Tsunekazu Yamano

Trophic effect of multiple growth factors in amniotic fluid or human milk on cultured human fetal small intestinal cells

Objectives To evaluate the role of growth factors in amniotic fluid and in human milk on gastrointestinal adaptation of the fetus and very low-birth-weight infants, the effects of these fluids and multiple growth factors were investigated in a human fetal small intestinal cell line (FHs 74 Int). Methods After FHs 74 Int cells were incubated with amniotic fluid, human milk, or recombinant growth factors, growth-promoting activity was measured by [3H]-thymidine incorporation into cells. Results Incubating cells with amniotic fluid or human milk promoted growth dose dependently. Genistein almost completely inhibited growth-promoting activity in amniotic fluid P = 0.002), and growth was partially inhibited by antibodies against epidermal growth factor (EGF) (P = 0.047), insulin-like growth factor-1 (IGF-1, P = 0.047), or fibroblast growth factor (FGF, P = 0.014). This activity in human milk was inhibited almost completely by genistein (P h 0.0001) and partially inhibited by antibodies against EGF (P = 0.036), IGF-1 (P = 0.009), FGF (P = 0.004), hepatocyte growth factor (HGF, P = 0.001), or transforming growth factor-&agr; (TGF-&agr;, P = 0.001). Although recombinant EGF, IGF-1, FGF, HGF, and TGF-&agr; elicited a synergistic trophic response on cultured cells, the response was much less than with amniotic fluid or with human milk. Conclusion In aminiotic fluid and in human milk, EGF, IGF-1, FGF, HGF, and TGF-&agr; have a strong trophic effect on immature intestinal cells and may be involved in perinatal gastrointestinal adaptation.

Aristaless -related homeobox gene disruption leads to abnormal distribution of GABAergic interneurons in human neocortex: evidence based on a case of X-linked lissencephaly with abnormal genitalia (XLAG)

X-linked lissencephaly with abnormal genitalia (XLAG) is a rare disorder caused by mutations in the aristaless-related homeobox (ARX) gene, located on Xp22.13. Arx-null mice show loss of tangential migration of GABAergic interneurons, presumably being related to caudal ganglionic eminence tangential migration. In the present study, we investigated a subpopulation of GABAergic interneurons in the brain of an infant with XLAG, who had a novel nonsense mutation of the ARX gene, compared with those of age-matched normal controls and Miller–Dieker syndrome. We performed immunocytochemistry for interneuron and migration markers. We found that glutamic acid decarboxylase (GAD)- and calretinin (CR)-containing cells were significantly reduced in the neocortex and located in the white matter and neocortical subventricular zone, while neuropeptide Y- or cholecystokinin-containing cells were normally distributed. Moreover, in the neocortical subventricular region, the GAD- and CR-containing cells expressed the radial migration marker Mash-1 as well as nestin. Our findings suggest that ARX protein controls not only the tangential migration of GABAergic interneurons from the ganglionic eminence, but also may serve to induce radial migration from the neocortical subventricular zone.

Gilbert syndrome caused by a homozygousmissense mutation (Tyr486Asp) of bilirubinUDP-glucuronosyltransferase gene

We report a case of Gilbert syndrome caused by a homozygous missense mutation (Tyr486Asp) of the bilirubin UDP-glucuronosyltransferase gene. Homozygous missense mutations of the gene have previously been recognized as responsible for Crigler-Najjar syndrome type II. We conclude that Gilbert syndrome in some patients results from homozygous missense mutations of the UDP-glucuronosyltransferase gene.

Clinico-pathological study on macular mutant mouse

SummaryThe macular mutant mouse was clinically and pathologically examined. The hemizygotes began to show white fur color and curly whiskers around postnatal day 3, then seizures and ataxia around day 8, while the normal littermates did not. The hemizygotes also increased weight gradually from birth to day 9, but then showed weight loss and died around day 15 with severe emaciation. These clinical features resembled those in Menkes kinky hair disease. There were no pathological changes in the cerebral cortex in the hemizygotes on day 7. On day 10, two to three clear vacuoles began to appear in a few neurons in the cerebrum. These neurons with vacuoles increased gradually in number and degenerative neurons were also observed by day 14. Ultrastructurally, they corresponded to giant abnormal mitochondria with an electron-lucent matrix and short peripherally located cristae. Other abnormal mitochondria, which were characterized by an electron-dense matrix with tubular or vesicular cristae, were also observed in the cerebral cortical neurons.

Development of corticospinal tract fibers and their plasticity I: Quantitative analysis of the developing corticospinal tract in mice

This study was undertaken to elucidate ultrastructurally and quantitatively the development of the corticospinal tract (CST) axons of mouse at the intumescence level of the cervical cord. An anterograde HRP study showed that the CST was located at the ventral one-third of the dorsal funiculus, and a few HRP-positive fibers were noted at the medialmost part of the ipsilateral anterior funiculus. Ultrastructurally, the CST was composed of unmyelinated axons, growth cones and a few degenerating axons until postnatal day 10 (P10), then the axons in CST gradually increased in size. The number of axons constituting the right CST was calculated at different days of age. The total numbers of axons at P0, P4, P14, P21 and P56 were 2.3 x 10(4), 6.2 x 10(4), 10.4 x 10(4), 7.1 x 10(4) and 3.5 x 10(4), respectively. These results indicate that the number of CST axons at the cervical intumescence of mouse becomes maximum at P14, and then decreases rapidly to reach the adult level of 3.5 x 10(4) (at P56), about 68% of them thus being lost.

