Biomedical subjects

K Miyamoto

Publications and source records attributed to K Miyamoto.

At least 91 records · Page 5Linked to original sources

PubMed2001

Effect of meropenem on disposition kinetics of valproate and its metabolites in rabbits.

PURPOSE: We investigated the effect of meropenem (MEPM) on the disposition kinetics of valproate (VPA) and its metabolites in rabbits. METHODS: Rabbits were given 75 mg/kg VPA intravenously with or without 300 mg/kg MEPM. RESULTS: The plamsa total clearance of VPA was significantly increased to about 1.5 times the control (6.09 mL/min/kg vs. 4.28 mL/min/kg) by MEPM (P < 0.05). The values of the area under the plasma concentration-time curve (AUC) of 2-en-VPA, a product of beta-oxidation, and VPA-glucuronide (VPA-G) were significantly decreased to about 55% and 78% of the control, respectively (P < 0.05). The cumulative urinary excretions of VPA in the control and MEPM-treated groups were 0.54% and 0.62% of the dose, respectively, whereas those of VPA-G were 45.6% and 62.5%, respectively. The urinary excretion of VPA-G was significantly increased by MEPM (P < 0.05). Further, in the case of 33.8 mg/kg VPA-G administered intravenously the AUC value of VPA-G was unchanged by MEPM, whereas that of the generated VPA was significantly decreased to about half of the control. CONCLUSIONS: The increase of the total clearance of VPA caused by MEPM appears to be a consequence of increased renal clearance of VPA-G, as well as suppression of VPA-G hydrolysis in the liver.

Animals↗

PubMed2001

Solution structures of the cytoplasmic linkers between segments S4 and S5 (S4-S5) in domains III and IV of human brain sodium channels in SDS micelles.

The two cytoplasmic linkers connecting segment S4 and segment S5 (S4-S5 linker) of both domain III (III/S4-S5) and IV (IV/S4-S5) of the sodium channel alpha-subunit are considered to work as a hydrophobic receptor for the inactivation particle because of the three hydrophobic amino acids of Ile-Phe-Met (IFM motif) in the III-IV linker of the sodium channel alpha-subunit. To date, the solution structures of the peptides related to III/S4-S5 (MP-D3: A1325-M1338) and IV/S4-S5 (MP-D4: T1648-L1666) of human brain sodium channels have been investigated using CD and (1)H NMR spectroscopies. SDS micelles were employed as a solvent. The micelles mimic either biological membranes or the interior of a protein and can be a relevant environment at the inactivated state of the channels. It was found that the secondary structures of both MP-D3 and MP-D4 assume alpha-helical conformations around the N-terminal half-side of the sequences, i.e. the residues between V1326 and L1331 in MP-D3 and between L1650 and S1656 in MP-D4. Residue A1329 in MP-D3, which is considered to interact with F1489 of the IFM motif, was found to be located within the alpha-helix. Residues F1651, M1654, M1655, L1657 and A1669 in MP-D4, which also play an important role in inactivation, formed a hydrophobic cluster on one side of the helix. This cluster was concluded to interact with the hydrophobic cluster due to the III-IV linker before the inactivation gate closes.

Amino Acid Motifs↗

PubMed2001

Decreased plasma activity of antithrombin or protein C is not due to consumption coagulopathy in septic patients with disseminated intravascular coagulation.

We investigated whether depressed plasma antithrombin and protein C activity, considered as a specific finding of disseminated intravascular coagulation (DIC), is due to consumption coagulopathy in septic patients with DIC. An analysis of hemostatic parameters was performed in 139 septic patients (68 with DIC and 71 without DIC). Plasma activity of antithrombin and protein C tended to be significantly decreased in septic patients with DIC but not in those without DIC (p < 0.001). However, when the septic patients were classified into three groups according to the albumin (or choline esterase) level, no significant differences in antithrombin activity or protein C activity were observed between the patients with and without DIC in any of the subgroups. Notably, neither the plasma activity of antithrombin nor protein C was decreased even in septic patients with DIC who had normal plasma levels of albumin (or choline esterase). No significant correlation was observed between plasma levels of thrombin-antithrombin complex (TAT) and antithrombin activity, or between plasma levels of TAT and protein C activity either in septic patients with DIC or without DIC. It is reasonable to conclude that the markedly reduced plasma activity of antithrombin and protein C is not due to consumption coagulopathy in septic patients with DIC.

Aged↗

PubMed2001

Inhibitory effect of interleukin 3 on early development of human B-lymphopoiesis.

