[Lymphangiomyomatosis pathologically diagnosed by surgery for pneumothorax].

We report a case of incipient pulmonary lymphangioleiomyomatosis (LAM) diagnosed by histopathological examination of excised lung. A 28-year-old woman was referred to our hospital because of recurrent left pneumothorax. Computed tomography showed no abnormality except for small bullae in the right middle lobe. She underwent video-assisted thoracoscopic surgery and we excised the apex of the left lung showing hypertrophic pleura. Microscopic examinations of the surgical specimen revealed multiple focal accumulations of small spindle-shaped cells stained positively with anti-HMB-45 antibody, specific for LAM. Furthermore, among these multiple lesions, vascular invasion of HMB-45 positive cells were observed, which demonstrates invasive and metastatic potential of LAM cells as previously reported. This case implicates a need for a careful pathological examination of excised specimens in female cases of surgically treated pneumothorax even though pre-operation or macroscopic examination shows no specific findings.

Highly repetitive DNA families restricted to germ cells in a Japanese hagfish (Eptatretus burgeri): a hierarchical and mosaic structure in eliminated chromosomes.

It is known that in eight hagfishes chromosome elimination occurs during early embryogenesis. The eliminated chromosomes are mostly C-band positive, so that none of the somatic cells have any C-band-positive chromatin. Recently, some highly repetitive DNA sequences have been reported as eliminated elements in these hagfishes based on molecular biological methods. However, no germline-restricted repetitive DNA have been directly isolated from the Japanese hagfish Eptatretus burgeri, from which approximately 21% of the total DNA is eliminated from presumptive somatic cells. Through electrophoretic investigation after digestion with restriction endonucleases, two DNA families that are restricted to germline DNA were isolated. Molecular cloning and sequence analysis revealed that these families are composed of closely related sequences of 64 and 57bp in length, respectively. Southern blot hybridization revealed that the two DNA families are restricted to germline DNA and were thus named EEEb1 and EEEb2, respectively. Moreover, these eliminated elements were highly and tandemly repeated, and it is predicted that they might amplify by saltatory replication and have evolved in a concerted manner. By densitometric scanning, EEEb1 and EEEb2 were found to amount to make up approximately 18.5 and 0.024% of the total germline genomic DNA, accounting for 88.6% of the total eliminated DNA. A fluorescence in situ hybridization experiment demonstrated that EEEb1 is located on all C-band-positive chromosomes that are limited to germ cells, suggesting that EEEb1 is the primary component of eliminated DNA of E. burgeri.

Depolarization-dependent survival of cultured mouse cerebellar granule neurons is strain-restrained.

Cerebellar granule neurons (CGNs) isolated from the rat are often used as a model system for the analysis of activity-dependent survival of neurons. These cells do not survive in culture without addition of a depolarizing agent (KCl or glutamate) to the medium. However, it has been reported that mouse CGN behave differently. Here we found that the requirement for depolarization for the survival of the mouse CGN was strain-dependent, which may be important for future analyses using transgenic animals. CGNs from the Balb/C mouse could survive without KCl addition, whereas CGNs from the C57Bl/6 mouse could not (similar to the rat CGN). The survival-promoting activity of the Balb/C mouse CGNs was transferable by coculturing. However, the medium of the Balb/C mouse CGN culture was ineffective, suggesting that the neurotrophic substance that is supposed to be released is labile. Although we are yet to specify the substance, it might be independent of neurotrophins. It is necessary to select the strain of mouse in the production of transgenic animals for the analyses of activity-dependent neuronal survival.

Increased exocytotic capability of rat cerebellar granule neurons cultured under depolarizing conditions.

To obtain insights into the mechanisms underlying activity-dependent survival of neurons, we surveyed various indices of cellular activity in rat cerebellar granule neurons cultured under conditions advantageous and disadvantageous for survival. Previously, we reported that the turnover of Ca2+ (both influx and efflux) is activated in raised K+-cultures (survival condition), although the cytoplasmic Ca2+ concentration is not affected. We also reported that endocytotic activity was high in the high K+-cultures. In the present study, we used the release of FM1-43 dye [N-(3-triethylammoniumpropyl)-4-(4-dibutylamino)styryl)py ridium bromide] to determine the exocytotic capabilities of neurons cultured in normal K+ (death condition), high K+ (survival condition) and brain-derived neurotrophic factor-supplemented (survival condition) media. The FM1-43 releases triggered by K+-induced depolarization and glutamate exposure were significantly higher in the high K+-cultures than in normal K+-cultures. Interestingly, the neurons whose survival was supported by brain-derived neurotrophic factor did not show high exocytotic capability, indicating that the high exocytotic capability is not a mere result of viability. However, the number of synaptic sites per cell (as monitored by synaptophysin immunopositivity) was unaffected by culture conditions. The present results suggest that an enhanced exocytotic activity supported by a strengthened exocytotic capability may underlie the high viability of rat cerebellar granule neurons cultured under depolarizing conditions.

