Biomedical subjects

M Maki

Publications and source records attributed to M Maki.

At least 91 records · Page 5Linked to original sources

PubMed1994

Unphosphorylated and tyrosine-phosphorylated forms of a focal adhesion protein, paxillin, are substrates for calpain II in vitro: implications for the possible involvement of calpain II in mitosis-specific degradation of paxillin.

Cell-to-substratum adhesion becomes weakened during mitosis of the cell cycle in fibroblasts. The level of one focal adhesion protein, paxillin, is greatly reduced in mitotic-arrested cells. We show here the possible involvement of calpain II, known to be localized in focal adhesion plaques, in the degradation of paxillin. Paxillin is tyrosine-phosphorylated during interphase of the cell cycle by protein tyrosine kinases (PTK) such as c-Src and Csk, and becomes dephosphorylated during mitosis. Our data, however, indicate that tyrosine phosphorylation of paxillin does not affect the rate of paxillin degradation by calpain in vitro.

3T3 Cells↗

PubMed1994

Amino-terminal conserved region in proteinase inhibitor domain of calpastatin potentiates its calpain inhibitory activity by interacting with calmodulin-like domain of the proteinase.

Calpastatin is a widely distributed endogenous inhibitor protein specifically acting on calpain (Ca(2+)-dependent proteinase) and is known to interact with the calmodulin-like domain (CaMLD) of the proteinase in a Ca(2+)-dependent fashion. The calpastatin molecule consists of four inhibitory domains (domains 1-4) with mutually homologous sequences in three regions designated as A, B, and C. Acidic amphiphilic alpha-helical motifs are found in both regions A and C. We investigated the correlation between the calpain inhibition potency and the ability of calpastatin to bind to recombinant CaMLD of the mu-calpain large subunit using various mutant proteins of pig calpastatin domain 1 expressed in Escherichia coli. Substitution of conserved Leu-161 with Pro in region A caused a reduction in activity of both calpain inhibition and CaMLD binding. Additional substitution of Leu-236 with Pro in region C further decreased the calpain inhibitory activity and caused a loss of CaMLD binding ability. The effects of mutation in region C alone on the above activities were smaller than those in region A. Although a mutant of deletion in the entire region B had no calpain inhibitory activity, it retained the CaMLD binding ability. On the other hand, although a region B oligopeptide had a moderate inhibitory activity, it had no CaMLD binding ability. These results suggest that region A has a role in potentiating the inhibitory activity of calpastatin by interacting with CaMLD of calpain to form a tighter complex where region B exerts the inhibitory function.

Amino Acid Sequence↗

PubMed1994

Real-time-analysis of the calcium-dependent interaction between calmodulin and a synthetic oligopeptide of calcineurin by a surface plasmon resonance biosensor.

The calcium-dependent interaction between calmodulin (CaM) and the synthetic oligopeptide of a predicted CaM-binding region of human calcineurin A-2 was analysed with an automated surface plasmon resonance biosensor, BIAcore. The oligopeptide was immobilized to a biosensor chip via the amino-terminal cysteine residue by a thiol-disulphide exchange method. The biosensor chip was regenerated by an EGTA-containing buffer after each analysis. Kinetics experiments showed that CaM bound with a high affinity to the oligopeptide in a Ca(2+)-dependent manner. The estimated rate constants of association (kass) and dissociation (kdiss) were 2.3 x 10(5) M-1.s-1 and 3.9 x 10(-3)s-1, respectively. The ratio of kdiss/kass, 1.7 x 10(-8) M, was in good agreement with the dissociation constant (Kd) of 2.4 x 10(-8) M determined from the equilibrium phase.

Amino Acid Sequence↗

PubMed1994

A new function of calphobindin I (annexin V). Promotion of both migration and urokinase-type plasminogen activator activity of normal human keratinocytes.

Calphobindin I (CPB-I, annexin V) is an anticoagulant protein purified from human placenta; it is a member of the annexin family that binds phospholipids in a calcium-dependent manner. In this study, we discovered and examined a new function of CPB-I: promotion of both the migration and the urokinase-type plasminogen activator (uPA) activity of normal human keratinocytes (NHK). While the treatment of NHK with a 10-micrograms/ml concentration of CPB-I for 24 h or 48 h caused an approximate 30% increase in the migration of NHK (compared with the no treatment), migration was inhibited when anti-CPB-I monoclonal antibodies (i.e. A46 and A180) were added along with the CPB-I. Moreover, while the treatment of NHK with a CPB-I concentration greater than 10 micrograms/ml caused a significant increase in the activity of secreted uPA, reflected in an approximately 40% increase in cell migration, uPA activity was inhibited both by cycloheximide and by monoclonal antibodies. This significant increase of secreted uPA was seen 8 h after the addition of CPB-I. Specific binding of CPB-I to NHK had a Kd value of 95.2 nM (equivalent to a CPB-I had no effect on NHK proliferation. Furthermore, CPB-I enhanced reepithelialization when it was applied locally twice a day to full-thickness cutaneous wounds made in male rats. Our results show that, during an injury, CPB-I helps reepithelialization through the promotion of both uPA synthesis and migration of keratinocytes without stimulating their proliferation.

