Estrogen inhibits bone resorption by directly inducing apoptosis of the bone-resorbing osteoclasts.

Estrogen deficiency causes bone loss, which can be prevented by estrogen replacement therapy. Using a recently developed technique for isolation of highly purified mammalian osteoclasts, we showed that 17 beta-estradiol (E2) was able to directly inhibit osteoclastic bone resorption. At concentrations effective for inhibiting bone resorption, E2 also directly induced osteoclast apoptosis in a dose- and time-dependent manner. ICI164,384 and tamoxifen, as pure and partial antagonists, respectively, completely or partially blocked the effect of E2 on both inhibition of osteoclastic bone resorption and induction of osteoclast apoptosis. These data suggest that the protective effects of estrogen against postmenopausal osteoporosis are mediated in part by the direct induction of apoptosis of the bone-resorbing osteoclasts by an estrogen receptor- mediated mechanism.

Role of osteopontin in bone remodeling caused by mechanical stress.

Changes in the number and proportion of osteopontin mRNA (Opn) expressing osteocytes and osteoclasts caused by the mechanical stress applied during experimental tooth movement were examined in the present study. Opn expression was detected in the osteocytes on the pressure side at the early stage, and gradually spread to those on the tension side and also to the osteoblasts and bone-lining cells in the alveolar bone. Only 3.3% of the osteocytes located on the pressure side expressed Opn in the interradicular septum of control rats; in contrast, the value was increased to 87.5% at 48 h after the initiation of tooth movement. These results indicate that these cells responded to mechanical stress loaded on the bone with expression of the osteopontin gene. Following the increased expression of Opn in these cells, a 17-fold greater number of osteoclasts compared with the control and numerous resorption pits were observed on the pressure side of the alveolar bone. Injection of arginine-glycine-aspartic acid-serine peptide but not that of arginine-glycine-glutamic acid-serine peptide strongly inhibited the increase in the number of osteoclasts. Furthermore, an in vitro migration assay demonstrated the chemotactic activity of osteopontin (OPN) on the precursor of osteoclasts. Our study strongly suggests that OPN is an important factor triggering bone remodeling caused by mechanical stress.

Mouse osteoblastic cells (MC3T3-E1) at different stages of differentiation have opposite effects on osteoclastic cell formation.

Using our new culture system for multinucleate cells (MNCs) that have many characteristics of osteoclasts, we examined the effects of factors produced by osteoblastic cells on osteoclastic cell formation. Conditioned medium (CM) from undifferentiated osteoblastic MC3T3-E1 cells during their growth phase inhibited MNC formation in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. Diluted CM (1:81) from differentiated cells obtained after cultivation for more than 20 days stimulated MNC formation, but at lower dilutions inhibited their formation. Dialyzed CM (greater than 2000 mol wt) from the differentiated cells was more stimulatory than undialyzed CM and showed no inhibitory effect on MNC formation. The inhibitory effect was observed with filtered (less than 3000 mol wt) CMs and was specific for osteoblastic cell CM. Prostaglandin E2 (PGE2) was detected in the CM from undifferentiated or differentiated MC3T3-E1 cells at concentrations (317 +/- 66 and 1287 +/- 179 pg/ml, respectively) sufficient to inhibit MNC formation, and this inhibition was partially abolished with CM (at 3-fold dilution) in indomethacin-treated cells (PGE2, less than 20 pg/ml), suggesting PGE2-mediated inhibition of MNC formation and the presence of another factor(s) besides PGE2 that influenced MNC formation. In contrast to day 3 CM plus 1,25-(OH)2D3, day 60 CM plus 1,25-(OH)2D3 induced MNC formation even in the absence of GM-CSF, and this induction was inhibited by an antibody to GM-CSF. Secondary colony formation assays showed the presence of a GM-CSF-like factor in the day 60 CM. These findings indicate that osteoblastic cells are involved in the process of osteoclastic cell formation, with at least two soluble factors produced by osteoblasts, a GM-CSF-like factor, which is stimulatory, and PGE2, which is inhibitory. The effects of CMs also differed depending on the stage of osteoblast differentiation.

Biological activity assessment of 1 alpha,25-dihydroxyvitamin D3-26,23-lactone and its intermediate metabolites in vivo and in vitro.

