Isolation and characterization of dental epithelial cells derived from amelogenesis imperfecta rat.

OBJECTIVE: Disruption of the third zinc finger domain of specificity protein 6 (SP6) presents an enamel-specific defect in a rat model of amelogenesis imperfecta (AMI rats). To understand the molecular basis of amelogenesis imperfecta caused by the Sp6 mutation, we established and characterized AMI-derived rat dental epithelial (ARE) cells. MATERIALS AND METHODS: ARE cell clones were isolated from the mandibular incisors of AMI rats, and amelogenesis-related gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Localization of wild-type SP6 (SP6WT) and mutant-type SP6 (SP6AMI) was analyzed by immunocytochemistry. SP6 transcriptional activity was monitored by rho-associated protein kinase 1 (Rock1) promoter activity with its specific binding to the promoter region in dental (G5 and ARE) and non-dental (COS-7) epithelial cells. RESULTS: Isolated ARE cells were varied in morphology and gene expression. Both SP6WT and SP6AMI were mainly detected in nuclei. The promoter analysis revealed that SP6WT and SP6AMI enhanced Rock1 promoter activity in G5 cells but that enhancement by SP6AMI was weaker, whereas no enhancement was observed in the ARE and COS-7 cells, even though SP6WT and SP6AMI bound to the promoter in all instances. CONCLUSION: ARE cell clones can provide a useful in vitro model to study the mechanism of SP6-mediated amelogenesis imperfecta.

GLCCI1 variant accelerates pulmonary function decline in patients with asthma receiving inhaled corticosteroids.

BACKGROUND: In steroid-naive patients with asthma, several gene variants are associated with a short-term response to inhaled corticosteroid (ICS) treatment; this has mostly been observed in Caucasians. However, not many studies have been conducted for other ethnicities. Here, we aimed to determine the relationship between the annual decline in forced expiratory flow volume in one second (FEV1 ) and the variant of the glucocorticoid-induced transcript 1 gene (GLCCI1) in Japanese patients with asthma receiving long-term ICS treatment, taking into account the effect of high serum periostin levels, a known association factor of pulmonary function decline and a marker of refractory eosinophilic/Th2 inflammation. METHODS: In this study, 224 patients with asthma receiving ICS treatment for at least 4 years were enrolled. The effects of single-nucleotide polymorphisms (SNPs) in GLCCI1, stress-induced phosphoprotein 1 (STIP1), and T gene on the decline in FEV1 of 30 ml/year or greater were determined. RESULTS: Besides the known contributing factors, that is, the most intensive treatment step, ex-smoking, and high serum periostin levels (≥95 ng/ml), the GG genotype of GLCCI1 rs37973, and not other SNPs, was independently associated with a decline in FEV1 of 30 ml/year or greater. When patients were stratified according to their serum periostin levels, the GG genotype of rs37973 was significantly associated with blood eosinophilia (≥250/µl) in the high serum periostin group. CONCLUSIONS: A GLCCI1 variant is a risk factor of pulmonary function decline in Japanese patients with asthma receiving long-term ICS treatment. Thus, GLCCI1 may be associated with response to ICS across ethnicities.

Long-term effects of post-ischaemic oestrogen on brain injury in a rat transient forebrain ischaemia model.

BACKGROUND: The current study was conducted to compare the effects of post-treatment with oestrogen on histological and neurological outcomes after short (7-day) and long (28-day) recovery periods in rats subjected to transient forebrain ischaemia. METHODS: Male Sprague-Dawley rats were randomly assigned to one of five groups: vehicle (7-day recovery period), vehicle (28-day recovery period), oestrogen (17ß-estradiol 200 µg/kg, 7-day), oestrogen (17ß-estradiol 200 µg /kg, 28-day), or sham surgical (n = 8 in each group). After forebrain ischaemia was induced with bilateral carotid artery occlusion and haemorrhagic hypotension (mean arterial pressure = 40 mmHg) for 10 min, the brain was reperfused for 7 or 28 days. Either 17ß-estradiol or vehicle was injected intravenously during the initial 2 min of reperfusion. To evaluate histological damage, the number of intact neurons per 1 mm in the hippocampal CA1 subfield was counted at 7 or 28 days after transient forebrain ischaemia. RESULTS: At 7 days after ischaemia, the number of intact neurons in the hippocampal CA1 subfield was significantly greater in the oestrogen group [57.5 (46.5)/mm: median (interquartile range)] than in the vehicle group [10 (19.5) /mm; P = 0.014]. However, there was no difference between groups at 28 days after ischaemia [vehicle: 11 (20)/mm vs. oestrogen: 6 (11)/mm]. The neurological deficit scores in the oestrogen and vehicle groups were not different from the sham group at any point post-ischaemia. CONCLUSION: The current study indicates that post-ischaemic administration of oestrogen provided short-term but not long-term neuroprotective effects in transient forebrain ischaemia in rats.

