<p>During brain activity neurons release the major excitatory transmitter glutamate, which is taken up by astrocytes and converted to glutamine. Glutamine returns to neurons for re-conversion to glutamate. This glutamate-glutamine cycle is energy demanding. Glutamate turnover in injured brain was studied using an animal iron-induced posttraumatic epilepsy model and using neurointensive care data from 33 patients with spontaneous subarachnoid hemorrhage (SAH). Immunoblotting revealed that the functional form of the major astrocytic glutamate uptake protein GLT-1 was decreased 1-5 days following a cortical epileptogenic iron-injection, presumably due to oxidation-induced aggregation. Using microdialysis it was shown that the GLT-1 decrease was associated with increased interstitial glutamate levels and decreased interstitial glutamine levels. The results indicate a possible posttraumatic and post-stroke epileptogenic mechanism. Analysing 3600 microdialysis hours from patients it was found that the interstitial lactate/pyruvate (L/P) ratio correlate with the glutamine/glutamate ratio (r =-0.66). This correlation was as strong as the correlation between L/P and glutamate (r=0.68) and between lactate and glutamate (r=0.65). Pyruvate and glutamine correlated linearly (r=0.52). Energy failure periods, defined as L/P>40, were associated with high interstitial glutamate levels. Glutamine increased or decreased during energy failure periods depending on pyruvate. Energy failure periods were clinically associated with delayed ischemic neurological deficits (DIND) or development of radiologically verified infarcts, confirming that L/P>40 is a pathological microdialysis pattern that can predict ischemic deterioration after SAH. DIND-associated microdialysis patterns were L/P elevations and surges in interstitial glutamine. Glutamine and pyruvate correlated with the cerebral perfusion pressure (r=0.25, r=0.24). Glutamine and the glutamine/glutamate ratio correlated with the intracranial pressure (r=-0.29, r=0.40). Glutamine surges appeared upon substantial lowering of the intracranial pressure by increased cerebrospinal fluid drainage. Increased interstitial glutamine and pyruvate levels may reflect augmented astrocytic glycolysis in recovering brain tissue with increased energy demand due to a high glutamate-glutamine turnover.</p>

The neurological wake-up test, NWT, is a clinical monitoring tool that can be used to evaluate the level of consciousness in patients with traumatic brain injury (TBI) and subarachnoid haemorrhage (SAH) during neurocritical care (NCC). Since patients with severe TBI or SAH are often treated with mechanical ventilation and sedation, the NWT requires that the continuous sedation is interrupted. However, interruption of continuous sedation may induce a stress response and the use of the NWT in NCC is controversial. The effects of the NWT on intracranial pressure (ICP) and cerebral perfusion pressure (CPP) were evaluated in 21 patients with TBI or SAH. Compared to baseline when the patients were sedated with continuous propofol sedation, the NWT resulted in increased ICP and CPP (p&lt;0.05). Next, the effects of the NWT on the stress hormones adrenocorticotrophic hormone (ACTH), cortisol, epinephrine and norepinephrine were evaluated in 24 patients. Compared to baseline, the NWT caused a mild stress response resulting in increased levels of all evaluated stress hormones (p&lt;0.05). To compare the use of routine NCC monitoring tools, the choice of sedation and analgesia and the frequency of NWT in Scandinavian NCC units, a questionnaire was used. The results showed that all 16 Scandinavian NCC units routinely use ICP and CPP monitoring and propofol and midazolam were primary choices for patient sedation in an equal number of NCC units. In 2009, the NWT was not routinely used in eight NCC units whereas others used the test up to six times daily. Finally, intracerebral microdialysis (MD), brain tissue oxygenation (PbtiO2) and jugular bulb oxygenation (SjvO2) were used in 17 TBI patients to evaluate the effect of the NWT procedure on focal neurochemistry and cerebral oxygenation. The NWT did not negatively alter interstitial markers of energy metabolism or cerebral oxygenation. In conclusion, the NWT induced a mild stress response in patients with TBI or SAH that did not result in a detectable, significant secondary insult to the injured brain. These results suggest that the NWT may safely be used as a clinical monitoring tool in the NCC of severe TBI and SAH in a majority of patients.

