X-irradiation causes a prolonged reduction in cell proliferation in the dentate gyrus of adult rats.

The effects of X-irradiation on proliferating cells in the dentate subgranular zone were assessed in young adult Fisher 344 rats exposed to a range of X-ray doses and followed for up to 120 days. Apoptosis was quantified using morphology and end-labeling immunohistochemistry, and cell proliferation was detected using antibodies against the thymidine analog BrdU and the cyclin-dependent kinase p34(cdc2). Radiation-induced apoptosis occurred rapidly, with maximum morphological and end-labeling changes observed 3-6h after irradiation. Twenty-four hours after irradiation cell proliferation was significantly reduced relative to sham-irradiated controls. The number of apoptotic nuclei increased rapidly with radiation dose, reaching a plateau at about 3Gy. The maximum number of apoptotic nuclei was substantially higher than the number of proliferating cells, suggesting that non-proliferating as well as proliferating cells in the subgranular zone were sensitive to irradiation. Subgranular zone cell proliferation was significantly reduced relative to age-matched controls 120 days after doses of 5Gy or higher. These findings suggest that neural precursor cells of the dentate gyrus are very sensitive to irradiation and are not capable of repopulating the subgranular zone at least up to 120 days after irradiation. This may help explain, in part, how ionizing irradiation induces cognitive impairments in animals and humans.

Screening of Brazilian fruit aromas using solid-phase microextraction-gas chromatography-mass spectrometry.

Manual headspace solid-phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) was used for the qualitative analysis of the aromas of four native Brazilian fruits: cupuassu (Theobroma grandiflorum, Spreng.), cajá (Spondias lutea. L.), siriguela (Spondias purpurea, L.) and graviola (Anona reticulata, L). Industrialized pulps of these fruits were used as samples, and extractions with SPME fibers coated with polydimethylsiloxane, polyacrylate, Carbowax and Carboxen were carried out. The analytes identified included several alcohols, esters, carbonyl compounds and terpernoids. The highest amounts extracted, evaluated from the sum of peak areas, were achieved using the Carboxen fiber.

Inhibition of dentate granule cell neurogenesis with brain irradiation does not prevent seizure-induced mossy fiber synaptic reorganization in the rat.

Aberrant reorganization of dentate granule cell axons, the mossy fibers, occurs in human temporal lobe epilepsy and rodent epilepsy models. Whether this plasticity results from the remodeling of preexisting mossy fibers or instead reflects an abnormality of developing dentate granule cells is unknown. Because these neurons continue to be generated in the adult rodent and their production increases after seizures, mossy fibers that arise from either developing or mature granule cells are potential substrates for this network plasticity. Therefore, to determine whether seizure-induced, mossy fiber synaptic reorganization arises from either developing or mature granule cell populations, we used low-dose, whole-brain x-irradiation to eliminate proliferating dentate granule cell progenitors in adult rats. A single dose of 5 Gy irradiation blocked cell proliferation and eliminated putative progenitor cells in the dentate subgranular proliferative zone. Irradiation 1 d before pilocarpine-induced status epilepticus significantly attenuated dentate granule cell neurogenesis after seizures. Two irradiations, 1 d before and 4 d after status epilepticus, essentially abolished dentate granule cell neurogenesis but failed to prevent mossy fiber reorganization in the dentate molecular layer. These results indicate that dentate granule cell neurogenesis in the mature hippocampal formation is vulnerable to the effects of low-dose ionizing irradiation. Furthermore, the development of aberrant mossy fiber remodeling in the absence of neurogenesis suggests that mature dentate granule cells contribute substantially to seizure-induced network reorganization.

Long-term impairment of subependymal repopulation following damage by ionizing irradiation.

In the mammalian brain, the subependyma (SE) contains stem cells capable of producing neurons and glia. In normal brain these stem cells are responsible, in part, for maintaining the morphologic and functional integrity of the SE; what role the cells of the SE play in brain injury has not yet been elucidated. The present study was designed to determine the long-term regenerative potential of the rat SE after significant depletion of stem cells. Ionizing irradiation was used to deplete cells of the SE and subsequent cellular responses were quantified using immunohistochemical analyses on formalin-fixed, paraffin-embedded tissues. A histomorphometric approach was used to quantify total cell number, number of proliferating cells, number of immature neurons, astrocytes, and undifferentiated components of the SE. Because there are no markers specific for stem cells, we used a repopulation assay as an indirect measure of stem cell response after injury. Our data showed clear radiation dose-dependencies in our quantitative endpoints, implying that there was progressively more stem cell damage with increasing radiation dose. Repopulation of the SE in terms of total cell number, number of proliferating cells and numbers of immature neurons was impaired in a dose-dependent fashion up to 180 days after treatment. These data suggest that after irradiation, surviving stem cells are unable to regenerate the SE. This inability to regenerate after stem cell damage/depletion could have important implications with respect to the normal function of the SE and the function of the SE after brain injury.

