PubMed Health⌕ Search

Biomedical subjects

A Suwabe

Publications and source records attributed to A Suwabe.

At least 19 recordsLinked to original sources

[Evaluation of dysmorphic red cells in the urinary sediment].

The usefulness of the morphologic examination of urinary red cells was first described by Birch and Fairley, who reported that the dysmorphic red cells(morphologically variable) were the marker for glomerular bleeding and the isomorphic red cells(morphologically uniform) for non-glomerular bleeding. They also noted that healthy individuals had dysmorphic cells, indicating a glomerular source. The relation between red blood cell morphology and the origin of hematuria has been confirmed subsequently by numerous clinical studies. Although the reports varied on the ratio(10-100%) of dysmorphic cells, their findings have been supported by many investigators. In 1991 Köhler noted that acanthocytes(AC) were the most characteristic red cell type for glomerular bleeding and that acanthocyturia > or = 5% was a good predictive marker for glomerular bleeding. Moreover the red cell was easily recognized. On the other hand, in 1993 Fairley and Birch reported that in glomerulonephritis, erythrocytes varied markedly in size, shape, and hemoglobin content, and that the pattern of morphology indicates the source of bleeding, not the morphology of individual cells. Both observations were confirmed in our in vitro experiments. To clarify the causative mechanism of acanthocyturia, the normal washed erythrocytes were successively exposed to two kinds of solutions simulating conditions in the distal tubules(105 mOsm, pH 5.5, NaCl 37 mmol/l, KH2PO4 1.0 mmol/l, Urea 0.9 g/l) and the collecting tubules(a mixture of NaCl and KH2PO4 solutions with the osmolality between 299 and 1192 mOsm). Various degrees of hemolytic process appeared in the first solution. However, AC appeared only in the second solutions with 9 mmol/l of KH2PO4 and 390 mOsm or more osmolality or with 725 mOsm and 4.1 mmol/l or more of KH2PO4. This result supported the nephron passage theory in the formation of AC in glomerular diseases.

Acanthocytes↗

Effect of a 5-HT(1A) receptor agonist, flesinoxan, on the extracellular noradrenaline level in the hippocampus and on the locomotor activity of rats.

We have studied effects of 5-hydroxytryptamine 1A (5-HT(1A)) receptor-selective compounds on the extracellular noradrenaline (NA) level in the hippocampus of rats using microdialysis and on their locomotor activity. A selective 5-HT(1A) receptor agonist, flesinoxan (5 mg/kg, i.p.) increased the extracellular NA level in the hippocampus, and increased the locomotor activity. Both responses were blocked by pretreatment with a 5-HT(1A) receptor antagonist, WAY100635 (1 mg/kg, i.p.) and an alpha(2) adrenoceptor agonist, clonidine (50 microg/kg, i.p.). Bilateral intrahippocampal injection of flesinoxan (200 nmol in 2 microl, respectively) increased the locomotor activity of rats and the intrahippocampal perfusion of flesinoxan (1 mM, 2 microl/min) increased the extracellular NA level in the hippocampus. Bilateral intrahippocampal injections of a small amount of WAY100635 (0.1 nmol in 2 microl, respectively) blocked the flesinoxan (5 mg/kg, i.p.)-induced hyperactivity. Flesinoxan (5 mg/kg, i.p.) did not significantly influence the level of serotonin or its major metabolite in the hippocampus, or dopamine or its metabolites in the striatum. In conclusion, these behavioral as well as pharmacological results indicate that postsynaptic 5-HT(1A) receptor activation by flesinoxan increase the extracellular NA level in the hippocampus, which may be the cause of the increase of the locomotor activity.

Animals↗

Nifedipine suppresses neointimal thickening by its inhibitory effect on vascular smooth muscle cell growth via a MEK-ERK pathway coupling with Pyk2.

