Neutrophils are thought to play a key role in tissue injury. We investigated the role of human neutrophils in the induction of injury to the human pulmonary artery endothelial cells and the effect of erythromycin on neutrophil-induced endothelial cell damage. Incubation of unstimulated neutrophils with endothelial cells increased the release of lactate dehydrogenase (LDH) activity and preloaded fura-2 from endothelial cells. When neutrophils were activated by phorbol myristate acetate, the release of LDH and fura-2 was enhanced further. Superoxide dismutase partially inhibited the release of LDH and fura-2 induced by neutrophils, whereas erythromycin markedly inhibited the release of endothelial cell LDH and fura-2 induced by neutrophils. These results suggest that endothelial cell injury is, at least in part, mediated by the action of superoxide and that erythromycin protects against neutrophil-induced endothelial cell injury.

Anti-Bacterial Agents↗

PubMed1997

KL-6, a human MUC1 mucin, is chemotactic for human fibroblasts.

KL-6 in serum and bronchoalveolar lavage fluid has been reported to be a sensitive marker indicating the activity of fibrosing lung diseases. The molecule is clustered in MUC1 mucin according to the findings of immunohistochemical and cytometric studies. To elucidate the pathogenic role of KL-6 in fibrosing lung disease, we characterized its biochemical properties and examined whether purified KL-6 is chemotactic for human fibroblasts in vitro using modified Boyden chambers. Biochemical properties of purified KL-6 were similar to those of other MUC1 mucins previously reported. KL-6 promoted the migration of 5 of 5 human lung fibroblasts and 3 of 4 human skin fibroblasts. Checkerboard analysis revealed that KL-6 was chemotactic as well as chemokinetic. Though platelet-derived growth factor, fibroblast growth factor, or fibronectin were also chemotactic for fibroblasts in the experimental system, only fibronectin augmented KL-6-induced chemotaxis. These observations indicate that KL-6 is one of the chemotactic factors for most fibroblasts and that the increased KL-6 in the epithelial lining fluid in small airways may cause the intra-alveolar fibrosis in fibrosing lung diseases.

Antigens↗

PubMed1997

Unfused C2C12 mouse skeletal muscle cells express neurofilament 140K protein.

Expression of neurofilament 140K protein in C2C12 mouse skeletal muscle cells was studied. Immunofluorescence and immunoblot analyses revealed that NF140K was expressed at the proliferative stage and was colocalized with the muscle-specific intermediate protein desmin. As muscle cell differentiation proceeded, the number of NF140K-positive cells decreased whereas the number of cells expressing muscle-specific marker proteins such as sarcomeric myosin heavy chain and troponin-T increased. Down-regulation of NF140K and upregulation of a myogenic regulatory gene, the myogenin gene, started simultaneously. In differentiated muscle cell cultures, unfused cells residing between myotubes remained NF140K-positive. NF140K and desmin double-positive cells were also found in a primary culture of adult mouse skeletal muscle cells. The results suggest that NF140K may be a unique marker for uncommitted myoblasts.

Animals↗

PubMed1997

Sex difference in the metabolism of pirmenol in rats.

The in vitro metabolism of pirmenol (cis-alpha-[3-(2,6-dimethyl-1-piperidinyl)propyl]-alpha-phenyl-2-pyri dinemethanol) and glucuronidation of its metabolites, a 4-hydroxylated derivative of pirmenol (M3) and 3-methylether of M3 (M5), were investigated using a hepatic 9000 x g supernatant and microsomes, respectively, of female and male rats in order to elucidate the higher urinary excretion of M3G and M5G (glucuronides of M3 and M5, respectively) in females previously observed in in vivo metabolism. Pirmenol delta1' iminium ion (M2) and M3 were formed by the oxidation of pirmenol in both sexes; however, M2 was the main metabolite in males, while M2 and M3 were formed at nearly the same level in females. On glucuronidation of M3 and M5, the Vmax values of both compounds were higher in female rats, consistent with the results in vivo. In addition, the sex difference in the urinary excretion ratio of M5G to M3G (1.1 in female, 2.5 in male) might reflect the lower availability of M3 for glucuronidation in male rats in vivo. The chromatographic separation of diastereomers of M5G was also described.

Animals↗

PubMed1997

[SMON and pharmacokinetics of chinoform with special reference to animal species difference].