Combined brachytherapy and external beam radiotherapy in normal monkey brains--experimental radiation necrosis evaluated by sequential magnetic resonance imaging.

Radiation-induced brain damage was evaluated using sequential magnetic resonance (MR) imaging in monkeys more than 1 year after either brachytherapy or combined brachytherapy and external beam radiotherapy (EBRT). MR images were obtained 1 week, 1 month, 3 months, and subsequently every 3 months after brachytherapy, and the volume of the lesions was measured. In all four monkeys receiving only brachytherapy and three of the four animals receiving combined brachytherapy and EBRT, MR imaging revealed only transient extensive edema 1 week after treatment and ring enhancement, which was maximal 3 months after treatment, surrounding the implantation site. In one of the four animals undergoing combined brachytherapy and EBRT, MR images obtained 9 months after treatment showed an irregularly extending enhanced lesion with edema. MR images obtained 15 months after brachytherapy in this monkey showed the lesion extended into the contralateral hemisphere through the corpus callosum. Necropsy revealed severe radiation necrosis. This animal developed a very similar MR imaging presentation to that often encountered in human brains after combined brachytherapy and EBRT.

Stereotactic brachytherapy for a cystic metastatic brain tumor in the midbrain. Case report.

The authors report a rare case of a cystic metastasis in the midbrain that was successfully treated by brachytherapy following stereotactic biopsy and aspiration of the intratumoral cyst. Stereotactic aspiration of cystic lesions can lead to clinical improvement and brachytherapy prevents cyst recurrence. A 46-year-old man was referred to the authors' institution with a 2-month history of a left hemisensory disturbance and a 1-month history of progressive hemiparesis. Magnetic resonance (MR) imaging revealed a ring-enhancing cystic mass in the midbrain. On the basis of this imaging study, a differential diagnosis that included brainstem abscess, glioma, and metastatic tumor was made. Magnetic resonance imaging-guided stereotactic biopsy and aspiration of the intratumoral cyst were performed, yielding 5 ml of yellowish-white fluid. Histological examination provided a diagnosis of adenocarcinoma. During the surgery, a catheter through which brachytherapy would be delivered was inserted at a predetermined target. The patient's left hemiparesis and sensory disturbance were markedly improved and brachytherapy was begun 2 days postoperatively. Three radioactive isotopes composed of iridium-192 were implanted to irradiate the tumor tissue. The total dose at the tumor periphery was 30 Gy, which was administered over 100 hours. External-beam radiotherapy (20 Gy) was added after completion of the brachytherapy. At discharge from the hospital, the patient was alert and all his neurological symptoms had resolved. Follow-up MR imaging revealed stabilization of the cyst and no recurrence of the tumor. The patient is alive and well 18 months following the brachytherapy. This case suggests that brachytherapy can delay cyst recurrence, suppress tumor growth, and prolong survival in patients with cystic brainstem metastasis.

Proton MR spectroscopy of delayed cerebral radiation in monkeys and humans after brachytherapy.

PURPOSE: To determine whether radiation necrosis can be differentiated from residual/recurrent tumor by proton MR spectroscopy. METHODS: We studied the effects of interstitial brachytherapy on the brains of healthy monkeys and in humans with glioblastoma multiforme. The effects of radiation therapy on normal brain tissue in monkeys were assessed with sequential proton MR spectroscopic studies 1 week to 6 months after brachytherapy. Proton MR spectroscopy was also performed in five patients with residual/recurrent glioblastoma multiforme (three of whom had radiation necrosis after brachytherapy), seven patients with newly diagnosed untreated glioblastoma multiforme, and 16 healthy volunteers, who served as a control group. RESULTS: In monkeys, the ratio of N-acetylaspartate (NAA) to creatine-phosphocreatine (Cr) and the ratio of choline-containing compounds (Cho) to Cr of the reference point were significantly lower 1 week after brachytherapy than before treatment. The ratio of NAA to Cho of the irradiated area tended to be higher 1 week after brachytherapy than before irradiation. These peak metabolic ratios showed characteristic changes 6 months after treatment. In two of three monkeys, lipid signal was elevated 6 months after irradiation. In the clinical study, the ratio of NAA to Cho in the area of radiation necrosis was significantly different from that in glioblastoma multiforme when compared with the contralateral hemisphere after irradiation. In addition, lipid signal was detected in all patients with radiation necrosis. CONCLUSION: It might be possible to use proton MR spectroscopy to differentiate radiation necrosis from residual/recurrent glioblastoma multiforme on the basis of comparisons with the contralateral hemisphere after radiation therapy.