The aim of this study was to determine whether nifedipine could suppress an atherogenic process such as balloon-injured intimal thickening in vivo and the proliferation of vascular smooth muscle cells (VSMC) in vitro. First, we examined the in vivo effect of nifedipine to determine whether it could suppress intimal thickening induced by balloon catheterization. Sprague-Dawley (SD) rats were divided into three groups (L, nifedipine 0.3 mg kg(-1) day(-1); H, nifedipine 3 mg kg(-1) day(-1); C, no nifedipine), and Alzet((R)) osmotic pumps were implanted in their backs for continuous administration. The neointimal layers were completely occupied by proliferated VSMC, and the area ratios of neointima/media treated with nifedipine significantly decreased dose-dependently compared to those of the control. Neither blood pressure nor lipid levels changed among the three groups. We next evaluated the in vitro effect of nifedipine on the proliferation of cultured rat VSMC. Nifedipine decreased the values of [(3)H]-thymidine incorporation and total cellular protein content as well as the levels of phosphorylated extracellular signal-regulated protein kinase (ERK) 1/2, mitogen-activated protein kinase kinase (MEK) 1/2, and even the phosphorylation of Pyk2, in dose-dependent fashions. In addition, nifedipine suppressed the levels of proliferative cell nuclear antigen (PCNA) dose-dependently in both VSMC and balloon-injured thoracic aortae. These results indicate that nifedipine has an inhibitory effect on intimal thickening by attenuating intimal VSMC proliferation, suggesting that nifedipine could be effective for preventing the progression of atherosclerotic plaque as in restenosis after angioplasty.

Actins↗

[The examinations for diffuse lung diseases].

Diffuse lung diseases show an abnormal shadow that is widely scattered on the bilateral lung fields in the chest X ray view and includes many respiratory diseases such as the infectious or the non-infectious disease; neoplasms. Among these, idiopathic pulmonary fibrosis(IPF) has been studied extensively because of its high frequency and difficulty of treatment. IPF is defined by the respiratory functions, the radiological findings, which depend on HRCT, and the histopathological evaluation by surgical lung biopsy. In particular, the histopathological appearance of usual interstitial pneumonia(UIP) is essential for the diagnosis of IPF. Most serum examinations such as angiotensin-converting enzyme, anti-nuclear antibodies are applied to rule out other diffuse lung diseases. SP-D or KL-6, which is the marker of the type II epithelial cells, is thought to be very useful for revealing the disease activity, but since it is not increased in the early stages of IPF, it is not applied in the diagnosis of IPF. The definitive serum examinations for the diagnosis or determinations of the therapeutic effect or prognosis of IPF have not been established. Easier, more useful and critical examinations including genetic diagnosis are required to manage patients with IPF.

Aged↗

[Approach to examining hypercalcemia in the clinical laboratory].

The investigation of hypercalcemia is carried out routinely in our laboratory to detect primary hyperparathyroidism (PHPT). In a retrospective study, seven patients with PHPT and fifty-one patients with non-parathyroid hypercalcemia (NPHC) were chosen in a particular year. To obtain a screening index for PHPT, discriminant analysis, using a stepwise variable select method, was applied to eight biochemical parameters in these patients. A discriminant function (F1) was derived from three biochemical parameters and then another discriminant function (F2) was also derived from three biochemical parameters in the F1-positive patients. In combination of these two functions (F1 and F2), the final sensitivity was 100% and specificity was 98% in diagnosing PHPT. This screening method was tested prospectively in fifty-six consecutive specimens of hypercalcemia (PHPT 4, NPHC 52) over the following six months. The result was also satisfactory with a sensitivity of 100% and specificity of 98%. It was proven that our screening method using discriminant functions (F1 and F2) was very useful for diagnosing patients with PHPT from the survey of hypercalcemia. Among these patients with hypercalcemia, the high ratio (54%) of those with malignancy was remarkable. This interesting result required us to investigate potential hypercalcemia, since the serum calcium concentration was masked by a lower level of serum albumin, which was frequently seen in these malignant patients. As the next step, we tried to adjust the serum calcium concentration based on the serum albumin concentration. A formula for adjusting the calcium concentration was derived from a linear structural relationship between calcium and albumin in 6,821 specimens within a +/- 2.5 second principal component score in 7,021 consecutive specimens in whom both calcium and albumin were measured in a particular year; Adjusted Calcium = Calcium - Albumin + 4. After adjustment using this formula, the calcium concentrations were elevated above the upper limit of the reference interval in 320 of 5,203 specimens (6%) within the reference interval and elevated to the reference interval in 1,390 of 1,579 specimens (88%) below a lower limit of reference interval. A prospective study was performed over the following three months. Fifty patients with hypercalcemia were screened using this formula. It was a surprise that thirty-one patients (62%) showed abnormal values after adjustment. These results suggest that calcium adjustment is necessary for interpreting the calcium concentration of patients with a reduced albumin concentration such as patients with malignancy.

