Novel growth factor supporting survival of murine primordial germ cells: evidence from conditioned medium of ter fetal gonadal somatic cells.

The ter (teratoma, chromosome 18) mutation causes a deficiency of primordial germ cells (PGCs) in ter/ter embryos from the ter congenic mouse strain at 8.0 days post coitum (dpc). In order to analyse the function of the ter gene, here we examined effects of conditioned medium (CM) from 14.5 dpc testicular and ovarian somatic cells of +/+, +/ter, or ter/ter genotype on mouse PGCs "mixed-cultured" with own somatic cells on feeder cells. The results showed that +/+ and +/ter CM supported survival in 9.5 and 11.5 dpc ICR PGCs but ter/ter CM did not rescue TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling)-positive apoptosis in the PGCs though it did not affect 5-bromo-2-deoxyuridine incorporation in PGCs. This supportive substance in +/+ CM, not ter/ter CM, was characterized as soluble, heat labile, and larger than 30 kDa. We also found that several known growth factors for PGCs and their receptors were expressed in ter/ter testes as well as +/+ testes, suggesting the ter function is independent. Thus, it was concluded that fetal gonadal somatic cells express a novel PGC growth factor (designated as TER Factor) supporting survival of PGCs not somatic cells and that the PGC deficiency in ter/ter testes is caused by a loss of this factor.

[Ruptured traumatic aneurysms of the peripheral anterior cerebral artery: study of delayed hemorrhage after closed head injury].

We report three cases of ruptured traumatic aneurysms of the peripheral anterior cerebral artery after closed head injury. These cases were all young men with closed head injury due to traffic accidents. Consciousness level on admission was coma in all three cases. Case 1 was a 19-year-old man with interhemispheric hematoma on initial CT, then 7 days later his consciousness cleared. However, 14 days later he suddenly lapsed into a deep coma with a severe frontal hemorrhage. Case 2 was a 13-year-old boy. Plain skull films demonstrated a frontal depressed fracture, but CT scan showed no bleeding. Four days later his consciousness cleared but 11 days after trauma, he lapsed into a deep coma with a frontal hemorrhage. Case 3 was a 22-year-old man. Initial CT showed a slight ventricular hemorrhage. Fourteen days later, his consciousness had almost cleared, but then he lapsed into a deep coma with a large frontal hemorrhage 11 weeks after the trauma. These patients all died within a few days after intracranial bleeding. All patients underwent cerebral angiography but none of them showed filling defect. Autopsy was performed and ruptured aneurysms were found on the distal anterior cerebral artery that had no relation to the branch of bifurcation. Histological examination demonstrated a lack of elastic lamina and media in all of these three cases, so each of them was a victim of so-called false aneurysm. Twenty reported cases of ruptured traumatic aneurysms of the peripheral cerebral artery with delayed hemorrhage after closed head injury were reviewed. Factors in the traumatic aneurysm showed no relation to the duration of disturbed consciousness. Within one month, delayed hemorrhage due to ruptured traumatic aneurysm occurred. None of the delayed hemorrhages involved subarachnoid hemorrhage. Subdural hematoma was seen in the distal middle cerebral artery and frontal hemorrhage was found in the distal anterior cerebral artery. We consider that frontal hemorrhage is a predictive finding for the type of delayed hemorrhage due to traumatic aneurysm in the distal anterior cerebral artery.

[Penetrating injury of the head, neck and chest by a nail-gun: a case report].

A very rare case of directly penetrating injury of the head, neck and chest caused by a nail-gun was reported. A 56-year-old male was admitted to our critical care center due to chest injury. On admission, he showed numbness of the hand & leg, left hemiparesis and hypalgesia. Physical examination disclosed three nails on the left anterior chest, but other wounds or nails were not found. Chest films showed three nails penetrating the lung without reaching the heart, but other nails were found by skull and neck films. One nail had penetrated the cervical canal at the C1 level through the posterolateral cervical region. Two other nails were demonstrated at the right temporal and the left frontal region, Computed tomography revealed no massive cerebral hemorrhage and cerebral angiography showed no extravasation and no passage through main vessels. Emergency surgery was performed uneventfully and the nails in the chest, neck and head were totally removed. He was discharged one month after surgery. Some injuries caused by a nail-gun have been reported in the world literature but in Japan this multiple injury case was the first reported. Since nail-gun injuries can cause multiple damage, systemic X-ray examination was very important.

