Fetal tissue transplants in animal models of Huntington's disease: the effects on damaged neuronal circuitry and behavioral deficits.

Accumulating evidence indicates that grafts of embryonic neurons achieve the anatomical and functional reconstruction of damaged neuronal circuitry. The restorative capacity of grafted embryonic neural tissue is most illustrated by studies with striatal tissue transplantation in animals with striatal lesions. Striatal neurons implanted into the lesioned striatum receive some of the major striatal afferents such as the nigrostriatal dopaminergic inputs and the gluatmatergic afferents from the neocortex and thalamus. The grafted neurons also send efferents to the primary striatal targets, including the globus pallidus (GP, the rodent homologue of the external segment of the globus pallidus) and the entopeduncular nucleus (EP, the rodent homologue of the internal segment of the globus pallidus). These anatomical connections provide the reversal of the lesion-induced alterations in neuronal activities of primary and secondary striatal targets. Furthermore, intrastriatal striatal grafts improve motor and cognitive deficits seen in animals with striatal lesions. Since the grafts affect motor and cognitive behaviors that are critically dependent on the integrity of neuronal circuits of the basal ganglia, the graft-mediated recovery in these behavioral deficits is most likely attributable to the functional reconstruction of the damaged neuronal circuits. The fact that the extent of the behavioral recovery is positively correlated to the amount of grafted neurons surviving in the striatum encourages this view. Based on the animal studies, embryonic striatal tissue grafting could be a viable strategy to alleviate motor and cognitive disorders seen in patients with Huntington's disease where massive degeneration of striatal neurons occurs.

Freeze-drying synthesis of an amorphous Zn(2+) complex and its transformation to a 2-D coordination framework in the solid state.

An amorphous and metastable precursor for a Zn two-dimensional coordination framework was synthesised via freeze drying. The precursor comprises randomly packed discrete clusters of a Zn complex. The amorphous-to-crystalline framework transformation, which was triggered by the gentle application of heat or pressure, was accompanied by a change in the coordination geometry of the Zn(2+) ions from tetrahedral to octahedral symmetry.

Generation of dopaminergic neurons in the adult brain from mesencephalic precursor cells labeled with a nestin-GFP transgene.

Mesencephalic precursor cells may one day provide dopaminergic neurons for the treatment of Parkinson's disease. However, the generation of dopaminergic neurons from mesencephalic precursors has been difficult to follow, partly because an appropriate means for recognizing mesencephalic ventricular zone precursors has not been available. To visualize and isolate mesencephalic precursor cells from a mixed population, we used transgenic mice and rats carrying green fluorescent protein (GFP) cDNA under the control of the nestin enhancer. nestin-driven GFP was detected in the mesencephalic ventricular zone, and it colocalized with specific markers for neural precursor cells. In addition, data from flow-cytometry indicated that Prominin/CD133, a cell-surface marker for ventricular zone cells, was expressed specifically in these GFP-positive (GFP(+)) cells. After sorting by fluorescence-activated cell sorting, the GFP(+) cells proliferated in vitro and expressed precursor cell markers but not neuronal markers. Using clonogenic sphere formation assays, we showed that this sorted population was enriched in multipotent precursor cells that could differentiate into both neurons and glia. Importantly, many neurons generated from nestin-GFP-sorted mesencephalic precursors developed a dopaminergic phenotype in vitro. Finally, nestin-GFP(+) cells were transplanted into the striatum of a rat model of Parkinson's disease. Bromodeoxyuridine-tyrosine hydroxylase double-labeling revealed that the transplanted cells generated new dopaminergic neurons within the host striatum. The implanted cells were able to restore dopaminergic function in the host striatum, as assessed by a behavioral measure: recovery from amphetamine-induced rotation. Together, these findings indicate that precursor cells harvested from the embryonic ventral mesencephalon can generate dopaminergic neurons able to restore function to the chemically denervated adult striatum.

Endovascular treatment of unruptured cerebral aneurysms.

