Abnormal neutrophil scattergram obtained using Pentra MS CRP in the patients with myelodysplastic syndrome showing dysgranulopoiesis.

INTRODUCTION: Pentra MS CRP is an automated hematology analyzer capable of cytochemistry using Chlorazol black E, a lipid-staining agent, for white blood cell (WBC) differentials. Pentra MS CRP displays a WBC scattergram according to the cell volume obtained using flow impedance and light absorbance reflecting the Chlorazol black E (CBE)-positive lipid content. METHOD: Neutrophil scattergrams obtained using Pentra MS CRP were compared between 5 patients with myelodysplastic syndrome (MDS) and normal controls. Sudan black B (SBB)-staining patterns of peripheral blood neutrophils were subdivided into four types (types I, II, III, and VI) based on their staining intensity and scored by counting 200 cells. Such SBB scores were also compared between the two groups. RESULTS: Neutrophil scattergrams deviated downward in the MDS group, suggesting the decreased CBE positivity that seemed reflect the reduction of the lipid content in dysplastic neutrophils. SBB scores determined in this study were also decreased in the MDS group when compared with those in normal controls. CONCLUSION: Pentra MS CRP might rapidly generate useful information on dysplastic neutrophils in patients with MDS based on its cytochemistry for WBC differentials during routine laboratory hematology.

Chicken Eyes absent 2 gene: isolation and expression pattern during development.

In all vertebrates studied (human, mouse, chicken), there are at least three genes related to Drosophila eyes absent (eya) gene. The chicken Eyes absent 2 (Eya2) cDNA was isolated from 14 day embryonic chicken lenses, and a complete open reading frame encoding a 59 kDa protein was elucidated. The chicken Eya2 protein is moderately conserved and 78-82% identical to the mouse and human Eya2. The Eya2 gene demonstrated a dynamic expression pattern in different tissues of diverse embryological origin. Expression of Eya2 was first detected at Hamburger and Hamilton stage 9 in the foregut. At later stages of development, Eya2 mRNA was detected in neural crest derivatives (dorsal root ganglia, branchial arches and cranial nerve ganglia). In the cranial placodes, expression of Eya2 was first detected in the nasal pit at stage 13. In the eye, expression of Eya2 was first convincingly detected in neural retina at stage 24 (day 4). The highest level of Eya2 mRNA in the lens was detected around day 9. Eya2 is also expressed in the cornea and iris. Therefore, chicken Eya2, as well as mouse Eya2, is expressed relatively early in the nasal (but not in the lens) placode and may mediate induction of the nasal placode. Expression of Eya2 in the wing and limb buds is consistent with its proposed role in the patterning of limb connective tissues.

Plasma-free and sulfoconjugated MHPG in major depressive disorders: differences between responders to treatment and nonresponders.

The plasma levels of free and sulfoconjugated forms of 3-methoxy-4-hydroxyphenylglycol (MHPG) were examined before and after treatment in 16 patients with unipolar major depressive disorders without melancholia. The patients were treated with intravenous administration of clomipramine for 4 weeks. Seven depressive disorder patients who showed marked improvement (the improvement group) revealed significant reduction in their plasma sulfoconjugated MHPG levels. In 6 depressive disorder patients who showed no improvement (the no-improvement group), the plasma sulfoconjugated MHPG levels showed no significant change after treatment. The remaining 3 patients, who showed ambiguous change after treatment, were excluded from the analysis. Levels of plasma-free MHPG showed significant change after treatment in neither the improvement group nor in the no-improvement group. It is suggested that levels of plasma sulfoconjugated MHPG may serve as an indicator of brain noradrenergic activity.

Immunohistochemical demonstration of DNA polymerase alpha in human brain-tumor cells.

