Restoration of cellular function of mesenchymal stem cells from a hypophosphatasia patient.

Mesenchymal stem cells (MSCs) can differentiate into multiple cell lineages and are used for regenerative treatments for a variety of diseases. However, the patient's cells cannot be used to treat genetic diseases. Allogeneic cells can serve as an alternative but long-term survival is uncertain. Our experience of allo-transplantation to a patient with hypophosphatasia, which is caused by mutations of the tissue non-specific alkaline phosphatase (TNSALP) gene resulting in low serum alkaline phosphatase (ALP) activity and skeletal deformity, did not improve these clinical characteristics. Therefore, we sought to use autologous MSCs for the treatment of hypophosphatasia. MSCs derived from the patient's bone marrow had a similar profile when compared with well-reported MSCs. However, the MSCs had extremely low ALP activity and could not produce a mineralized bone matrix even under the osteogenic culture conditions. We therefore transduced a retroviral vector with TNSALP promoter-driven TNSALP gene in the MSCs. In the culture condition, the MSCs had about 7-fold higher ALP activity than did mock-transduced MSCs, and showed mineralization as well as bone-specific markers. Furthermore, the MSCs, but not mock-transduced MSCs, newly formed bone at the frequency of 50% in nude rats. Transplantation of the TNSALP-transduced autologous MSCs might become a new therapy for hypophosphatasia.

Proliferation of peritoneal mast cells in the skin of W/Wv mice that genetically lack mast cells.

Presence of mast cell precursors in the mouse peritoneal cavity was demonstrated, and the precursors were characterized. When a cell suspension, containing mast cell precursor(s), was directly injected into the skin of genetically mast cell-deficient WBB6F1 (WB X C57BL/6)-W/Wv mice, a cluster composed of approximately 2,000 mast cells appeared at the injection site. By determining the proportion of injection sites at which the mast cell cluster appeared, the concentration of mast cell precursors can be calculated by limiting dilution analysis. The concentration in the peritoneal cavity was about five times as great as the concentration in the bone marrow. Although peritoneal mast cell precursors were shown to originate from the bone marrow, physical characterization revealed that the peritoneal precursors differed from the marrow precursors. The peritoneal precursors were less susceptible to irradiation than the marrow precursors; the former were heavier than the latter. When a 95% pure mast cell suspension was prepared from the peritoneal cells by the removal of phagocytes and the density gradient centrifugation, 1 out of 16 cells had the potentiality to make a mast cell cluster in the skin of the W/Wv mice. Moreover, when a single mast cell was identified under the phase contrast microscope and picked up with the micromanipulator, 1 out of 17 mast cells made the cluster. This indicated that some peritoneal mast cells kept extensive proliferative potentiality even after morphological differentiation. In other words, some peritoneal mast cells themselves may function as the committed precursors.

Pleiotropic effects of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors: candidate mechanisms for anti-lipid deposition in blood vessels.

The 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) are considered first-line therapeutic agents for the prevention of coronary heart disease and atherosclerotic disorders related to hypercholesterolemia. Statins inhibit lipid deposition in the aortic endothelium. Although it has been accepted that the statins are potent inhibitors of cholesterol biosynthesis in the liver and that they lower circulating cholesterol levels, several cholesterol-independent (pleiotropic) effects have been reported. The cholesterol-independent effects of statins involve normalization of the nitric oxide (NO)-NO synthase system, anti-inflammatory effects through the inhibition of cytokine/chemokine production, inhibition of vascular smooth muscle cell proliferation and migration, and inhibition of platelet thrombus formation/reduction of the thrombotic response. Some pleiotropic effects of statins may depend on the inhibition of the biosynthesis of farnesyl- and geranylgeranyl-nonsterol compounds from mevalonate in the cells. The Rho/Rho kinase pathway and the phospatidylinositol-3 kinase/Akt pathway mediate the pleiotropic effects of statins. As variations occur in absorption, metabolism, and excretion mechanisms due to the characteristics of specific statins including their hydrophilicity and lipophilicity, there are differences in the transfer mechanisms of statins into tissues. However, the pleiotropic effects occur regardless of statin hydrophilicity and lipophilicity. This review summarizes the pleiotropic effects of statins on lipid deposition in blood vessels.