76 consecutive patients with 78 unruptured cerebral aneurysms underwent endovascular therapy from July 1999 to May 2004 in our institute. For the wide-necked aneurysms, the remodeling technique, double microcatheter technique, or stent-assisted coil embolization was used, while a parent artery occlusion or covered stent was applied for the giant or fusiform aneurysms. Immediate angiographical results demonstrated 33 complete occlusions, 26 neck remnants, and 14 dome fillings. Four cases were treated with parent occlusion or stenting only, and one case was not treated with embolization but with clipping due to the rupture of the aneurysm during coil embolization. Immediate angiographic findings demonstrated that in aneurysms between 5 to 10 mm, the rate of complete occlusion was 48%, that of neck remnants 33%, and that of dome fillings 27%. In aneurysms between 11 to 25 mm, the rate of complete occlusion was 14%, that of neck remnants 28%, and that of dome fillings was 58%. In the angiographic follow-up results, all aneurysms smaller than 5 mm showed complete occlusion. In aneurysms between 5 to 10 mm, 74% of the aneurysms showed complete occlusion, and 21% showed neck remnants, and 5% showed dome filling. In aneurysms between 10 to 24 mm, 25% showed complete occlusion, while 75% showed dome filling. The overall mortality rate was 0% and the morbidity rate was 3.7% (2 major strokes, 1 minor stroke) at 30-days after embolization. In the clinical follow-up study, one case of a large basilar tip aneurysm caused a fatal rupture 28 months after the initial embolization. Endovascular therapy was performed on the unruptured aneurysms and was found to be an acceptable treatment, except for durability in cases of large aneurysms.

Synthesis of magnesium ZIF-8 from Mg(BH4)2.

Porous Mg(2-methyl imidazolate)2 (Mg-ZIF-8) was synthesised from Mg(BH4)2 as a precursor under an Ar atmosphere. It possesses an uncommon tetrahedral Mg(2+)-N coordination geometry that is stabilised by the formation of a framework, and it exhibits a Brunauer-Emmett-Teller surface area greater than 1800 m(2) g(-1).

Glass formation via structural fragmentation of a 2D coordination network.

The structure of a glass obtained by the melt quenching of a two-dimensional (2D) coordination network was examined. X-ray analyses disclosed a 2D-to-0D structural transformation before and after glass formation. The mechanism is unique to coordination compounds, as it is characterized by labile and flexible coordination bonds.

Innervation of the cerebral veins as compared with the cerebral arteries: a histochemical and electron microscopic study.

The distribution of nerve fibers in the cerebral veins was studied by catecholamine fluorescence simultaneously with acetylcholinesterase (AChE) histochemistry. A comparison of nerve fibers in the cerebral arteries was made. The ultrastructure of terminal boutons in the veins fixed with potassium permanganate was also studied. In the adventitia of the cerebral artery, green fluorescent aminergic fibers and brownish AChE-reactive (probably cholinergic) fibers were observed. In contrast, the cerebral venous system showed no AChE-positive fibers. Catecholamine fluorescent varicose fibers were detected in the dural sinus, the internal cerebral vein, and the superficial vein of Labbé. The highest density of aminergic fibers was found in the dural sinus and the second highest in the internal cerebral vein. Most of the terminal boutons in the adventitia of the cerebral veins were found adjacent to a muscle-like cell and showed only cored vesicles under electron microscopy. Results of our study suggest that the cerebral venous system has a neurogenic innervation, mainly from aminergic fibers, which is different from the neurogenic supply to the cerebral arterial system.

Immunohistochemical demonstration of vasopressin nerve fibers in the cerebral artery.

Vasopressin-immunoreactive nerve fibers were demonstrated in the cerebral pial arteries by peroxidase immunohistochemistry. In the large pial artery (proximal part of the middle cerebral artery), they ran longitudinally to the long axis of the vessel. They ran in a spiral pattern in the distal part of the middle cerebral artery. Even in small arteries, vasopressin nerve fibers were found arranged in a longitudinal fashion. The present morphological data suggest that vasopressin nerve fibers in the cerebral artery may play a role in cerebral circulation.

A light and electron microscopic immunohistochemical study of vasoactive intestinal polypeptide- and substance P-containing nerve fibers along the cerebral blood vessels: comparison with aminergic and cholinergic nerve fibers.

Vasoactive intestinal polypeptide (VIP)- and substance P-containing nerve fibers were observed in the cerebral blood vessels using an immunohistochemical technique. VIP-containing nerve fibers distributed in a spiral pattern, similar to that of muscle cells. Under electron microscopic observation, VIP-immunoreactive terminals lay close to a muscle cell in the inner layer of the adventitia. In contrast, substance P-containing nerve fibers showed a meshwork pattern in the outer layer of the adventitia. Using both acetylcholinesterase (AChE) staining and VIP immunohistochemistry, AChE-positive and VIP-immunoreactive nerve fibers revealed almost the same distribution in the same specimen. The present data suggest that VIP-containing nerve fibers may play a role in the smooth muscle control of the blood vessels, whereas substance P-containing nerve fibers may not take part in muscle control.

