Identification of regulatory sequences and binding proteins in the type II sodium/phosphate cotransporter NPT2 gene responsive to dietary phosphate.

Dietary phosphate (P(i)) is a most important regulator for renal P(i) reabsorption. The type II sodium-dependent phosphate (Na/P(i)) cotransporters (NPT2) are located at the apical membranes of renal proximal tubular cells and major functional transporters associated with renal P(i) reabsorption. The consumption of a low-P(i) diet induces the synthesis of NPT2, whereas a high P(i) diet decreases it. The molecular mechanisms of regulation by dietary P(i) are not yet known. In this report, in weaning mice fed a low-P(i) diet for 4 days, the NPT2 mRNA level was increased 1.8-fold compared with mice fed a normal P(i) diet. This increase was due to an elevation of the transcriptional activity. In the NPT2 gene promoter, the DNA footprint analysis showed that six regions were masked by the binding protein, but at the position -1010 to -985 upstream of the transcription start site, the binding clearly responded to the levels of dietary P(i). The phosphate response element (PRE) of the NPT2 gene was found to consist of the motif related to the E box, 5'-CACGTG-3'. A yeast one-hybrid system was used to clone a transcription factor that binds to the PRE sequences in the proximal promoter of the NPT2 gene. Two cDNA clones that encoded protein of the mouse transcription factor muE3 (TFE3) were isolated. This is a DNA-binding protein that activates transcription through the muE3 site of the immunoglobulin heavy chain enhancer. TFE3 antibody completely inhibited the binding to the PRE. The coexpression of TFE3 in COS-7 cells transfected with the NPT2 gene promoter markedly stimulated the transcriptional activity. The feeding of a low P(i) diet significantly increased the amount of TFE3 mRNA in the kidney. These findings suggest that TFE3 may participate in the transcriptional regulation of the NPT2 gene by dietary P(i).

Oligodendroglioma of the "widespread" type in the spinal cord.

The authors present clinical, radiological, and pathological features of the "widespread" type of primary spinal oligodendroglioma arising in a 12-year-old boy. The initial symptoms were gait disturbance, hypesthesia of the lower trunk, and scoliosis. Gadolinium-enhanced magnetic resonance imaging revealed an intramedullary tumor with syringomyelia. The tumor spread rapidly in a "widespread" pattern within the spinal cord between C7 and T12. One year after surgery the patient had made a poor neurological recovery despite the two-stage tumor resection. Only five cases of widespread tumors, including our case, have been described with discussion of their clinical manifestations.

An angiomyoma with t(X;10)(q22;q23.2).

A t(X;10)(q22;q23.2) translocation was detected as the only chromosomal aberration in primary short-term cultured cells from an angiomyoma of a 58-year-old woman; 6p, 13q, and 21q rearrangements have been previously demonstrated by others in two cases of angiomyoma. This type of translocation has not been reported in other benign tumors including leiomyomas and angiomyomas, although it has been detected in an ependymoma. This is thought to be a third case of angiomyoma showing karyotypic abnormalities.

Periosteal osteosarcoma of the femur with bone marrow involvement: a case report.

Periosteal osteosarcoma is an exceedingly rare type of chondroblastic osteosarcoma, showing rather better prognosis, and secondary bone marrow involvement is unusual. A case of a 22 year old male with periosteal osteosarcoma of the right femur with an associated bone marrow lesion is presented. The juxtacortical tumor, 16 x 11 x 9 cm, was located on the bone cortex of the upper diaphysis and extended into the surrounding soft tissues. A minimal bone marrow lesion was present, although the bone cortex was quite intact. Microscopically, the tumor consisted exclusively of atypical chondroblastic cells with a small osteoblastic area. The bone marrow lesion, interestingly, contained both multiple nodules of well-differentiated chondrosarcomatous components and a few demarcated foci of atypical spindle cells producing a fine osteoid matrix. It was reasonable to conclude, therefore, that this tumor was a periosteal osteosarcoma with an unusual secondary bone marrow lesion rather than a conventional (central) chondroblastic osteosarcoma with soft tissue invasion. The patient's good prognosis with no tumor recurrence or metastasis during more than 7 years follow-up after surgery supports this conclusion.

Effect of reperfusion by direct percutaneous transluminal coronary angioplasty on ventricular late potentials in cases of total coronary occlusion at initial coronary arteriography.