Animals↗

PubMed1994

Analysis of calcium-dependent interaction between amino-terminal conserved region of calpastatin functional domain and calmodulin-like domain of mu-calpain large subunit.

Calpain requires Ca2+ both for proteolysis of its substrates and for interaction with its endogenous inhibitor, calpastatin. Although calmodulin-like domains (CaMLDs) of large and small subunits of calpain have been suggested to be the sites for Ca(2+)-dependent interaction with calpastatin, specificity and molecular basis of the interaction have remained unclear. We investigated the interaction between the CaMLD of human mu-calpain large subunit expressed in Escherichia coli and a 19-residue synthetic oligopeptide corresponding to the region A (the amino-terminal conserved acidic region) of one of the four repetitive functional domains of calpastatin. The recombinant CaMLD bound to the oligopeptide immobilized on Sepharose beads in a Ca(2+)-dependent manner. The CaMLD failed in binding to a mutant oligopeptide with one amino acid substitution. Enhancement of fluorescence intensity of a hydrophobic probe, 2-(p-toluidino)naphthalene-6-sulfonate, was observed upon incubating with the CaMLD and further increased by Ca2+. The Ca(2+)-dependent enhancement of fluorescence intensity was strongly suppressed by the wild type oligopeptide, but not by the mutant one. Kinetic experiments were performed with BIAcore where binding of the CaMLD to the oligopeptide immobilized on a biosensor chip was detected as real time signals of surface plasmon resonance. The determined dissociation constant (KD) was 3.1 x 10(-9) M. These results suggest that the region A of calpastatin binds to the CaMLD in a specific manner similar to interactions between calmodulin-binding peptides and calmodulin where hydrophobic properties are known to be important.

Amino Acid Sequence↗

PubMed1994

[Therapeutic effectiveness of 131I-MIBG on malignant pheochromocytoma--results of long-term follow-up].

The therapeutic response of 131I-MIBG was evaluated in 4 patients with malignant pheochromocytoma who had been treated with 131I-MIBG and followed-up over 5 years. The patients were 2 men and 2 women with ages ranging from 41 to 69 years old (mean 53 years). The primary tumors in 3 of 4 patients had been resected four to eight years before 131I-MIBG treatment. One patient was diagnosed as adrenal pheochromocytoma, and two were retroperitoneal paraganglioma. And in one patient, the resection of primary mediastinal tumor was not performed due to the adhesion to pericardium but the diagnosis of paraganglioma was obtained by biopsy of bone lesion. All patients showed the clear accumulation of 131I-MIBG in tumor on scintigraphy. The number of doses of 131I-MIBG ranged from one to three times with 3.7 GBq per administration and a cumulative activity from 3.7 to 11.1 GBq. Treatment effect was obvious in one patient with lung, bone, and lymph node metastases whose cumulated absorbed dose with 11.1 GBq of 131I-MIBG exceeded over 150 Gy. At the present time, the duration of survival since the beginning of initial 131I-MIBG therapy is over 5 yrs. The other three patients, however, showed little effects, and died with the disease in 2.6 to 4.1 years after the initial 131I-MIBG therapy. 131I-MIBG will become a promising agent for therapy in patients with malignant pheochromocytoma with high degree of accumulation.

3-Iodobenzylguanidine↗

PubMed1994

[Evaluation of myocardial uptake of beta-methyl-(123I)-iodophenylpentadecanoic acid (123I-BMIPP)].

To evaluate the myocardial uptake of beta-methyl-(123I)-iodophenylpentadecanoic acid (123I-BMIPP), nineteen patients with ischemic heart disease including left ventricular hypertrophy (mean age 63 +/- 7.8, 14 males and 5 females) underwent BMIPP myocardial scintigraphy. Myocardial uptake (MU) of BMIPP to the total injected dose was calculated from anterior view of the planar image in all subjects, and was compared with plasma glucose (BS), triglyceride (TG), and free fatty acid (FFA). It was also compared with left ventricular mass (LVM) calculated with echocardiography. MU was not related to BS, TG, and FFA, however had the positive correlation with LVM (r = 0.676, p < 0.01). Myocardial uptake per left ventricular mass (MU/LVM) had the negative correlation with LVM (r = -0.671, p < 0.01). Further studies for the significance of MU/LVM will be required.