The proliferative capacity of brain-tumor cells was analyzed in vitro and in situ using monoclonal antibody (MAb) against deoxyribonucleic acid (DNA) polymerase alpha. For the in vitro studies, two cultured human glioma cell lines were investigated using MAb against DNA polymerase alpha, the MAb Ki-67, a serum against proliferating cell nuclear antigen (PCNA/cyclin), bromodeoxyuridine (BUdR), and an anti-BUdR MAb. During exponential growth of the cells, the percentage of polymerase alpha-positive cells (the "polymerase alpha score") ranged from 72.0% to 77.1%, the Ki-67-positive cells (the "Ki-67 score") ranged from 43.4% to 59.4%, the PCNA/cyclin-positive cells from 30.9% to 41.4%, and the BUdR labeling index from 28.6% to 39.3%. For the in situ studies, tissue from 60 human brain tumors and from two normal human brains was investigated and the polymerase alpha scores and Ki-67 scores were compared. In normal brain tissue, no immunostaining was found by either method. In brain tumors, both the polymerase alpha scores and the Ki-67 scores correlated with the histological grade of malignancy. Polymerase alpha scores were generally higher than Ki-67 scores in the same specimen, especially in malignant brain tumors. These findings suggest that immunostaining of DNA polymerase alpha is a convenient and important new method by which to estimate the cellular proliferation rate of brain tumors. Polymerase alpha scores may be closer to the growth fraction of the individual tumor than the MAb Ki-67 or other scores.

Sex differences in appetitive learning of mice.

Learning abilities of male and female ddY mice were compared in two appetitive tasks (a lever-press task and an 8-arm radial maze task), and two avoidance tasks (a shuttle box task and a light-dark discrimination T-maze task). In the two types of appetitive learning, male mice were significantly superior to female mice. Sex differences were particularly apparent in the acquisition process. In contrast, there was no significant sex difference in learning of the two avoidance tasks. A sex difference in appetitive learning was not found in juvenile mice prior to sexual maturation, and the mice which had established a lever-press response as juveniles did not show any significant difference in the performance level when tested as adults. Thus, a sex difference appeared only in the acquisition stage of adult mice. These results suggest that there exists a sex difference in motivation level for hunting food but not for feeding, and that it causes a sex difference in appetitive learning.

Insertion of stabilizing loci in vectors of T7 RNA polymerase-mediated Escherichia coli expression systems: a case study on the plasmids involving foreign phospholipase D gene.

Plasmids carrying stabilizing loci were used in the expression of phospholipase D (PLD) gene fused with pelB signal sequence by a recombinant strain of Escherichia coli BL21 (DE3) using T7 RNA polymerase mediated expression system. By checking the living cell number and the percentage of the plasmid-bearing cells, it was found that the plasmids involving PLD gene were not stable under noninduced conditions and that, after the induction, the number of plasmid-bearing cells were rapidly decreased to almost zero. Then, a biologically stabilizing locus such as par B, ccd, or par was inserted into the plasmids. The newly constructed plasmids were maintained very stably in the recombinant cells until the cells were induced. However, after the induction, almost all the recombinant cells were rapidly killed due to highly toxic PLD. Using the best one of the stabilized PLD-expressing plasmids, PLD production was improved 2-fold.

Impairment of acquisition of learning by inhibitors of RNA or protein synthesis.

The effects of inhibitors of RNA or protein synthesis (cycloheximide, puromycin and actinomycin D) on acquisition of learning in mice were investigated using various types of shock avoidance tasks. Mice having uniform and superior learning ability were used as subjects. Drugs were injected into the lateral ventricle 1 hr before training for avoidance tasks in three different shapes of maze and in a one-way shuttle box and for those based on light-dark discrimination. In all cases of training, the drugs were found to impair the acquisition of learning. The impairment was pronounced in a relatively complicated task such as H-maze learning as compared to a simple task such as T-maze learning. The drugs never affected the performance of pretrained animals. During the discussion, the acquisition of learning would be related to some specific process of RNA and protein synthesis in the brain.

Long-term observation of a case of pulmonary arteriovenous fistula with pulmonary hypertension.

A case of pulmonary arteriovenous fistula (PAVF) with pulmonary hypertension (PH) occurring in an adult woman is described. Resection of PAVF was not performed and she has been followed up for 5 years, receiving repeated right cardiac catheterization. We discuss the causal relationship of PH and the development of PAVF.

Preretinal membrane formation in rabbit eyes after intravitreal transplantation of cultured astrocytes.

