CPB. Scavenging properties of the CPB circuit contribute very much to patient safety regarding reduction of venous air. Awareness and education create the possibilities for further reduction of GME during cardiopulmonary bypass.

Does heparin rebound lead to postoperative blood loss in patients undergoing cardiac surgery with cardiopulmonary bypass?

BACKGROUND: Heparin rebound is a common observed phenomenon after cardiac surgery with CPB and is associated with increased postoperative blood loss. However, the administration of extra protamine may lead to increased blood loss as well. Therefore, we want to investigate the relation between heparin rebound and postoperative blood loss and the necessity to provide extra protamine to reverse heparin rebound. METHODS: We searched PubMed, Cochrane, EMBASE, Google Scholar and Web of Science to review the question: "Does heparin rebound lead to postoperative blood loss in patients undergoing cardiac surgery with cardiopulmonary bypass." Combination of search words were framed within four major categories: heparin rebound, blood loss, cardiac surgery and cardiopulmonary bypass. All studies that met our question were included. Quality assessment was performed using the Cochrane risk of bias (RoB2) tool for randomized controlled trials and the risk of bias in non-randomized studies of intervention (ROBINS-I) for non-randomised trials. RESULTS: 4 randomized and 17 non-randomized studies were included. The mean incidence of heparin rebound was 40%. The postoperative heparin levels, due to heparin rebound, were often below or equal to 0.2 IU/mL. We could not demonstrate an association between heparin rebound and postoperative blood loss or transfusion requirements. However the quality of evidence was poor due to a broad variety of definitions of heparin rebound, measured by various coagulation tests and studies with small sample sizes. CONCLUSION: The influence of heparin rebound on postoperative bleeding seems to be negligible, but might get significant in conjunction with incomplete heparin reversal or other coagulopathies. For that reason, it might be useful to get a picture of the entire coagulation spectrum after cardiac surgery, as can be done by the use of a viscoelastic test in conjunction with an aggregometry test.

The influence of gaseous microemboli on various biomarkers after minimized cardiopulmonary bypass.

INTRODUCTION: Gaseous microemboli that originate from the cardiopulmonary bypass circuit may contribute to adverse outcome after cardiac surgery. We prospectively evaluated the influence of gaseous microemboli on the release of various biomarkers after use of a minimally invasive extracorporeal technology system. METHODS: In 70 patients undergoing coronary artery bypass grafting with minimized cardiopulmonary bypass, gaseous microemboli were measured intraoperatively with a bubble counter. Intra- and postoperative biomarker levels for inflammatory response (interleukin-6, C5b-9), endothelial damage (von Willebrand factor, soluble vascular cell adhesion molecule-1), oxidative stress (malondialdehyde, 8-isoprostane, neuroketal), and neurological injury (neuron-specific enolase, brain-type fatty acid-binding protein) were analyzed using immune assay techniques. The relationship between gaseous microemboli number or volume and the incremental area under the curve (iAUC24h) or peak change for the biomarkers was calculated. RESULTS: All biomarkers except for malondialdehyde increased at least temporarily after coronary artery bypass grafting with a minimally invasive extracorporeal technology system. The median total gaseous microemboli number was 6,174 (interquartile range: 3,507-10,531) and the median total gaseous microemboli volume was 4.31 µL (interquartile range: 2.71-8.50). There were no significant correlations between total gaseous microemboli number or volume and iAUC24h or peak change for any of the biomarkers. After controlling for the variance of possible other predictor variables, multiple linear regression analysis showed no association between gaseous microemboli parameters and release of biomarkers. CONCLUSION: This study showed no evidence that gaseous microemboli contribute to increased biomarker levels after coronary artery bypass grafting with cardiopulmonary bypass. A reason for the absence of damage by gaseous microemboli may be the relative and considerably small amount of gaseous microemboli entering the patients in this study.

Carbon Dioxide Flush of an Integrated Minimized Perfusion Circuit Prior to Priming Prevents Spontaneous Air Release Into the Arterial Line During Clinical Use.