The relationship between the time required for reperfusion and the incidence of late potentials was studied in 94 patients who survived a first acute myocardial infarction (AMI) and who showed total occlusion of the infarct-related artery at an initial coronary arteriography. Sixty-three patients who successfully underwent direct percutaneous transluminal coronary angioplasty (PTCA), and 31 who were treated conventionally (controls), underwent signal-averaged electrocardiography. Direct PTCA patients were classified into 5 groups according to the time required for reperfusion: < or = 4, 4-6, 6-8, 8-10, and > or = 10 h. The incidence of late potentials in these groups was 8%, 12%, 14%, 33%, and 43%, respectively, and 48% in the controls. Late potentials were recorded more frequently as the period until successful reperfusion increased: the incidence of late potentials was significantly lower in the < or = 4 and 4-6 h groups than in the controls (p < 0.005 and p < 0.05, respectively). Therefore, reperfusion achieved within 6 h reduced the incidence of late potentials in AMI patients and may be effective for preventing malignant ventricular arrhythmias.

Establishment and characterization of a new human clear-cell sarcoma cell-line, HS-MM.

We have established and morphologically characterized a new human clear-cell sarcoma cell-line, HS-MM, from the pleural effusion of a 39-year-old man with pulmonary metastasis derived from the primary popliteal tumour. The HS-MM cells were round or spindle-shaped, with round nuclei containing extremely prominent nucleoli. Light microscopically, the heterotransplanted nude mouse tumours showed essentially the same features as those of the original sarcoma, revealing an alveolar pattern of tumour cells with abundant clear cytoplasm. Both in vitro and in vivo, the cells reacted with anti-S-100 protein and melanoma-specific HMB 45 antibodies by immunohistochemistry. Ultrastructurally, the cells contained round euchromatin-rich nuclei with large nucleoli revealing conspicuous nucleolonema, and a large amount of glycogen and a few lysosomal dense bodies, but no premelanosomes in their cytoplasm. The HS-MM cell line was thus fully proven to exhibit the unique characteristics of a clear sarcoma both in vitro and in vivo, being also compatible with an amelanotic melanoma. This cell line will therefore be extremely useful for clinicopathological and histogenetic studies on clear cell sarcomas.

Establishment and characterization of a new human synovial sarcoma cell line, HS-SY-II.

BACKGROUND: The line of differentiation in synovial sarcoma still remains controversial. Thus far, only a few human synovial sarcoma cell lines have been described. However, their morphologic characteristics have not been fully established. EXPERIMENTAL DESIGN: We established a new synovial sarcoma cell line (HS-SY-II) from pleural effusion with lung metastasis in a typical example of the monophasic spindle cell type. The HS-SY-II cells, in vitro and in vivo, were examined by light microscopy, immunohistochemistry, electron microscopy, and cytogenetics. RESULTS: The HS-SY-II cells showed a hypertriploid karyotype with complex chromosome abnormalities including pathognomonic t(X;18)(p11;q11), and have been stably maintained for more than 40 months in vitro, showing rather small spindle or polygonal shape without conspicuous pleomorphism. Histologic features of initially and serially transplanted tumors in nude mice were essentially the same as those of the original sarcoma, corresponding to the monophasic spindle cell variant with a prominent palisading pattern and calcified foci in parts. The HS-SY-II cells in vitro and in vivo similarly expressed vimentin and cytokeratin by immunohistochemistry, and also exhibit the same ultrastructural features such as irregularly shaped nuclei with prominent nucleoli, many paranuclearly running intermediate filaments, and filopodia-like processes. CONCLUSIONS: This HS-SY-II cell line retaining the distinct morphological characteristics as the monophasic spindle cell type of synovial sarcoma therefore will be extremely useful for various pathomorphologic investigations on synovial sarcoma.

[The influence of atrioventricular asynchronous contraction on left and right ventricular performance].

To investigate the influence of atrioventricular asynchronous contraction on left and right ventricular performance, pulsed Doppler echocardiographic studies were performed in 10 patients who received permanent pacemaker (VVI mode), but without significant heart disease except for complete heart block. After setting the pacing rate at 40 per min, the performance was analyzed during the patient's own slow ventricular rate. Flow velocity patterns at the left (LVOT) and right ventricular outflow tracts (RVOT) were recorded by pulsed Doppler echocardiography, and ejection time (EjT), acceleration time (AcT), peak velocity (PV) and flow velocity integral (FVI), which is proportional to stroke volume, were measured for each outflow tract. When the patient's own atrial contraction occurred during ventricular systole, EjT, AcT, PV and FVI of flow at the LVOT and EjT, AcT and FVI of flow at the RVOT were decreased. Percent change of the FVI of flow at the RVOT (-34.6%) was significantly greater than that of flow at the LVOT (-16.2%, p < 0.01). These results indicate that the loss of right ventricular performance might play a prominent role in the genesis of the hemodynamic deterioration with atrioventricular asynchronous contraction.