Neural activation of the brain with hemodynamic insufficiency.

Little is known about how ischemia affects hemodynamic responses to neural activation in the brain. We compare the effects of a motor activation task and a cerebral vasodilating agent, acetazolamide (ACZ), on regional cerebral blood flow (rCBF) in primary sensorimotor cortex (PSM) in six patients with major cerebral artery steno-occlusive lesions without paresis of the upper extremities. Quantitative rCBF was measured in all patients using H2(15)O autoradiographic method and positron emission tomography. The CBF was determined at rest, during a bimanual motor activation task, and 10 minutes after ACZ administration. With bimanual motor activation, rCBF increased significantly in both PSM compared with at rest (P < 0.01 on lesion side, and P < 0.02 on contralateral side). However, rCBF did not increase after ACZ injection in the PSM on the lesion side, whereas rCBF increased significantly in the contralateral PSM after ACZ injection compared with the level at rest. This result suggests that despite a decreased hemodynamic reserve, there is a nearly normal flow response to neural activation, indicating that the mechanism of vasodilation responsible for perfusion change is different for acetazolamide and neural activation. The relations among neural activation, hemodynamic status, and cerebral metabolism in the ischemic stroke patients are discussed.

Effects of delayed treatment with nebracetam on neurotransmitters in brain regions after microsphere embolism in rats.

1. The effects of delayed treatment with nebracetam, a novel nootropic drug, on neurotransmitters of brain regions were examined in rats with microsphere embolism-induced cerebral ischaemia. 2. Cerebral ischaemia was induced by administration of 900 microspheres (48 microns) into the internal carotid artery. The rats with stroke-like symptoms were treated p.o. with 30 mg kg-1 nebracetam twice daily. The levels of acetylcholine, dopamine, noradrenaline, 5-hydroxytryptamine (5-HT) and their metabolites in the cerebral cortex, striatum and hippocampus of animals with microsphere embolism were determined by high performance liquid chromatography (h.p.l.c.) on the 3rd and 7th days after the operation. 3. Although the microsphere embolism induced significant changes in most of the neurotransmitters and some of their metabolites in the brain regions, the delayed treatment with nebracetam partially restored only the hippocampal 5-HT and the striatal dopamine metabolite contents on the 3rd day. 4. The hippocampal in vivo 5-HT synthesis, but not the striatal dopamine synthesis, was attenuated in rats with microsphere embolism on the 3rd day, but was restored by treatment with nebracetam. In vivo striatal dopamine turnover rate of the rats with microsphere embolism was inhibited on the 3rd day irrespective of treatment with nebracetam. 5. The present study provides evidence for a possible action of nebracetam on 5-HT metabolism in the ischaemic brain.

Monozygotic twins of different apparent sex.

We report on twins of unlike sex who shared a 45,X/46,X,+mar karyotype. The mar chromosome was found to be Yq- by DNA analysis. Marker studies, including 8 VNTR loci, yielded a probability of monozygosity of 0.99999996.

Phenotypes associated with replacement of His by Arg in the Pendred syndrome gene.

BACKGROUND: Pendred syndrome is often associated with inner ear malformations, especially enlarged vestibular aqueduct (EVA). Recently, mutations in the Pendred syndrome gene (PDS) have been reported in patients with EVA, in addition to those with classical Pendred syndrome. OBJECTIVE: The aim of this study was to investigate the genotype-phenotype correlations of PDS. METHODS: Each of the 21 exons and flanking splice regions of PDS was analysed by direct DNA sequencing in nine patients with EVA; allele-specific amplification was performed to confirm the mutation. Genetic analyses were compared with thyroid function tests, perchlorate discharge tests, thyroid volume and pure-tone audiogram. Magnetic resonance imaging was used to determine the volume of the endolymphatic duct and sac of each patient. RESULTS: A missense mutation, H723R, was identified in the homozygous state in three patients and in the heterozygous state in another three. Although none of the patients had goitre, increased serum thyroglobulin and an abnormal degree of iodide release were correlated with the number of mutant alleles identified. However, there was no relationship between the degree of hearing loss and the number of mutant alleles. CONCLUSION: The present study reveals that the number of mutant alleles correlates with the degree of subclinical thyroid abnormality, but not with the degree of hearing loss in Japanese patients with the PDS missense mutation H723R.