Recently, an oxygenator with an integrated centrifugal blood pump (IP) was designed to minimize priming volume and to reduce blood foreign surface contact even further. The use of this oxygenator with or without integrated arterial filter was compared with a conventional oxygenator and nonintegrated centrifugal pump. To compare the air removal characteristics 60 patients undergoing coronary artery bypass grafting were alternately assigned into one of three groups to be perfused with a minimized extracorporeal circuit either with the conventional oxygenator, the oxygenator with IP, or the oxygenator with IP plus integrated arterial filter (IAF). Air entering and leaving the three devices was measured accurately with a bubble counter during cardiopulmonary bypass. No significant differences between all groups were detected, considering air entering the devices. Our major finding was that in both integrated devices groups incidental spontaneous release of air into the arterial line in approximately 40% of the patients was observed. Here, detectable bolus air (>500 µm) was shown in the arterial line, whereas in the minimal extracorporeal circulation circuit (MECC) group this phenomenon was not present. We decided to conduct an amendment of the initial design with METC-approval. Ten patients were assigned to be perfused with an oxygenator with IP and IAF. Importantly, the integrated perfusion systems used in these patients were flushed with carbon dioxide (CO2 ) prior to priming of the systems. In the group with CO2 flush no spontaneous air release was observed in all cases and this was significantly different from the initial study with the group with the integrated device and IAF. This suggests that air spilling may be caused by residual air in the integrated device. In conclusion, integration of a blood pump may cause spontaneous release of large air bubbles (>500 µm) into the arterial line, despite the presence of an integrated arterial filter. CO2 flushing of an integrated cardiopulmonary bypass system prior to priming may prevent spontaneous air release and is strongly recommended to secure patient safety.

Clinical evaluation of the air-handling properties of contemporary oxygenators with integrated arterial filter.

Gaseous microemboli (GME) may originate from the extracorporeal circuit and enter the arterial circulation of the patient. GME are thought to contribute to cerebral deficit and to adverse outcome after cardiac surgery. The arterial filter is a specially designed component for removing both gaseous and solid microemboli. Integration of an arterial filter with an oxygenator is a contemporary concept, reducing both prime volume and foreign surface area. This study aims to determine the air-handling properties of four contemporary oxygenator devices with an integrated arterial filter. Two oxygenator devices, the Capiox FX25 and the Fusion, showed significant increased volume of GME reduction rates (95.03 ± 3.13% and 95.74 ± 2.69%, respectively) compared with both the Quadrox-IF (85.23 ± 5.84%) and the Inspire 6F M (84.41 ± 12.93%). Notably, both the Quadrox-IF and the Inspire 6F M as well as the Capiox FX 25 and the Fusion showed very similar characteristics in volume and number reduction rates and in detailed distribution properties. The Capiox FX25 and the Fusion devices showed significantly increased number and volume reduction rates compared with the Quadrox-IF and the Inspire 6F M devices. Despite the large differences in design of all four devices, our study results suggest that the oxygenator devices can be subdivided into two groups based on their fibre design, which results in screen filter (Quadrox-IF and Inspire 6F M) and depth filter (Capiox FX25 and Fusion) properties. Depth filter properties, as present in the Capiox FX25 and Fusion devices, reduced fractionation of air and may ameliorate GME removal.

The Val158Met polymorphism of the COMT gene is associated with increased pain sensitivity in morphine-treated patients undergoing a painful procedure after cardiac surgery.

AIMS: The catechol-O-methyltransferase (COMT) Val158Met polymorphism affected pain sensitivity of healthy volunteers upon application of experimental pain stimuli. The relevance of these findings in morphine-treated postoperative cardiac patients undergoing painful healthcare procedures is unknown; therefore, the aim of this study was to investigate whether the COMT Val158Met polymorphism increases pain sensitivity in morphine-treated patients undergoing an unavoidable painful routine procedure after cardiac surgery. METHODS: One hundred and seventeen postoperative cardiac patients in the intensive care unit were genotyped for the COMT Val158Met polymorphism. All patients were treated with continuous morphine infusions for pain at rest, and received a bolus of morphine (2.5 or 7.5 mg) before a painful procedure (turning and/or chest drain removal) on the first postoperative day. Numerical rating scale (NRS) scores were evaluated at the following four time points: at baseline (at rest), and before, during and after the painful procedure. RESULTS: Overall mean NRS scores were significantly higher in patients carrying the Met-variant allele. During the painful procedure, the mean NRS score was significantly higher for Met/Met patients compared with Val/Met and Val/Val patients (mean NRS 3.4 ± 2.8, 2.7 ± 2.4 and 1.7 ± 1.7, respectively; P = 0.04). In Met/Met patients, the increase in NRS scores during the painful procedure compared with the baseline NRS score was clinically relevant (ΔNRS ≥ 1.3) and statistically significant and appeared to be independent of sex and the morphine bolus dose. CONCLUSIONS: Our results show that the COMT Val158Met polymorphism contributes to variability in pain sensitivity after cardiac surgery of morphine-treated patients in the intensive care unit, because Met-allele carriers were more sensitive to overall pain and procedure-related pain.

