46

Sequential Observation of Hydrocephalic Pro­cess in the Infants Associated with Myelome­ningocele by Volume-tric Calculation from CT Scans Syun Yamasaki, MD, Hiroatsu Hojo, MD, Kenkichi Kataoka, MD, and Shozo Nakano, MD Departments of Neurosurgery (SY) and Pediatric Neurology (HH, KK, SN), Shizuoka Children's Hospi· tal, Shizuoka

For the purpose of recognizing the hydrocephalic pro­cess, the ventricular volumes of hydrocephalic infants associated with myelomeningocele were calculated from sequential CT scans, and were compared with area and linear measurements of ventricles of CT slices. The regions labeled as ventricle were measured by planimetry to give ventricular volume for that slice. The calculated total volume was simply the sum of ventricular volume of all CT slices. The calculated volumes of six neonates were 8-106 ml (average 42 ml). In four of these neonates who showed moderate or marked grade of ventricular dilatation were per­formed the CSF shunts before the repairs of myelome­ningocele, and the other two infants who had the ventricular volumes more than about three times larger than that of neonatal period were operated at the age of one month. In preshunt period, the calculated ventricular volume was nearly correlated the ventricle­brain ratio and the occipital cerebroventricular index, but its correlation was not recognized in postshunt period. The sequential change of the calculated ven­tricular volumes is considered to indicate the hydro­cephalic; process. It is desirable to recognize the volume change of ventricles by means of volumetry from CT scans in hydrocephalic infants associated with myelomeningocele.

Key words: Hydrocephalus, myelomeningocele, CT scan, ventricular volume.

47

Biochemical and Cellular Studies in Tuberous Sclerosis. II. Proline and Hydroxyproline in Urine, Tissues and Cultured Skin Fibroblasts Harumi Tanaka, MD, and Masataka Arima, MD Division of Child Neurology, National Center for Ner­vous, Mental and Muscular Disorders, Kodaira, Tokyo

In a previous study we suggested an increased collagen synthesis in tuberous sclerosis (TS). We present here that contents of hydroxyproline (H) in affected tissues and urine are high in TS, and that the cultured skin fibroblasts (F) from TS show distinct characters under low or high serum- or proline-containing media.

Materials and Methods TS tissues of 5 patients were obtained from necropsy or plastic surgery. Urine were collected from 10 pa­tients with TS and 20 controls. Determination of H was carried out by the method of Bergman et al. F of 6 strains from TS (4 strains from adenoma sebaceum, 2 strains from fibrosis) and 7 strains from controls were maintained in McCoy's 5a medium with proline or Dulbecco's medium without proline (D) containing 17% fetal calf serum. The protein content of F was determined by the method of Lowry et al., under different experimental conditions with addition of proline and low or high concentrations of fetal calf serum. F were stained by Gomori's one step trichrome (G), PAS-alcian blue and Sudan black Bmethods.

Results 1) Tumors in kidney, pancreas, lung and heart except for brain contained more H than surrounding normal tissues. Collagen sheet in TS skin and fibrosis showed the same H content as control skin. 2) Ratios of H to creatinine in urine of TS were all higher than those in age-matched control with or without treatment of anticonvulsant, except for muscle disease. 3) F from adenoma cultured in D with low serum showed higher protein contents than control skin F. When proline was added to D with 17% serum, protein content per flask in F from adenoma was higher than that in con­trol F. 4) F from adenoma stained by G revealed more brownish F than control, especially F from fibrosis stained black. The cultured F from TS patients tended to contain many large cells.

Conclusion Based on higher content of H in urine and tissues of patients with TS, an increase in collagen synthesis may exist in TS. Furthermore it appears that F from TS grow in medium containing relatively few serum fac­tors. Last the difference between TS and control cells as to the staining color of F suggests that TS cells possess proline-dependent nature.

References Tanaka H, Arima M, et al. Brain Dev 1979;1:218 (Abst). Bergman I, Loxley R. Anal Chem 1963;35:1961.

Key words: Tuberous sclerosis, hydroxyproline, collagen, fibroblasts, adenoma sebaceum.

Brain & Development, Vol 3, No 2,1981 181

48

Mucopolysaccharides in the Skin Lesions from Patients with Tuberous Sclerosis Norio Sakuragawa, MD, and Masataka, Arima, MD Division of Child Neurology, National Center for Nervous, Mental and Muscular Disorders, Kodaira, Tokyo

Fischer et al described recently the increased contents of acidic glycosaminoglycans (GAG) in an angio­fibroma on the face from a patient with tuberous sclerosis. A content of total carbohydrate was also increased in the GAG fraction of the tumor. We deter­mined the GAG contents and its individual composi­tion in the angiofibroma and shagreen patch from two different patients.