The protective effect of dexamethasone against radiation damage induced by interstitial irradiation in normal monkey brain.

OBJECTIVE: The protective effect of dexamethasone against radiation damage is unclear. We examined the effect of early treatment of high-dose dexamethasone on iridium-192-induced damage to normal brain tissue. METHODS: Brain damage induced by interstitial irradiation with iridium-192 was evaluated with sequential magnetic resonance imaging and proton magnetic resonance spectroscopy in 11 adult monkeys, with or without short-term high-dose dexamethasone treatment. Dexamethasone (1 mg/kg of body weight/d) was administered intramuscularly to five irradiated animals every 24 hours, beginning 2 days before and ending 7 days after irradiation. Magnetic resonance imaging and proton magnetic resonance spectroscopy were performed 1 week, 1 month, and 3 months after irradiation. RESULTS: Magnetic resonance imaging performed 1 week after irradiation revealed marked edema in five nontreated animals. In dexamethasone-treated animals, the volume of edema was reduced significantly, compared to that of nontreated animals, 1 week and 1 month after irradiation. The volume of ring enhancement in dexamethasone-treated animals was also reduced significantly, compared to that of nontreated animals, 3 months after the irradiation. Proton magnetic resonance spectroscopy spectra revealed that N-acetylaspartate and choline peaks were reduced 1 week after irradiation in both groups. However, there were no statistically significant differences between the two groups at any time points. CONCLUSION: These results suggest that dexamethasone treatment may have an antiedema effect at an early stage and may modify subsequent development of vascular and inflammatory changes but may have no effect of preventing radiation-induced necrosis and the reduction of N-acetylaspartate after brachytherapy.

Apoptosis in the subependyma of young adult rats after single and fractionated doses of X-rays.

Ionizing radiation is commonly used in the treatment of brain tumors but can cause significant damage to surrounding normal brain. The pathogenesis of this damage is uncertain, and understanding the response of potential target cell populations may provide information useful for developing strategies to optimize therapeutic irradiation. In the mammalian forebrain, the subependyma is a mitotically active area that is a source of oligodendrocytes and astrocytes, and it has been hypothesized that depletion of cells from this region could play a role in radiation-induced white matter injury. Using a distinct morphological pattern of nuclear fragmentation and an immunohistochemical method to specifically label the 3'-hydroxyl termini of DNA strand breaks (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling), we quantified apoptosis in the subependyma in the young adult rat brain after single and fractionated doses of X-rays. Significant increases in apoptotic index (percentage of cells showing apoptosis) were detected 3 h after irradiation, and the peak apoptotic index was detected at 6 h. Six h after irradiation, the dose response for apoptosis was characterized by a steep increase in apoptotic index between 0.5 and 2.0 Gy and a plateau from 2-30 Gy. The fraction of cells susceptible to apoptosis was estimated to be about 40%, and treatment of rats with cycloheximide inhibited apoptosis. When daily 1.5-Gy fractions of X-rays were administered, the first three fractions were equally effective at decreasing the cell population via apoptosis. There was no additional apoptosis or decrease in cellularity in spite of one to four additional doses of X-rays. Those data suggested some input of cells into the subependymal population during fractionated treatment, and subsequent studies showed that there was a significant rise in 5-bromo-2' deoxyuridine labeling index 2-3 days after irradiation, indicating increased cellular proliferation. The proliferative response after depletion of cells via apoptosis may represent the recruitment of a relatively quiescent stem cell population. It is possible that the radiation response of subependymal stem cells and not the apoptotic-sensitive population per se are critical elements in the response of the brain to radiation injury.

Effect of adjuvant iridium-192 brachytherapy on the survival of patients with malignant gliomas.