Prolonged coma due to amitriptyline overdose and genetic polymorphism: a case report.

BACKGROUND: Reduced consciousness has a wide variety of possible causes, not infrequently being toxic in nature. An intoxication might be obvious, but in this paper an unexpected case with a tricyclic antidepressant is presented. CASE PRESENTATION: A 76-year-old caucasian female was found unconscious. Primary diagnostic evaluation, including a negative drugs of abuse test, did not give direction to any clear cause. Yet an intraventricular conductive disorder with widening of the QRS complex and electroencephalogram abnormalities did suggest a possible drug effect. Heteroanamnestic information led to the suspicion of an amitriptyline intoxication, which was confirmed by further laboratory analysis. The patient remained comatose for several days. High concentrations of amitriptyline indicated a large overdose of amitriptyline and, in combination with a cytochrome P450 2D6 poor metabolizer status, could explain the long persistence of her comatose state. CONCLUSION: We present a tricyclic antidepressant intoxication, where the patient is thought to have taken a large amount of amitriptyline at once, which, in combination with a cytochrome P450 2D6 poor metabolizer status, led to an unusual long persistence of her coma.

Air removal efficiency of a venous bubble trap in a minimal extracorporeal circuit during coronary artery bypass grafting.

The use of minimized extracorporeal circuits (MECC) in cardiac surgery is expanding. These circuits eliminate volume storage and bubble trap reservoirs to minimize the circuit. However, this may increase the risk of gaseous micro emboli (GME). To reduce this risk, a venous bubble trap was designed. This study was performed to evaluate if incorporation of a venous bubble trap in a MECC system as compared to our standard minimized extracorporeal circuit without venous bubble trap reduces gaseous micro emboli during cardiopulmonary bypass (CPB). Forty patients were randomly assigned to be perfused either with or without an integrated venous bubble trap. After preliminary evaluation of the data of 23 patients, the study was terminated prior to study completion. The quantity and volume of GME were significantly lower in patients perfused with a venous bubble trap compared to patients perfused without a venous bubble trap. The present study demonstrates that a MECC system with a venous bubble trap significantly reduces the volume of GME and strongly reduces the quantity of large GME (>500 µm). Therefore, the use of a venous bubble trap in a MECC system is warranted.

Exogenous natural surfactant for treatment of acute lung injury and the acute respiratory distress syndrome.

RATIONALE: Compositional changes in surfactant and/or decreased surfactant content of the lungs are common features in patients with acute respiratory failure. Instillation of exogenous surfactant into the lungs of neonates with respiratory distress syndrome or pediatric patients with acute respiratory distress syndrome (ARDS) has resulted in improved survival. OBJECTIVES: We conducted this trial to determine whether the instillation of exogenous surfactant would improve the Day 28 outcome of adult patients with acute lung injury (ALI) or ARDS. METHODS: A total of 418 patients with ALI and ARDS were included in an international, multicenter, stratified, randomized, controlled, open, parallel-group study. We randomly assigned 418 patients to receive usual care either with or without instillation of exogenous natural porcine surfactant HL 10 as large boluses. MEASUREMENTS AND MAIN RESULTS: The primary endpoint was death rate before or on Day 28. Secondary endpoints were adverse event and death rate on day 180. The 28-day death rate in the usual care group was 24.5% compared with 28.8% in the HL 10 group. The estimated odds ratio for death at Day 28 in the usual care group versus the HL 10 group was 0.75 (95% CI, 0.48-1.18; P = 0.22). The most common adverse events related to HL 10 administration were temporary hypoxemia defined as oxygen saturation less than 88% (51.9% in HL 10 group vs. 25.2% in usual care) and hypotension defined as mean arterial blood pressure less than 60 mm Hg (34.1% in HL 10 group vs. 17.1% in usual care). CONCLUSIONS: In this study, instillation of a large bolus of exogenous natural porcine surfactant HL 10 into patients with acute lung injury and ARDS did not improve outcome and showed a trend toward increased mortality and adverse effects. Clinical trial registered with www.clinicaltrials.gov (NCT 00742482).