[The mechanism of cardiorespiratory arrest due to subarachnoid hemorrhage].

This report describes the clinical course of patients with sudden cardiorespiratory arrest (CRA) due to subarachnoid hemorrhage (SAH). We have seen fifteen patients of SAH that presented initially as CRA. All of them were diagnosed as SAH by CT scan. The patients were divided into two groups; one group (early DOA group) included 11 patients, who had been recognized as CRA within 60 minutes from the onset of SAH, the other group (late DOA group) consisted of 4 patients, who developed CRA more than 60 minutes after the initial onset. The major mechanism leading to delayed CRA in the late DOA group appeared to have been from brain stem herniation, but another mechanism appeared to be involved in sudden CRA in the early DOA group. Sixty percent of our patients with CRA due to SAH had a low serum potassium concentration, though hypokalemia was observed in only 4 out of 100 patients with CRA due to diseases other than SAH. These facts suggest that sympathetic hyperstimulation might result not only from stress but also from a disorder of the central autonomic nervous system. We speculate that the mechanism leading to early CRA after SAH appears to result from a disorder of the central autonomic nerve system.

[Cisplatin-etoposide chemotherapy of an embryonal carcinoma arising in the basal ganglia of the cerebrum: a case report].

Cases of embryonal carcinoma arising in the basal ganglia are rarely reported. According to the literature available, only 5 cases of embryonal carcinoma, arising from the basal ganglia, have been reported to date. This paper reports one such case we recently encountered. The patient was a 15-year-old boy. He was first admitted to another hospital because of occasional headache and vomiting. During the hospital stay, CT scans revealed abnormalities. For this reason, the patient was referred to our critical care center. Upon admission to our center, a physical examination revealed no abnormalities, but neurological examination disclosed left hemiparesis. CT scans revealed a large mass lesion of a low to high density in the right basal ganglia, accompanied by midline shift and ventricular dilatation. Elevation of human chorionic gonadotrophin (HCG) and alpha-fetoprotein (AFP) in both serum and cerebrospinal fluid (CSF) was observed. The tumor with multiple cysts was removed totally by craniotomy. The removed tissue was rated histopathologically as mixed-type germ cell tumor composed of germinoma and embryonal carcinoma. The removed tumor cells were found immunohistologically to contain HCG and AFP. Postoperative CT scans showed complete disappearance of the tumor. Taking into account a recent report that a combined cisplatin and etoposide therapy (PE chemotherapy) was effective in treating intracranial germ cell tumors, we used this chemotherapy for postoperative management of this patient. The patient underwent 3 cycles of PE chemotherapy during the 3 months after surgery. The elevated HCG and AFP levels in serum and CSF returned to their normal range within 2 months after surgery. CT and tumor markers revealed no signs of recurrence.(ABSTRACT TRUNCATED AT 250 WORDS)

A new type of defect in the gene for bilirubin uridine 5'-diphosphate-glucuronosyltransferase in a patient with Crigler-Najjar syndrome type I.