Alpha fetoprotein-producing acinar cell carcinoma of the pancreas showing multiple lines of differentiation.

An alpha fetoprotein (AFP)-producing tumour occurring in the head of the pancreas of a 30-year-old woman is reported. Histological examination revealed a markedly solid proliferation of tumour cells with prominent nucleoli and occasional luminal structures, some of which contained mucinous material stained with mucicarmine and alcian blue. No squamoid corpuscles were recognized. Immunohistochemistry showed intense positivity for lipase trypsin, and AFP basically, and single cells were also positive for carcino-embryonic antigen, CA19-9, synaptophysin and neuron-specific enolase. Pancreatic hormone-positive cells were absent. Electron microscopical examination revealed numerous granules of variable sizes in the tumour cells, which were considered to be zymogen. The tumour is an acinar cell carcinoma with multi-directional differentiation including the ability to produce AFP. Among AFP-positive pancreatic tumours, acinar cell carcinoma and pancreatoblastoma seem to be the most frequent.

Centrally administered ouabain aggravates central sleep apneas.

The presence of endogenous digitalis-like factors in the central nervous system suggests their functional significance in the central nervous system. Three-day infusions of three-stepped doses of the digitalis agent ouabain (1-100 ng.kg body wt-1.h-1) into the lateral cerebral ventricle of freely moving rats caused a dose-dependent increase in the number of central-apneic episodes during rapid-eye-movement sleep without affecting the time spent in rapid-eye-movement sleep or basic respiratory rate. These results suggest that endogenous digitalis-like factors may be involved in the genesis of central sleep apneas.

Surgical treatment of large ventricular septal defect with pulmonary hypertension in the first 24 months of life.

A total of 74 patients under 24 months of age with large ventricular septal defects (VSD) and pulmonary hypertension were subjected to surgical treatment from 1969 through 1975. Emergency pulmonary artery (PA) banding was performed in 13 patients during the first year of life with 1 death from postoperative respiratory failure. Primary closure of the VSD was performed in 61 patients using simple hypothermia and short-term coronary perfusion, with an operative mortality of 1.6%. There were no late deaths or neurological disturbances. Normal hemodynamic data were obtained in all 7 patients who underwent postoperative cardiac catheterization from one month to five years after the primary correction. It is concluded that primary closure of a VSD in infancy is reasonable and that PA banding is indicated only for those patients less than 6 months old with a complicated defect or in an emergency situation.

Hemi-parkinsonism in multiple system atrophy: a PET and MRI study.

We selected 6 patients presenting with hemi-parkinsonism from a total of 20 patients with probable multiple system atrophy (MSA) and studied their nigrostriatal lesions using magnetic resonance (MR) imaging and positron emission tomography (PET) with 18F-labeled 2-deoxy-2-fluoro-D-glucose (FDG). T2 weighted MR images demonstrated a decreased signal intensity in the putamen of all patients. This decreased signal was more intense in the nucleus contralateral to the affected body side in 5 patients. A decreased signal in the substantia nigra was found, expanding more on the contralateral side in 3 patients. T1-weighted images showed that the contralateral putamen was smaller in size than the ipsilateral. These findings indicated that the iron deposit and the neuronal cell loss in the degenerative process were more remarkable in the contralateral nuclei. FDG uptake in 5 patients had likewise declined more in the contralateral than in the ipsilateral putamen. The study shows that these patients have the nigrostriatal lesions as described in previous reports on MSA and that an asymmetric lesion relating to clinical signs is present in the nigrostriatal system. When a patient presents with hemi-parkinsonism alone, MR imaging and PET/FDG are useful for the clinical diagnosis of MSA.

Parieto-occipital glucose hypometabolism in Parkinson's disease with autonomic failure.