Aged↗

PubMed1994

[Effect of calcium antagonist (benidipine) and alpha 1 blocker (urapidil) on renal hemodynamic responses to two acute environmental stresses in spontaneously hypertensive rats (SHR)].

Renal hemodynamic responses were studied in spontaneously hypertensive rats (SHR) and Wistar-Kyoto Rats (WKY) to two acute stresses: environmental stress (foot shock (FS) and air jet (AS)). The effects of calcium channel blocker (benidipine) and alpha 1 blocker (urapidil) on these responses were studied using an ultrasonic pulsed Doppler flowmeter. The increments in mean arterial pressure (MAP) and renal vascular resistance (RVR) were greater in SHR during both stresses. On the other hand, the decrease in the renal blood flow (RBF) with these stresses almost disappeared with renal denervation. These renal hemodynamic responses in SHR disappeared with alpha 1 blocker (urapidil), but not with calcium channel blocker (benidipine). The sympathetic nervous system became hyperactive in SHR under environmental stress, Which induced specific renal hemodynamic change. These results suggest that investigations on essential hypertension should focus on clarifying not only systemic hypertensive reaction, but also changes in renal hemodynamics. Furthermore, it is necessary for antihypertensive therapy to take these hemodynamic changes into consideration.

Acute Disease↗

PubMed1994

[Quantitative analysis of 123I-metaiodobenzylguanidine myocardial scintigraphy by myocardial uptake using a phantom].

To evaluate the quantitative analysis of 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy, total injected dose measured by first pass (FP) method (TFP) was compared with that measured by phantom method using an acrylic phantom in 45 patients with cardiac disease. Heart per mediastinum ratio (H/M) was compared to myocardial uptake calculated with TFP. The total injected dose measured using the phantom in which the syringe was set in depth of 3.5 cm (Tpham) was correlated with TFP (r = 0.73, p = 0.0001). When Tpham was corrected by body weight (c-Tpham), c-Tpham showed better correlation with TFP. MU calculated by TFP (MU-FP) was well correlated with MU by c-Tpham (MU-pham) (r = 0.94, p = 0.001). These results indicate that phantom method is sufficient to substitute for FP method. Though H/M was correlated with MU-FP (p < 0.001), the interpatient variation was relatively large. Then the analysis by H/M is insufficient to substitute for the myocardial uptake. It is though to be enough to use the phantom method on daily routine work, since this method is accurate and easy to quantitate the myocardial uptake of MIBG taking a short time.

3-Iodobenzylguanidine↗

PubMed1993

Clinical evaluation of low-molecular-weight heparin (FR-860) on disseminated intravascular coagulation (DIC)--a multicenter co-operative double-blind trial in comparison with heparin.

This study was evaluated the effectiveness, safety and utility of FR-860 and to compare those with heparin in patients with Disseminated intravascular coagulation (DIC). A diagnosis of DIC was made based on the criteria proposed by the Research Committee on DIC in the Ministry of Health and Welfare of Japan. FR-860 (FR group, 75 anti-factor Xa international units/kg/day) and Heparin (HP group, 240 units/kg/day) were administered for 5 days by continuous intravenous infusion. The total number of enrolled patients was 126 cases, and after excluding 1 case a total of 125 cases. Moderate or higher improvement of bleeding symptoms was 33.3% in the FR group and 18.5% in the HP group. On the organic symptoms, FR group showed a significantly higher improvement rate than the HP group, 20.5% and 8.2% respectively. On the overall efficacy of cases with pretreatment plasma AT III levels of less than 21 mg/dl or less than 70%. FR group showed a significantly higher improvement rate than the HP group. The safety rate of FR-860 (93.4%) was a significantly higher than that of the HP group (79.7%). Our report demonstrates that FR-860, as a therapeutic agent for the treatment of patients with DIC, is significantly higher safety and clinical utility as compared with heparin.

Adult↗

PubMed1993

Molecular diversity of calpastatin in human erythroid cells.