Improved analgesia after the realisation of a pain management programme in ICU patients after cardiac surgery.

BACKGROUND AND OBJECTIVE: Although clinical guidelines recommend systematic evaluation of pain in ICU patients, we know little about the effects from such systematic pain evaluation. This study aims to quantify the effect of a pain management programme in the ICU. METHODS: In this prospective two-phase study, pain levels scored by ICU patients after cardiac surgery through sternotomy were compared before and after the implementation of a pain management programme. The pain management programme consisted of a three-fold strategy; all staff was trained in assessing pain and in providing adequate analgesia, a new patient data management system obliged nurses to ask patients for their pain score three times a day and the preferred analgesic treatment was optimised. The numeric rating scale (NRS 0-10) was used by 190 patients. A NRS at least 4 was considered unacceptable. A generalised linear mixed-effects model was used for analysing repeated measurements data. RESULTS: The occurrence of unacceptable pain (NRS > or = 4) was significantly lower in the intervention group [odds ratio 2.54 (95% confidence interval 1.22-5.65; P = 0.01) for the control group]. Patients in the intervention group received significantly more morphine (29.3 vs. 22.6 mg a day, P<0.01), with higher morphine amounts administered to patients with higher NRS scores (P = 0.01). In the control group, no such relationship was observed (P = 0.66). There was no difference in length of stay in the ICU or in ventilation time. CONCLUSION: The intervention programme successfully reduced the occurrence of unacceptable pain. Further improvement of pain management should focus on the prevention of pain.

Atracurium during induced hyperthermia.

During hyperthermic intraperitoneal chemotherapy (HIPEC), we observed a partial recovery from neuromuscular block in a hyperthermic patient after hours of monitored adequate surgical relaxation and continuous infusion of atracurium during normothermia. This recovery is indicative of the higher clearance of atracurium during hyperthermia. This case report emphasizes the clinical relevance of the well-known temperature dependence of the Hofmann elimination of atracurium. Moreover, this report illustrates the importance of monitoring muscle relaxation during HIPEC. Clinicians should be aware that the usual continuous infusion rate of atracurium at 0.3 mg.kg(-1).h(-1) may be inadequate in hyperthermic patients.

Inflammatory response and optimalisation of perioperative fluid administration during hyperthermic intraoperative intraperitoneal chemotherapy surgery.

BACKGROUND: Surgical cytoreduction and simultaneous hyperthermic intraoperative intraperitoneal chemotherapy (HIPEC) for peritoneal carcinomatosis has a high incidence of postoperative complications. Inadequate intraoperative volume therapy is a known risk factor for the development of postoperative complications. Another possible risk factor is the inflammatory response due to surgery and HIPEC. The aim of this observational pilot study was to monitor fluid intake in the first 24 hours peri- and postoperative by using a non-invasive cardiac output indicator. Furthermore, we measured circulating cytokines and evaluated the possible relation of these changes of inflammatory response with the non-invasive monitored fluid management. METHODS: Twenty-four patients undergoing cytoreductive surgery and HIPEC for peritoneal carcinomatosis were included. Patients were randomised into either a liberal fluid management group using intra-arterial blood pressure and central venous pressure measurement or a restrictive group by using intra-arterial blood pressure and central venous pressure measurement with FloTrac/Vigileo monitoring. Cytokines were measured with multiplex immunoassays. RESULTS: We found no difference in the amount of fluid administration in patients undergoing HIPEC surgery with FloTrac/Vigileo monitoring compared to standard care. Furthermore, there was no difference in mortality, ICU and hospital length of stay between both groups. A severe inflammatory response was seen in all patients after the HIPEC procedure with a rapid increase of interleukins and C-reactive protein (CRP). There was however no difference between our intervention and control group in the severity of this reaction. Finally, we found no relation between the severity of the inflammatory response and mortality, or a composite end-point of mortality and severe complications within 30 days postoperative. CONCLUSIONS: FloTrac/Vigileo monitoring does not lead to a more restrictive fluid administration and does not influence short-term clinical course in patients undergoing HIPEC surgery. The procedure itself leads to a severe inflammatory response, which is not affected by the use of FloTrac/Vigileo. Our data do not support the use of FloTrac/Vigileo monitoring in patients undergoing HIPEC surgery concerning fluid restrictive management.