Interleukin 3 (IL-3) is known as a stimulator of proliferation and differentiation of non-lymphoid cells, but information about the activity of IL-3 in lymphopoiesis is limited. In the present study, we examined the effect of IL-3 on human B-lymphopoiesis using the co-culture system on a murine stromal cell line, MS-5, with stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF). Although a large number of CD45+CD19+ B cells were generated in the co-culture of cord blood CD34+ cells, the addition of IL-3 to the co-culture suppressed the B-cell generation in a dose-dependent manner. In the co-culture, CD34+IL-3 receptor alpha-chain (IL-3Ralpha)+ cells, but not IL-3Ralpha- cells, produced B cells, and IL-3 suppressed the B-cell generation from CD34+IL-3Ralpha+ cells, suggesting that IL-3 exerts an inhibitory effect through the binding to IL-3Ralpha on CD34+ cells. Because the delayed addition of IL-3 and the short-term exposure to IL-3 showed that IL-3 acted as an inhibitor at an early stage of B-cell development from CD34+ cells, before the generation of CD19+ B cells, the effect of IL-3 on the early development of B cells from immature CD34+CD38- cells was examined. Individual colonies, produced from clone-sorted CD34+CD38- cells by SCF, thrombopoietin and a complex of IL-6/soluble IL-6R with or without IL-3, were divided into two fractions and analysed for B-cell potential using co-culture on MS-5 with SCF and G-CSF, and haematopoietic potential using methylcellulose clonal culture. Although some colonies cultured without IL-3 showed both B-cell and haematopoietic potential, all the colonies cultured with IL-3 showed only haematopoietic potential. These results indicate that IL-3 has an inhibitory effect on the B-cell generation from human lymphohaematopoietic progenitor cells.

Animals↗

PubMed2001

Embryological consideration of drainage of the left testicular vein into the ipsilateral renal vein: analysis of cases of a double inferior vena cava.

The right gonadal vein (GV=testicular vein in men, ovarian vein in women) usually drains into the inferior vena cava (IVC) while the left gonadal vein drains into the left renal vein (RV). This anatomical difference induces relatively weak haemodynamics in the left testicular vein (TV) and is considered to be a cause of a left varicocele. In textbooks on embryology, it has been documented that bilateral supracardinal veins (=origin of right and left IVC) and the subcardinal sinus (=origin of RVs and GVs) symmetrically develop during early embryogenesis. However, persistence and regression of the right and left supracardinal veins, respectively, results in drainage of the left GV into the ipsilateral RV. A double IVC (DIVC) commonly originates from a failure of disappearance of the left supracardinal vein. Although there have been a considerable number of case reports on DIVC, little attention has been paid to the anatomy of the left GV in such cases. We report here an autopsy case, a 72-year-old Japanese man, with a DIVC. This case belongs to type BC of McClure and Butler's classification. In this case, it was observed that the right TV drained into the confluence of the right IVC with the ipsilateral RV, while the left TV drained into the left RV in spite of the presence of the left IVC. This case indicates that the embryonic anastomosis point between the subcardinal sinus and the supracardinal vein on the left side is different from that on the right side. Statistical analysis of many case reports of DIVC also suggests that the bilateral supracardinal veins tend to asymmetrically anastomose with the subcardinal sinus during embryogenesis. These data imply that drainage of the left GV into the ipsilateral RV leads to regression of the left supracardinal vein but also to asymmetrical anastomosis between the supracardinal veins and the subcardinal sinus.

Aged↗

PubMed2001

Doppler index and plasma level of atrial natriuretic hormone are improved by optimizing atrioventricular delay in atrioventricular block patients with implanted DDD pacemakers.

Doppler index is the sum of isovolumetric contraction time and isovolumetric relaxation time divided by ejection time and has clinical value as an index of combined systolic and diastolic myocardial performance. This crossover study compared the Doppler index and atrial natriuretic hormone (atrial natriuretic peptide) [ANP] between optimal (AV) delay and prolonged AV delay in patients with DDD pacemakers. The study included 14 patients (6 men, 8 women, age 78.4+/-9.3 [SD] years) with AV block with an implanted DDD pacemaker. AV delay was prolonged in a 25-ms, stepwise fashion starting from 125 ms to 250 ms. Pacing rate was set at 70 beats/min. Cardiac output (CO) was assessed by pulsed Doppler echocardiography, and optimal AV delay was defined as the AV delay at which CO was maximum, and an AV delay setting of 250 ms as prolonged AV delay. Plasma level of ANP and Doppler index determined by echocardiography were measured 1 week after programming. AV delay was switched to another AV delay and measurements were repeated after 1 week. Optimal AV delay was 159+/-19 ms. Doppler index was significantly lower at optimal AV delay than at prolonged AV delay (0.68+/-0.26 vs 0.92+/-0.30, P < 0.05). The plasma ANP level was significantly lower at optimal AV delay than at prolonged AV delay (29.0+/-30.7 vs 52.6+/-44.9 pg/mL, P < 0.05). In conclusion, the Doppler index and the plasma ANP level were significantly lower at optimal AV delay than at prolonged AV delay. This study shows the importance of the optimal AV delay setting in patients with an implanted DDD pacemaker, the Doppler index and plasma ANP levels are good indicators for optimizing AV delay.