According to differences in mobility on SDS-polyacrylamide gel electrophoresis, calpastatins (inhibitor proteins of the calcium-dependent proteinase calpain) are classified into the tissue type (100-120 kDa) and the erythrocyte type (70 kDa), which lacks the amino-terminal domains (domains L and 1). We investigated the molecular diversity of calpastatin in human hematopoietic cells by Western-blot analysis and by the reverse-transcription-polymerase-chain reaction method. While the mononuclear and polymorphonuclear cells in peripheral blood showed the tissue type (110 and 114 kDa), a cell line of erythroid cells (JK-1) showed both the tissue type (110 kDa) and the erythrocyte type (70 kDa) at approximately equal ratios. When the lysate of JK-1 cells was incubated in the presence of ATP, the 110-kDa form was degraded much faster than the 70-kDa form. In human erythrocytes, the 110-kDa form was identified as the tissue type by an antibody recognizing domain L, and this form was also present in addition to the predominant 70-kDA form. JK-1 cells, as well as nucleated cells in peripheral blood, contained calpastatin mRNA with exon-3-deleted. Glioblastoma and fibroblast cell lines expressed the nondeleted calpastatin mRNA in addition to the deletion type, and they showed bands corresponding to 117 kDa as well as 110 and 114 kDa. The 117-kDa band was detectable by an anti-exon 3 peptide antibody. These results suggest that diversity among the tissue type calpastatins is caused by both alternative splicing and post-translational processing whereas the apparent conversion from the tissue type to the erythrocyte type is caused by proteolytic processing.

Adenosine Triphosphate↗

PubMed1993

Requirement of different subdomains of calpastatin for calpain inhibition and for binding to calmodulin-like domains.

Calpain requires Ca2+ for both proteolysis of its substrates and interaction with its endogenous inhibitor, calpastatin. The mechanism of inhibition of calpain by calpastatin has remained unsolved, although Nishimura and Goll [J. Biol. Chem. 266, 11842-11850 (1991)] reported that autolyzed calpain fragments containing calmodulin-like domains (CaMLDs) bound to an immobilized calpastatin column. We investigated the correlation between CaMLD-binding and calpain inhibition using immobilized columns of gene-engineered CaMLDs derived from the human mu-calpain large subunit and various recombinant calpastatin mutants. Among the four internally repetitive inhibitory domains of calpastatin, each having conserved regions A, B, and C, only domains 1 and 4 showed the binding activity. The region B deletion mutant of domain 1, retaining the CaMLD-binding ability, no longer had the calpain inhibition activity, and became susceptible to proteolysis. In contrast, a synthetic oligopeptide of region B with moderate calpain inhibition activity did not bind to the column. Domain 3 acquired the binding ability on substitution of region A with that of domain 1. These results suggest that calpain inhibition and binding to the CaMLDs are not correlated or mediated by different subdomains of calpastatin.

Amino Acid Sequence↗

PubMed1993

[Clinical utility of thallium-201 chloride in the diagnosis of parathyroid glands in patients with hyperparathyroidism].

Clinical utility for detection of hyperfunctioning parathyroid glands in patients with primary and secondary hyperparathyroidism (PHPT, SHPT) using 201TlCl and 99mTcO4-images was estimated in 50 patients including 18 PHPT and 32 SHPT (male; n = 31, female; n = 19, 51.5 +/- 11.1 yr). Detection of 94 hyperfunctioning parathyroid glands in 34 cases were achieved correctly by 201TlCl and 99mTcO4- images in agreement with surgical findings. Detectability of hyperfunctioning parathyroid glands was 100% in more than 1.0 g, 62.0% in more than 0.5 g and 41.5% in less than 0.5 g respectively. Ectopic parathyroid glands in two cases and a metastasis to pretracheal lymph node in one case were clearly demonstrated. Our results show that parathyroid scintigraphy using 201TlCl and 99mTcO4- as well as 131I or 123I is useful and safe method for preoperative detection of hyperfunctioning parathyroid glands.

Adult↗

PubMed1993

[Radionuclide therapy of thyroid disease--radioactive iodine therapy].

Radioiodine, 131I, has been used for treatment of hyperthyroidism and metastases of well differentiated thyroid carcinoma since 1942. The use of 131I to treat hyperthyroidism and metastatic foci of thyroid cancer is one of the least invasive and most effective methods since the ionization produced by beta rays of 131I and it's high target-to-nontarget ratio are suitable. The feature of therapy with 131I on hyperthyroidism has a slow improvement of symptom, a high incidence of hypothyroidism, and an exact therapeutic effect. And, there is no significant difference between the patients with late-onset hypothyroidism and euthyroidism after 131I-therapy in the pretreatment factors, such as thyroid hormone levels, thyroid weight, 131I administration dose, and absorbed dose and also in the course of follow-up studies. On the treatment of metastases of thyroid cancer with 131I, it is very effective for the patients with fine or occult type of pulmonary metastases on chest XP, who are younger than 40 years old. On the other hand, the response to bone metastases of 131I therapy is limited to only a part of tumors and/or temporally in most cases though high grade of 131I accumulation is seen on scintigram. Both surgery and a curative dose of external irradiation should be combined with radioisotope therapy for the patients of bone metastases.