Comparative anatomy of the distribution of catecholamines within the inferior olivary complex from teleosts to primates.

The distribution of catecholamine (CA) in the inferior olivary complex (IO) of various vertebrate (from fish to monkey) was investigated by means of the histofluorescence technique. In addition, using rats, a further attempt was made to elucidate the origins of CA in the IO. The IO of the lower vertebrates (from fish to birds) was in general poorly innervated by the CA neuron system. IO in the lower mammals, such as insectivora and bats, contained only a few CA nerve terminals, while that in the higher mammals such as rat, guinea pig, rabbit, cat, and monkey revealed quite a number. In these animals, species-species patterns of CA nerve terminals were found. In the rat, the highest concentration was observed in the dorsal lamella of the principal nucleus and in guinea pig ventral lamella. In the rabbit and cat, maximum CA nerve terminals were detected in the dorsal accessory nucleus, while in the monkey, they were detected in the medial accessory nucleus. The retrograde tracer technique of horseradish peroxidase (HRP) suggested that the main source of the abundant CA terminals in IO of the rat might be A1, A2, and A3 noradrenaline neurons, though not locus coeruleus and not dopaminergic ones.

Thiamin and pyridoxine requirements during intravenous hyperalimentation.

Studies were undertaken to determine rational dosages of vitamin B1 and B6 during long-term intravenous hyperalimentation, using more sensitive techniques than formerly used to evaluate B1 and B6 status. A standard vitamin combination, type A, (usually commercially available products) has been used up to now because of convenience, disregarding the effects of long-term administration. This combination lacks biotin, folic acid, and vitamin E and contains from 10 to 100 times the dietary allowances of such vitamins as B1, B2, B6, B12, and C. In response to the possibility of vitamin overdose, two new vitamin combinations, type B (from commercial products) and type C (a convenient and easily administered combination produced at the hospital) were developed in order to provide the normal dietary allowances and at the same time eliminate any harmful side-effects. From the results obtained, 5 mg/day for thiamin HCl and 3 mg/day for pyridoxine HCl in type B and type C were found to be a sufficient and safe level as opposed to 55 mg/day for thiamin HCl and 102 mg/day for pyridoxine HCl in type A.

Embryonic striatal grafts restore neuronal activity of the globus pallidus in a rodent model of Huntington's disease.

It has been demonstrated in rats that embryonic striatal grafts placed in the excitotoxically lesioned striatum establish neuronal connections with the host globus pallidus. In order to determine whether the morphologically verified connections between the grafts and host are functional, the present study investigated the effects of embryonic striatal grafts on changes in the neuronal activity of the globus pallidus in rats with quinolinic acid-induced striatal lesions. The activity of pallidal neurons was determined by use of quantitative cytochrome oxidase histochemistry and an electrophysiological technique. Striatal lesions induced an increase in both the cytochrome oxidase activity and the spontaneous firing rate of the globus pallidus ipsilateral to the lesions. Grafts derived from the lateral ganglionic eminence, but not the medial ganglionic eminence, reversed the lesion-induced increase in the cytochrome oxidase activity of the globus pallidus with concomitant reduction of apomorphine-induced rotational asymmetry. The lateral ganglionic eminence grafts also attenuate the increase in the firing rate of pallidal neurons in rats with striatal lesions. The present results provide evidence that striatal lesions lead to the loss of a tonic inhibitory input to the globus pallidus with consequent increase in the activity of pallidal neurons, and that intrastriatal striatal grafts reverse the altered activity of pallidal neurons. The findings strongly suggest that embryonic striatal grafts functionally repair the damaged striatopallidal pathway.

Expression of c-fos, heat shock protein 70, neurotrophins, and cyclooxygenase-2 mRNA in response to focal cerebral ischemia/reperfusion in rats and their modification by magnesium sulfate.