To investigate the characteristics of regional cerebral metabolism in a subgroup of patients with Parkinson's disease and autonomic failure, we studied seven patients with Parkinson's disease with autonomic failure (PA group), 11 patients with Parkinson's disease without apparent autonomic failure (PD group), and nine normal controls using fluoro-deoxyglucose positron emission tomography (FDG-PET). To determine differences in metabolic distribution among these groups, regional relative glucose metabolic rates (RGMR), which were normalized with cerebellar values, were calculated and age-adjusted covariance analyses were done. When compared with that of controls. RGMR in the cerebral cortex of the PA group was markedly reduced in the occipital cortex (P<0.001), inferior parietal cortex (P<0.005) and superior parietal cortex (P<0.005), but without a decrease in the sensory motor and medial temporal cortices, putamen and thalamus. In contrast, the PD group did not show significant focal hypometabolic distribution. Our findings raise the possibility that Parkinson's disease with autonomic failure may overlap with the features of dementia with Lewy bodies.

Experience with the ultrasonic surgical aspirator in a cavernous hemangioma of the cavernous sinus.

A patient with a cavernous hemangioma of the cavernous sinus was operated upon using the Cavitron Ultrasonic Surgical Aspirator (CUSA). Intracapsular subtotal removal of the tumor was accomplished efficiently with the CUSA. At the end of the procedure, however, the CUSA penetrated not only the capsule of the tumor, but also the wall of the internal carotid artery. Advantages and disadvantages of CUSA surgery for cavernous hemangiomas of the cavernous sinus are discussed.

Comparison of vasodilatory effect of carbon dioxide inhalation and intravenous acetazolamide on brain vasculature using positron emission tomography.

Carbon dioxide (CO2) and acetazolamide are increasingly being used as vasodilators to detect cerebrovascular reserve capacity in patients of chronic cerebrovascular disease. The functional cerebrovascular reserve or ability of cerebral vessels to lower their resistance in response to decrease in cerebral perfusion pressure is expressed as change in cerebral blood flow from baseline under a vasodilatory stimuli. Theoretically a vasodilator causing maximum vasodilation, and thereby expressing complete reserve capacity would be more suitable for such a purpose. We quantitatively compared the vasodilating effect of 5% CO2 inhalation and 1 g of intravenous acetazolamide by positron emission tomography. Cerebrovascular reserve was quantified in six patients with chronic cerebrovascular disease in the same sitting, using oxygen-15 labeled water (H2(15)O) positron emission tomography at rest, during 5% CO2 inhalation and after 1 g intravenous acetazolamide. A significant linear correlation in both nonlesion hemisphere (r = 0.701, p < 0.001) and in lesion hemisphere (r = 0.626, p < 0.005) was found between CO2 and acetazolamide for cerebrovascular reserve capacity. This correlation improved by considering cerebrovascular reserve per unit change in arterial carbon dioxide (r = 0.744, p < 0.001 in nonlesion hemisphere and r = 0.721, p < 0.001 in lesion hemisphere). The quantitative value of global reserve capacity was different by CO2 stimuli (5.2%) and acetazolamide (49.7%). Though a similar vasodilatory response is elicited by both vasodilators, acetazolamide seems to be more potent and therefore should be preferred to detect patients with exhausted cerebrovascular reserve capacity.

Rapidly progressive pulmonary alveolar proteinosis in a patient with chronic myelogenous leukemia.

A 46-year-old man with chronic myelogenous leukemia (CML) was admitted to our hospital because of high fever. The chest radiographs showed bilateral groundglass-like infiltrates in the perihilar region. Transbronchial lung biopsy (TBLB) and autopsy revealed PAS-positive granular materials characteristic of pulmonary alveolar proteinosis (PAP). He had received interferon-alpha since the time of CML diagnosis. Busulfan had never been administered. Altered cell-mediated immunity was thought to be closely related to the development of PAP.

Recurrent malignant glioma: detection with 131I labeled monoclonal antibody G-22, positron emission tomography and magnetic resonance imaging.

A 45-year-old man with suspected recurrent malignant glioma was evaluated by magnetic resonance imaging (MRI), positron emission tomography (PET) and 131I labeled monoclonal antibody G-22 (G-22) scan. Following Gadolinium-DTPA, a TI-weighted spin echo image (TR 500 msec, TE 20 msec) demonstrated a large mass with an irregular margin in the left temporo-parietal area. An 18F labeled fluorodeoxyglucose PET study demonstrated marked accumulations in the left temporo-parietal area. Serial 131I-G-22 scintigraphy was obtained for a week after the injection. The uptake was most increased on the 2nd day after the injection. 131I G-22 was specific for tumor-associated antigens.