The biological activity of 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], 23(S)25(R)-1 alpha,25(OH)2D3-26,23-lactone, and three intermediate metabolites of the lactone in vivo and in vitro was comparatively examined. The three intermediate metabolites, 1 alpha,25(R)26(OH)3D3, 1 alpha,23(S)25(R)26(OH)4D3, and 23(S)25(R)-1 alpha,25(OH)2D3-26,23-lactol, stimulated increases, as did 1 alpha,25(OH)2D3, in intestinal calcium transport and serum calcium level in vitamin D-deficient rats fed a low-calcium diet. On the other hand, 23(S)25(R)-1 alpha,25(OH)2D3-26,23-lactone increased the calcium transport but decreased the serum calcium level. 1 alpha,25(OH)2D3,23(S)25(R)-Lactone and the other three metabolites stimulated multinucleate cell formation from hematopoietic blast cells in a manner correlated with their binding affinities for the 1 alpha,25(OH)2D3 receptor. But 23(S)25(R)-lactone did not show any inhibitory effect on the multinucleate cell formation induced by 1 alpha,25(OH)2D3 in contrast to the results obtained from unfractionated marrow cultures. Conditioned medium obtained from 23(S)25(R)-lactone-treated MC3T3-E1 cells inhibited the formation, probably by the action of some inhibitory factors elaborated by the cells treated with the lactone, whereas conditioned medium obtained from 1 alpha,25(OH)2D3 or other metabolite-treated MC3T3-E1 cells stimulated the formation. These findings suggest that 23(S)25(R)-1 alpha,25(OH)2D3-26,23-lactone might inhibit bone resorption through an inhibition of osteoclastic cell formation and that other vitamin D3 metabolites stimulate bone resorption by development of new osteoclastic cells in addition to indirect osteoclast activation.

Inhibitory mechanisms of H(+)-ATPase inhibitor bafilomycin A1 and carbonic anhydrase II inhibitor acetazolamide on experimental bone resorption.

The effects of the vacuolar-type H(+)-ATPase inhibitor bafilomycin A1 (baf.A1) and the carbonic anhydrase II inhibitor acetazolamide (AZ) on bone resorption and procathepsin L secretion of rat osteoclasts were investigated using the bone slice assay method, pit formation test. Baf.A1 completely suppressed osteoclastic bone resorption stimulated by parathyroid hormone (PTH), but did not affect procathepsin L secretion, while AZ suppressed both bone resorption and procathepsin L secretion. These findings suggest that bone resorption by procathepsin L secretion and its processing are regulated by proton production and proton secretion.

Bronchoscopic and angiographic comparison of bronchial arterial lesions in patients with hemoptysis.

In seven patients with nonmalignant disease, we bronchoscopically observed various types of bronchial arterial lesions that may have caused hemorrhage. Five of the seven showed a bulging lesion, and the other two demonstrated an intrabronchial mass. We also examined these seven patients using selective bronchial arteriography. Herein we report our comparative study of the bronchoscopic findings and the bronchial arteriograms of these bronchial arterial lesions. The bulge observed in bronchoscopy corresponded either to an aneurysm or to a hypervascular area in the bronchial arteriogram. The mass lesions corresponded to a hypervascular area or a focal dilatation in the bronchial arteriogram. The intrabronchial lesions observed bronchoscopically either disappeared or were significantly diminished by bronchial arterial embolization for management of the hemorrhage. A histologic examination in two patients who underwent surgery revealed vascular lesions corresponding to the intrabronchial lesions in bronchoscopy. The results of this comparative study have important application in the bronchoscopic examination of bronchial arterial lesions in patients with hemoptysis.

Clinical pharmacology and toxicity of low daily administration of oral etoposide in advanced lung cancer patients.

In this study, pharmacokinetics, clinical effects, and toxicity of daily oral etoposide were studied in 12 patients with advanced lung cancers. Administration schedule was 25 mg/body everyday of a 4-week cycle or 50 mg (25 mg twice)/body for 14 consecutive days of a 4-week cycle. The pharmacokinetic parameters of both groups in serum were as follows: 1) mean peak plasma concentration of 0.91 +/- 0.18 micrograms/mL (25 mg) and 1.24 +/- 0.12 micrograms/mL (25 mg twice); 2) elimination half-lives of 6.48 +/- 1.09 hours (25 mg) and 3.64 +/- 0.27 hours (25 mg twice); and 3) the area under the plasma concentration curve of 7.09 +/- 1.26 micrograms.hr/mL (25 mg) and 8.67 +/- 0.77 micrograms.hr/mL (25 mg twice). There was a significant difference between those two schedules in terms of the plasma concentration and the duration at a low concentration (greater than 1 micrograms/mL) of etoposide. Low daily administration of oral etoposide is shown to be well tolerated and easy on the patient. More studies are needed to study whether prolonged schedules of lower doses of etoposide could result in improved prognosis.