The marginal area surrounding a region of ischemic brain tissue, designated as the penumbra, is of interest as a potential area for the rescue of neurons from cell death. Despite its clinical importance, relatively little is known about the molecular events leading to changes in brain cells in the penumbra following ischemia. In the first part of this study, we used in situ hybridization to investigate the temporal and spatial expression of c-fos, heat shock protein 70 (HSP70), neurotrophins and inducible cyclooxygenase-2 (COX-2) in the rat brain following a 2-h occlusion of the middle cerebral artery (MCA) with reperfusion. In the penumbra and surrounding cortex, upregulation of c-fos, brain-derived neurotrophic factor (BDNF), and COX-2 mRNAs was observed, while expression of HSP70 mRNA was restricted to the penumbra. This spatial discrepancy of mRNA expression suggests that different mechanisms are involved in the regulation of c-fos/BDNF/COX-2 and HSP70 expression. Intravenous infusion of magnesium sulfate (25 mg/kg) decreased both the infarct volume and upregulation of these mRNAs, suggesting its therapeutic potential.

Skin lesions during intravenous hyperalimentation: zinc deficiency.

Of 54 patients who received prolonged intravenous hyperalimentation during the past 4 years, six developed progressive, characteristic skin lesions. These cutaneous manifestations frequently were accompained by abdominal symptoms resembling those of acrodermatitis enteropathica in many respects. In the first four of these six patients, skin eruptions disappeared promptly after the initiation of oral feeding, which led us to assume that these skin manifestations are due to some unknown nutrient deficiency. Our experience in the remaining two patients enabled us to presume the underlying pathology to be zinc deficiency.

Total parenteral nutrition in familial hypercholesterolemia: importance of caloric control.

Effects of total parenteral nutrition (TPN) on serum cholesterol levels were examined in four patients with homozygous and three patients with heterozygous familial hypercholesterolemia and 13 patients without hypercholesterolemia. Fat-free TPN was administered in average doses of 26.5 cal/kg/day (43.6 cal/kg/day in a child) for 5 to 14 weeks. The serum cholesterol level decreased markedly by 47.5% and 58.6% in homozygous and heterozygous cases, respectively. Despite restriction in calories of TPN, protein nutrition was maintained in all patients. It became apparent that in familial hypercholesterolemia, whether the heterozygous or homozygous form, TPN performed at a well-adjusted caloric intake provided an adequate control of the serum cholesterol level as far as TPN was continued. In patients without hypercholesterolemia under TPN, serum cholesterol levels varied correlatively with the amount of calories to be administered. Compared with values before TPN, serum cholesterol level as far as TPN was continued. In patients without hypercholesterolemia under TPN, serum cholesterol levels varied correlatively with the amount of calories to be administered. Compared with values before TPN, serum cholesterol levels at the fourth week of TPN were elevated or maintained in patients receiving more than 40 cal/kg/day, while it was reduced in patients given less than 40 cal/kg/day. There was a statistically significant correlation between daily caloric intake per body weight and change of serum cholesterol level at the fourth week of TPN. This indicates that caloric control is of importance in TPN-induced change in serum cholesterol levels.

Adriamycin-lipiodol suspension for i.a. chemotherapy of hepatocellular carcinoma.

Physicochemical properties of two types of adriamycin preparation, suspensions and emulsions prepared for i.a. chemotherapy of hepatocellular carcinoma, were investigated. A suspension was prepared by dispersing adriamycin directly into the lipid contrast medium, Lipiodol, whereas an emulsion was obtained by emulsifying an aqueous solution of adriamycin into Lipiodol. The dispersibility of the drug in each preparation was examined microscopically. The chemical stability of and drug release from the preparation were determined by high-performance liquid chromatography and spectrophotometry, respectively. The suspension was then given to ten patients with primary hepatocellular carcinoma. The suspension maintained good dispersibility without coagulation of drug particles, whereas coalescence of aqueous droplets and the resultant phase separation occurred 4 h after preparation of the emulsion. Both preparations maintained the initial drug content for at least 1 week at room temperature. The release of adriamycin was more prolonged in the suspension than in the emulsion. After i.a. administration of the suspension, a selective accumulation of Lipiodol in the tumor and decrease in serum alpha-fetoprotein (AFP) levels were found in most patients. A significant amount of adriamycin was still detected in hepatic specimens resected from two patients 1 and 2 months after treatment. These findings suggest that the adriamycin-Lipiodol suspension may be a useful preparation for targeting chemotherapy to hepatocellular carcinoma.

Metabolic inhibition enhances selective toxicity of L-DOPA toward mesencephalic dopamine neurons in vitro.