The experimental reproduction of SMON using several kinds of animals given a prolonged administration of chinoform has been carried out by many investigators because of the importance to solve the problem of etiology in the SMON. In these experiments, it is demonstrated that a marked species difference was observed in the relationship between the doses given to animals and the frequency of the onset of neurologic symptoms. Although dogs were accepted as the most suitable animal model for SMON, pathological changes in the peripheral nerve of the dog were not observed. The blood level or tissue distribution of chinoform after oral, intravenous or intraperitoneal administration of the drug differed in the animal species. Thus, it is considered that the species difference in the onset of neurologic symptoms is principally caused by the difference in pharmacokinetics of chinoform in each animal. Moreover, for the onset of neurologic symptoms in animals, perhaps it is necessary to maintain the level of unconjugated chinoform in the nerve tissues around several to over ten micrograms/ml for three or four weeks as well as that in SMON patients while the neurologic symptoms or pathological changes do not appear in some kinds of animals at these levels. In a study on the cellular toxicity of chinoform, many other problems remain to be solved although degeneration or uncoupling on oxidative phosphorylation in mitochondria of the axons by chinoform and lipid peroxidation of the membrane by chinoform-ferric chelate have been already shown.

Amebicides↗

PubMed1997

Interaction of orally administered 5-[3-[((2S)-1,4-benzodioxan-2-ylmethyl)amino]propoxy]-1,3-benzodioxole (MKC-242) with 5-HT1A receptors in rat brain.

The present study was carried out to clarify whether orally administered 5-{3-[((2S)-1,4-benzodioxan-2-ylmethyl)amino]propoxy}-1,3-be nzodioxole (MKC-242), a serotonin1A (5-HT1A)-receptor agonist having potent anxiolytic-like and antidepressant-like effects in animal models, binds to 5-HT1A receptors in rat brain. Quantitative autoradiography showed that orally administered MKC-242 (0.1-0.5 mg/kg) caused a significant decrease in 3[H]8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) binding in the hippocampus and dorsal raphe nucleus sections. The decrease in the binding by MKC-242 was observed up to 4 hr after administration, and the effective doses were similar to those in its anxiolytic-like effect in the animal models. Repeated treatment of MKC-242 (0.5 mg/kg/day, p.o.) or buspirone (30 mg/kg/day, p.o.) for 2 weeks did not affect [3H]8-OH-DPAT binding in both sections. These results suggest that orally administered MKC-242 at the low doses that do not show 5-HT1A-receptor-mediated in vivo responses such as the hypothermic effect, adrenocortical effect and the decrease in 5-HT turnover passes the blood-brain barrier and subsequently binds to 5-HT1A receptors in rat brain. In addition, they indicate that repeated stimulation of the receptors by the agonists does not affect the number of the binding sites.

5,7-Dihydroxytryptamine↗

PubMed1997

Modification of the response of a quiescent cell population within a murine solid tumour to boron neutron capture irradiation: studies with nicotinamide and hyperthermia.

C3H/He mice bearing SCC VII tumours received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps, to label all proliferating (P) cells. 20 min after intraperitoneal injection of sodium borocaptate-10B (BSH), or 3 h after oral administration of dl-p-boronophenylalanine-10B (BPA), the tumours were irradiated with thermal neutrons. To modify the uptake dose of 10B, nicotinamide (NA) was intraperitoneally injected 60 min before the administration of 10B-compounds and/or the tumours were heated to 41.5 degrees C for 20 min immediately before irradiation. After irradiation, the tumours were excised, minced and trypsinized. The tumour cell suspensions were then incubated with cytochalasin-B (a cytokinesis-blocker). The micronucleus (MN) frequency in cells not BrdU-labelled (quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. With or without the administration of 10B-compounds, the sensitivity of Q cells was lower than that of total (P + Q) tumour cells. With thermal neutron irradiation in the presence of either BPA or BSH, the MN frequency in each cell population was increased. A greater increase in the MN frequency of total tumour cells was observed after thermal neutron irradiation in the presence of BPA than in the presence of BSH. The distribution of 10B from BPA into tumour cells was thought to be more dependent on the uptake ability of the tumour cells than that from BSH. Sufficient quantity of 10B from these two 10B-compounds to cause a highly lethal event inside the cancer cell with thermal neutron irradiation could not be delivered to Q cells. When NA and/or heat treatment were combined with 10B-compound administration, NA increased MN frequency in the BSH treated total cells, and heat treatment elevated MN frequency in Q cells. From the viewpoint of cell kill effect, the combined treatment with nicotinamide and heat treatment was more useful than treatment with either nicotinamide or heat treatment alone, not only in the total tumour cells but also in the Q cells.

Animals↗

PubMed1997

Tumor neutrophil elastase is closely associated with the direct extension of non-small cell lung cancer into the aorta.

