Thalamotomy and pallidotomy constitute the two principal neurosurgical treatments for Parkinson's disease. Both the thalamus nuclei and the internal globus pallidus structures are found very close to the internal capsule, a critical structure that must be avoided in performing the lesion. This forces the lesion of parts of the thalamus and the globus pallidus mediale to be achieved with a spatial precision on the order of 2mm. The leukotome is a neurosurgical tool consisting of a 20cm rigid shaft with a thin metallic wire attached to its tip that can be taken in and out of the body of the shaft. It is used as a knife to perform the excision. / Some critical basal ganglia structures show no anatomical differences at all on MRI scans, even though they have different functionality. In order to provide the neurosurgeon with this missing information, we have developed a deformable volumetric atlas of the basal ganglions from the cryogenic slices atlas of Shaltenbrand and Wahren. The atlas can be non-linearly deformed to fit a patient's MRI. / We have developed a visualization platform for pre-operative and intra-operative utilization of the atlas and of the MRI datasets. Any number of datasets can be superimposed (MRI, atlases, virtual lesions...). Merging of datasets with different extents and different resolutions is supported. The platform includes 3D visualization tools as well. Graphical tools allow the neurosurgeon to see projections of the leukotome from any point of view over the MRI data and the atlas. The software is able to create models of lesions that can be compared with the MRI data and the atlas. It can also suggest an operation protocol (how to use the leukotome) if a target volume is given. This target volume can easily be drawn on the atlas by the neurosurgeon.

When the Y chromosome of a Mus. musculus domesticus mouse strain is placed onto the C57BL/6J(B6) background, half of the XY (B6.Y$ rm sp{Dom})$ progeny develop as females, which ovulate very few eggs and fail to develop zygotes beyond the two-cell stage. In the present study, oocyte-cumulus complexes were isolated from ovaries of juvenile B6.Y$ rm sp{Dom}$ females, as well as XX and XO females for comparison, and matured and fertilized in vitro. The results indicate that (1) In vitro procedure yielded far more mature oocytes than through ovulation; (2) The oocytes underwent apparently normal nuclear maturation, but failed to develop after fertilization; and (3) Addition of FSH during oocyte maturation increased the percentage of fertilization and the first cell cleavage, but not further development. The oocytes from XO ovaries were comparable with those from XX rather than XY ovaries, suggesting that the presence of the Y chromosome is responsible for the infertility of the B6.Y$ rm sp{Dom}$ female.

Marrow stromal cells (MSCs) are pluripotent stem cells which may participate in the repair of damaged organs. We tested the hypothesis that MSCs can be recruited to the heart upon myocardial infarction, and play pathophysiological roles in the subsequent healing and adaptation process. An animal model was created with labeled MSCs in its bone marrow and then a myocardial infarction produced. Donor MSCs from isogenic Lewis rats were harvested, multiplied in culture and labelled. Labelled cells were intravenously injected into the recipient rats and one week later upon the engraftment of these labelled MSCs within the bone marrow, rats underwent a coronary artery ligation or sham operation. The hearts were removed at various time points and the presence of labelled cells in the heart was confirmed and their phenotypes identified. We confirmed the presence of labelled cells in the rat bone marrows and the presence of labelled cells in the infarcted myocardium at all time points studied, but not in the normal hearts. There was evidence for myogenic differentiation with some labelled cells expressing smooth muscle/myofibroblast phenotypes and appearing to participate in vasculogenesis. Our evidence is consistent with the hypothesis that myocardial infarction may send a signal to recruit MSCs to the injured heart, where they undergo milieu-dependent differentiation. The ability of these cells to express various phenotypes may allow them to participate in the pathophysiology of post-infarct remodeling and angiogenesis. Therapeutic implantation of MSCs thus may further enhance such effects.