Crigler-Najjar syndrome (CN) type I, which is characterized by the complete absence of bilirubin uridine 5'-diphosphate-glucuronosyltransferase (UGT) activity, is inherited as an autosomal recessive trait associated with unconjugated hyperbilirubinemia. Phenobarbital has no effect on the bilirubin concentration in the serum of patients with CN type I. Recently, cDNA for two human liver bilirubin UGT (UGT1A and UGT1D) were isolated, and their genetic organization was determined. The UGT1A (UGT1*1) and UGT1D (UGT1*4) genes each have a unique exon 1, whereas exons 2-5 are common to both genes. It has been predicted that some defect in the exons common to both genes is responsible for the absence of UGT1A and UGT1D activities in CN type I, and five cases with such a mutation have been reported. We describe here a new type of defect in the gene for bilirubin UGT in a patient with CN type I, namely, an abnormality in the exon 1 that is characteristic of the UGT1A gene. This mutation is a single nucleotide substitution, that is, C is changed to A at base position 840 in UGT1A cDNA, and this change results in a stop codon. Our patient is homozygous for the defect, and his nonconsanguineous parents and elder brother, who are clinically normal, are heterozygous for the defective allele. No mutation was detected in any exons of the UGT1D gene. Our results suggest that a homozygous nonsense or deletion mutation is detected not only in the exons common to UGT1A and UGT1D genes but also in unique exon 1 of UGT1A in CN type I.

[A case of cardiogenic shock caused by Vibrio parahaemolyticus].

A case of a 53 year old healthy female complaining of diarrhea and abdominal pain after taking raw fish is presented. She immediately went into shock and unconsciousness. Central venous pressure was 8 cmH2O and her ECG showed a first-degree AV block and ST-T changes in almost all leads. After mechanical ventilation and administration of dopamine, dobutamine, cefotiam, ciprofloxacin, she became alert and recovered from her critical condition. V. parahaemolyticus which produces thermostable direct hemolysin (TDH) was cultured from the feces on admission. Kanagawa phenomenon was positive. Arterial blood culture was negative and the titer of serum endotoxin was low. The diagnosis of cardiogenic shock due to exotoxin produced by V. parahaemolyticus was made. Serological examination by ELISA showed elevation of IgG class antibody against TDH and TRH (TDH related hemolysin). And antibody against TDH was normalized after 180 days. By review of literature, there are some case reports of cardiogenic shock complicated with V. parahaemolyticus infection, but few showed elevation of antibody against TDH and TRH in the serum of the survived patient.

Identification of defect in the genes for bilirubin UDP-glucuronosyl-transferase in a patient with Crigler-Najjar syndrome type II.

Crigler-Najjar syndrome (CN) type II is characterized by severe chronic nonhemolytic unconjugated hyperbilirubinemia due to reduced hepatic bilirubin UDP-glucuronosyl-transferase (UGT) activity. Two bilirubin UGT isozymes, UGT1A and UGT1D, have been identified. We analyzed the DNA sequence of the bilirubin UGT genes in a 5-year-old Japanese male patient with CN type II, who had consanguineous parents. Point mutations were found on exons 1 of the UGT1A and UGT1D genes. The abnormalities were single nucleotide substitutions of G by A and of T by C at base position 211 of UGT1A cDNA and at base position 395 of the UGT1D, respectively. We found another single nucleotide substitution of T by G on exon 5 common to both genes at base position 1456 of the UGT1A cDNA or 1459 of the UGT1D cDNA. These three mutations result in changes of glycine to arginine and of tyrosine to aspartic acid at amino acid positions 71 and 486 of the UGT1A protein, and of leucine to proline and of tyrosine to aspartic acid at amino acid positions 132 and 487 of the UGT1D protein, respectively. Our patient was homozygous for all defects and his parents and elder brother were heterozygous for all defective alleles. The findings suggest that the CN Type II is inherited as an autosomal recessive trait.

Suprasellar embryonal carcinoma which developed ten years after local radiation therapy for pineal germinoma.

We recently encountered a case of suprasellar embryonal carcinoma which developed 10 years after local radiation therapy for pineal germinoma. This 29-year-old male was admitted because of abnormal behavior, disturbed consciousness, and increased intracranial pressure. Computed tomography and magnetic resonance imaging disclosed a tumor spreading from the suprasellar region to the third ventricle and the anterior horn of the left lateral ventricle, accompanied by fresh hemorrhage within the tumor. He underwent subtotal removal of the tumor, followed by chemotherapy with cisplatin and etoposide. The tumor was histologically rated as embryonal carcinoma with differentiation into yolk sac tumor and choriocarcinoma.