OBJECTIVE: Neutrophil elastase (NE) is the only neutral protease that is able to degrade insoluble elastin and other extracellular matrix constituents, and thus, may be involved in tumor invasion and metastasis. Using a highly specific and sensitive enzyme immunoassay (EIA), we recently demonstrated that immunoreactive (ir)-NE is produced by non-small cell lung cancer cell lines. We have measured the ir-NE concentration in non-small cell lung cancer tumor extracts and have evaluated its association with disease stage. METHODS: We measured the ir-NE concentration in 144 non-small cell lung cancer tumor extracts using EIA, which permits the rapid measurement of both the free and alpha1-protease inhibitor (alpha1-PI) complexed form of ir-NE. In 15 clinical T4 (cT4) patients, we also determined the concentration of free ir-NE in tumor extracts using a kit that detects only NE complexed with alpha1-PI and subtracting that value from the total NE concentration. RESULTS: ir-NE was detected in tumor extracts from 115 of 144 patients, ranging from 0.21 to 23.35 microg/100 mg protein. When the 144 specimens were grouped according to the clinical stage of disease, the ir-NE concentration (mean+/-SE) was significantly higher in those with cT4 disease (n=15; 7.90+/-1.88 microg/100 mg protein) than in those with cT1 (n=29; 1.27+/-0.27; p<0.001), cT2 (n=64; 1.18+/-0.17; p<0.001), or cT3 disease (n=26; 1.99+/-0.38; p<0.003). There was no significant association between the ir-NE concentration and cN-factor or any other clinical features. When the ir-NE concentration in the tumor extracts of the cT4 patients was compared with respect to the tumor invasion sites, the ir-NE level was significantly higher in those with surgical T4 (sT4) disease with aortic invasion (n=4; 17.4+/-3.10) than in those who were down-staged postoperatively (n=5; 4.9+/-1.33; p=0.005) or those with sT4 disease with involvement of other sites (n=6; 4.07+/-1.83; p=0.004). Similar results were observed for the free form of ir-NE. CONCLUSIONS: These data suggest that NE may be involved in tumor progression of non-small cell lung cancer. Since the aorta is one of the richest sources of polymeric and insoluble elastin, this enzyme may play an active role in the direct extension of the tumor into the aorta.

Adult↗

PubMed1997

Expression of prolactin gene in mouse placenta during late pregnancy: detection of mRNA and its translation product.

To examine the existence of PRL messenger ribonucleic acid (mRNA) in the mouse placenta during late pregnancy, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot analysis were carried out followed by nucleotide sequence analysis of cDNA. Total RNA extracted from each tissue was reverse-transcribed, followed by PCR with two oligonucleotide primers specific or a part of mouse PRL (mPRL) cDNA. An amplified RT-PCR product of predicted size was detected in all samples from the placenta of days 16 and 18 pregnant mice. This product was specifically hybridized with a probe overlapping an entire sequence of mPRL cDNA in Southern blot analysis. Nucleotide sequence analysis also provided evidence that the amplified cDNA had a nucleotide sequence completely identical to the mPRL cDNA sequence reported previously. Furthermore, mPRL with a slightly bigger molecular weight than that of pituitary PRL was detected in the placenta of days 12, 14, 16 and 18 pregnancy by immunoblot analysis. These results suggest that PRL mRNA and its translation product are synthesized in mouse placenta during late pregnancy.

Animals↗

PubMed1997

Starvation-induced increase in the parathyroid hormone/PTH-related protein receptor mRNA of bone and kidney in sham-operated and thyroparathyroidectomized rats.

Parathyroid hormone (PTH) acts on bone and kidneys by binding to PTH/PTH-related protein (PTHrP) receptors and regulating calcium (Ca) and phosphorus (P) homeostasis. PTH/PTHrP receptor mRNA was expressed at high levels in PTH target tissues such as the kidneys and bone including the calvaria, femur, and tibia. Because short-term starvation influences Ca and P ion homeostasis, we measured changes in PTH/PTHrP receptor mRNA expression in the bone and kidneys. Food deprivation for 3 days decreased the serum Ca and P concentrations, and reinstitution of feeding for 2 days normalized the serum Ca level and significantly increased the serum P level. Concomitantly, rat immunoreactive PTH (riPTH) was increased during starvation and returned to the control level after 2 days of subsequent feeding. Serum 1 alpha, 25-dihydroxyvitamin D3 (1,25(OH)2D3) concentrations did not significantly change during starvation and subsequent feeding. Starvation up-regulated PTH/PTHrP receptor mRNA expression in both bone and kidney. The effects of food deprivation on the receptor transcript abundance were greater in bone (threefold increase compared with control) than in the kidney (1.8-fold increase), whereas the mRNA level increase by food deprivation was more rapid in the kidneys than in bone. The PTH-induced adenylyl cyclase activity of renal membranes increased in starvation. Feeding after starvation normalized the mRNA levels in both tissues. Serum PTH depression, initiated by thyroparathyroidectomy, did not affect PTH/PTHrP receptor mRNA levels in bone and kidney in rats that were fed or starved for 3 days. The abundance of receptor mRNA in bone and kidney was significantly lower in fed rats given either corticosterone or vehicle than in starved rats. These data indicate that starvation induces PTH/PTHrP receptor mRNA expression in bone and kidney, independently of serum PTH and corticosterone concentrations. The factors leading to up-regulated receptor mRNA induced by starvation remain unknown.