The effect of iridium-192 brachytherapy (BRTX) on the survival of patients with malignant gliomas was evaluated in 83 patients with malignant gliomas (42 astrocytoma grade III, 41 glioblastoma multiforme) over a period of 8.5 years. Fifty patients (Group 1) received only standard external beam radiotherapy (EBRT) (mean dose 51.5 +/- 12.4 Gy in 2.0 Gy fractions), and 33 patients (Group 2) received EBRT (mean dose 51.0 +/- 10.8 Gy) combined with BRTX (mean dose 50.2 +/- 13.2 Gy, dose rate of 0.3-0.4 Gy/hr). The median survival periods for patients in Groups 1 and 2 were 12.2 and 23.7 months, respectively (p = 0.0145). The median survival for 17 patients in Group 2 with glioblastoma multiforme was 21.9 months. Using BRTX as an adjuvant to EBRT appeared to confer survival benefits compared to only EBRT (p = 0.0284). Univariate and multivariate analysis identified the variables of histological diagnosis, location, Karnofsky performance status, and BRTX as relevant risk factors for survival time (p < 0.05 for each factor). Among these factors, BRTX was the most important for prolonging survival (p = 0.0015). Adjuvant iridium-192 BRTX and conventional EBRT appears to greatly improve the survival time of patients with malignant gliomas compared to only EBRT and may be the treatment of choice in selected patients with tumors located in deep-seated or eloquent areas.

[Usefulness of neoadjuvant brachytherapy in the treatment of pineoblastoma: a case report].

The treatment strategy and prognosis of pineal cell tumors are still subjects of debate because of their rarity and the mixture of pineoblastoma and pineocytoma as components. Pineoblastoma is believed to be the more malignant tumor and total gross resection of this tumor is very difficult because of its invasive tendency and location. Although the effectiveness of external irradiation and chemotherapy has been reported, the outlook for patients with this tumor is extremely poor. We treated a case of pineoblastoma with a single total resection nine months after interstitial irradiation and chemotherapy. In this case, brachytherapy was successful as neoadjuvant therapy for decreasing the tumor's volume and clarifying its boundary. The patient was a 36-year-old woman who had complained of occipital headache for about a month. On admission, neurological examination revealed bilateral papilloedema but, otherwise, there were no deficits. A magnetic resonance image (MRI) showed a homogeneously enhanced tumor in the pineal region and obstructive hydrocephalus. Two weeks after ventriculo-peritoneal shunting, a stereotactic needle biopsy was performed using a BRW MRI-guided stereotactic apparatus, and three catheters for interstitial brachytherapy were implanted into the tumor through the biopsy tract. Iridium-192 seeds were inserted through the catheters and kept there for 8 days to irradiate 40 Gy at the tumor periphery. Four courses of chemotherapy with carboplatin (400 mg/m2) and VP-16 (400 mg/m2) were administered after brachytherapy. Repeat MRI scans every month showed a gradual regression of tumor volume. However, the tumor did not disappear and no further decrease in tumor volume was observed 8 months after brachytherapy. A suboccipital craniotomy was performed 9 months after brachytherapy and en bloc tumor resection was achieved readily via an infratentorial supracerebellar approach. After receiving another course of chemotherapy and external irradiation at 40 Gy, the patient was discharged without any symptoms and no residual tumor or recurrence was observed for 24 months after brachytherapy and 15 months after surgery. The clinical course of this case suggests that a combination of stereotactic biopsy and brachytherapy with chemotherapy followed by surgery may be a good strategy for the treatment of pineal cell tumors regardless of their histopathological features.

[The use of twist drill craniostomy in stereotactic surgery for brain tumors].

A twist drill craniostomy is a rapid and relatively simple technique for perforating the skull in order to gain access to the epidural and subdural spaces and the brain. Fifty-eight patients underwent 173 twist drill craniostomies in the stereotactic procedures for the diagnosis and the treatment of brain tumors (brain tumor biopsy, implanting radioactive sources and placement of the catheters for navigation surgery) from September, 1992, through to May, 1995, at our institute. The technique was compared with standard burr hole craniostomy used in 42 patients with brain tumor. In the twist drill craniostomy, the scalp was penetrated directly by hand twist drill (2.7 mm in diameter) and scalp bleeding was able to be be controlled by injecting local anesthetic with epinephrine subcutaneously. The safety stop on the drill was set in advance based on the expected thickness of the skull at its penetration point to provide control of penetration depth as it passed through the skull hole and dura. A hole in the skull was made as the drill was advanced until a change in resistance indicated that the inner table of the skull had been penetrated and the dura lacerated. The cannula with stylet was then inserted through the guide tube to assure the penetration of the skull and dura. As the cannula penetrated the dura, a loss of resistance was noted as it proceeded through the dura toward the pial surface, The time needed in this procedure was less than 3 minutes. Associated with this procedure, there were no complications such as bleeding, or infection in 173 twist drill craniostomies in the 58 patients. Our experience with this procedure proved it to be simple, efficient, safe and superior to conventional burr hole craniostomy.