Adult↗

Artificial surfactant (Surfactant TA) modulates adherence and superoxide production of neutrophils.

Neutrophils cause lung injuries by releasing proteases and active oxygen radicals in patients with acute respiratory distress syndrome (ARDS). Artificial surfactant is used to replace native surfactant whose functions are deteriorated by serum-derived inhibitors in these patients. We investigated potential interactions between exogenous surfactant (Surfactant TA) and neutrophils in in vivo and in vitro experimental models. Neutrophil alveolitis was induced in hamster lungs by the intratracheal administration of bleomycin (5 mg/kg) on Day 0. Some of the animals were followed by replacement with Surfactant TA (5 and 10 mg/100 g body weight) on Day 1. Alveolar cells were harvested by lung lavage on Day 2. The numbers of the neutrophils obtained from the lungs treated with bleomycin and Surfactant TA were unchanged, but the superoxide production from these cells was significantly decreased when compared with control animals (no Surfactant TA). From the in vitro experiments, Surfactant TA was shown to inhibit adherence and superoxide production of human neutrophils. These effects were derived from the heat-resistant components of Surfactant TA and were mimicked by treatment with liposomes of dipalmitoyl phosphatidylcholine. Surfactant-TA-treated neutrophils were demonstrated to have picnotic nuclei and to express Fas antigens, which were characteristic of apoptotic cells. These results suggest that exogenous Surfactant TA may play an important role not only in improving surfactant functions but in preventing neutrophils from further activation, probably through enhancing apoptosis.

1,2-Dipalmitoylphosphatidylcholine↗

[The effects of surfactant-TA on bleomycin-induced lung injury and lung fibroblast proliferation].

We investigated whether artificial surfactant modified pulmonary fibrosis following the intratracheal administration of bleomycin (BLM 3.75 mg/kg) in rats. Twenty-four hours after the administration of bleomycin, the lungs were washed twice with surfactant-TA (S-TA, 1 mg/ml, 6 ml x 2 times), and replaced with 0.5 ml of 10 mg/ml S-TA. Fourteen days later the animals were sacrificed and the lungs were assayed for collagen content and pathologic changes. BLM-treated rats showed both increases in wet lung weights and lung collagen content, and a loss in body weights (due to lung injury and fibrosis), but these changes were significantly inhibited by the intratracheal administration of S-TA. In vitro studies demonstrated that S-TA inhibited lung fibroblast proliferation in a dose dependent manner (0.01-0.5 mg/ml). This inhibition was also seen in native rat lung surfactant and its protein and lipid components. However, S-TA, did not change the number of rat type II pneumocutes in culture, and likely protected type II cells from dedifferentiation. S-TA inhibited BLM-induced pulmonary fibrosis, probably by directly inhibiting lung fibroblast proliferation.

Animals↗

Calcium dependent association of surfactant protein A with pulmonary surfactant: application to simple surfactant protein A purification.

Surfactant protein A (SP-A) is an abundant lipoprotein component of pulmonary surfactant that plays multiple roles in surfactant homeostasis within the lung. A simple and rapid purification procedure for SP-A is described. Purified surfactant is washed by centrifugation with Ca2+ containing buffer to remove residual soluble proteins. Following the Ca2+ buffer wash, the surfactant pellet is washed in buffer containing EGTA and Mg2+ which releases the bound SP-A in almost pure form. Subsequent chromatography of the SP-A on Sephacryl S-500 yields homogeneous preparations of the protein. The SP-A purified using this procedure requires no exposure to either detergents or organic solvents to remove lipid. SP-A prepared by this new method inhibits lipid secretion from alveolar type II cells as effectively as SP-A prepared by other methods. In addition, the SP-A depleted surfactant produced in the first step of this procedure is capable of binding exogenous SP-A in a time dependent, saturable and Ca2+ dependent manner.