Short latency somatosensory evoked potential in children.

The short latency somatosensory evoked potential was studied in 90 normal children of 1 month to 16 years old and 7 adults. Somatosensory stimuli were delivered through a disc electrode placed over the median nerve at the wrist joint. The uniform recording sites used were the central region of the scalp, and the seventh cervical spine or Erb's point. Reference electrodes were placed on the hand contralateral to the median nerve stimulated. Three positive peaks (P1, P2 and P3) and one negative peak (N1) were consistently recorded, a further positive peak (P4) after N1 was not always observed. The latency of each peak per 1 m body length decreased with age until 2 or 5 years of age. The latency of each peak after 2 years of age was positively correlated with the body length and arm length. The value of P1 peak latency per 1 m body length reaches adult values at an earlier rate than the value of P3 peak latency and P2-P3 latency per 1 m body length. This suggests that central lemmiscal pathways mature at a slower rate than peripheral nerve fibers. The wave form pattern of the short latency somatosensory evoked potential changed to the adult pattern at 10 years of age. The peak latency of P4 during deep sleep was slightly prolonged. In recording on infants during sleep, the EEG should be monitored to determine the stage of sleep.

Postural effects on behavioral states of newborn infants--a sleep polygraphic study.

Physiological and behavioral correlates of neonates in the supine and prone positions were examined. Polygrams were recorded in 10 newborn infants in the prone and supine positions. Newborn infants slept more in the prone position than in the supine, and quiet sleep was significantly more in the prone position. Gross movement, jerky movement and twitch movement were less in the prone than the supine position. There was no difference in localized movement or tremor-like movement in the two positions. Respiration was more regular in the prone than the supine position. Sleep apnea (greater than or equal to 6 seconds) was less in the prone position. The pulse rate during quiet sleep was higher in the prone position.

A case of lissencephaly (agyria) syndrome with apnea--polygraphic study.

A 4-month-old infant with lissencephaly syndrome who was diagnosed on the basis of clinical symptoms and computed tomography experienced four apnea attacks during three polysomnographies. Apnea attacks were of two types. Type I apnea attacks were thought to be sleep apnea. The cause is unknown. Apnea was followed by tonic seizures due to hypoxia. Type II apnea attacks were thought to be epileptic apneic attacks, because they proceeded from the epileptic discharges in the right anterior temporal region (lead of electrooculogram). The origin of Type II apnea attacks was focused in the limbic system.

Increased body movements during sleep in Gilles de la Tourette syndrome.

Overnight sleep polygrams were recorded form 9 patients with Gilles de la Tourette syndrome (GTS). Six of 9 patients had abnormal electroencephalograms, but no specific abnormalities were detected. Body movements and twitch movements during sleep were analyzed. At all stages of sleep, body movements during sleep were more frequent in cases of GTS than those in normal controls. Twitch movements in stage REM of sleep were significantly increased in GTS. These results are consistent with the idea that GTS is due to an imbalance between the central neurotransmitters, catecholamine and serotonin.

Influence of ACTH therapy on overnight sleep polygrams in infantile spasms.

Overnight sleep polygrams were recorded before and during therapy in nine patients with infantile spasms. Results showed that ACTH therapy increased the waking time and decreased rapid eye movement sleep. Thus it caused sleep disturbance in patients with infantile spasms. During ACTH therapy the number of rapid eye movements/min and the pulse rate decreased significantly. Body movements/min also decreased, but not significantly. These results suggest that ACTH therapy may inhibit functions of the central nervous system. The respiratory rate increased during ACTH and clonazepam therapy, probably in association with the decrease or the absence of seizures. These findings indicate the necessity for further studies on whether ACTH therapy is really of value in patients with infantile spasms, and show that if ACTH is given, the period of therapy should be as short as possible.

Denaturing gradient gel electrophoresis analyses of microbial community from field-scale composter.

The diversity of microbial community during the decomposition of waste in a field-scale composter (Hazaka system) was investigated by denaturing gradient gel electrophoresis (DGGE). The composter operates at a high temperature through a self-heating system, creating a thermophilic (60-76 degrees C) stage during the initial phase and a mesophilic (45 degrees C) stage towards the later phase of the composting period. The pH of the system (pH 7.75-8.10) did not vary significantly during the process while moisture content was reduced from 48.8% to 25.1%. DGGE and 16S rDNA analyses showed that the following genera were found throughout the process: Propionibacterium sp., Methylobacterium sp., Pseudomonas sp., and Bradyrhizobium sp. Different Bacillus spp. thrive at the thermophilic or the mesophilic stage while Clostridium sp. was only found at the initial phase of the process. Staphylococcus sp. and Caulobacter sp. or Brevundimonas sp. existed during the later phase of the composting period.

