Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2008. / Härtill 5 uppsatser.

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2010. / Härtill 4 uppsatser.

Alzheimer's disease (AD) is the most common cause of dementia; a problem that is growing in size and cost as the population ages. Early soluble aggregates composed misfolded 'amyloid' peptide sequences have been implicated as key to the initiation and onset of AD pathology, although little is definitively known as to when and how these assemblies form or interact to instigate pathology. The primary focus of this study was to evaluate whether L-Tryptophanol (Trol) signal, which has been shown to be induced via soluble amyloid species, increases with AD severity in a range of ex vivo human samples. Testing of this hypothesis was carried out in several stages: Initially synthetic versions of the amyloid beta (Aβ) peptide were tested in vitro to corroborate Trols propensity to associate to amyloid assemblies and allow for method development. Next, a range of brain lysates from several transgenic mouse lines and aged human AD cases and controls were assessed using the reporter. These experiments demonstrated Trols sensitivity to Aβ and tau, and provided compelling evidence that Trol signal tracks disease progression in brain lysates. During the final stage of testing cerebrospinal fluid (CSF) from AD and Parkinson's disease (PD) patients, and blood plasma samples from PD patients was evaluated. Results from this phase of testing indicated that Trol was able to detect differences in sample composition between healthy and diseased individuals, however differences were not clear cut and could have been affected by confounding factors. Overall, the data presented here suggest that Trol may be able to track disease progression in amyloidopathies when implemented in brain lysates. However, further testing is required to completely validate this finding. These findings highlight the potential of simple techniques for amyloid detection to aid within the diagnosis, evaluation of disease progression and study of AD and other neurodegenerative diseases.

Thesis (Master's Diploma(Technology (Medical Technology))-- Cape Technikon, 1997 / Meningitis in children is a common and serious disease. Bacterial and tuberculous meningitis often lead to neurological complications. A sensitive marker to predict brain damage in children with meningitis could be of great importance. Frithz F et aI, 1982 suggested that increased adenylate kinase values could indeed be used as a marker for brain damage. Adenylate kinase (AK) is an enzyme present in brain tissue. Low concentrations are present in normal cerebrospinal fluid (CSF) « 1 uti). Increased concentrations were found in cases of ischemic brain damage (Frithz et aI, 1982), malignant brain tumours (Ronquist G et aI, 1977) and bacterial meningitis. As AK has a low molecular weight (22,00 Daltons), in comparison to other kinases (40,000 Daltons) it is one of the first enzymes that can be detected in the CSF after brain damage and it can thus be used as a reliable marker for brain cell damage. The aim of this study was to quantify the AK values in CSF of children with bacterial and tuberculous meningitis and to evaluate their use to predict the neurological outcome in children with bacterial and tuberculous meningitis. Eighty eight children with tuberculous meningitis (TBM) and thirty three children with bacterial meningitis were included in the study. Sixty children with suspected meningitis but who were later diagnosed with urinary tract infections, gasto-enteritis, bronchitis, febrile convulsions or other non-neurological infections were used as controls. The results showed raised AK values in the CSF of children with bacterial- and TB meningitis. There was a statistically significant difference of AK values between stage III and II TBM AK values in patients at week 1 after diagnosis (p=0,03). There was also a statistically significant correlation between CSF AK values and lactate concentrations (P=0,001) which reflected hypoxic brain metabolism. Although AK values did not always correlate directly with the patients’ clinical outcome, there is proof that increased AK values in CSF can be used to predict neurological outcome.

The lactate level assesment in various body fluids plays an important role in the diagnostics of mitochondrial disorders in children. However, the interpretation of lactate level is often difficult due to its unspecificity and variability even in particular mitochondrial disorders. Three specific aims have been stated in this PhD Thesis: 1. To analyse the role of lactate examination in the differential diagnosis between children with mitochondrial disorders and children with other diseases. 2. To study the lactate level differences in various mitochondrial syndromes. 3. To characterise the clinical and laboratory data of neonates with mitochondrial disorders and to suggest new diagnostic algorhytms. Clinical and laboratory data from patients hospitalized in the Department of Pediatrics were collected. Laboratory methods were provided in the cooperation with the Mitochondrial laboratory of the Department of Pediatrics and Institute of Inherited Metabolic Disorders. The study with lactate levels in 107 patients documented that brief seizures lasting less than 2 minutes did not increase lactate concentration in the CSF. CSF-lactate was a relialable marker in differential diagnosis in the children with mitochondrial disorders against children with epilepsy. 2. The severity of particular phenotype is more...