Adult↗

PubMed1993

Long cellular repeats flanking a defective HTLV-I provirus: implication for site-targeted integration.

Retroviruses generally integrate as proviruses which are flanked by long-terminal repeats (LTRs) on both 5' and 3' ends. Since these LTRs are required for the efficient integration mediated by the viral integrase, it is believed that defective proviruses with a single LTR are normally formed by deletion after integration. However, we found no deletion of cellular sequences around the integration site of such a defective HTLV-1. Rather, we identified 99 bp-long direct repeats adjacent to both ends of the defective provirus. The repeated cellular sequences contained a potential poly(A) signal followed by a retroviral primer-binding-site-like sequence. The presence of the direct repeats of cellular sequences can be explained by the integration of the defective virus through homologous recombination between cellular and viral read-through sequences.

Base Sequence↗

PubMed1993

[Recent advances in diagnosis of disseminated intravascular coagulation in obstetrics and gynecology].

Since obstetrical DIC is an acute serious situation, complicated time-consuming tests do not help to make a prompt diagnosis of DIC. Lowered ESR, prolonged bleeding time and defective patterns of thrombelastography are the suggestive findings of DIC, which are available at the bed-side. Obstetrics-specific DIC scoring system, in which high scores were given for clinical items, rather than laboratory tests, showed a good correlation with the regular DIC score in Japan. Since blood samples obtained from pregnant or parturient women showed different patterns of coagulation and fibrinolysis from the nonpregnant condition, attention should be paid to diagnose obstetrical DIC as accurately as possible.

Biomarkers↗

PubMed1993

[131I-therapy of differentiated thyroid carcinoma with distant metastases--relation between absorbed dose by quantitative SPECT and outcome of the patients in thyroid carcinoma].

The correlation of absorbed doses of tumors in 18 patients of differentiated thyroid carcinoma with distant metastases, who were treated by 131I and followed over 5 years, with their outcome were analyzed and the clinical significance of determination of absorbed dose was discussed. Radioactivities of 131I in the tumors were measured by using SPECT at the time of therapy. Absorbed dose was calculated based on the MIRD equation. Outcome of 8 patients were evaluated as good and their absorbed dose was 10-630 Gy with 2-22 g of tumor volume, 1.2-3.5 days of effective half life (EHL) and follow-up term was 8.6 +/- 0.9 years. The absorbed dose of 10 patients whose outcome were evaluated as poor, was 5-81 Gy with 7-215 g of tumor volume, 1.3-5.3 days of EHL and follow-up term was 5.6 +/- 2.5 years. The initial treatment seemed to be important for 131I therapy, since the absorbed doses in the following therapy became reduced. When the absorbed dose of the tumor exceeded over 94 Gy at initial treatment, good clinical courses were obtained. These results indicate that the quantitative SPECT for 131I therapy is clinically valid and that the calculated absorbed doses correlate well with outcome of the patients.

Adenocarcinoma↗

PubMed1992

Expression of calpain II gene in human hematopoietic system cells infected with human T-cell leukemia virus type I.

We examined the distribution of calpains I and II in human hematopoietic system cell lines by Western and Northern blot analyses and enzyme activity assay. Expression of calpain I, a low Ca(2+)-requiring cysteine protease, was observed in all human T-cell lines tested. By contrast, expression of calpain II, a high Ca(2+)-requiring form, in human T-cells was closely correlated with human T-cell leukemia virus type I (HTLV-I) infection, which is known to result in the expression of adult T-cell leukemia-associated antigens, interleukin-2 (IL-2) receptor alpha, and Ca(2+)-dependent cell proliferation. Specific expression of calpain II in HTLV-I-infected cells occurred at the mRNA level. Furthermore, expression of calpain II in human natural killer-like cells was augmented by HTLV-I pX gene transfection. In HTLV-I-infected cells, the trans-acting transcriptional activation of the long terminal repeat and control elements for the IL-2 receptor alpha, c-fos, and granulocyte-macrophage colony-stimulating factor genes by the Tax from the pX gene is already known. Our results suggest that the similar trans-activation occurs to the calpain II gene in HTLV-I-infected hematopoietic system cells.

Blotting, Western↗