[Image guided stereotactic biopsy for brain tumors: experience of 71 cases].

Despite advances in the neuroimaging of the brain, an accurate diagnosis of intrinsic lesion of the brain requires tissue sampling and histological verification. Seventy-one patients with intraparenchymal lesion of the brain underwent CT or MRI-directed stereotactic biopsy at Okayama University Hospital between June, 1987 and March, 1995. There were 32 men and 39 women whose ages ranged between 7 and 78 years (mean 46.1 years). All patients underwent preoperative cerebral angiography, high resolution contrast enhanced CT and MRI. The lesions were located in the hemisphere in 40 cases, the thalamus or basal ganglia in 14, the midline (corpus callosum or ventricle) in 11, the pineal region in 4, the suprasellar in one and multiple sites in one. A Brown-Roberts-Wells (BRW) CT-directed stereotactic system was used for biopsy under CT guidance. For MRI-directed biopsies a prototype modification of the BRW frame was employed. Target localization was performed using either CT or MRI. Usually one or two targets within the lesion were chosen and target coordinates were calculated using the CT or MRI scan soft ware. Positive diagnosis was obtained in 67 cases and the accuracy of the histological diagnosis was 94.4%. There were 53 gliomas, 4 metastasis, 5 germinomas, 3 malignant lymphomas, one pineoblastoma, one infarction and 4 negative biopsies. Bleeding as a complication due to stereotactic intervention occurred in one patient (1.4%). To patients with potentially inoperable lesions or lesions which might be best treated by chemotherapy or irradiation, modern techniques of neurosurgery now offer the option of precise stereotactic biopsy through small twist-drill burr holes as opposed to open biopsy.(ABSTRACT TRUNCATED AT 250 WORDS)

[Synergistic effect of carboplatin and hyperthermia in rat and human glioma cell lines].

To assess the interaction of carboplatin and hyperthermia in vitro, the thermochemosensitivities of three glioma cell lines, C6 rat glioma cell line, human glioma cell lines T98G and KMG4, were examined by 3-(4,5-dimethylthiazol-2-yl) -2,5 diphenyltetrazolium bromide (MTT) assay. The cell survival of each cell line decreased according to increasing CBDCA concentration and temperature. With a certain CBDCA concentration, the cell survival at following temperature was significantly decreased from that at 37 degrees: 43 degrees C and 44 degrees C for C6 cells (2.5 micrograms/ml); 41 degrees C, 42 degrees C, 43 degrees C and 44 degrees C for C6 cells (128 micrograms/ml); 42 degrees C, 43 degrees C and 44 degrees C for KMG4 cells (8 micrograms/ml) (CBDCA concentration within parentheses). It is generally considered that the highest tolerable temperature of normal brain is 42 degrees C for 60 minutes, while under 43 degrees C, there is a possibility that a sufficient tumoricidal effect might not be obtained. This study revealed enhanced cytotoxicity of CBDCA with hyperthermia at the temperature lower than 42 degrees C and suggests the possibility to gain increased tumoricidal effect without injuring normal brain by hyperthermia at the normal-tissue-tolerant temperature with systemic administration of relatively lower dose of CBDCA.

Enhancement effects of barbital on the teratogenicity of aminopyrine.

Aminopyrine and its compound with barbital have been used in humans as analgesics and antipyretics. A compound, pyrabital (2 molecules of aminopyrine and 1 molecule of barbital) given daily on Days 9, 10 and 11 of gestation produced significant yields of fetal deaths and malformations in ICR/Jcl mice. Most malformations induced were ruptured omphaloceles (eventration of the abdominal viscera), which were associated with malrotation of the intestine, cleft palates, and tail anomalies, finger and toe anomalies. Aminopyrine also induced significant yields of fetal deaths and malformations. However, the incidence of fetal deaths and malformations induced by a dose of pyrabital was significantly higher than that by an equivalent dose of aminopyrine which was contained in pyrabital. When aminopyrine (0.21 mg/g) and barbital (0.09 mg/g) were given in 2 separate injections to pregnant mice, teratogenicity was approximately equal to that by the equivalent dose of pyrabital (0.3 mg/g). Consequently, potent teratogenicity of pyrabital is not caused by the compound, but only by the coexistence of barbital and aminopyrine. Such enhancement effects of barbital may be due to the induction of enzymes responsible for transforming aminopyrine to teratogenic forms, because pretreatment with barbital and phenobarbital similarly enhanced embryotoxicity of aminopyrine.