The effect of a new calcium antagonist, TA3090 (clentiazem), on experimental transient focal cerebral ischemia in cats.

BACKGROUND AND PURPOSE: TA3090 (Clentiazem) has been shown to have cerebrovascular protective properties in three experimental studies. An in vivo investigation was undertaken to determine its effects on pial arteries and cerebral blood flow and its therapeutic value in transient focal cerebral ischemia. METHODS: This experiment was divided into two protocols. In the first, 200 or 400 micrograms/kg per hour TA3090 was administered continuously for 3 hours in cats without ischemic insult (n = 6 for each group). The effects on pial arteries and cerebral blood flow were estimated. In the second protocol, 400 micrograms/kg per hour TA3090 (treated group, n = 14) or physiological saline (control group, n = 10) was administered 5 minutes before 1 hour of middle cerebral artery occlusion in cats. The effects on the pial arteries and cerebral blood flow were observed continuously, followed by autoradiography for a quantitative measurement of cerebral blood flow 5 hours after middle cerebral artery recirculation. The volumes of the cerebral edema and infarct were estimated by planimetry from cerebral preparations made for histological examination. RESULTS: Pial arteries dilated by up to approximately 10% in the 400-micrograms group and 3% in the 200-micrograms group 30 minutes after administration of TA3090. Increases in cerebral blood flow of about 10% in the 400-micrograms group and 2% in the 200-micrograms group were demonstrated with laser Doppler flowmetry. In the second protocol, dilatation of pial arteries was significantly smaller during and after the ischemic insult in the treated group compared with the control group (p < 0.01). Cerebral blood flow decreased less significantly during ischemia (p < 0.01 at the end of ischemia) and increased less significantly after ischemia (p < 0.01 at the end) in the treated group compared with the control group. Autoradiography showed a more remarkable increase in cerebral blood flow due to luxury perfusion in the cerebral cortex, which was mainly perfused by the middle cerebral artery on the affected side in the control group (p < 0.01). Cerebral blood flow in the cerebral cortex, thalamus, and caudate nucleus on the contralateral side of the treated group increased by about 20% more than that of the control group (p < 0.05). Cerebral edema and infarction were much smaller in the treated group than in the control group (p < 0.01). CONCLUSIONS: 1) TA3090 dilates pial arteries and increases cerebral blood flow in normal brain regions in a dose-response fashion; 2) in ischemic regions compared with those in untreated animals, TA3090 results in a lesser reduction of cerebral blood flow during ischemia and in a lesser degree of hyperemia during reperfusion; 3) TA3090 is associated with less pial artery dilatation during ischemia, presumably due to improved collateral flow; and 4) the improved hemodynamic state with TA3090 is associated with significant reduction of cerebral edema and infarct size.

[Experimental study on brain reversibility following temporary regional cerebral ischemia; from the point of view of vessel behavior and histological changes].

Using a middle cerebral artery occlusion model in adult cats, we investigated the ischemic time threshold for inflicting damage on cerebral vessel behavior and brain tissue. With the transorbital approach, a middle cerebral artery (MCA) was exposed and temporarily obstructed by a Zen's clip. Animals were divided into ten groups (each group: N = 8) according to the ischemic time of 10, 20, 30 minutes, 1, 2, 4, 6, 8, 12, 24 hours. Five hours after the recanalization, Evans' blue dye was injected intravenously and after further 30 minutes, the brain was fixed with transaortal perfusion for histological examinations. A cranial window was made above the ectosylvian gyrus which has poor anastomosis. The pial vessel behavior was observed through the cranial window and evaluated using an intravital microscope and a videoangiometer. Considering the correlation between functional change of vessel behavior and pathological change in the brain, the threshold time of ischemia inflicting irreversible damage was estimated. In the 30 minute ischemia group, deterioration of vessel behavior began to be observed as well as extravasation of Evans' blue dye. As the ischemic duration became longer, infarction and hemorrhage, which showed close correlation, increased. When the duration of ischemia was 6 hours, the infarcted area was significantly larger than that of the 4-hour ischemia group. Intracranial pressure (ICP) markedly increased as soon as recanalization took place. We think this phenomenon is due mainly to swelling. About three hours after recanalization, ICP further increased due to severe vasogenic edema in addition to the brain swelling.(ABSTRACT TRUNCATED AT 250 WORDS)

[The effect of repeated occlusions of the middle cerebral artery on pial arterial behavior and cerebral structure].