Animals↗

[Effects of surfactant TA on adherence and structure of human peripheral blood neutrophils].

Artificial surfactant is used to treat patients with the adult respiratory distress syndrome; we studied its effects on neutrophil function. Neutrophils were isolated from healthy volunteers and their adherence to plastic surfaces was used as an index of their function. Surfactant TA (S-TA, 0.16-5 mg/mL) dose-dependently inhibited neutrophil adherence stimulated by n-formyl-methionyl-leucyl-phenyl-alanine, phorbol myristate acetate, and tumor necrosis factor. This inhibition was observed whether untreated S-TA or heated S-TA was used. Electron microscopy revealed an increase in the number of apoptotic neutrophils with pyknotic nuclei and smooth cell surfaces after treatment with S-TA which suggests that neutrophils apoptosis was increased. The number of apoptotic neutrophils increased with increasing time of incubation with S-TA, and was also high in the presence of G-CSF, which inhibits neutrophil apoptosis. These results show that S-TA can inhibit neutrophil function, and they suggest that S-TA therapy for the adult respiratory distress syndrome not only corrects the surface-tension abnormality, but can also inhibit infiltration of neutrophils.

Apoptosis↗

[Effects of erythromycin on surfactant synthesis, secretion, and recycling by rat alveolar type II cells].

Effects of erythromycin on surfactant synthesis, secretion, and recycling by rat alveolar type II cells were examined. Type II cells were isolated from rat lungs and incubated with 3H-choline, a precursor of surfactant. Synthesis was quantified from the amount of 3H-phosphatidylcholine and from the distribution to lamellar body fractions as measured with sucrose density gradients. Secretion was quantified from the percent of synthesized 3H-phosphatidylcholine released into the medium over 3 hr. Recycling was quantified from the percent uptake over 1 h of 3H-labeled synthetic liposomes. Type II cells incubated with erythromycin (5-50 micrograms/ml) for 22 hrs secreted less surfactant in response to PMA and ATP than did control cells. Erythromycin did not affect synthesis or recycling of surfactant by type II cells. These results show that erythromycin inhibits surfactant secretion from type II cells, and suggest that erythromycin may be effective in surfactant-excess states such as alveolar proteinosis.

Animals↗

[Inhibitory effect of rat alveolar type II cells on production of superoxide by neutrophils].

Neutrophils induce lung injury by releasing superoxides (O2-) and elastases. Alveoli contain anti-neutrophil agents such as surfactant. We studied the effects of alveolar type II cells on neutrophil functions. Neutrophils were isolated from human venous blood and their O2- production was measured by the cytochrome C reduction method. Type II cells and fibroblasts were isolated from rat lungs. Neutrophil O2-production was inhibited in the presence of type II cells. This effect was specific to type II cells: fibroblasts caused no inhibition. The inhibition did not require cell-to-cell contact, and it was abolished by the nitric oxide synthase inhibitor L-NAME. These results show that type II cells can inhibit neutrophil O2-production, and suggest that anti-neutrophil activity in alveoli may involve both surfactant and type II cells.

Animals↗

[Roles of surfactant protein A (SP-A) and phospholipids on regulation of surfactant secretion from alveolar type II epithelial cells].

Surfactant protein A (SP-A) is known to inhibit surfactant secretion from pulmonary alveolar type II epithelial cells (type II cells). It is unknown whether SP-A acts as an inhibitor in the presence of lipids since SP-A mostly exists complexed with surfactant lipids. In this paper we focused on this issue using purified surfactant, purified SP-A and synthesized lipid liposomes. Surfactant secretion from type II cells was evaluated as TPA-stimulated secretion of radiolabeled lipids. The results demonstrated that 1) the inhibition of lipiD secretion by purified surfactant depended on its lipid components, not on the SP-A component, 2) the inhibitory effect of SP-A purified from surfactant was reversed in the presence of lipid liposomes, and 3) SP-A free from surfactant was unable to inhibit lipid secretion. We conclude from these results that SP-A can potentially inhibit surfactant secretion, but surfactant secretion is mainly regulated by surfactant lipids.