Hospital-onset tuberculosis in compromised host.

A total of 21 patients developed active tuberculosis (TB) during hospitalization. Active TB was identified by bacteria-positive, biopsy or autopsy. Infection was confirmed to the lung, pleura, lymph node and miliary lesions and 7 patients had open tuberculosis. In half of the patients, chest X-ray films demonstrated unusual findings in adult tuberculosis: lower lung field pneumonia and miliary pattern. All the patients suffered from severe underlying diseases and an intensive therapy with steroid, immunosuppressive agents, antitumor drugs, radiation and operation was found as predisposing factors for TB occurrence. Nine patients recovered from current infection with anti-tuberculosis drugs; 14 patients died and TB directly caused death in 8 patients. These data strongly suggest that TB is one of the most important infections in compromised hosts. We emphasize that this infection presents a serious clinical problem in a general hospital today.

Hyperexcitability of cervical motor neurons during neck flexion in patients with Hirayama disease.

We recorded the motor evoked potentials (MEPs) from the affected thenar muscles during neck flexion in 4 patients in the early progressive phase of Hirayama disease. The MEP size significantly increased during neck flexion in patients compared with normal controls. In 2 patients who were treated with a neck collar, the intrinsic muscle atrophy gradually recovered together with increased grip power, and the time course of changes in MEP during neck flexion became normal after treatment with a neck collar for 7-16 months. The technique described here would be useful for evaluating the vulnerability of cervical motor neurons and the therapeutic effect of the neck collar in Hirayama disease.

[A case of Pasteurella multocida infection in bronchiectasis].

Pasteurella multocide (P. multocida), a small gram-negative bacillus, has been known to be the causative agent of hemorrhagic septicemia in animals. P. multocida infection in human was reported as skin abscess and/or septicemia after an animal bit or scratch. Pulmonary infections of P. multocida have been developed in the patients with chronic pulmonary diseases such as bronchiectasis. In Japan, however, P. multocida respiratory tract infections are rare. In this report, a 80-year-old female with bronchiectasis was admitted on August, 1985. She had a productive cough, hemosputum, and a low grade fever. The chest X-P on admission showed an atelectasis of the left middle lobe and severe bronchiectatic changes of the left lower lobe. P. multocida was isolated from her sputa. The chemotherapy of CTM resulted in clinical improvement. On May 1988, she complained of a productive cough and a low grade fever again. P. multocida was isolated from the sputum on several occasions in significant numbers (1 x 10(8)/ml). Recently, the cases of the chronic respiratory diseases have been increasing. We think, P. multocida is important and should be considered as a pathogen in the care of chronic pulmonary diseases.

[A study of neutrophil functions in patients with chronic respiratory tract diseases].

We tested blood neutrophil functions in the patients with chronic respiratory tract diseases to study the mechanism of susceptibility to bacterial infections. Peripheral blood neutrophils were obtained from 15 healthy subjects and 14 patients including diffuse panbronchiolitis, bronchiectasis, chronic emphysema and chronic bronchitis. Seven patients suffered from the chronic P. aeruginosa infection. Firstly, neutrophil chemotaxis was determined by the method of Boyden Chamber assays using FMLP as a neutrophil chemoattractant. The number of migrated neutrophils were 239.2 +/- 65.6 cells/50 HPF in the patients group and 256.6 +/- 49.0 cells/50 HPF in the control group. Secondly, neutrophil phagocytosis against P. aeruginosa, E. coli and K. pneumoniae was determined by phagocytic activity (PA) and phagocytic index (PI). PA against each bacteria was 44.1 +/- 13.2% (P. aeruginosa), 44.8 +/- 12.3% (E. coli) and 35.8 +/- 13.6% (K. pneumoniae) in the patients group and 42.3 +/- 10.6% (P. aeruginosa), 43.0 +/- 11.9% (E. coli) and 36.3 +/- 16.0% (K. pneumoniae) in the control group. PI against each bacteria was 2.2 +/- 0.6 (P. aeruginosa), 2.1 +/- 0.3 (E. coli) and 2.6 +/- 0.9 (K. pneumoniae) in the patients group and 2.2 +/- 0.6 (P. aeruginosa), 2.2 +/- 0.4 (E. coli) and 2.5 +/- 0.6 (K. pneumoniae) in the control group. Thirdly, neutrophil bacteriocidal activity was determined by superoxide production and intracellular killing efficiency. Superoxide produced from OPZ-triggered neutrophils was 17.8 +/- 6.5 nmol/3.5 X 10(6) cells/20 min in the patients group and 20.2 +/- 5.8 nmol/3.5 X 10(6) cells/20 min in the control group, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[A clinical study of pulmonary tuberculosis in lung cancer patient].