Temporary clipping of the major arterial trunk is a very important maneuver to control excessive unexpected bleeding during a neurosurgical operation, but repeated temporary clippings sometimes give rise to severe neurological deficits after surgery. In clinical practice, a major stroke can occur after many transient ischemic attacks without distinct angiographic occlusion. To confirm and explain these clinical experiences, the present study was performed. First, 20-min, 30-min and 1-h occlusion of the middle cerebral artery was performed in each of 5 cats, and pial arterial behavior, cerebral edema and infarction were observed. In the 20-min occlusion group, no abnormal change was found 5 hours after recirculation. In the 30-min occlusion group, cerebral edema was present in 10.5 +/- 4.2% of the hemisphere, but no infarction was observed, and pial arterial caliber remained in a 10% dilated state throughout the experimental periods. In the 1-h occlusion group, cerebral edema was present in 41.2 +/- 7.5% of the hemisphere and infarction was found in 34.5 +/- 9.5%. Pial arteries returned to a 20% dilated state but redilated by 45% at the end of experiment. As the second experiment, three 20-min occlusions at 1-h interval and two 30-min occlusions at 1-h interval were performed in each 10 cats. Pial arteries had dilated by 40% after release of the last occlusion in both groups. The extent of cerebral edema was 19.5 +/- 8.1% of hemisphere in the 20-min occlusions group and 36.6 +/- 9.7% in the 30 min occlusions group.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of repeated temporary clipping of the middle cerebral artery on pial arterial diameter, regional cerebral blood flow, and brain structure in cats.

Temporary clipping of the major arterial trunk is an important maneuver to control excessive unexpected bleeding in neurosurgical operations; however, repeated temporary clipping can give rise to severe neurological deficits after surgery. The present study was performed to confirm and explain these clinical findings. Initially, a single 20-minute or 1-hour occlusion of the middle cerebral artery was performed in each of 5 cats. Pial arterial diameter was determined by video imaging, regional cerebral blood flow was measured by autoradiography, and cerebral edema and infarction were observed. In the 20-minute occlusion group, no abnormal changes were found 5 hours after recirculation. In the 1-hour occlusion group, pial arteries were dilated by 45%, and regional cerebral blood flow increased to more than twice the resting cortical values. The extent of cerebral edema was 41.2 +/- 7.5% (SE) and infarction was 34.5 +/- 9.5% (SE) of the hemisphere. In the second experiment, three 20-minute occlusions of the middle cerebral artery in a 1-hour interval were performed in 20 cats. In 10 of them, thiopental (40 mg/kg) was used to protect the brain. In the group without barbiturate treatment, pial arteries were dilated by 40% at the end of experiment, regional cerebral blood flow decreased to about 70% compared with single 20-minute occlusion, cerebral edema was 19.5 +/- 8.1% (SE), and infarction was 8.1 +/- 3.7% (SE) of the hemisphere. In the treated group, these were only trivial changes. The effect of repeated clipping may cumulatively cause brain damage, and barbiturates should be used whenever repeated clipping is necessary.

[The influence of volume expansion with induced hypertension on vessel reactivities, blood-brain-barrier, cerebral edema and infarction].