The human progranulin gene encodes a 593 amino acid secreted glycoprotein, progranulin (also called PC cell-derived growth factor, acrogranin or granulin/epithelin precursor). The biological activities of this protein were poorly defined at the beginning of the project. Here, we determined that the progranulin gene is widely expressed in vivo and strongly associated with immune, neuronal and highly proliferative epithelial cells. It is not expressed in fibroblasts and endothelial cells in vivo unless these cells are stimulated by, for example, tissue damage and this expression correlates temporally with the healing process. Thus progranuhn is induclbly expressed in mesenchymal cells but is a constitutive product in proliferative epithelial cells. These studies defined a physiological context for investigating the roles of progranulin in the body. To correlate the expression studies with function, we assessed the ability of progranulin to regulate wound repair and epithelial growth. Progranulin enhances the growth and motility of fibroblasts and endothelial cells, and promotes the formation of tubule-like structures by endothelial cells on Matrigel, suggesting that progranulin may accelerate fibroplasia and serve as a pro-angiogenic factor in the proper environment. These effects are mediated through the activation of the p44/42 MAP kinase, PI-3 kinase and focal adhesion kinase (FAK) pathways, as inlubition of the MAP kinase and PI-3 kinase pathways by PD98059 and wortmannin blocked progranulin-induced cell migration, and progranulin was found to hyper-phosphorylate FAK in endothelial cells. The association of strong progranulin expression with highly proliferative epithelial cells suggests that progranulin may take part in epithelial homeostasis. To investigate this hypothesis, we either overexpressed or downregulated the progranulin gene in SW-13 and MDCK cells. SW-13 cells are derived from human adrenal cancer, but their growth resembles non-transformed epithel

Recent medical and surgical advances have greatly enhanced the survival of children with congenital heart defects that were once considered lethal or extremely debilitating. Consequently there now is a shift in concern from the integrity of the heart to that of the brain. Studies that describe the spectrum of developmental disabilities are lacking in these new survivors. / The primary objective of this doctoral thesis was to characterize the extent and nature of neurodevelopmental impairments and disabilities in young infants with congenital heart defects, using a range of standardized developmental and functional measures, as well as electrophysiological techniques. A high prevalence of acute neurologic, motor, and global developmental impairments were present in our cohort at the time of surgery, and persisted well after surgical intervention. Moreover, functional difficulties and greater dependence in self-care and mobility were also common in this population. Somatosensory evoked potential abnormalities in newborns were found to be very predictive of developmental delays. Multiple markers (i.e. medical, surgical, developmental, and environmental) of brain injury were identified that appear to collectively mediate the outcome of this vulnerable group of children. / Clearly, this is a high-risk population that would benefit from routine developmental screening by rehabilitation specialists. This would facilitate early identification of those at risk and permit the institution of early intervention programs to maximize outcome and minimize burden of care.

The role of perivascular nerve fibers, smooth muscle cells and endothelium in the regulation of human and bovine cerebral artery contractility was investigated in vitro and the effect of cold-storage on vessel function was determined. Stimulation of perivascular nerve endings over a range of transmural electric field stimulation produced frequency- and duration-dependent responses in arteries of both species. The pharmacological properties of the cerebral vessels were studied using 17 different vasoactive agents which acted directly on the smooth muscle and/or on the endothelium. Parallel studies on human and bovine vessels indicate that the cow may be a very useful source of tissue for further experimentation. Cold-storage of cerebral arteries did not significantly alter vessel viability or responsiveness to vasoactive compounds, although a reduction in sensitivity to transmural electric field stimulation in human middle and posterior arteries was observed.

Amyloidosis is a chronic degenerative disease that is characterized by the deposition of fibrous $ beta$-pleated amyloid proteins in various soft tissues and organs (Glenner 1980, Kisilevsky 1983 & 1987). Sporadic form of Alzheimer's disease is one of the most common forms of systemic amyloidosis, with the current prevalence of approximately 4 million cases in the United States and 6 to 8 million cases worldwide (Kisilevsky 1987, Coleman 1990). / Amyloidosis can be experimentally induced in animals. In the inflammation-associated mouse model of amyloidosis, the preamyloid phase, which usually lasts for a number of weeks, is shortened to 24 to 48 hr in the recipient mice upon the administration of amyloid enhancing factor (AEF). AEF activity is present in a much higher level in amyloidotic animals, as well as the amyloidotic tissues in humans (Werdlin and Ranlov 1966, Kisilevsky 1983, Ali-Khan et al. 1988, Alizadeh-Khiavi et al. 1988). / Using brain extracts from Alzheimer patients and liver and spleen extracts from our well-characterized alveolar hydatid cyst (AHC)-mouse model of amyloidosis (Ali-Khan et al. 1983b, Alkarmi and Ali-Khan 1984, Abankwa and Ali-Khan 1988a & 1988b, Alizadeh-Khiavi and Ali-Khan 1988, Du and Ali-Khan 1990) I have been able to purify ubiquitin (UB) and demonstrate its AEF activity in mice (Alizadeh-Khiavi and Ali-Khan 1990, Alizadeh-Khiavi et al. 1990a & 1990b). UB, a stress protein universally found in all eukaryotes, can function as a sentinel-like molecule in the degradation of short-lived or abnormal proteins (Rechsteiner 1987). At a level as low as 10 $ mu$g, the tissue derived UB shows potent AEF activity in the mouse bioassay. The fact that recombinant wild type ubiquitin (R-UB) possess AEF activity, that the AEF activity of both crude and R-UB is abolished by anti-ubiquitin antibody, and that the same antibody inhibits the in vitro amyloidogenic activity by ubiquitin-rich macrophages, further establishes the role of ubiquitin in amyloidogenesis. Using immunohistochemistry and immuno-electron microscopy I have confirmed the cell origin of AEF/ubiquitin, localized its presence in the intracellular compartments of polymorphonuclear leukocytes and macrophages, and demonstrated its binding to murine AA amyloid.