Animals↗

[Spontaneously healed P. carinii pneumonia in the course of steroid therapy for interstitial pneumonia--many trophozoites in bronchoalveolar lavage].

A case in which P. carinii was observed in bronchoalveolar lavage fluid during steroid therapy for interstitial pneumonia in a 63-year-old man is reported, he had received steroid therapy for interstitial pneumonia of unknown origin. Three weeks later, he developed acute pneumonia with Streptococcus pneumoniae, and simultaneously P. carinii was detected in the bronchoalveolar lavage fluid. Both the pneumonic shadows and P. carinii disappeared following intravenous infusion of penicillin and rapid reduction of steroid. Electron microscopic analysis of P. carinii demonstrated numerous tubular expansions and endogenies of P. carinii, suggesting that P. carinii was growing in the intra-alveolar spaces. Phospholipid analysis demonstrated a transient increase in total phospholipid content during P. carinii pneumonia, suggesting that P. carinii can affect surfactant metabolism.

Bronchoalveolar Lavage Fluid↗

Pulmonary surfactant secretion is regulated by the physical state of extracellular phosphatidylcholine.

Pulmonary alveolar type II cells synthesize, secrete, and recycle the components of pulmonary surfactant. In this report we present evidence that dipalmitoylphosphatidylcholine is a potent inhibitor of surfactant lipid secretion by type II cells. Monoenoic and dienoic phosphatidylcholines with fatty acids of 16 or 18 carbons are ineffective as inhibitors of surfactant lipid secretion. In contrast, disaturated phosphatidylcholines, with either symmetric or asymmetric pairs of fatty acids of 14, 16, or 18 carbons, exhibit inhibition of surfactant secretion that correlates extremely well with the phase transition temperature (Tc) of the phospholipid. The inhibitory activity of dipalmitoylphosphatidylcholine is not dependent upon lipid stereochemistry. N-Methylated derivatives of dipalmitoylphosphatidylethanolamine are significantly less effective than phosphatidylcholine as inhibitors. Phosphatidylcholines below their phase transition temperature are inhibitors of surfactant secretion, whereas those above their phase transition temperature are either ineffective or weakly inhibitory. The phase transition dependence of inhibition is observed when type II cells are incubated at 37 degrees C with different species of phosphatidylcholine. In addition, if type II cells are stimulated to secrete at different temperatures the efficacy of a given phospholipid as an inhibitor is dependent on its relationship to Tc (i.e. dipalmitoylphosphatidylcholine with a Tc of 41 degrees C significantly inhibits secretion at 37 degrees C but not at 42 degrees C). Inhibition of surfactant secretion by dipalmitoylphosphatidylcholine is abrogated when it is incorporated into the same liposome with dioleoylphosphatidylcholine as a 50:50 mixture. In contrast, the simultaneous addition of two separate populations of liposomes, one composed of dipalmitoylphosphatidylcholine and the other composed of dioleoylphosphatidylcholine, does not significantly alter the inhibitory activity found with dipalmitoylphosphatidylcholine alone. These data provide compelling evidence that the physical state of phosphatidylcholine can regulate surfactant secretion from alveolar type II cells and suggest a unique mechanism for regulating exocytosis in the alveolus of the lung.

1,2-Dipalmitoylphosphatidylcholine↗

Alveolar type II cells isolated after silica-induced lung injury in rats have increased surfactant protein A (SP-A) receptor activity.

We examined surfactant secretion and its regulation by surfactant protein A (SP-A) in alveolar type II cells isolated from silica-treated rats to determine the role of SP-A-mediated regulatory control of phospholipid secretion in the pathogenesis of silica-induced alveolar proteinosis. Type II cells were isolated at weekly intervals for 28 d after silica or saline instillation. The maximum total binding of [125I]SP-A (internalized and surface-bound SP-A) to type II cells increased with time after silica instillation and, at 21 d, was 4-fold greater than that of type II cells isolated from saline-treated rats (272.8 +/- 42.5 and 65.4 +/- 9.8 ng/10(5) cells, respectively; P less than 0.05). Type II cells isolated from silica-treated rats showed a 2-fold increased surface binding and a 3-fold increased internalization compared to control cells. The receptor affinity for SP-A was the same for type II cells isolated from silica- and saline-treated animals. Type II cells isolated 14 d after silica instillation were separated into normotrophic and hypertrophic populations by centrifugal elutriation. Hypertrophic cells showed significantly elevated maximum total binding compared to normotrophic cells. The secretion of [3H]phosphatidylcholine [( 3H]PC) by type II cells from silica- and saline-treated animals was also compared. Type II cells from silica-treated animals showed lower basal and tetradecanoyl phorbol acetate (TPA)-stimulated [3H]PC secretion than cells from saline-treated animals at each time point after instillation. SP-A inhibited TPA-stimulated [3H]PC secretion similarly in type II cells isolated after either silica or saline instillation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Temporal segregation of surfactant secretion and lamellar body biogenesis in primary cultures of rat alveolar type II cells.