Astrocytes were cultured from the cerebral cortices of newborn rabbits and were so confirmed by staining of the glial fibrillary acidic protein (GFAP). In Experiment I, the astrocytes of newborn rabbits of both sexes from one mother were pooled and suspended in a culture medium, and approximately 1-2 X 10(4) cells were injected into the vitreous cavity of one eye and the culture medium alone was injected into the fellow eye of the mother rabbit. In Experiment II, the astrocytes cultured from the cerebral cortex of a single newborn rabbit of known sex were injected into both eyes of the mother or father rabbit. In the 24 eyes receiving the astrocyte injection, 22 eyes developed preretinal membrane in 2.5 to 14 weeks: 11 eyes developed traction retinal detachment. In the 12 control eyes receiving the culture medium injection, preretinal membrane was seen only in one eye. The preretinal membrane consisted predominantly of spindle-shaped cells positively stained by the peroxidase-antiperoxidase method for GFAP staining. Electron microscopy showed that the major component cells of the preretinal membrane were astrocytes with characteristic intermediate filaments; no retinal pigment epithelial cells were observed. In Experiment II, the average frequency of sex chromatin detection was 4.6% and 55.2% in the male and female astrocyte cultures, respectively. Three groups of experiments were made, A) the female astrocytes injected into the mother's eyes, B) the male astrocytes injected into the mother's eyes and C) the female astrocytes injected into the father's eyes. The sex chromatin was found in the cells of the preretinal membrane at the average frequencies of 63.5%, 34.4% and 55.5% in Groups A, B and C, respectively. It was concluded that the astrocytes injected into the vitreous cavity proliferated and formed the preretinal membrane to a degree that caused traction retinal detachment, without involvement of the retinal pigment epithelial cells. It was also thought that the astrocytes from the retina of the adult host eyes participated to some extent in the preretinal membrane formation.

Creating notifications tailored for individual workers by using a database: a conceptual framework and its application.

Although prevention strategies to assure good mental health in the workplace have become important, worker notification in studies using psychometric tests has rarely been discussed. Until now it has been difficult to prepare a large number of notifications that are tailored well for individual workers. We have developed a system to create notifications that explain the results for individual workers in plain language using a relational database (RDB). First, scores for each test were divided into categories and the workers' data was classified. Then, explanations were written for each category. RDB software read component files into a database file, integrated all data, and printed out notifications. After the system was developed, we realized its high potential for use in occupational health care, as well as in many other fields. We also report an example of its application.

The Japanese adaptation of the STAI Form Y in Japanese working adults--the presence or absence of anxiety.

Symptom endorsements and response patterns of 1,862 Japanese adult workers (1,509 males and 353 females) to the Japanese adaptation of the State-Trait Anxiety Inventory Form Y (STAI-JY) items, were examined in this study. The mean STAI-JY State and Trait anxiety scores of Japanese workers were substantially higher than those of American workers reported in the Manual, due primarily to the much higher scores of Japanese workers in responding to the anxiety-absent items. The correlations between the State and Trait anxiety-present scales and those of their anxiety-absent scales' counterparts were higher than those between the State anxiety-present and -absent scales and those of their Trait scales' counterparts. These findings suggested that responses to anxiety-present and -absent items should be considered independently in scoring the STAI-JY scales in Japanese working adults.

Thermal damage threshold of brain tissue--histological study of heated normal monkey brains.

The thermal damage threshold of normal brain tissue was evaluated from immediate and delayed histological changes caused by hyperthermia treatment of normal monkey (Macaca fuscata) brains. A 2450 MHz microwave antenna and an antenna cooling system devised by our group were used for interstitial hyperthermia treatment. The antenna within the cooling system was inserted through a small craniectomy under general anesthesia. The temperature at a reference point, 4 mm radially away from the surface of the cooling system, was maintained at 42, 43, 44, 45, or 46 degrees C for 60 minutes. Eighteen animals were treated and sacrificed immediately after the treatment, while nine animals were treated and sacrificed 7 days after the treatment. The histological changes were studied microscopically on sections stained with HE or Kluver-Barrera's method. The non-survival experiment demonstrated that areas heated at 44 degrees C or below showed no obvious irreversible changes. The survival experiment showed areas heated at 44 degrees C or above developed coagulative necrosis. These histological findings indicate that thermal damage occurs in normal brain tissue after heating at 44 degrees C or above for 60 minutes, suggesting that the safety limit for brain hyperthermia is 43 degrees C for 60 minutes.

Preliminary results of interstitial 192Ir brachytherapy for malignant gliomas.