Influence of passive and active pendular head rotation on horizontal optokinetic nystagmus.

The influence of pendular head rotation on optokinetic nystagmus was examined using a vestibulo-optic stimulator (pendular rotating chair with an optic cylinder) to study passive head rotation, and an optic cylinder which was rotated by a motor fixed to the head to study active head rotation. Pendular head rotation and optic stimuli were simultaneously and independently applied horizontally. The optic cylinder consisted of 12 vertical stripes rotating at a uniform velocity of 30 degrees/s or 90 degrees/s. Passive pendular head rotation was applied at a frequency of 0.1 Hz and a peak angular velocity of 30 degrees/s. Active head rotation was applied for a period of approximately 10 s, and at an amplitude of approximately 50 degrees. Optokinetic nystagmus was enhanced when the head was rotated in the opposite direction to the optic cylinder. However, when the head and the optic cylinder were rotated in the same direction, optokinetic nystagmus was inhibited. There was little difference between the effects of passive and active head rotation on enhancement. However, during active head rotation, optokinetic nystagmus was less inhibited than during passive head rotation.

Measurement during convulsions of guanidino compound levels in cerebrospinal fluid collected with a catheter inserted into the cisterna magna of rabbits.

An easy method for collecting cerebrospinal fluid (CSF) was developed using a catheter inserted into the cisterna magna of rabbits. Levels of guanidino compounds in CSF collected by this method were measured. Levels of guanidinoacetic acid and creatinine increased at the onset of a pentylenetetrazol (PTZ)-induced convulsion. These levels normalized 2 h after the convulsion. The arginine level started to decrease 2 h after the convulsion, continued to decrease gradually and normalized 4 days after the convulsion. These results suggest that guanidinoacetic acid and creatinine are related to the initiation of PTZ-induced convulsions, and that 4 days are required for the metabolism of guanidino compounds to return to normal after a convulsion in rabbits.

Determination of neurotransmitter release into the caudate nucleus during convulsions induced by pentylenetetrazole using in vivo differential pulse voltammetry.

In vivo differential pulse voltammetry using an electrochemically treated carbon fiber electrode was applied to the investigation of the in vivo release of indoleamine and catecholamine within the caudate nucleus of freely moving and immobilized rats during convulsions induced by pentylenetetrazole (PTZ). Two distinct oxidation peaks, on at 130 mV (3,4-dihydroxyphenylacetic acid (DOPAC] and the other at 300 mV (5-hydroxyindoleacetic acid), were observed in voltammograms obtained from the caudate nucleus. In the caudate nucleus of freely moving rats, the in vivo oxidation current that peaked at 300 mV showed almost no change during and after tonic convulsions induced by 60 mg/kg of PTZ i.p. During tonic convulsions, the amplitude of the DOPAC oxidation peak significantly decreased to 75% of the peak height recorded prior to the injection of PTZ, and the minimum lasted for about 30 min; then the mean curve slowly recovered to the control level within 60 min. These results suggest that the release of dopamine (DA) in the caudate nucleus of freely moving rats decreased during tonic convulsions induced by PTZ. In another experiment, the EEGs of immobilized rats were recorded simultaneously, and the changes in the EEG pattern were used as an index of convulsion. In voltammograms from the caudate nucleus of immobilized rat, the peak height of the 130-mV oxidation potential significantly increased during ictal seizures. The increase lasted for 3-6 min after the ictal seizures. The severe electrical activity of the brain during ictal seizures interfered with the recording of some voltammograms.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between sibship size and neurotic symptoms of anxiety, hypochondriasis and depression in Japanese neurotic children and adolescents.

Two hundred and sixty-eight Japanese aged 19 or below were studied retrospectively from their medical records to determine the relationship between the sibship size (number of siblings in the family) and the appearance of symptoms of anxiety, hypochondriasis and depression during the time period I (1955 + 1960 + 1965) and the time period II (1970 + 1975 + 1978). The results showed that only sons and either sex of a large sibship size (4 or more) possessed these three symptoms to a larger extent than only daughters and either sex of a medium sibship size (2 or 3) during both time periods. These three adult-type neurotic symptoms were thought to be manifested among the children and adolescents who communicated primarily with adults and/or those who were required by their parents to exhibit adult-type behavior whether in traditional or in modern Japanese families.

Rubeomycin, a new anthracycline antibiotic complex. I. Taxonomy of producing organism, isolation, characterization and biological activities of rubeomycin A, A1, B and B1.

A new antibiotic complex has been obtained from the cultures of an actinomycete, strain FA-1180, isolated from a soil sample collected at lake side of Biwa in Japan. On the basis of taxonomic studies the producing microorganism is designated as Actinomadura roseoviolacea var. biwakoensis nov. var. The antibiotic complex belongs to the class of anthracycline glycoside antibiotics. All components form deep red fine needles on crystallization; components are named rubeomycin A, A1, B and B1. These components exhibit activity against Gram-positive bacteria as well as Yoshida sarcoma cell in vitro. These components are also effective on P388 leukemia.