Recent in vitro studies have described the toxicity of levodopa (L-DOPA) to dopamine (DA) neurons. We investigated whether metabolic inhibition with rotenone, an inhibitor of complex I of the mitochondrial respiratory chain, may enhance the toxicity of L-DOPA toward DA neurons in mesencephalic cultures. The uptakes of DA and GABA were determined to evaluate the functional and morphological integrity of DA and non-DA neurons, respectively. Pretreatment with rotenone significantly augmented the toxic effect of L-DOPA on DA neurons. Interestingly, prior metabolic inhibition with rotenone rendered DA cells susceptible to a dose (5 microM) of L-DOPA that otherwise exhibited no toxic effect. DA uptake was more intensely attenuated than GABA uptake after the combined exposure to rotenone and L-DOPA. This was confirmed by cell survival estimation showing that tyrosine hydroxylase-positive DA cells are more vulnerable to the sequential exposure to the drugs than total cells. The selective toxic effect of L-DOPA on rotenone-pretreated DA neurons was significantly blocked by antioxidants, but not antagonists of NMDA or non-NMDA glutamate receptors. This indicates that oxidative stress play a central role in mediating the selective damage of DA cells in the present experimental paradigm. Our results raise the possibility that long-term L-DOPA treatment could accelerate the progression of degeneration of DA neurons in patients with Parkinson's disease where potential energy failure due to mitochondrial defects has been demonstrated to take place.

The distribution of adrenergic receptors in cerebral blood vessels: an autoradiographic study.

The first morphological evidence of the existence of adrenergic receptors (alpha 1, alpha 2 and beta) within the vascular walls of the central nervous system were presented using the in vitro receptor autoradiographic technique. In the rat pial arteries all three types of adrenergic receptors were demonstrated, whereas the human pial arteries failed to show significant autoradiographic grains of alpha 1 type of adrenergic receptors indicating a considerable inter-species difference in the distribution of adrenergic receptors. alpha 2 and beta receptors in human pial arteries were found not only in the arterial smooth muscle layers but also in the endothelial layers. This suggests a possibility that circulating sympathomimetic agents play some role in controlling the tone or permeability of vascular walls within the central nervous system. A distinct distribution of alpha 1 receptors in cortical layer IV where the vascular plexus was richest may suggest a relation of alpha 1 receptors and blood flow of brain parenchyma.

Dephosphorylation-induced decrease of anti-apoptotic function of Bcl-2 in neuronally differentiated P19 cells following ischemic insults.

It is known that Bcl-2 has a protective effect against neuronal ischemia. Some reports speculate anti-apoptotic function of Bcl-2 depends not on the expression level but on the phosphorylation state. We found induction of apoptosis and CPP32 activation by energy impairment (3-nitropropionic acid (3-NP)-treatment or glucose-deprivation) in the neuronally differentiated P19 cells. Time course study of cell viability following ischemic insults showed that the number of viable cells decreased along with the increase in the amount of dephosphorylated Bcl-2 without obvious quantitative alteration of the protein. Then, we generated differentiated P19 cells overexpressing wild-type Bcl-2 (P19/wt. Bcl-2) or phosphorylation-negative Bcl-2 mutant (P19/mut.Bcl-2), in which alanine was substituted for serine 70. When the cell viability was examined within 24 h, P19/mut.Bcl-2 was more vulnerable to energy impairment as compared with P19/wt.Bcl-2. In addition, overexpression of wild-type Bcl-2 inhibited DNA laddering and CPP32 activation induced by the insults, while that of mutant Bcl-2 did not. These findings suggest that the phosphorylation state, as well as the expression level, of Bcl-2 plays an important role to modulate its protective effect against ischemic insults.

Unilateral spatial neglect due to a haemorrhagic contusion in the right frontal lobe.

We report two cases of unilateral spatial neglect associated with an isolated right frontal lobe lesion. Case 1 was a 59-year-old, right-handed man, who developed a left hemiplegia, disorientation, and frontal lobe neglect associated with a haemorrhagic contusion following a head injury. Case 2 was a 55-year-old, right-handed man, who also developed disorientation and frontal lobe neglect secondary to a haemorrhagic contusion following a head injury. 99mTc HM-PAO SPECT revealed an isolated reduction in the regional cerebral blood flow (CBF) around the haematoma in the frontal lobe; blood flow to remaining parts of the brain was normal. Damage to the right frontal lobes of these patients was confirmed as being the cause of the unilateral spatial neglect in accordance with the results of CBF studies.