Twenty-six patients with recurrent or unremovable malignant gliomas were treated by interstitial brachytherapy with iridium-192 seeds. Stereotactic implantation of the afterloading catheters using the Brown-Roberts-Wells computed tomography (CT)-guided stereotactic system was performed in 24 patients and surgical implantation in two patients with pontine glioma. The response to therapy was measured by serial CT, magnetic resonance imaging, and clinical examination. Tumor regression was seen in 17 patients 1-3 months after implantation. Tumor progression was seen in only three patients. After interstitial brachytherapy, the most commonly observed CT finding was central low density. Median survival time was 18 months after implantation. Autopsies in five patients revealed the delayed effects of radiation injury such as typical vascular changes, microcalcification, and coagulative necrosis in the implant area and tumor recurrence at the periphery. The results suggest that brachytherapy is not curative but prolonged the median survival time by 6 months.

[Immunohistochemical study for choroid plexus papillomas and ependymomas].

An immunohistochemical study of glial fibrillary acidic protein (GFAP), S-100 protein, cytokeratin (CKER), epithelial membrane antigen (EMA), and transthyretin (TTR) was carried out on 11 cases of choroid plexus papilloma (CPP) and 19 of ependymoma, using the peroxidase antiperoxidase technique. Among the 11 cases of CPP, all 11 were positive for CKER, EMA, and TTR, 10 for S-100 protein, and five for GFAP. Most of the GFAP-positive papilloma cells were simultaneously positive for CKER. However, these GFAP-positive cells were negative for TTR. Among the 19 cases of ependymoma, 16 were positive for GFAP, 17 for S-100 protein, three for CKER, eight for EMA, but none for TTR. Some GFAP-positive cells were also stained for CKER. However, TTR was not found in any of the ependymal cells. These findings suggested that CPP cells which show ependymal or glial differentiation lost the ability to synthesize TTR which is known to be synthesized in the epithelial cells of the choroid plexus. The more GFAP-positive cells present in a CPP, fewer TTR-positive cells are present. Though CPPs are usually easily distinguishable from ependymomas, occasional doubt arises concerning the differential diagnosis between CPP and papillary ependymoma. TTR can be a very useful diagnostic marker of CPP.

[A study on the usefulness of stress assessment based on the THP health measurement--comparison with the general health questionnaire].

Although the "Total Health Promotion Plan (THP)" drawn up by the Labor Ministry includes a self-administered stress checklist (THP-SC) as a health measurement, THP-SC has not been utilized in the way that it had been planned. Hence, we studied the usefulness of this list by comparing it with the General Health Questionnaire (GHQ). We made a questionnaire that included both THP-SC and the 60-item GHQ, and collected answers from 261 white-collar workers (male, average age 43.7). The GHQ was evaluated using the Goldberg's method. When analyzing THP-SC, the number of answers indicating stress was counted, and the respondents were divided into two groups based on the cut-off points of 75 percentile or 90 percentile. In the 60-item GHQ, 48 respondents out of 243 with valid answers had a higher score than the cut-off point, and in the 12-item GHQ, 77 out of 256 had a higher score. The average number of THP-SC was 5.67 +/- 3.19. Concerning the relationship with GHQ, many items of the 'A' section of THP-SC were significant. The comparison also suggested that four or more items checked in 'A' section or seven or more items checked in the whole list would mean that a respondent may be under stress.

Reliability of the State-Trait Anxiety Inventory, Form Y in Japanese samples.

The internal consistency of the State-Trait Anxiety Inventory, Form Y was examined using data collected from Japanese participants by five diverse surveys, in which one included American university students. Cronbach coefficient alpha was calculated separately for state and trait items as well as for anxiety-present and absent items. The internal consistency was higher for the anxiety-absent items than those of the state and trait anxiety items, but this tendency was not clear for the anxiety-present items. The trait anxiety items showed the lowest internal consistency for all Japanese groups, whereas the anxiety-present items showed the lowest alpha for American university students. It can be considered that this difference might induce the difference in two--factor structure between Japanese and people in Western countries.

Positive and negative affect in the factor structure of the State-Trait Anxiety Inventory for Japanese workers.

The factor structure of Form Y of the State-Trait Anxiety Inventory (STAI-Y) was examined with 1,862 Japanese adult workers (1,509 men, 353 women). The initial principal component analysis extracted three factors based on the scree test. All 20 state (S-Anxiety) and 20 trait (T-Anxiety) items had dominant salient loadings on the first factor, which reflected "over-all anxiety." The three factors identified by an oblique (promax) rotation were labeled "anxiety-absent," "state anxiety-present," and "trait anxiety-present." All 20 items with dominant salient loadings on the first oblique factor were clearly grouped on the basis of their content, indicating the absence of anxiety. The second and third oblique factors, defined almost entirely by the STAI-Y anxiety-present items, clearly reflected the state-trait distinction in this sample of Japanese workers.