During brain activity neurons release the major excitatory transmitter glutamate, which is taken up by astrocytes and converted to glutamine. Glutamine returns to neurons for re-conversion to glutamate. This glutamate-glutamine cycle is energy demanding. Glutamate turnover in injured brain was studied using an animal iron-induced posttraumatic epilepsy model and using neurointensive care data from 33 patients with spontaneous subarachnoid hemorrhage (SAH). Immunoblotting revealed that the functional form of the major astrocytic glutamate uptake protein GLT-1 was decreased 1-5 days following a cortical epileptogenic iron-injection, presumably due to oxidation-induced aggregation. Using microdialysis it was shown that the GLT-1 decrease was associated with increased interstitial glutamate levels and decreased interstitial glutamine levels. The results indicate a possible posttraumatic and post-stroke epileptogenic mechanism. Analysing 3600 microdialysis hours from patients it was found that the interstitial lactate/pyruvate (L/P) ratio correlate with the glutamine/glutamate ratio (r =-0.66). This correlation was as strong as the correlation between L/P and glutamate (r=0.68) and between lactate and glutamate (r=0.65). Pyruvate and glutamine correlated linearly (r=0.52). Energy failure periods, defined as L/P&gt;40, were associated with high interstitial glutamate levels. Glutamine increased or decreased during energy failure periods depending on pyruvate. Energy failure periods were clinically associated with delayed ischemic neurological deficits (DIND) or development of radiologically verified infarcts, confirming that L/P&gt;40 is a pathological microdialysis pattern that can predict ischemic deterioration after SAH. DIND-associated microdialysis patterns were L/P elevations and surges in interstitial glutamine. Glutamine and pyruvate correlated with the cerebral perfusion pressure (r=0.25, r=0.24). Glutamine and the glutamine/glutamate ratio correlated with the intracranial pressure (r=-0.29, r=0.40). Glutamine surges appeared upon substantial lowering of the intracranial pressure by increased cerebrospinal fluid drainage. Increased interstitial glutamine and pyruvate levels may reflect augmented astrocytic glycolysis in recovering brain tissue with increased energy demand due to a high glutamate-glutamine turnover.

There is a paucity of studies, clinical and experimental, attesting to the benefit of cerebral perfusion pressure (CPP) directed pressor therapy following traumatic brain injury (TBI). The current study evaluates this therapy in a swine model of TBI and hypotension. Forty-five anesthetized and ventilated swine received TBI followed by a 45% blood volume bleed. After 1 h, all animals were resuscitated with 0.9% sodium chloride equal to three times the shed blood volume. The experimental group (PHE) received phenylephrine to maintain CPP > 80 mm Hg; the control group (SAL) did not. Outcomes in the first phase (n = 33) of the study were as follows: cerebro-venous oxygen saturation (S cvO2), cerebro-vascular carbon dioxide reactivity (δScvO2), and brain structural damage (β-amyloid precursor protein [βAPP] immunoreactivity). In the second phase (n = 12) of the study, extravascular blood free water (EVBFW) was measured in the brain and lung. After resuscitation, intracranial and mean arterial pressures were >15 and >80 mm Hg, respectively, in both groups. CPP declined to 64 ± 5 mm Hg in the SAL group, despite fluid supplements. CPP was maintained at >80 mm Hg with pressors in the PHE group. PHE animals maintained better ScvO2 (p < 0.05 at 180, 210, 240, 270, and 300 min post-TBI). At baseline, 5% CO2 evoked a 16 ± 4% increase in ScvO2, indicating cerebral vasodilatation and luxury perfusion. By 240 min, this response was absent in SAL animals and preserved in PHE animals (p < 0.05). Brain EVBFW was higher in SAL animals; however, lung EVBFW was higher in PHE animals. There was no difference in βAPP immunoreactivity between the SAL and PHE groups (p > 0.05). In this swine model of TBI and hypotension, CPP directed pressor therapy improved brain oxygenation and maintained cerebro-vascular CO2 reactivity. Brain edema was lower, but lung edema was greater, suggesting a higher propensity for pulmonary complications.