High- and moderately high-methionine uptake demonstrated by PET in a patient with a subacute cerebral infarction.

In patients with cerebral tumors, high accumulations of L-methyl-11C-methionine (11C-Met) have been reported in some cases of cerebral ischemic disease, but no high accumulations of 11C-Met in areas where only transient arterial occlusions are most likely to occur have been reported. Herein we present a case of a high accumulation of 11C-Met in an area of frontal interhemispheric cerebral infarction and a moderately high accumulation with an unclear margin in a distant frontal convexity area. A craniotomy revealed a subacute stage of cerebral infarction in the interhemispheric lesion, and an ischemic change in the distant convexity area. Sixteen months after onset, CT scans demonstrated an infarction area in the interhemispheric lesion only, and no atrophic changes were observed in the distant convexity area indicating that no serious tissue damage had occurred.

Studies on the delayed neurotoxicity of organophosphorus compounds- (I).

Delayed neurotoxicity experiments of an organophosphorus compound, TOCP, on hens and quails were carried out. The animals were orally exposed with TOCP in dose of 400 mg/kg, and maintained for 25 and 50 days under observation, respectively. They were sacrificed and dissected at different periods after the exposure, and the histopathological examinations were made on those animals. During the periods of 50 days, no abnormal symptoms except for acute poisoning were noted in the quails. As to the hens, neurological disorders were observed from around 12 days of the experiment. In hens, the morphological alteration in the earlier stage of the experiment was perivascular cuffing of small round cells seen in the cerebrum, cerebellum and spinal cord. The major alteration was degeneration of axons and myelin in the white matter of the spinal cord. A minimal degree of these changes was found even in the hens in which the clinical signs of neurotoxicity had not yet been observed. Later the process expanded to the whole spinal cord as well as the sciatic nerve. The electron microscopy performed after 15 days revealed moderate change of myelin and axons. From these results, it was concluded that the clinical signs as well as the morphological changes were closely related to the exposure of the compound, TOCP.

Studies on the delayed neurotoxicity of organophosphorus compounds--(II).

Delayed neurotoxicity experiment was carried out on an organophosphorus compound (TOCP) in hens. Fifteen hens, 1.9 kg of their average body weight and 20 months of age, were orally administered with TOCP in a dose of 400 mg/kg body weight. Two animals out of 15 were sacrificed after 7 days to examine alterations in the nervous system under electron microscope. On the remaining 13 animals, histopathological examinations were carried out after 35 days. During the course of the experiment, progressive neuropathy developed in the animals at 10 to 12 days after the exposure to TOCP. Examinations of the sciatic nerve under electron microscope revealed slight axonal degeneration while the myelin sheath was found to be rather intact. On the animals after the observations periods of 35 days, marked axonal and myelin degeneration was observed in the anterior and posterior funiculus, and in the spinocerebellar tracts of the spinal cord as well as in the cerebellar peduncle and in the white matter of the cerebellum associated with glial proliferation. The localization and degree of these changes were considered to be "Dying Back", showing systemic neuropathy. Moreover, the morphological alterations of the nerve cells were observed in the Purkinje cells, in the cerebellar nucleus, gracilis nucleus, and anterior horn cells of the lumbar spinal cords, characterized by loss of nerve cells and/or tigrolysis. Muscular lesions showed small group atrophy, corresponding to Type I fiber atrophy.

Studies on the delayed neurotoxicity of organophosphorus compounds--(III).

TOCP (Tri-orthocresyl phosphate), an organophosphorus compound, has been implicated in producing neuropathy in the male S. D. rats. Repeated subcutaneous doses of TOCP (600 mg/kg) for up to 6 weeks produced ataxia, most striking at 50 days after final injection, followed by gradual recovery. Ultrastructurally, the internal structure of affected nerve fibers was primarily composed of altered smooth endoplasmic reticulum, tubular membrane system, and mitochondria, although myelin sheath was found to be essentially normal. In the histopathological examination, axonal and myelin degeneration was disclosed in the gracile nucleus and in the gracile fasciculus of the cords as well as in the sciatic nerves. The localization and degree of these changes were considered to be "dying back", showing systemic neuropathy. In addition, muscular lesion showed small group atrophy, corresponding to Type I fiber atrophy.