Pulmonary alveolar type II cells synthesize and secrete the phospholipids of surfactant. However, type II cells isolated from adult rat lungs rapidly lose their characteristic morphology and differentiated functions (such as surfactant-specific phospholipid and protein biosynthesis) when maintained on tissue culture plastic. In this study, phospholipid secretion and its regulation by type II cells grown on tissue culture plastic were examined up to 8 days after isolation. Type II cells were preincubated with [3H]choline for varying 24-h periods during culture prior to examining phosphatidylcholine ([3H]PtdCho) secretion. Type II cells cultured for 4 days and incubated with [3H]choline 24 h before the secretion experiment failed to show significant basal and tetradecanoyl phorbol acetate (TPA, 100 nM)-stimulated [3H]PtdCho secretion (basal, 0.29 +/- 0.01%; TPA, 0.48 +/- 0.04%). In contrast, type II cells incubated with [3H]choline for the first 24 h during culture and then cultured for 3 more days showed significant [3H]PtdCho secretion (basal, 1.27 +/- 0.19%; TPA, 6.24 +/- 0.82%). Subcellular fractionation of type II cells revealed that [3H]choline was incorporated into phosphatidylcholines in a lamellar body-enriched fraction during the first 24 h of culture but that the assimilation of phosphatidylcholine into the lamellar body fraction progressively declined with increasing time in culture. Radiolabel incorporated into the lamellar body fraction labeled during the first 24 h of culture was detectable for up to 8 days in culture. The [3H]PtdCho incorporated into the lamellar body during the first 24 h of culture was lost gradually over 8 days, suggesting the continuous secretion or turnover of the lamellar bodies during culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Expression and characterization of rat surfactant protein A synthesized in Chinese hamster ovary cells.

Rat surfactant protein A (SP-A) was expressed in a Chinese hamster ovary (CHO-K1) cell line and characterized for biologic activity using assays for receptor binding and modulation of phospholipid secretion from isolated type II cells. The CHO-K1 cell line was cotransfected with separate plasmids encoding for the rat SP-A, dihydrofolate reductase and neomycin phosphotransferase, respectively. Antibiotic (Geneticin-G418)-resistant transformants were screened by ELISA for the secretion of recombinant SP-A into the media. Northern analysis of the transfected cell lines demonstrated the expression of both 1.6 kb and 0.9 kb mRNA species for SP-A, consistent with the proposed differential polyadenylation of the primary transcript. Amplification with methotrexate resulted in a dose-dependent increase in mRNA for SP-A and a 20-fold increase in the production of recombinant SP-A relative to untreated cells. Maximum production of SP-A was 370 micrograms of SP-A/l of media in a 4-day incubation. Recombinant SP-A was purified from the serum-free media of large scale cultures of transfected, amplified CHO cells by affinity chromatography on mannose-Sepharose. The recombinant SP-A migrated similarly to native SP-A by NaDodSO4-PAGE analysis under reducing and nonreducing conditions and under reducing conditions after digestion with N-glycanase. Recombinant SP-A effectively competed with 125I-native SP-A for binding to the high affinity receptor for SP-A on isolated plasma membranes from rat alveolar type II cells. The recombinant SP-A was as effective as native SP-A in the inhibition of secretion of phospholipid from isolated type II cells. We conclude that recombinant rat SP-A produced in Chinese hamster ovary cells is physically and functionally similar to native rat SP-A.

Animals↗