Cerebral microaneurysms found incidentally during aneurysm surgery.

Not uncommonly, cerebral microaneurysms are found incidentally during surgery for another previously diagnosed cerebral aneurysm(s). The frequency and angiographic characteristics of such incidental microaneurysms are retrospectively summarized. Seventeen patients were identified as harboring incidental microaneurysms, comprising 4.9% of the whole series. The middle cerebral artery (MCA) was the most frequent location (seven cases, 41%) of these microaneurysms. There was a tendency for MCA microaneurysms to be contiguous to a previously known, larger aneurysm at the same location. Neurosurgeons as well as interventional neuroradiologists should be aware of the possible presence of these incidental microaneurysms while treating patients with a cerebral aneurysm(s). Although the actual clinical implications of these incidental microaneurysms have not been elucidated, the few additional risks to patients already surgically exposed for the treatment of another aneurysm, along with the possible benefit of preventing their rupture and growth, would justify the surgical treatment of these microaneurysms.

Potential fascial dome made by the upper leaf of the phreno-esophageal membrane.

We describe the configuration and size of the artificial fascial dome created in 57 cadavers. This dome protrudes into the thoracic cavity from the esophageal hiatus. This dome was a potential space realized by finger dissection (i.e., a specific but common surgical procedure during surgery of the upper part of the stomach). The vagus nerves penetrated the top of the dome and ran down along the esophagus. The height of the ventral wall of the dome ranged from 10-60 mm, while the dorsal wall was 10-40 mm longer than the ventral one since the dorsal wall attached to the lower, dorsal limb of the esophageal hiatus. Accordingly, the dorsal wall separated the "thoracic" aorta from the "abdominal" esophagus. We considered that the upper leaf of the phreno-esophageal membrane forms the fascial dome, although the lower leaf of the membrane was not identified in this study. According to the results, we proposed a schematic representation of the phreno-esophageal membrane.

[Clinical utility and safety of diagnostic thoracoscopy].

The increasing use of thoracoscopy performed under local anesthesia has made contributions to the diagnosis of pleural disease with effusion. During the past 7 years, we have performed 100 such thoracoscopy procedures using a flexible fiberoptic bronchoscope. On the basis of our clinical findings, we are able to discuss the utility and safety of this procedure. The causes of pleural effusion were carcinomatous pleurisy in 72 cases, tuberculosis pleurisy in 15 cases, infection without tuberculosis in 4 cases, malignant pleural mesothelioma in 8 cases and one case of asbestosis. The success rate of thoracoscopic pleural biopsies were 97% for carcinomatous pleurisy, 100% for malignant pleural mesothelioma and 86% for tuberculosis pleurisy. This procedure was performed with no serious effect on blood pressure, oxygen saturation, monitored ECG or BGA data, and with no serious complications. Therefore, we concluded that this method is very useful for the diagnosis of pleural effusions and has few complications.

[Clinical significance of serum KL-6 in pulmonary sarcoidosis].

We investigated whether the level of serum KL-6 could be an activity marker for pulmonary sarcoidosis. In 33 patients with pulmonary sarcoidosis, the relationships between serum KL-6 levels and diagnostic imaging, serum angiotensin-converting enzyme (ACE) levels, serum lysozyme levels, steroid therapy, and prognosis were evaluated. There were no significant differences in the level of serum KL-6 when the patients were divided on the basis of radiographic findings, but the level of serum KL-6 was markedly elevated in some patients with stage-II pulmonary sarcoidosis. There was a significant correlation between serum KL-6 levels and the following two parameters: serum ACE and lysozyme levels. Among patients with a high initial level of serum KL-6, pulmonary sarcoidosis tended to become exacerbated within one year. Steroid therapy significantly decreased the level of serum KL-6, suggesting that the level of serum KL-6 could be an activity indicator for pulmonary sarcoidosis. Immunohistochemical staining by anti-KL-6 antibody revealed that KL-6 was localized in proliferating type-II alveolar epithelial cells.