We have reviewed 442 patients with lung cancer. There were 323 male patients with a mean age of 65.8 yr and 119 female patients with a mean age of 66.0 yr. Histological types of lung cancer were squamous cell carcinoma 177 patients (40.0%), adenocarcinoma 167 (37.8%), small cell carcinoma 75 (17.0%), large cell carcinoma 17 (3.8%) and undifferentiated carcinoma 6 (1.4%). When lung cancer was diagnosed, 55 patients (12.5%) showed tuberculous lesions on the initial chest X-ray film. The majority of these tuberculous lesions were old changes, but only one with cavitary lesion was confirmed to be active. Although there was no statistical significance, the incidence of squamous cell carcinoma was higher in the 55 patients with tuberculous lesions than in the remaining 387 patients without ones. In the present study, five patients developed active pulmonary tuberculosis while on a therapy of lung cancer. All of these five patients were male and they had advanced lung cancer on admission. The diagnosis of active tuberculosis was made by autopsy in two patients and by culture--positive after death in two. Only one patient was identified to have active tuberculosis by broncho-alveolar lavage. Three of 5 patients showed old tuberculous lesions on the initial chest X-ray film, but all of five patients showed caseoinfiltrative shadows when active tuberculosis occurred. In addition, the prevalence of active pulmonary tuberculosis was significantly higher in the patients with old tuberculous lesions (5.6%: 3 out of 54 patients) than in those without ones (0.52%: 2 out of 387 patients). Finally, all of the five patients died. The causes of death were lung cancer in three patients, both pulmonary tuberculosis and lung cancer in one and pulmonary tuberculosis in one.

[Clinical significance of serum CA125 in patients with tuberculous pleurisy].

We measured CA125 levels of the sera and pleural effusions in both patients with tuberculous pleurisy (TB) and with benign non-tuberculous pleurisy (non-TB). In all the TB patients, serum CA125 levels were increased (78 to 370 U/ml, mean +/- SD = 167.3 +/- 96.8 U/ml, n = 8), and were significantly higher than those in non-TB patients (167.3 +/- 96.8 U/ml v.s. 36.9 +/- 18.4 U/ml, p less than 0.01). Neoplastic diseases or gynecological disorders were not found in these patients. On the other hand, either CA125 or LDH levels of pleural effusions were not significantly different between these two groups. Although adenosine deaminase (ADA) levels in pleural effusions were also significantly higher in the TB patients (p less than 0.05), there were no correlation between serum CA125 and ADA levels in pleural effusions. Serial measurement of serum CA125 levels in the TB patients revealed that serum CA125 levels were markedly decreased one to two months after anti-tuberculous therapy (172.6 +/- 103.3 U/ml to 23.3 +/- 9.9 U/ml, p less than 0.01). It is suggested that the measurement of serum CA125 in patients with tuberculous pleurisy is useful as an indicator of disease activity.

[Rapid and complete hematopoietic reconstitution following peripheral blood stem cell autotransplantation in a child with T cell acute lymphoblastic leukemia].

Rapid and complete hematopoietic reconstitution was achieved in a child with T cell acute lymphoblastic leukemia who was autografted with peripheral blood stem cells (PBSC). A large number of PBSC was collected by two courses of 3-4 hour-lasting lymphopheresis during early remission induced by the second-line chemotherapy and then cryopreserved in liquid nitrogen. A myeloid progenitor cell dose of 203 X 10(4) CFU-GM/kg body weight was reinfused to the patient following marrow-ablative chemotherapy (MCNU 600 mg/m2, cytosine arabinoside 6 g/m2, etoposide 300 mg/m2, cyclophosphamide 160 mg/kg). Neutrophil count reached 0.5 X 10(9)/l by day + 7 and platelet count reached 20 X 10(9)/l by day + 9. Thereafter, white blood cell count continued to increase and reached a maximum of 38 X 10(9)/l on day + 14. Thus, the rapid recovery of hematopoiesis minimised marrow aplasia-related risks. This approach of stem cell rescue operation can be applied to the treatment of children with cancer, who otherwise have no hope to be cured, as an alternative to bone marrow transplantation.