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We examined the extracellular vesicle (EV) and RNA composition of pooled normal cerebrospinal fluid (CSF) samples and CSF from five major neurological disorders: Alzheimer's disease (AD), Parkinson's disease (PD), low-grade glioma (LGG), glioblastoma multiforme (GBM), and subarachnoid haemorrhage (SAH), representing neurodegenerative disease, cancer, and severe acute brain injury. We evaluated: (I) size and quantity of EVs by nanoparticle tracking analysis (NTA) and vesicle flow cytometry (VFC), (II) RNA yield and purity using four RNA isolation kits, (III) replication of RNA yields within and between laboratories, and (IV) composition of total and EV RNAs by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and RNA sequencing (RNASeq). The CSF contained similar to 106 EVs/mu L by NTA and VFC. Brain tumour and SAH CSF contained more EVs and RNA relative to normal, AD, and PD. RT-qPCR and RNASeq identified disease-related populations of microRNAs and messenger RNAs (mRNAs) relative to normal CSF, in both total and EV fractions. This work presents relevant measures selected to inform the design of subsequent replicative CSF studies. The range of neurological diseases highlights variations in total and EV RNA content due to disease or collection site, revealing critical considerations guiding the selection of appropriate approaches and controls for CSF studies.

The lactate level assesment in various body fluids plays an important role in the diagnostics of mitochondrial disorders in children. However, the interpretation of lactate level is often difficult due to its unspecificity and variability even in particular mitochondrial disorders. Three specific aims have been stated in this PhD Thesis: 1. To analyse the role of lactate examination in the differential diagnosis between children with mitochondrial disorders and children with other diseases. 2. To study the lactate level differences in various mitochondrial syndromes. 3. To characterise the clinical and laboratory data of neonates with mitochondrial disorders and to suggest new diagnostic algorhytms. Clinical and laboratory data from patients hospitalized in the Department of Pediatrics were collected. Laboratory methods were provided in the cooperation with the Mitochondrial laboratory of the Department of Pediatrics and Institute of Inherited Metabolic Disorders. The study with lactate levels in 107 patients documented that brief seizures lasting less than 2 minutes did not increase lactate concentration in the CSF. CSF-lactate was a relialable marker in differential diagnosis in the children with mitochondrial disorders against children with epilepsy. 2. The severity of particular phenotype is more...

Previous studies showed that memantine inhibits tau hyperphosphorylation in vitro. In this study, phosphorylated tau (P-tau) and total tau (T-tau) were measured before and after 6 month treatment with memantine in 12 subjects ranging from normal cognition with subjective memory complaints, through mild cognitive impairment to mild Alzheimer's disease. Thirteen non-treated individuals served as controls. Treatment was associated with a reduction of P-tau in subjects with normal cognition. No treatment effects were seen among impaired individuals, suggesting that longer treatment time may be necessary to achieve biomarker effect in this group.

Cerebrospinal fluid (CSF) analysis is a standard part of a diagnostic evaluation. Commonly evaluated components include the cell count, protein concentration, glucose, and cytology. CSF analysis can be diagnostic in some diseases such as fungal infections and CNS lymphoma. Often, CSF analysis is not specific, but more information can be obtained. Matrix Metalloproteinases (MMPs) are enzymes that have been found in human CSF. They are calcium and zinc dependent endoproteinases with overlapping substrates. They hydrolyze at least one component of tissue extracellular matrix (ECM), such as collagen or elastin. They are important in normal physiologic processes such as angiogenesis, reproduction and wound healing. One class of MMPs, the gelatinases, degrade gelatins and type IV collagen include MMP 2 and MMP 9. MMPs are important in many pathological processes that involve unregulated matrix destruction such as arthritis, neoplasia and CNS diseases. MMP2 is known to be constituitively produced in CSF while MMP 9 is present only in certain pathologic conditions such as multiple sclerosis, neoplasia and inflammatory diseases. We hypothesize that MMP2 is present in normal canine CSF while MMP 9 is absent. Cerebrospinal fluid samples were taken from 23 normal dogs that were being used for other research purposes. Each CSF sample was evaluated immediately for red blood cells (RBCs), white blood cells (WBCs), protein, and glucose, and then stored at -70°C. Cytological examination was also performed. CSF samples were considered normal if the protein was less than 25 mg/dl, WBCs were less than 6 µl, and RBCs were less than 25 µl. Each dog was euthanized and the brains processed for routine histopathology. MMP analysis was done using gelatin zymography and an enzyme linked immunosorbent assay (ELISA). Bands of enzyme activity were visible following staining due to enzyme degradation of the gelatin. A commercially available polyclonal sandwich ELISA was used to identify the pro form of MMP2. The mean WBC count for the CSF samples was 0.96 WBC/ml with a range of 0-3 WBC/ml. The mean protein was 12 mg/dl, with a range of 8-17 mg/dl. The mean RBC count was 3.65 RBC/ml with a range of 0-21 RBC/ml. All normal samples of CSF contained a band of clearing that corresponded to the human commercial standard of proMMP2. No other major bands of clearing were noted on normal samples. The commercial human standards also contained ProMMP2. Other bands were present, but were faint and variable. Using a polyclonal antibody based sandwich ELISA, with samples run in triplicate, the mean pro MMP 2 levels were determined to be 5.61 ng/ml with a range of 3.36 - 10.83 ng/ml. We conclude that normal CSF values are narrower than what has been previously reported for protein concentration and WBC count. Also, the pro form of MMP 2 is present in normal canine CSF based on results of gelatin zymography and ELISA. / Master of Science