[A study on the mechanism of enhanced diuresis following direct hemoperfusion with polymyxin B-immobilized fiber].

In the present study, we applied direct hemoperfusion with polymyxin B-immobilized fiber(PMX-DHP) to patients who developed endotoxin shock after laparotomy, and examined the influence of PMX-DHP on the kidney function. Seven patients were enrolled in this study, whose conditions were matched to the following criteria: 1) endotoxin shock was highly suspected, 2) blood pressure became stable before PMX-DHP was indicated, 3) renal function(demonstrated with creatinine clearance(CCr) and fractional excretion of sodium (FENa)) was proven before the surgery. All patients underwent emergency surgery in Fuji City General Hospital because of perforative peritonitis. A 2-hour session of PMX-DHP was performed on the day of the laparotomy and the second 2-hour treatment was performed the following day. Urine was collected at 2 hours before starting PMX, during the treatment, and 2 hours after PMX-DHP, and urine volume(U-Vol), sodium and creatinine levels of urine were monitored. Sodium and creatinine levels in the serum were measured at the start and end of the PMX-DHP session. Average atrial natriuretic polypeptide (ANP) was obtained using a total of 8 samples from the 14 treatment sessions. Parameters of hemodynamics such as pulmonary capillary wedge pressure(PCWP) were monitored at the start and end of PMX-DHP session. Urine volume increased significantly during and after PMX-DHP. The change in urine volume correlated significantly with the change in CCr during PMX-DHP, and with the change in FENa after PMX-DHP. The change in FENa was significantly correlated with the changes in hemodynamic factors such as PCWP and with the change in serum ANP, but no significant correlation was observed between the change of CCr and the other parameters. In conclusion, the early increase in urine volume with PMX-DHP treatment might be attributable to the increase in glomerular filtration independently of systemic hemodynamic factors.

Three-dimensional reconstruction of transcription termination factor rho: orientation of the N-terminal domain and visualization of an RNA-binding site.

The Escherichia coli rho transcription termination protein is a hexameric helicase, and is believed to function by separating an RNA-DNA hybrid. Unlike hexameric DNA helicases, where a single strand of DNA passes through the central channel, it has been proposed that the RNA wraps around the outside of the ring. We have generated a three-dimensional reconstruction of rho, and localized a tRNA molecule bound to the primary RNA-binding site to the outside of the ring. An atomic structure of the N-terminal domain of rho fits into our reconstruction uniquely, with the residues involved in RNA-binding on the outside of the ring. Although rho shares a common structural core with the F1-ATPase and other hexameric helicases, there has been a divergence in function due to rho's N-terminal domain, which has no homology to other helicases.

Usefulness of serum carboxy-terminal telopeptide of type I collagen (ICTP) as a marker of bone metastasis from lung cancer.

BACKGROUND: Serum pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP) is a metabolite of type I collagen comprising 90% or more of organic substances in bone. Its usefulness as a marker of bone metastasis from malignant tumors is expected. METHOD: We measured ICTP to evaluate its clinical usefulness for diagnosis of bone metastasis in 140 patients with lung cancer. For comparison, serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA 21-1), gastrin-releasing peptide precursor (ProGRP), alkaline phosphatase and calcium were simultaneously measured. ICTP was measured by double-antibody radioimmunoassay. RESULTS: ICTP was significantly higher in patients with bone metastasis from lung cancer than in the group without bone metastasis, patients with other pulmonary diseases or healthy control subjects and showed excellent sensitivity and specificity, indicating that this marker is highly useful for complementary diagnosis of bone metastasis from lung cancer. Moreover, the survival duration was significantly shorter in the ICTP-positive group than in the ICTP-negative group, suggesting that ICTP can be a prognostic factor in lung cancer. CONCLUSION: It is suggested that measurement of ICTP is worthwhile as a serological diagnostic method of bone metastasis from lung cancer. Moreover, since repeated measurements are possible, this measure was considered very helpful in complementary diagnosis of bone metastasis and also as a standard to determine the timing of examinations such as bone scintigraphy.

Synthesis and biological evaluation of novel 9-functional heterocyclic coupled 7-deoxy-9-dihydropaclitaxel analogue.