Small pieces of angiofibromas and a shagreen patch were obtaine<,i from patients with TS for a cosmetic surgery. Control skin from individuals, aged 14-64 years, were available for a purpose of amputation of a lower leg or from abdomen at autopsy. Control skins were separated into epidermis and dermis by the ex­tension methods of Van Scott. Angiofibroma were subjected to analyses as a whole. A block of the shagreen patch were separated into epidermis, central and peripheral areas of a tumor. Acetone-ether dried and defatted samples were digested with papain and pronase, followed by alkaline treatment and dialysis. Dializants were precipitated by 2.5% CPC and its supernatant (CPC non-precipitable GAG) was subjected to the further purification to get the more purified GAG. GAG contents were determined by the carba­zole method. One and two-dimensional electrophoresis on cellulose acetate strips for determination of in­dividual GAG was carried out.

Water contents of control skins were 66.5-67.9% in both layer. It was increased in angiofibroma, while decreased in the shagreen patch. CPC precipitable GAG in the control epidermis was 708 ± 332 j.lg uronic acidjg dry wt and 993 ± 509 in control dermis. Angio­fibroma from TS patients contained 980-1,370 J.Lg uronic acidjg dry wt. Shagreen patch showed lower contents of uronic acid (320-390 j.lg uronic acidjg dry wt) than that of controls. One dimensional elec­trophoresis of the purified GAG from the control skin revealed the presence of hyaluronic acid, derma tan sulfate and possibly a small amount of chondroitin 4 and 6 sulfate. Hyaluronic acid was not detectable in the shagreen patch, confirmed by the two-dimensional electrophoresis. CPC non-precipitable GAG was de­creased in the central area of the shagreen patch (7 j.lg uronic acidjg dry wt) but increased in the peri­pheral area (50 j.lg uronic acidjg dry wt). Control dermis from amputated sample contained 18.9 j.lg uronic acidJg dry wt.

We interpret these data to suggest that the de­creased contents of GAG in the shagreen patch, especially in hyaluronic acid, may reflect the bio­chemical changes of GAG metabolism in the tumorous lesions of TS.

Reference Fischer MH, et al. J Ment Def Res 1977;21:251.

Key words: Glycosaminoglycans, tuberous sclerosis, shagreen patch, angiofibroma.

182 Brain & Development, Vol 3, No 2,1981

49

Four Cases of Bourneville-Pringle Disease Asso­ciated with Brain Tumor Tadashi Tsuchida, MD, Kenichi Kamata, MD, Masa­haru Kawamata, MD, and Kohei Okada, MD Department of Neurosurgery, Brain Research Institute, Niigata University, Niigata

It is well known that brain tumors are seldom seen in patients with Bourneville-Pringle disease (BPD). There have, however, been isolated cases reported in this country, in contrast to the numerous reports of brain tumors in patients with von Recklinghausen disease. The authors have encountered 4 cases of BPD associ­ated with brain tumor. The clinical features, radiolo­gical and pathological findings, and our experience in treating these 4 patients are summarized in this report.

Materials and Results Age of patients ranged from 3 to 19; 3 were boys and one was a girl. Seizures were the first neurological sign in all 4 patients. Only one patient had a family history of cutaneous manifestations. At the time their brain tumors were diagnosed, physical examination of the patients showed following; hemangiomatous eruption (adenoma sebaceum) was present in all patients, the so-called "shagreen" patch on the trunk in 3, a white leaf-shaped macula on the limbs ih 2, a cafe-au-lait spot on the back in one, and Koenen tumors at the toenails in one. Three patients had papilledema and one had several nodules in the retina. Ventriculo­graphy was used to locate 1 of the 4 tumors. In each case the tumor was in the region of foramen Monro and was associated with ventricular dilatation. Angio­graphy, which was performed in 3 cases, showed moderate vascularization. In 3 cases CT scans were done and revealed well circumscribed tumors as the high density area which were markedly enhanced by contrast medium. Three patients underwent cranio­tomy. After a shunt operation, total removal was carried out in 2 cases, one by transventricular and one by transcallosal approach. Both of them are doing well postoperatively. However, one patient whose tumor was partially excised in 1967 died in the 4th year after surgery. One patient is being traced with anticonvul­sants because of the small size of the tumor and the lack of IICP signs. Histological studies were done in 3 cases. In one case, the tuber of the cortex lacked the usual neuronal lamination. The pathological study of 3 cases with ventricular tumors showed variations in each case; plump cell astrocytoma, hemangioma with­out glial components, and so-called astroblastoma.

Conclusion The brain tumors in 4 patients suffering from BPD who all had typical cutaneous manifestations were situated near the foramen Monro. In CT scans all of them were contrast enhanced. We advocate that tumors in the lateral ventricle of BPD patients should be totally excised.