[Interstitial brachytherapy for malignant gliomas using the Brown-Roberts-Wells (BRW) stereotactic system].

Eleven patients harboring recurrent or deep-seated malignant gliomas were treated by interstitial brachytherapy with 192Ir seed assembly, between June 1987 and September 1989. Implantations for the afterloaded catheter were performed using the Brown-Roberts-Wells (BRW) CT guided stereotactic system. The number of seeds and the distribution of the implants were chosen in such a way that the minimum tumor dose of 30-50Gy could be delivered to the surface or 1cm beyond the rim of the contrast enhancement. The radioactive sources were held in the afterloaded catheters that were removed after the desired dose had been delivered. Response to therapy was measured by serial CT scans and clinical examination. Tumor regressions were seen by CT scans made 2 or 3 months after implantation. One tumor showed complete regression (CR), four showed partial response (PR), one showed minor response (MR) and 5 showed no change (NC). Overall response rate was 54%. Six patients died 3 to 18 months following implantation, and five are still alive 7 to 27 months after implantation. No complications such as infection or hemorrhage were observed during the treatment. A patient harboring large (6.5cm in diameter) recurrent glioblastoma in the rt. parietal robe required a craniotomy due to the mass growing for one and half month after implantation, and radiation necrosis of the entire tumor mass was documented. The technique of stereotactic interstitial implantation was clinically well tolerated and easily reproducible and our preliminary results seemed encouraging. Technical improvement to achieve an adequate isodose distribution to cover the tumor volume might lead to improved survival rates.

[Study on the extent of peritumoral edema in glioma: its correlation with the proliferative potential and tumor-infiltrating mononuclear cells].

The correlation between the extent of peritumoral edema and the proliferative potential or the infiltration of mononuclear cells was studied in 17 gliomas. The peritumoral edema was evaluated on contrast enhanced CT scan as the ratio of the low density area around the tumor to the enhanced high density area. The proliferative potential and the infiltration of mononuclear cells into the tumor were investigated immunohistochemically using monoclonal antibody (MAb) against DNA polymerase alpha and anti-Leu MAb's respectively. There was a significant correlation between the extent of the peritumoral edema and the percentage of DNA polymerase alpha positive cells. The degree of the infiltration of mononuclear cells into the tumor tissue also correlated with the extent of peritumoral edema. In gliomas with high proliferative potential and/or severe infiltration of mononuclear cells, the peritumoral edema may be aggravated by disruption of the blood-brain-barrier and increased vascular permeability.

[Basal ganglia germinoma treated with interstitial brachytherapy; case report].

A case of primary intracranial germinoma in the left basal ganglia treated with interstitial brachytherapy was reported. A 15-year-old boy was referred to our hospital for evaluation of right hemiparesis. A CT scan showed a slightly hyperdense mass with multiple cystic low density in the left basal ganglia. The mass was heterogeneously enhanced after intravenous administration of contrast material. T1 weighted image showed a slightly hyperintense mass with cystic components and the mass was heterogeneously enhanced with Gd-DTPA. T2 weighted MR image showed a mixed intensity mass and peritumoral edema. Stereotactic needle biopsy and implantation of 3 catheters for interstitial brachytherapy were performed simultaneously using BRW CT guided stereotactic apparatus. After the histological diagnosis was confirmed to be two cell pattern germinoma, 9 iridium-192 seeds were inserted into the catheters and maintained for 10 days to give 35Gy of irradiation at the tumor periphery. Subsequent CT scans showed marked tumor regression and the clinical symptoms were improved. Germinoma originating in the basal ganglia is rare and hard to diagnose previous to biopsy. Histological confirmation is essential before initiation of treatment because germinoma is commonly thought to be radiosensitive tumor. The interstitial brachytherapy enables selective irradiation of the tumor and actually causes no complications such as bone marrow suppression or cerebral atrophy. The neuroradiological findings, especially of CT scan and MRI, were presented and the strategy for treatment of germinoma in basal ganglia was discussed.