Comparison of different pain scoring systems in critically ill patients in a general ICU.

BACKGROUND: Pain in critically ill patients in the intensive care unit (ICU) is common. However, pain assessment in critically ill patients often is complicated because these patients are unable to communicate effectively. Therefore, we designed a study (a) to determine the inter-rater reliability of the Numerical Rating Scale (NRS) and the Behavioral Pain Scale (BPS), (b) to compare pain scores of different observers and the patient, and (c) to compare NRS, BPS, and the Visual Analog Scale (VAS) for measuring pain in patients in the ICU. METHODS: We performed a prospective observational study in 113 non-paralyzed critically ill patients. The attending nurses, two researchers, and the patient (when possible) obtained 371 independent observation series of NRS, BPS, and VAS. Data analyses were performed on the sample size of patients (n = 113). RESULTS: Inter-rater reliability of the NRS and BPS proved to be adequate (kappa = 0.71 and 0.67, respectively). The level of agreement within one scale point between NRS rated by the patient and NRS scored by attending nurses was 73%. However, high patient scores (NRS > or = 4) were underestimated by nurses (patients 33% versus nurses 18%). In responsive patients, a high correlation between NRS and VAS was found (rs = 0.84, P < 0.001). In ventilated patients, a moderate positive correlation was found between the NRS and the BPS (rs = 0.55, P < 0.001). However, whereas 6% of the observations were NRS of greater than or equal to 4, BPS scores were all very low (median 3.0, range 3.0 to 5.0). CONCLUSION: The different scales show a high reliability, but observer-based evaluation often underestimates the pain, particularly in the case of high NRS values (> or = 4) rated by the patient. Therefore, whenever this is possible, ICU patients should rate their pain. In unresponsive patients, primarily the attending nurse involved in daily care should score the patient's pain. In ventilated patients, the BPS should be used only in conjunction with the NRS nurse to measure pain levels in the absence of painful stimuli.

Intraoperative endobronchial tube displacement requiring unplanned extracorporeal circulation support.

We describe the adoption of extracorporeal circulation (ECC) as temporary pulmonary support in a patient operated on for a broncho-pleural fistula. Complex thoracic surgical procedures can require one-lung ventilation with double-lumen or selective bronchial tubes. Intraoperative tube displacement can cause severe desaturation and hypoxia, finally resulting in severe cerebral or multiorgan damages. Cardiopulmonary bypass, being able to temporarily take over the gas exchange function of the lung, is the only chance to support pulmonary function in cases such as this one. We suggest performing complex thoracic surgical procedures in centers with the availability and experience of ECC techniques.

Effect of Oxygenator Size on Air Removal Characteristics: A Clinical Evaluation.

During cardiopulmonary bypass (CPB), gaseous microemboli (GME) are released into the patients' arterial bloodstream. Gaseous microemboli may contribute to the adverse outcome after cardiac surgery. Recently, two oxygenator models with or without integrated arterial filter (IAF) were designed and only differ in size, leading to a change of 20% in surface area of the hollow fibers and 25% in blood velocities. The aim of this study was to assess the air removal characteristics of the inspire oxygenators with or without IAF. Sixty-eight patients were randomly assigned to four different groups: optimized adult and full adult and an additional IAF. Gaseous microemboli reduction rates were measured with a bubble counter. The number of GME reduction rates showed no differences. However, both models reduced significantly less volume of GME (optimized adult: 40.6% and full adult: 50.3%) compared with both models with IAF (88.7% and 88.5%, respectively). No significant differences of reduction rates were found between both devices without IAF and also not between both models with IAF. In conclusion, the larger inspire oxygenator tends to remove more GME. No effect from size of oxygenator device with integrated screen filter on GME reduction was observed. The inspire oxygenators with IAF may be considered as an adequate GME filter.