Key words: Bourneville·Pringle disease, brain tumor, surgical therapy, tuberous sclerosis.

50

A Case of Unilateral Facial Hemangioma, Cere­brovascular Abnormalities and Hypoplasia of Cerebellum Yuji Mizuno, MD, Toru Kurokawa, MD, Seiichiro Takaki, MD, and Yuji Numaguchi, MD Departments of Pediatrics (YM, ST, TK) and Radiolo­gy (YN), Faculty of Medicine, Kyushu University, Fukuoka

Facial hemangioma is frequently accompanied by intracranial abnormalities. This is the case of a 5-year­old girl with left facial strawberry mark hemangioma, cerebrovascular abnormalities and hypoplasia of cere­bellum.

Case Report She had no complaints other than the facial hemangi­oma. On neurological examinations, however, ab­normalities were seen on the left side; i.e., internal strabismus, visual impairment, disturbance of eye movement as well as optic atrophy of the left eye, and left cerebellar signs. A skull X-ray disclosed bUlging of inion and thinning of the occipital bone, but there were no calcifications. Brain computed tomography revealed a cystic lesion with CSF density in the left posterior fossa and vascular lesions enhanced in the region of bilateral middle cerebral arteries. RI cis­ternography and right vertebral angiography showed that the cystic lesion was a large cisterna magna ac­companied with hypoplasia of left cerebellum. A cere­bral angiography revealed stenosis of the left internal carotid artery, a saccular aneurysm at the syphon of the same artery, and occlusion of the basilar artery.

Discussion There are several neurocutaneous syndromes with facial hemangioma: i.e., Sturge-Weber syndrome, Bon­net-Duchaume-Blanc syndrome and Wyburn-Masson syndrome. However, the cerebrovascular abnormalities characteristic in these syndromes are angiomatous mal­formations, not stenosis nor occlusion. In 1978, Castroviejo briefly described seven cases of facial and scalp capillary hemangioma associated with vascular and nonvascular intracranial malformations, one of which resembles ours. This case only being sum­marized, its identity to ours is uncertain. Our case may be a new neurocutaneous syndrome which pre­viously has not been fully described.

Conclusion The present case of a 5-year-old girl with the left facial hemangioma associated with cerebrovascular abnor­malities and ipsilateral cerebellar hypoplasia represents a new syndrome.

Key words: Facial hemangioma, cerebrovascular ab­normalities, cerebellar hypoplasia, neurocutaneous syndrome.

51

Effect of Neonatal Hypoxia on Brain Develop­ment. II. Catch-Up Growth after Hypoxia Hiroshi Yoshioka, MD, Akira Yoshida, MD, Masaharu Ochi, MD, Masahiro Mino, MD, Kiminao Mizukawa, MD, Yasuo Kasubuchi, MD, and Tomoichi Kusunoki, MD Department of Pediatrics, Kyoto Prefectural Univer· sity of Medicine, Kyoto

In the previous report we have shown that neonatal hypoxia causes a reduced rate of cell proliferation with a delay in the G2-phase in the external granular layer of the 2-day-old mouse cerebella. This report describes a subsequent catch-up growth, with special reference to the cell proliferation by using autoradio­graphic technique.

Material and Methods Two-day-old mice, ICR-JCL strain, were put into a chamber for 30 minutes which was continuously flushed with 100% CO2 gas and almost equilibrated with 0% O2 , 100% CO2 , A cell-cycle study on the external matrix cells at the external granular layer of the cerebellum from 7-day-old CO2 exposed and control mice was carried out by means of 3H-thymi­dine autoradiography. On midsagittal sections of the CO2 exposed and control 20-day-old mouse cerebella, Purkinje cells were counted blindly.

Results The increase in brain weight of the CO2 exposed mice was temporarily stunted. The mean value of brain weights of the experimental animals was significantly less than that of age-matched control at 5 days (p<0.001) and at 10 days (p<0.005). Thereafter the brain weights of experimental animals rapidly increased and there was no significant difference in the brain weight at 20 days between two groups. Generation time of the external matrix cells at the external granular layer of the control mice was about 16 hours, whereas that of the CO2 exposed mice was about 14 hours. The durations of the So, G,-, G2-,

and M-phases in the control mice were about 9, 5, 1.5, and 0.5 hours, respectively, while those in the CO2

exposed were about 8.5, 3.5, 1.5, and 0.5, respective­ly. The mean number of Purkinje cells of the control animals was 867, whereas that of the experimental animals was 757 (p<0.005).

Conclusion Neonatal hypoxia had an adverse effect on neuronal proliferation, which reverted to normal spontaneously in the older animals. It also reduced Purkinje cell numbers.

Key words: Neonatal hypoxia, Purkinje cells, hypoxia, brain development, catch-up growth, autoradiography.

Brain & Development, Vol 3, No 2,1981 183