The original purpose of this study was to compare initial resuscitation of hemorrhagic hypotension after traumatic brain injury (TBI) with saline and shed blood. Based on those results, the protocol was modified and saline was compared to a blood substitute, diaspirin cross-linked hemoglobin (DCLHb). Two series of experiments were performed in anesthetized and mechanically ventilated (FiO2 = 0.4) pigs (35-45 kg). In Series 1, fluid percussion TBI (6-8 ATM) was followed by a 30% hemorrhage. At 120 min post-TBI, initial resuscitation consisted of either shed blood (n = 7) or a bolus of 3x shed blood volume as saline (n = 13). Saline supplements were then administered to all pigs to maintain a systolic arterial blood pressure (SAP) of >100 mmHg and a heart rate (HR) of <110 beats/min. In Series 2, TBI (4-5 ATM) was followed by a 35% hemorrhage. At 60 min post-TBI, initial resuscitation consisted of either 500 mL of DCLHb (n = 6) or 500 mL of saline (n = 5). This was followed by saline supplements to all pigs to maintain a SAP of >100 mmHg and a HR of <110 beats/min. In Series 1, most systemic markers of resuscitation (e.g., SAP, HR, cardiac output, filling pressures, lactate, etc.) were normalized, but there were 0/7 vs. 5/13 deaths within 5 h (P = 0.058) with blood vs. saline. At constant arterial O2 saturation (SaO2), mixed venous O2 saturation (SvO2), cerebral perfusion pressure (CPP), and cerebral venous O2 saturation (ScvO2) were all higher, intracranial pressure (ICP) was lower, and CO2 reactivity was preserved with blood vs. saline (all P < 0.05). In Series 2, SAP, ICP, CPP, and lactate were higher with DCLHb vs. saline (all P < 0.05). Cardiac output was lower even though filling pressure was markedly elevated with DCLHb vs. saline (both P < 0.05). Neither SvO2 nor cerebrovascular CO2 reactivity were improved, and ScvO2 was lower with DCLHb vs. saline (P < 0.05). All survived at least 72 h with neuropathologic changes that included sub-arachnoid hemorrhage, midline cerebellar necrosis, and diffuse axonal injury. These changes were similar with DCLHb vs. saline. Thus, whole blood was more effective than saline for resuscitation of TBI, whereas DCLHb was no more, and according to many variables, less effective than saline resuscitation. These experimental results are comparable to those in a recent multicenter trial using DCLHb for the treatment of severe traumatic shock. Further investigations in similar experimental models might provide some plausible explanations why DCLHb unexpectedly increased mortality in patients.

The Arterial Spin Labelling (ASL) method is a Magnetic Resonance technique used toquantify the cerebral perfusion. It has the big advantage to be non-invasive so doesn’tneed the injection of any contrast agent. But due to a relatively low Signal-to-NoiseRatio (SNR) of the signal acquired (only approximately 1% of the image intensity), ithas been hampered to be widely used in a clinical setting so far.The primary objective of this project is to make the method more robust by improvingthe quality of the images, the SNR, and by reducing the acquisition time. DifferentASL protocols with different sets of parameters have been investigated. The modificationsperformed on the protocol have been investigated by analyzing images acquired onhealthy volunteers. An optimized protocol leading to a good trade-off between the differentaspects of the method, has been suggested. It is characterized by a 3:43:44:0mm3with a two-segment acquisition.A more advanced ASL method implies the acquisition of images at different inversiontimes (TI), which is called the mutli-TI method. The influence of the range of TI used inthe method has been explored. An optimized TI range (from 410ms to 3860ms, sampledevery 150ms) has been suggested to make the ASL method as performant as possible.A numerical model and a fitting algorithm have been used to extract the informationon the perfusion from the images acquired. Different models have been investigated aswell as their influence on the reliability of the results.Finally, a criterion has been implemented to evaluate the reliability of the results sothat the clinician or the user of the method can figure out how much he can count onthe results provided by the method.

Cerebral small vessel disease (SVD) is identifiable by clinical, neuroimaging, neuropathological and cognitive findings. The aim of this thesis was to assess SVD and cerebral perfusion on magnetic resonance imaging (MRI) in a 75-year-old population and compare the findings with scars of myocardial infarctions, cardiovascular risk markers and cognitive function. In addition, the evolution of SVD over 5 years was studied. The study population included subjects from the Prospective Investigation of the Vasculature in Uppsala Seniors (PIVUS) study. The subjects had been chosen in a randomized manner from the register of the municipality. MRI of the brain and the heart, cognitive tests and blood tests for cardiovascular risk factors were performed in 406 subjects at age 75 years and 250 of them were re-examined 5 years later at the age of 80. Paper 1 showed that unrecognized myocardial infarctions (UMIs) were found in 120 subjects (30%) and recognized myocardial infarctions (RMIs) in 21 (5%). Men with RMIs displayed an increased prevalence of cortical and lacunar cerebral infarctions, whereas women with UMIs more frequently had cortical cerebral infarctions. Paper 2 showed that one or more brain infarcts were seen in 23% of the subjects (20% had only lacunar infarcts, 1% had only cortical infarcts and 2% had both). Hypertension and obesity were significantly associated with an increased risk of infarction.  The newer risk markers investigated were not significantly associated with brain infarcts. Paper 3 showed that MRI manifestations of SVD progressed over 5 years. Relative cerebral blood flow (rCBF) was not associated with WMH volume or progression of WMH volume. Paper 4 showed that moderate to severe WMHs and incident lacunar infarcts on brain MRI were associated with a mild impairment of executive function. In conclusion, this longitudinal population based study compares MRI manifestations of SVD with clinical data, providing knowledge that may be used in further investigations of preventive interventions and for identification of disease in early stages.