Complexes between C1q and C3 or C4: novel and specific markers for classical complement pathway activation.

Classical pathway activation is often assessed by measuring circulating levels of activated C4. However, this parameter does not discriminate between activation through the classical or the lectin pathway. We hypothesized that during classical pathway activation, complexes are formed between C1q and activated C4 or C3. Using ELISA, we investigated whether such complexes constitute specific markers for classical pathway activation. In vitro, C1q-C3d/C4d complexes were generated upon incubation of normal recalcified plasma with aggregated IgG or an anti-C1q mAb that activates C1 (mAb anti-C1q-130). In contrast, during incubation with C1s or trypsin, C1q-C3d/C4d complexes were not generated, which excludes an innocent bystander effect. Additionally, C1q-C3d/C4d complexes were not generated during activation of the alternative or the lectin pathway. Repeated freezing and thawing did not influence levels of C1q-C3d/C4d complexes in recalcified plasma. To measure C1q-complement complexes in plasma samples, we separated unbound complement proteins from C1q-C3d/C4d complexes in the samples prior to testing with ELISA. In samples from patients undergoing cardiopulmonary bypass surgery or suffering from rheumatoid arthritis, we found higher levels of C1q-C4 complexes than in samples from healthy individuals. We conclude that complexes between C1q and C4 or C3 are specific markers of classical complement pathway activation.

Vitamin B-6 vitamers in human plasma and cerebrospinal fluid.

BACKGROUND: Vitamin B-6 comprises a group of 6 interrelated vitamers and is essential for numerous physiologic processes, including brain functioning. Genetic disorders disrupting vitamin B-6 metabolism have severe clinical consequences. OBJECTIVE: To adequately diagnose known and novel disorders in vitamin B-6 metabolism, a reference set is required containing information on all vitamin B-6 vitamers in plasma and cerebrospinal fluid (CSF). DESIGN: Concentrations of vitamin B-6 vitamers in the plasma and CSF of 533 adult subjects were measured by ultra high-performance liquid chromatography-tandem mass spectrometry. RESULTS: The relative vitamin B-6 vitamer composition of plasma [pyridoxal phosphate (PLP) > pyridoxic acid (PA) > pyridoxal] differed from that of CSF (pyridoxal > PLP > PA > pyridoxamine). Sex influenced vitamin B-6 vitamer concentrations in plasma and CSF and should therefore be taken into account when interpreting vitamin B-6 vitamer concentrations. The strict ratios and strong correlations between vitamin B-6 vitamers point to a tight regulation of vitamin B-6 vitamer concentrations in blood and CSF. Given the unique design of this study, with simultaneously withdrawn blood and CSF from a large number of subjects, reliable CSF:plasma ratios and correlations of vitamin B-6 vitamers could be established. CONCLUSIONS: We provide an extensive reference set of vitamin B-6 vitamer concentrations in plasma and CSF. In addition to providing insight on the regulation of individual vitamers and their intercompartmental distribution, we anticipate that these data will prove to be a valuable reference set for the diagnosis and treatment of conditions associated with altered vitamin B-6 metabolism.

D-amino acid aberrations in cerebrospinal fluid and plasma of smokers.

The glutamatergic neurotransmission system and the N-methyl-D-aspartate receptor (NMDAR) have been implicated in smoking and alcohol consumption behavior. Preclinical studies have demonstrated that nicotine and ethanol influence NMDAR functionality, which may have a role in tendencies to consume these substances. Nonetheless, little is known about concentrations of NMDAR coagonists in the cerebrospinal fluid (CSF) and plasma of individuals who smoke or consume alcohol. Glycine and L- and D-stereoisomers of alanine, serine, and proline were therefore measured using ultra-high-performance liquid chromatography-tandem mass spectrometry in 403 healthy subjects. Nicotine and alcohol consumption were quantified using questionnaires. Possible differences in NMDAR coagonist concentrations in plasma and CSF were investigated using ANCOVA with age, body mass index, and storage duration as covariates. The significance threshold was Bonferroni corrected (α=0.00625). Compared with non-smokers, smokers displayed lower levels of D-proline in plasma (p=0.0027, Cohen's d=-0.41) and D-proline in CSF (p=0.0026, Cohen's d=-0.43). D-Serine in CSF was higher in smokers than in non-smokers (p=0.0052, Cohen's d=0.41). After subdividing participants based on smoking quantity, dose-dependent decreases were demonstrated in smokers for D-proline in plasma (F=5.65, p=0.0039) and D-proline in CSF (F=5.20, p=0.0060). No differences in NMDAR coagonist levels between alcohol consumption groups were detected. To our knowledge, this is the first report to implicate D-amino acids in smoking behavior of humans. Whether such concentration differences lie at the root of or result from smoking habits may be addressed in prospective studies.