Eighteen anesthetized cats were randomly divided into 3 groups. One group was treated with 10 ml/kg of 6% hydroxyethyl starch (HES). Another group was treated with 15 ml/kg, and there was a control group. There were six cats in each group. Cerebral pial arterial reactivities were measured by means of intravital microscopic observation through a cranial window. Middle cerebral artery (MCA) was clipped for 70 minutes using a transorbital approach. Five hours later, Evans blue dye was injected intravenously and then the animals were killed. Administration of HES or Ringer's solution (control) was started 15 minutes before clipping of MCA. Systemic arterial pressure was elevated by using dopamine at the same time as the MCA occlusion was carried out. Pial arteries dilated soon after the clipping of MCA in all three groups, but showed different patterns in each group after recirculation of MCA. In the control group, they returned to 20% of their dilated state first, but, after about 4 hours, they redilated by 45% at the end of experiment periods. In the group treated with 10 ml/kg of 6% HES, they remained in an almost resting state after MCA recirculation, but in the group with 15 ml/kg, they constricted by 10% at the end of the experiment periods. Regarding the Evans blue dye extravasation, cerebral edema and infarction, they were largest in the control group. But animals treated with 15 ml/kg of 6% HES showed significantly larger Evans blue dye extravasation and cerebral edema than that of animals with 10 ml/kg. However cerebral infarctions were almost the same sizes in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effect of calcium antagonist nimodipine and induced hypertension on cerebral vessel reactivities, cerebral blood flow, cerebral edema and infarction in cats with one hour middle-cerebral-artery occlusion].

Treatment with the calcium entry blocker nimodipine is recommended as effective therapy for cerebral ischemia due to cerebral vasospasm or cerebral thrombosis. On the other hand, treatment with induced hypertension is a widely accepted measure to reverse ischemic deficits caused by vasospasm. Thus, a combination of the two regimens--nimodipine and induced hypertension--may have real benefits for cerebral ischemia. But it is possible that the benefit of one is abolished by adverse effects of the other, or that a combination of both may not be as effective as the use of only one therapy. In order to investigate these problems, the effects of nimodipine and induced hypertension on cerebral vessel, cerebral blood flow, cerebral edema and cerebral infarction using a one hour middle-cerebral-artery occlusion model in cats. Twenty-one anesthetized cats were divided into a control group, the nimodipine-treated group, and the nimodipine-and-induced-hypertension group. There were seven cats in each group. Occlusion of the middle cerebral artery (MCA) was continued for one hour in each animal. Induced hypertension was a little higher than resting values, and it was continued for only one hour during MCA occlusion, brought on by instillation of dopamine. Cerebral pial arteries dilated much more prominently during and after the occlusion of MCA in the nimodipine-and-induced-hypertension group than other groups. Although cerebral blood flow in the nimodipine group, and the nimodipine-and-induced-hypertension group increased more in the non-ischemic hemisphere, the most remarkable increase was seen around the infarcted cortex in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

[Dermal patch anesthesia to prevent pain from dermal puncture--10% lidocaine aqueous gel with 3% glycyrrhetinic acid 3-0 hemiphthalate disodium].

The clinical efficacies of 10% lidocaine aqueous gel with and without 3% glycyrrhetinic acid 3-0 hemiphthalate disodium (GAHPh) applied as a skin patch for reduction of pain from venous cannulation were evaluated in a double blind study. Twenty-four adult patients, who were scheduled for surgery under general anesthesia, gave informed consent to participate in this study. The patients were allocated randomly into two groups: one for a dermal patch GAHPh (GAHPh group) and the other for a dermal patch without GAHPh (plain group). Approximately 0.3g of either gel with or without GAHPh, soaked in a round sponge (25mm in diameter, 1mm in thickness), was applied over the selected vein on the arm and was covered with an adhesive plastic film (Tegaderm). Pain score was graded by the number of painful spots out of the 5 tests in the treated skin area. In patients with a pain score under 1, venous cannulation was carried out without an intradermal injection of a local anesthetic and pain associated with the cannulation procedure was graded by patients on a scale of 5, where 0 = no pain, 1 = little pain, 2 = moderate pain, 3 = painful, 4 = very painful. The mean application time periods were 59.3 min for the GAHPh group and 60.3 min for the plain group. Transient local redness was observed in 8 patients after removal of the gels; 3 in the GAHPh group and 5 in the plain group. The mean pain score (1.3 +/- 1.5) in the GAHPh group, was significantly lower than that (2.5 +/- 1.7) in the plain group (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)