A esquizofrenia é um transtorno psiquiátrico incapacitante que afeta estimadamente 1% da população mundial. Delírios, alucinações, desorganização de pensamento e prejuízo cognitivo são as principais marcas da Esquizofrenia. Fisiologicamente, além de anormalidades funcionais e estruturais, alterações na atividade neuronal são reportadas. Como a atividade neuronal possui uma relação direta com o fluxo sanguíneo cerebral (CBF, Cerebral Blood Flow), a técnica de Imagens por Ressonância Magnética, denominada Marcação dos Spins Arteriais (ASL, Arterial Spin Labeling), que permite a obtenção de mapa quantitativo de CBF, é uma ferramenta útil na avaliação funcional cerebral. Além disso, a ASL pode ser usada na avaliação da conectividade funcional, que é eficiente na investigação de rupturas funcionais entre as regiões do cérebro. Comparando com um grupo de sujeitos saudáveis, os pacientes com esquizofrenia, recrutados no Hospital das Clínicas de Ribeirão Preto (HCFMRP), apresentaram redução de CBF em regiões bilaterais do polo frontal e giro frontal superior, giro frontal medial direito, partes triangular e opercular do giro frontal inferior direito, divisão posterior do giro supramarginal esquerdo, divisão superior e inferior do córtex occipital lateral esquerdo e polo occipital. A conectividade funcional, avaliada por três diferentes métodos (baseado em semente, análise de componentes independentes e teoria dos grafos), se apresentou prejudicada em regiões envolvendo funções motoras, sensoriais e cognitivas dos pacientes. Portanto, utilizando uma técnica de imagem completamente não invasiva, foi possível observar déficits de CBF e alterações na organização funcional do cérebro de pacientes com esquizofrenia, relacionados com os sintomas e características da psicopatologia. / Schizophrenia is a disabling psychiatric disorder that affects around 1% of the population worldwide. Delusions, hallucinations, disorganized thought, and cognitive deficits are the main features of schizophrenia. Physiologically, in addition to functional and structural abnormalities, changes in neuronal activity are reported. Since the Cerebral Blood Flow (CBF) is directly related with neuronal activity, the Magnetic Resonance Imaging (MRI) technique called Arterial Spin Labeling (ASL), which allows the quantification of CBF, is a useful tool in brain functional evaluation. In addition, ASL can be used to assess functional connectivity, which is efficient in investigating functional impairment between regions of the brain. Patients with Schizophrenia, recruited at the Clinical Hospital (HCFMRP), presented a reduction of CBF in bilateral regions of the frontal pole and superior frontal gyrus, right medial frontal gyrus, triangular and opercular parts of the right inferior frontal gyrus, posterior division of left supramarginal gyrus, superior and inferior division of left lateral occipital cortex and occipital pole. Functional connectivity, assessed by three different methods (seed-based, independent component analysis and graph theory), was impaired in regions involving patients\' motor, sensory and cognitive functions. Therefore, using a noninvasive imaging technique, it was possible to observe CBF deficits and alterations in the functional organization of the brain of schizophrenia patients, related to the symptoms and characteristics of the psychopathology.