A common variant in ERBB4 regulates GABA concentrations in human cerebrospinal fluid.

The neuregulin 1 (NRG1) receptor ErbB4 is involved in the development of cortical inhibitory GABAergic circuits and NRG1-ErbB4 signaling has been implicated in schizophrenia (SCZ). A magnetic resonance spectroscopy ((1)H-MRS) study has demonstrated that a single-nucleotide polymorphism in ERBB4, rs7598440, influences human cortical GABA concentrations. Other work has highlighted the significant impact of this genetic variant on expression of ERBB4 in the hippocampus and dorsolateral prefrontal cortex in human post mortem tissue. Our aim was to examine the association of rs7598440 with cerebrospinal fluid (CSF) GABA levels in healthy volunteers (n=155). We detected a significant dose-dependent association of the rs7598440 genotype with CSF GABA levels (G-allele standardized ß=-0.23; 95% CIs: -0.39 to -0.07; P=0.0066). GABA concentrations were highest in A homozygous, intermediate in heterozygous, and lowest in G homozygous subjects. When excluding subjects on psychotropic medication (three subjects using antidepressants), the results did not change (G-allele standardized ß=-0.23; 95% CIs: -0.40 to -0.07; P=0.0051). The explained variance in CSF GABA by rs7598440 in our model is 5.2% (P=0.004). The directionality of our findings agrees with the aforementioned (1)H-MRS and gene expression studies. Our observation therefore strengthens the evidence that the A-allele of rs7598440 in ERBB4 is associated with increased GABA concentrations in the human central nervous system (CNS). To our knowledge, our finding constitutes the first confirmation that CSF can be used to study genotype-phenotype correlations of GABA levels in the CNS. Such quantitative genetic analyses may be extrapolated to other CSF constituents relevant to SCZ in future studies.

Season of sampling and season of birth influence serotonin metabolite levels in human cerebrospinal fluid.

BACKGROUND: Animal studies have revealed seasonal patterns in cerebrospinal fluid (CSF) monoamine (MA) turnover. In humans, no study had systematically assessed seasonal patterns in CSF MA turnover in a large set of healthy adults. METHODOLOGY/PRINCIPAL FINDINGS: Standardized amounts of CSF were prospectively collected from 223 healthy individuals undergoing spinal anesthesia for minor surgical procedures. The metabolites of serotonin (5-hydroxyindoleacetic acid, 5-HIAA), dopamine (homovanillic acid, HVA) and norepinephrine (3-methoxy-4-hydroxyphenylglycol, MPHG) were measured using high performance liquid chromatography (HPLC). Concentration measurements by sampling and birth dates were modeled using a non-linear quantile cosine function and locally weighted scatterplot smoothing (LOESS, span = 0.75). The cosine model showed a unimodal season of sampling 5-HIAA zenith in April and a nadir in October (p-value of the amplitude of the cosine = 0.00050), with predicted maximum (PC(max)) and minimum (PC(min)) concentrations of 173 and 108 nmol/L, respectively, implying a 60% increase from trough to peak. Season of birth showed a unimodal 5-HIAA zenith in May and a nadir in November (p = 0.00339; PC(max) = 172 and PC(min) = 126). The non-parametric LOESS showed a similar pattern to the cosine in both season of sampling and season of birth models, validating the cosine model. A final model including both sampling and birth months demonstrated that both sampling and birth seasons were independent predictors of 5-HIAA concentrations. CONCLUSION: In subjects without mental illness, 5-HT turnover shows circannual variation by season of sampling as well as season of birth, with peaks in spring and troughs in fall.