Monoclonal antibodies (MoAbs) raised against human transitional cell carcinoma (TCC) of the urinary bladder, as well as MoAbs binding to carcinoembryonic antigen (CEA), major histocompatibility antigens and M344 (TCC-specific antigen), were used to study cellular heterogeneity in normal and neoplastic urothelium. Heterogeneity of antigen expression was related to differentiation, proliferative potential, tumor progression and clinical outcome. MoAb 5.48 binds preferentially to superficial cells of normal and neoplastic urothelium. The antigen contains terminal sialic acid, and is present on several glycoproteins. Antigen 5.48 expression is lost with increasing grade and stage, whereas expression suggests a good prognosis. In one TCC, antigen 5.48 expression was associated with loss of proliferative potential. Beta-2-microglobulin binds to the differentiated cells of normal and neoplastic urothelial tissues, but it was not associated with grade, stage or clinical outcome. CEA expression was associated with malignancy and a poor prognosis. M344 expression was associated with malignancy and a good prognosis.

Polyinosinic: polycytidylic acid (pI:C), an interferon (IFN) inducer, was used to investigate the role of IFN in the graft-versus-host reaction (GVHR), induced by injecting parental cells into F1 recipients and assessed for immunosuppression and pathological lesions. / PI:C-treatment of recipients, but not donors, before GVHR-induction suppressed the GVHR, an effect seen only with C57BL/6 (B6) and not A donor cells. Using fluorescein-labelled donor cells, pI:C-treatment was seen to cause a marked decrease in donor cell survival after 2 days. Elimination of donor cells was specific for the B6 donor, was associated with increased natural killer (NK) cell but not macrophage cytotoxic activity, was radioresistant and anti-asialo GM1 sensitive, evidence supporting NK-mediated rejection. / Plotting donor cell recovery against the number of cells injected into variously treated recipients indicated that in unstimulated mice a constant proportion of the injected cells were rejected, pI:C increasing that proportion, suggesting that pI:C changes F1 NK target repertoire. / PI:C-treatment after GVHR-induction increased the severity of the GVHR, especially soon after GVHR-induction, the effect waning afterwards. No strain dependence was observed. / These results demonstrate that IFN/IFN-activated cells play an important role in the regulation and in the immunosuppression/pathogenesis of a GVHR.

Background. Peripheral vein infusion thrombophlebitis (PVIT) is a complication of intravenous therapy. We hypothesized that inherited thrombophilia may increase the risk of PVIT. / Purpose. In preparation for a multi-center study of our hypothesis, we conducted a pilot study to estimate PVIT incidence, measure the prevalence of inherited thrombophilia, and pilot test the study procedures. / Methods. A prospective case-control study of 25 cases (patients with PVIT) matched on catheter duration to 25 controls. PVIT risk factors and inherited thrombophilia were assessed. / Results. PVIT incidence was 14 per 1000 catheter-days. There were no significant differences in the prevalence of the inherited thrombophilia disorders among cases and controls (32% vs. 48%). A previous history of PVIT was noted in 4 cases compared to 0 controls. Procedural problems included a high rate of non-consent and inadequate communication with the laboratory. / Conclusions. Though an association between PVIT and inherited thrombophilia was not shown, a previous history of PVIT among cases supports a biological predisposition to PVIT. Our pilot study did provide useful data on PVIT incidence and procedural issues used to design a more definitive study of inherited thrombophilia and PVIT.