A esquizofrenia é um transtorno psiquiátrico incapacitante que afeta estimadamente 1% da população mundial. Delírios, alucinações, desorganização de pensamento e prejuízo cognitivo são as principais marcas da Esquizofrenia. Fisiologicamente, além de anormalidades funcionais e estruturais, alterações na atividade neuronal são reportadas. Como a atividade neuronal possui uma relação direta com o fluxo sanguíneo cerebral (CBF, Cerebral Blood Flow), a técnica de Imagens por Ressonância Magnética, denominada Marcação dos Spins Arteriais (ASL, Arterial Spin Labeling), que permite a obtenção de mapa quantitativo de CBF, é uma ferramenta útil na avaliação funcional cerebral. Além disso, a ASL pode ser usada na avaliação da conectividade funcional, que é eficiente na investigação de rupturas funcionais entre as regiões do cérebro. Comparando com um grupo de sujeitos saudáveis, os pacientes com esquizofrenia, recrutados no Hospital das Clínicas de Ribeirão Preto (HCFMRP), apresentaram redução de CBF em regiões bilaterais do polo frontal e giro frontal superior, giro frontal medial direito, partes triangular e opercular do giro frontal inferior direito, divisão posterior do giro supramarginal esquerdo, divisão superior e inferior do córtex occipital lateral esquerdo e polo occipital. A conectividade funcional, avaliada por três diferentes métodos (baseado em semente, análise de componentes independentes e teoria dos grafos), se apresentou prejudicada em regiões envolvendo funções motoras, sensoriais e cognitivas dos pacientes. Portanto, utilizando uma técnica de imagem completamente não invasiva, foi possível observar déficits de CBF e alterações na organização funcional do cérebro de pacientes com esquizofrenia, relacionados com os sintomas e características da psicopatologia. / Schizophrenia is a disabling psychiatric disorder that affects around 1% of the population worldwide. Delusions, hallucinations, disorganized thought, and cognitive deficits are the main features of schizophrenia. Physiologically, in addition to functional and structural abnormalities, changes in neuronal activity are reported. Since the Cerebral Blood Flow (CBF) is directly related with neuronal activity, the Magnetic Resonance Imaging (MRI) technique called Arterial Spin Labeling (ASL), which allows the quantification of CBF, is a useful tool in brain functional evaluation. In addition, ASL can be used to assess functional connectivity, which is efficient in investigating functional impairment between regions of the brain. Patients with Schizophrenia, recruited at the Clinical Hospital (HCFMRP), presented a reduction of CBF in bilateral regions of the frontal pole and superior frontal gyrus, right medial frontal gyrus, triangular and opercular parts of the right inferior frontal gyrus, posterior division of left supramarginal gyrus, superior and inferior division of left lateral occipital cortex and occipital pole. Functional connectivity, assessed by three different methods (seed-based, independent component analysis and graph theory), was impaired in regions involving patients\' motor, sensory and cognitive functions. Therefore, using a noninvasive imaging technique, it was possible to observe CBF deficits and alterations in the functional organization of the brain of schizophrenia patients, related to the symptoms and characteristics of the psychopathology.

<p>Traumatic brain injury (TBI) is a major cause of death and disability. Intracranial secondary insults (e.g. intracranial haematoma, brain oedema) and systemic secondary insults (e.g. hypotension, hypoxaemia, hyperthermia) lead to secondary brain injury and affect outcome adversely. In order to minimise secondary insults and to improve outcome in TBI-patients, a secondary insult program and standardised neurointensive care (NIC) was implemented. The aim of this thesis was to describe patient outcome and to explore the occurrence and prognostic value of secondary insults after the implementation.</p><p>Favourable outcome was achieved in 79% and 6% died of the 154 adult TBI patients treated in the NIC unit 1996-97. In an earlier patient series from the department, 48% made a favourable outcome and 31% died. Hence, the outcome seems to have improved when NIC was standardised and dedicated to avoiding secondary insults. </p><p>Secondary insults counted manually from hourly recordings on surveillance charts did not hold any independent prognostic information. When utilising a computerised system, which enables minute-by-minute data collection, the proportion of monitoring time with systolic blood pressure > 160 mm Hg decreased the odds of favourable outcome independent of admission variables (odds ratio 0.66). Hyperthermia was related to unfavourable outcome. Hypertension was correlated to hyperthermia and may be a part of a hyperdynamic state aggravating brain oedema. </p><p>Increased proportion of monitoring time with cerebral perfusion pressure (CPP) < 60 mm Hg increased the odds of favourable outcome (odds ratio 1.59) in patients treated according to an intracranial pressure (ICP)-oriented protocol (Uppsala). In patients given a CPP-oriented treatment (Edinburgh), CPP <60 mm Hg was coupled to an unfavourable outcome. It was shown that pressure passive patients seem to benefit from an ICP-oriented protocol and pressure active patients from a CPP-oriented protocol. The overall outcome would improve if patients were given a treatment fit for their condition.</p>