Early ependymal changes in experimental hydrocephalus after mumps virus inoculation in hamsters

SummaryTo elucidate the pathogenesis of early ventricular dilatation in hydrocephalus, we examined early morphological changes in ependymal layers at the lateral ventricles in suckling hamsters without aqueductal stenosis 5 days after the intracerebral inoculation of mumps virus. Mumps virus antigen was detectable in all ependymal cells. The ependymal cilia had almost disappeared and only the microvilli remained. A number of supra-ependymal cells were also observed on the surface of the lateral ventricles. Transmission electron microscopy revealed intracytoplasmic viral-like inclusions in the infected ependymal cells. These results suggest that functional and morphological disturbances in infected ependymal cells may cause early ventricular dilatation before aqueductal stenosis occurs.

Reorganization of the corticospinal tract following neonatal unilateral cortical ablation in rats.

The corticospinal tract in the rat after neonatal ablation of the unilateral cerebral cortex was studied morphologically and histochemically using the retrograde and antegrade horseradish peroxidase (HRP) tracing methods. The normal corticospinal tract in the lumbar cord was composed of a number of small and some large axons. In the atrophic corticospinal tract related to the ablated cerebral cortex, the small axons were decreased in number two weeks after the operation. However, new myelinated small axons appeared around day 28 and their diameters increased gradually from after day 56 to day 84. The original large axon in the atrophic corticospinal tract was much more increased in size than that in the corticospinal tract of the non-operated-on control. When HRP was injected into the left cervical cord of the adult rat whose right cerebral cortex had been ablated during the neonatal period, a considerable number of HRP-labeled neurons was seen in the healthy left cerebral cortex. When the corticospinal tract was traced antegradely by injecting HRP into the healthy left cerebral cortex, an aberrant corticospinal tract reaching into the ipsilateral dorsal funiculus was observed. These results give a morphological basis for the well known fact that children who have had brain damage during the neonatal period and early infancy have the capacity for recovery of motor function.

Destruction of external granular layer and subsequent cerebellar abnormalities.

SummaryThis study was undertaken to elucidate the relationship between the time of destruction of the external granular layer and subsequent cerebellar abnormalities. Mice were injected s. c. with 30 mg/kg body weight (b. wt.) of cytosine arabinoside on days 2, 3, and 4, on days, 4, 5, and 6, on days 7, 8, and 9, and on days 10, 11, and 12, and designated as group I, II, III, and IV, respectively.In group I, disarrangement of Purkinje cells and heterotopic granule cells in the molecular layer were observed on all lobes of cerebellum. Heterotopic granule cells were seen on all lobes in group II, whereas disarrangement of Purkinje cells was observed only in the region from the anterior to middle lobes. In group III, heterotopic granule cells were limited to the area from anterior to middle lobes, but there was no disarrangement of Purkinje cells. Group IV cerebellum did not show abnormal cytoarchitecture.Golgi-Cox studies showed abnormal arborization of Purkinje cells in each experimental group. They were arbitrarily classified into inverted Purkinje cells, lying Purkinje cells, T-shaped Purkinje cells, and poorly arborized Purkinje cells. The earlier the postnatal treatment the more severe were the abnormalities of Purkinje cell dendrite.According to the electron-microscopic study, some glomerular synaptic complexes, which are normally confined to the internal granular layer, were observed even in the molecular layer in groups I, II, and III. Some of the Purkinje cell dentritic spines did not make synapses with parallel fibers in any of the experimental groups.The results indicate that severity of abnormal arborization of Purkinje cells is dependent on the period of destruction of the external granular layer. Formation of heterotopic granule cells was dependent on the destruction of the external granular layer up to day 10 after birth.

Asymptomatic cerebral infarction in Kawasaki disease

Meningitis, facial palsy, subdural effusion, and cerebral infarction have been reported to be some of the nervous system complications of Kawasaki disease which usually are clinically obvious. A 22-month-old boy with Kawasaki disease, who developed asymptomatic cerebral infarction, is reported. Hyperpyrexia persisted for 57 days; echocardiography and magnetic resonance imaging revealed the presence of aneurysms in the coronary, axillary, and internal iliac arteries. This study indicates that patients with Kawasaki disease may have silent cerebral infarction.

Connatal tuberculosis in an extremely low birth weight infant: case report and management of exposure to tuberculosis in a neonatal intensive care unit.

Abstract A case of connatal tuberculosis in an extremely low birth weight infant is reported. The patient was a female with a birth weight of 973 g born in the 27th week of pregnancy. She developed respiratory distress and signs of infection immediately after birth, which did not respond to mechanical ventilation, antibiotics, and corticosteroid therapy. Connatal tuberculosis was confirmed at 48 days of age by isolation of Mycobacterium tuberculosis from the infants tracheal aspirate and the mothers menstrual discharge. The infant died of respiratory failure at 90 days of age. Mantoux tuberculin skin tests (TST) were performed on 99 infants, 144 medical staff members, and two family members. TST conversion occurred in three medical staff members, and preventive therapy with isoniazid was initiated. Eight exposed infants had normal chest X-rays and negative gastric aspirates for acid-fast bacilli and all received preventive isoniazid therapy. No case of tuberculosis developed during the 2-year follow-up period. Conclusion Connatal tuberculosis should be considered in neonatal respiratory infection resistant to antibiotics. Prevention of transmission of tuberculosis on the neonatal intensive care unit by chemoprophylaxis is important.