[Ossifying fibroma in the cranial vault. Case report].

Ossifying fibroma is relatively common in the maxilla, but rare in the cranial vault. A 10-year-old girl was referred for painful swelling of the left temporal region. On admission, she presented no abnormal physical and neurological findings except for the painful swelling. Plain skull X-ray films showed a radiolucent lesion of the left temporal bone about 4 cm in diameter, with a hyperostotic area of the parietal side. Computed tomography scan using bone window level also showed an abnormal density lesion in the same site. Curettage of this tumor was performed from a cosmetic point of view and at the family's petition. Histological examination showed vascular fibrous tissue in which lamellar bone was surrounded by osteoblasts.

Morphological characterization of a newly established human osteosarcoma cell line, HS-Os-1, revealing its distinct osteoblastic nature.

A newly established human osteosarcoma cell line, HS-Os-1, from an osteoblastic tumor arising in the left humerus of an 11-year-old girl was morphologically characterized in vitro and in vivo. HS-Os-1 cells in a monolayer have been maintained for more than 2 years since the initial cultivation, and were round or polygonal in shape with marked pleomorphism. Their cytoplasm was strongly positive for specific markers of osteoblasts, such as alkaline phosphatase and osteocalcin. Tumors induced in nude mice by HS-Os-1 cell inoculation at passage 12 or 23 revealed typical histological features of osteoblastic osteosarcoma, similar to those observed in the original tumor, producing prominent osteoid matrix with calcification. Ultrastructurally, HS-Os-1 cells in vitro and tumor cells in vivo showed similar well-developed, markedly dilated rough endoplasmic reticulum, polysomes and microfilaments in their cytoplasm. Additionally, many collagen fibers associated with deposition of electron-dense material were detected in the stroma featuring osteoid matrix. Thus, the HS-Os-1 cell line was shown to exhibit its osteoblastic nature in vitro and in vivo, and therefore might become an extremely useful tool for various pathomorphological investigations on human osteosarcomas.

[Establishment and characterization of a human cell line, HS-MM, derived from a case of clear cell sarcoma].

The characteristics of a new human clear cell sarcoma (CCSa) cell line, HS-MM, established from the pleural effusion in a 39-year-old man with lung metastasis, have been morphologically studied in vitro and in vivo. HS-MM cells growing on a cover-slip were round or spindle in shape with round nuclei containing extremely prominent nucleoli. Heterotransplantation of the cells into nude mice was easily succeeded following tumor development. Light microscopically, HS-MM cells, both in vitro and in vivo, were positive for anti-S-100 protein and anti-melanoma specific antibodies with immunostain, but no melanin pigment was detected in them. Ultrastructurally, the cells had round euchromatin-rich nuclei with large nucleoli revealing conspicuous nucleolonema, and contained a few mitochondria, rough endoplasmic reticulum and lysosomal dense bodies, besides a large amount of glycogen, but no melanosome in their cytoplasm. HS-MM cells retained and fully expressed morphologically unique characteristics as a CCSa, compatible with amelanotic type of malignant melanoma also. This cell line, HS-MM, therefore, proves to be extremely useful for clinicopathological studies on a CCSa.

[Establishment and characterization of a cell line, HS-Os-1 derived from an osteoblastic type of human osteosarcoma].

A new human cell line, HS-Os-1, derived from a case of osteoblastic osteosarcoma arising in the humerus of an 11-year-old girl was established. Light microscopically, HS-Os-1 cells growing in a monolayer (in vitro) were pleomorphic, intermingled with a few multinucleated giant ones, and positive with alkaline phosphatase reaction. In the transplanted tumors in athymic nude mice (in vivo), atypical spindle or polygonal cells densely proliferated with prominent osteoid formation and even calcification. HS-Os-1 cells, both in vitro and in vivo, were mostly positive for vimentin and a few for S-100 protein. Ultrastructurally, HS-Os-1 cells in vitro and in vivo also revealed essentially the same features as the eccentrically located, euchromatin-rich nuclei with prominent nucleoli, a lot of well-developed, irregularly-dilated rough endoplasmic reticula, polysomes and microfilaments in the cytoplasm. Namely, HS-Os-1 cells fully expressed and possessed morphological characteristics as osteoblastic nature during the cultivation and heterotransplantation. This cell line, therefore, proved to be extremely useful to search for human osteosarcomas.