Adenylyl Cyclases↗

PubMed1997

Concealed left ventricular hypertrophy and diastolic dysfunction in hypertrophic cardiomyopathy in the presence of acute left ventricular volume overload. A case report.

We report a patient in whom hypertrophic cardiomyopathy, with both left ventricular hypertrophy and diastolic dysfunction, was masked by acute severe aortic regurgitation and marked left ventricular dilation. Upon admission, 1) two-dimensional echocardiogram of the left ventricle revealed a dynamic and flail vegetation on the aortic right coronary cusp and marked left ventricular dilation, 2) a massive aortic regurgitant signal was recorded by color Doppler flow imaging, and 3) transmitral flow velocity by pulsed Doppler echocardiogram revealed a pseudonormalization. However, symmetric hypertrophy of the left ventricular wall, a decrease in early diastolic wave and a compensatory increase in atrial systolic wave of the transmitral flow velocity appeared after successful aortic valve replacement.

Aortic Valve↗

PubMed1997

Transesophageal echocardiographic prediction of initially successful electrical cardioversion of isolated atrial fibrillation. Effects of left atrial appendage function.

Left atrial appendage (LAA) flow velocities prior to electrical cardioversion were recorded using transesophageal pulsed Doppler echocardiography to predict initially successful cardioversion of isolated atrial fibrillation (AF). Patients with AF were placed into either a success group (19 patients) in which sinus rhythm was maintained for at least 2 days or a failure group (12 patients). The duration of AF was shorter in the success group. The maximum left atrial diameter was the same for the two groups. The maximum LAA area was smaller in the success group. The maximum forward and backward LAA velocities were greater in the success group, as were the mean forward and backward LAA velocities. In the patients with mean LAA flow velocities greater than 19 cm/sec, the success of cardioversion could be predicted with high sensitivity (80%) and specificity (88%). We conclude that the duration of AF, the maximum LAA area, and LAA flow velocities prior to cardioversion predict the initial recovery of sinus rhythm for isolated AF.

Adult↗

PubMed1997

Restoration of disturbed tooth eruption in osteopetrotic (op/op) mice by injection of macrophage colony-stimulating factor.

Osteopetrotic (op/op) mice show severe osteosclerosis caused by an inherited deficiency of osteoclast and resultant failure of tooth eruption, which can be cured by the injection of macrophage colony-stimulating factor (M-CSF). The present study revealed that consecutive injections of M-CSF in these mutant mice brought about a recovery of bone resorption resulting in the resumption of growth of tooth root and periodontal ligament. Bone resorption at the inner surface of bony crypts was noted on the 5th day after the start of M-CSF injections. This activity was reduced with the progress of root and periodontal ligament formation, being confined to the basal and crestal portion of bony crypts by the 15th day of the experiment. Second molars emerged into the oral cavity on the 15th day, but no eruption of first molars was observed until the 20th day. Throughout the experiment, first molars exhibited appreciable root deformity, which was less severe in second molars. Delayed eruption of first molars was thought to be related to the severity of the disturbance of root formation.

Age Factors↗

PubMed1997

Sonic analysis for detection of cervical and intracranial vascular disease.

A sonic analysis system was developed for the detection of cervical and intracranial vascular disease. The system analyzes recorded sound signals converted to digital data, and plots the frequency, time interval after the QRS wave of the electrocardiogram, and amplitude on the graph using contour lines. Sonic analysis of 22 patients with and 23 patients without cerebrovascular disease identified a characteristic pattern of disease called the "circular pattern." Five of 10 patients with internal carotid artery stenosis, four of seven with cerebral aneurysms, three with cerebral arteriovenous malformation, and two of two with dural arteriovenous malformation showed the circular pattern. Only one of 23 control patients showed the circular pattern. This system is a promising method for cost-effective mass screening for the early detection of cerebrovascular disease.

Adult↗