Novel 9-functional heterocyclic coupled 7-deoxy-9-dihydropaclitaxel analogues 17 and 22-24 synthesized from a natural taxoid 5-cinnamoyltriacetyltaxicin-I (3) and their biological evaluation in tubulin assembly activity and cytotoxicity in vitro against several human tumor cell lines are first presented. The biologically tested results show that 17, 22 and 23 are inactive in tubulin assembly assay and have no more remarkable cytotoxicities against human tumor cell lines SK-OV3, WIDR and MCF-7, though 22 and 23 exhibit more potent cytotoxicity against human liver cancer and human esophagus cancer cell lines (BEL-7402 and ECa-109) than paclitaxel.

Incidence of acquired renal cysts in biopsy specimens.

AIMS: This study investigated whether or not acquired renal cysts develop in patients with mild chronic renal failure. METHODS: A retrospective study was carried out using renal biopsy specimens from 720 patients. A renal cyst was defined as a tubule dilated >200 microm. RESULTS: Renal cysts were found in 21 of 720 renal biopsy specimens. Serum creatinine of 21 patients with renal cysts was significantly higher than that of 699 patients without cysts (2.59 +/- 2.64 vs. 1.09 +/- 0.79 mg/dl) (p < 0.0001). Poor renal function (serum creatinine >1.6 mg/dl) reveals more cyst formation on biopsy specimens than good renal function (serum creatinine <1.5 mg/dl). Cysts were observed in 11 of 607 (1.8%) patients less than 50 years of age and in 10 of 113 (8.8%) patients over 51 years. To exclude simple cysts which are commonly observed in older subjects, 11 patients under 50 years of age were extensively examined. Mean serum creatinine was 2.98 +/- 3.06 mg/dl (0.7-10.4 mg/dl). These 11 patients revealed low creatinine clearance of 47.5 +/- 25.6 ml/min (5-71 ml/min). Creatinine clearances in 7 patients were 52-71 ml/min (serum creatinine 0.7-2.0 mg/ dl). One of 11 biopsy specimens with cysts was examined by immunohistochemistry on lectin. This specimen was positive for tetragonolobus lectin and negative for peanut lectin, suggesting that the epithelial cells lining the cyst were derived from proximal tubules, unlike those of simple cysts. CONCLUSION: These results suggest that low normal renal function such as creatinine clearances 52-71 ml/min due to nephron loss is sufficient to induce acquired cyst development in various renal diseases.

Postischemic hypothermia attenuates apoptotic cell death in transient focal ischemia in rats.

Hypothermia confers potent neuroprotection against ischemic injury. Attenuation of apoptosis by hypothermia can be one of the responsible mechanisms. In this study, in situ DNA nick-end labeling (TUNEL) and immunostaining of Bax protein were performed to evaluate the effect of postischemic hypothermia on apoptotic cell death, employing rodent transient focal ischemia. Animals received 1 hour of transient focal ischemia. Brain temperature was maintained at 37.5 +/- 0.5 degrees C during ischemia. Immediately after reperfusion, animals were assigned to either a normothermic or hypothermic group. In hypothermia, animals were cooled and brain temperature was lowered to 34.5 +/- 1.0 degrees C. Prolonged hypothermia was maintained for 16 hours and animals rewarmed. In both groups, TUNEL and immunostaining of Bax was performed. In normothermia, the number of TUNEL positive cells reached the peak at 2 days after ischemia and decreased gradually. In hypothermia, the peak was shifted to 3 days after ischemia. The number of TUNEL positive cells in hypothermia was persistently below that of normothermia. Similarly, in hypothermia, immunostaining of Bax showed attenuated immunoreactivity compared with that in normothermia. In conclusion, postischemic hypothermia reduced both the number of TUNEL positive cells and immunoreactivity of Bax, which may be one of the responsible mechanisms with which hypothermia exerts neuroprotection.

Cognitive function and language of a child with an arachnoid cyst in the left frontal fossa.

The authors inspected the cognitive function of a boy with a congenital arachnoid cyst in the left frontal fossa. Neuropsychological tests described his cognition, including language, memory, and frontal functions, as mildly retarded overall. The boy's verbal ability was delayed, but his frontal function was intact. We supposed that the cyst would not affect his cognitive function. SPECT images showed low perfusion in the left temporal lobe. Dysfunction of the left temporal lobe should cause delayed language leading to learning difficulty. Neuropsychological evaluation of cognitive function concurrent with rCBF imaging was necessary not only for describing the influence of the cyst but also in discriminating its psychological aspects.