[Changes in muscarinic cholinergic receptor and choline acetyltransferase in experimental ischemic brain].

Acetylcholine (ACh) is the neurotransmitter related to learning and memory. The activity of its metabolic enzyme - choline acetyltransferase (CAT) is found to be remarkably decreased at autopsy. We consider that there is some relationship between ACh and cerebrovascular dementia such as dementia in ischemia and forgetfulness after ischemia. So we studied the relationship between ischemia and cholinergic neuron. We examined changes of muscarinic cholinergic receptor (m-ChR) in experimental ischemia by binding assay and autoradiography, and performed immunohistochemical study of CAT that is, acetylcholine synthetase, by PAP method. We also studied delayed neuronal death from the aspect of cholinergic system because the hippocampus receives ACh pathway arising from the basal nucleus. Materials used include: Pulsinelli's 4 vessel occluded rats, Tamura's MCA occluded rats and forebrain ischemia mongolian gerbils. In ischemia, the number of m-ChR decreased and binding affinity of m-ChR increased, and recirculation caused increased the number of m-ChR. While m-ChR changed immediately after ischemia, it was not until the fourth day that CAT positive cells decreased in hippocampus. In other words, at first m-ChR in postsynaptic membrane changed in ischemia, and with the progress of neuronal damage, CAT also changed. After m-ChR decreased in the thalamus, stria terminalis and Meynert nucleus 1 day following ischemia, it decreased in hippocampus after 7 days. We can consider receptor changed corresponding to the pathway of cholinergic neuron. Our study suggested that the receptor change in cholinergic system plays some role in the delayed neuronal death in the hippocampus.

[A case of eclampsia with intraventricular hemorrhage].

Our case is a 31-year-old primipara who had undergone a cesarean section for multiple pregnancy and was referred to our clinic because of attack of eclampsia. On admission, her consciousness level was lethargic. Computed tomography on admission showed high density area indicating right paraventricular hemorrhage with ventricular perforation, and low density areas in bilateral basal ganglia. Examination of the cerebral angiogram done six days after onset of symptoms revealed multisegmental vasospasm in Willis' circle mainly. An emergent ventricular drainage was performed. Two weeks later, repeat angiogram revealed that spasm had almost disappeared. Repeat CT scan three months after the onset showed small low density area in the right caudate nucleus and disappearance of the low density areas in bilateral basal ganglia. Her clinical course was uneventful and she was discharged without neurological deficit at about three weeks after the onset. In spite of absence of subarachnoid clot on CT scan, remarkable multisegmental spasm was found on cerebral angiogram. We suspected that eclampsia may have been responsible for the spasm in this case. Bleeding was believed to have originated in the superolateral angle of the lateral ventricle. It was supposed that infarcted hemorrhage or bleeding from small artery due to changes in arterial or venous pressure might have occurred and penetrated into the lateral ventricle.

Further observations of the digital plethysmography in response to auditory stimuli and its clinical applications.

Further observations of the digital plethysmography with auditory stimuli and its clinical applications were performed in patients with vibration disease and heart diseases. The responsive pattern to the auditory stimuli in the digital plethysmogram could be faithfully reproducible if it elapses more than 5 minutes apart between the first stimulus and the second one. The responsive patterns were divided into four types: normal (N), hyperreactive (I and D) types and hyporeactive (P) type. The values of urinary catecholamine increased in parallel to the activity level of the autonomic nerve. The hyperreactive type (D) had the highest value of urinary catecholamine, and vice versa. With regard to the age, the elderly was prone to show hyporeactive (P) type, and the young tended to be hyperreactive (D) type. Psychologic factors were examined by Cornell medical index. Neurosis was not necessarily related to the hyperreactive type. In clinical applications, it was observed by this method that the autonomic imbalance in patients with vibration disease, angina pectoris, or hypertension was recovered by the treatment, and the abnormal types of the response recovered to N type. In conclusion, the digital plethysmography with auditory stimuli as one of the autonomic nerve function tests would be much useful and non-invasive method for observation of clinical course and decision of the severity.