A case of traumatic shock complicated by methamphetamine intoxication.

A case of a 38-year-old male with traumatic shock complicated by methamphetamine intoxication is presented. The patient was involved in an assault which resulted in cardiac tamponade and right ventricular outflow laceration. Pericardiocentesis was immediately performed. However, profound metabolic acidosis greatly in excess of that expected from the short duration of the shock was revealed by arterial blood gas analysis. Another cause of the metabolic acidosis was suspected. The patient subsequently admitted to intravenous use of methamphetamine. Following hemodynamic and metabolic stabilization by continuous pericardial drainage and intravenous administration of sodium bicarbonate, the patient underwent cardiac surgery. His postoperative course was uneventful. There is a substantial association between methamphetamine users and traumatic accidents. In such cases, early identification of drug use is important. Marked metabolic acidosis, which conflicts with the diagnosed cause of shock, may be a clinical clue to methamphetamine intoxication.

[Operation for left renal cell carcinoma complicated with acute pulmonary tumor embolism and cocurrent removal of tumor].

The prognosis of pulmonary tumor embolism is said to be poor and only a limited number of patients with this disease have survived. The patient was a 64-year-old male suffering from left renal cell carcinoma complicated with tumor extending from the left renal vein to the inferior vena cava. The patient underwent an operation for left renal cell carcinoma during which he developed tumor embolus to the pulmonary artery. The occurrence of the acute embolism was promptly detected and the removal of tumor was performed under cardiopulmonary bypass. The patient made good postoperative progress.

[A case of the pulmonary thrombosis caused by unilateral pulmonary artery occulusion test].

A 71-year-old male, diagnosed as lung cancer, underwent unilateral pulmonary occulusion test. Through the guidewire, 7.5 Fr thermodilution catheter with occlusion balloon was introduced to the left pulmonary artery from the right internal jugular vein. Heparinized physiological saline solution was injected into the distal site of the occulusion. The occulusion time was 15 minutes. Pulmonary artery pressure and wedge pressure were within normal range. Soon after the examination, the pulmonary arteriogram (PAG) showed the defect of the branch to the lingular segment and the lower lobe. We made a diagnosis of pulmonary thrombosis. Three days after the administration of urokinase and heparin, both pulmonary perfusion scintigram and PAG exhibited the reperfusion to these areas. After the thrombolytic therapy was accomplished, antithrombin III and protein C in the serum showed within normal range. It was possible that the damage on the intima due to the thermodilution catheter or the guidewire and the following blood congestion by the pulmonary artery occulusion caused the thrombosis.

Plasma cell granuloma extending from the extracranial to the intracranial space associated with Epstein-Barr virus infection.

A 70-year-old male presented with a plasma cell granuloma extending from the extracranial to the intracranial space. Findings of preoperative magnetic resonance imaging and intraoperative observation indicated that the lesion extended from the temporal muscle to the subarachnoid space, penetrating the frontal bone. The subarachnoid lesion was composed of neutrophils indicating the presence of acute or subacute inflammation. The final diagnosis of the resected tumor was plasma cell granuloma. High levels of antibodies against Epstein-Barr (EB) virus in the cerebrospinal fluid and the immunohistochemical demonstration of EB nuclear antigens in the plasma cell granuloma suggested that EB virus infection was associated with the development of plasma cell granuloma in this patient.

[Bronchiolitis obliterans preceding rheumatoid arthritis: effect of clarithromycin].

A 62-year-old man was referred to our department because of exertional dyspnea and a 6-year history of coughing and sputum production. He had never smoked, and had had an operation for chronic paranasal sinusitis. Coarse crackles and rhonchi were audible over both lower lung fields. The cold hemagglutinin titers were high. pulmonary function tests showed airflow obstruction, and a sputum culture revealed Hemophilus influenzae A chest X-ray film and a CT scan showed diffuse micronodular shadows in the centrilobular regions, mild ectasis of bronchioles mainly in the lower lung fields, and mild hyperinflation. A specimen of lung tissue was obtained by thoracoscopic biopsy, and histologic examination showed bronchiolitis obliterans, with bronchiolar narrowing or obliteration due to submucosal fibrosis and inflammation. Rheumatoid arthritis was diagnosed 14 months after the operation. The patient was treated with clarithromycin for 3 years. Respiratory symptoms were relieved and pulmonary function gradually improved.