Aged↗

PubMed2001

Combined use of sphenoidal electrodes and the dipole localization method for the identification of the mesial temporal focus.

UNLABELLED: We attempted to sub-classify four cases who show temporal spikes on standard scalp electroencephalogram (EEG), using sphenoidal electrodes and the dipole localization METHOD: In a case with mesial temporal epilepsy, spikes showed phase reversal in a sphenoidal electrode, and the spike dipoles were estimated to be in the mesial temporal lobe. In a case with lateral temporal epilepsy, spikes showed no phase reversal in a sphenoidal electrode, and the spike dipoles were estimated to be in the lateral temporal lobe. In two cases out of four, spikes showed phase reversal in sphenoidal electrodes, whilst the dipoles were estimated to be in the frontal lobe. Clinical features also suggested a diagnosis of frontal lobe epilepsy. In one of the two cases in which frontal lobe epilepsy was suspected, ictal dipoles as well as interictal spike dipoles indicated participation of the frontal lobe in the genesis of seizures. Nevertheless, only mesial temporal lobectomy was performed based on results obtained by invasive subdural electrodes. As a result, seizures were not controlled. Although sphenoidal electrodes were useful for differentiating between mesial and lateral temporal lobe foci, it is advisable to use them in combination with the dipole localization method to identify frontal lobe foci.

Action Potentials↗

PubMed2001

Clinical application of plasma clearance of iohexol on feline patients.

Glomerular filtration rate (GFR) was estimated by plasma clearance of iohexol (PCio) in 52 conscious cats presented for a variety of reasons to Angel Animal Hospital over a 2-year period. Cats were divided into four groups according to their clinical conditions and reasons for measuring PCio. The median PCio (ml/min/kg) was 3.68 in normal cats (NM), 2.39 in cats with suspected renal disease (SP), 1.35 in cats referred to confirm renal dysfunction (RD), and 0.84 in cats with apparent clinical signs of renal failure (RF). There was a significant difference between the results for each group. The respective medians of blood urea nitrogen (BUN) and plasma creatinine concentration (Pcr) (mg/dl) were 15 and 1.40 in NM cats, 21 and 1.71 in SP cats, 30 and 2.20 in RD cats, and 48 and 3.30 in RF cats. The reference values of BUN and Pcr were 21 +/- 7 mg/dl and 1.5 +/- 0.4 mg/dl (mean +/- SD). Diminished renal function could not be detected in SP cats by either BUN or Pcr, while a marked decrease of GFR was demonstrated before BUN and Pcr increased, indicating the insensitivity of BUN and Pcr in detecting renal dysfunction in cats. PCio can be performed non-invasively in conscious cats, which improves the veterinarian's ability to detect early stages of chronic renal disease.

Animals↗

PubMed2001

Matrix metalloproteinase-2 in synovial lavage fluid of patients with disorders of the temporomandibular joint.

We examined the matrix metalloproteinase-2 (MMP-2) activity in synovial lavage fluid of patients with disorders of the temporomandibular joint (TMJ) and explored the possible correlationship between MMP-2 activity and radiological changes. We studied 86 patients and 10 healthy volunteers. An arthrogram and a double contrast arthrotomogram were taken to evaluate intra-articular morphological changes. The patients were divided into three groups: no abnormality (n = 36), internal derangement (n = 39), and osteoarthritis (n = 11). Samples of synovial fluid were studied by gelatin zymography, and we sought a correlation between the band detected and radiological findings. ProMMP-2 was detected in all samples and active MMP-2 was detected in 9/36 with no abnormality, 14/39 with internal derangement and 5/11 with osteoarthritis. No active form of MMP-2 was detected in the control group. The incidence of active MMP-2 was high in the internal derangement group and highest in the osteoarthritis group, which suggests that active MMP-2 plays an important part in the development of conditions of the TMJ.

Adolescent↗

PubMed2001

Microcirculatory alterations in a Mongolian gerbil sinus-vein thrombosis model.

BACKGROUND AND PURPOSE: The pathophysiology of sinus vein thrombosis (SVT) is still controversial in patients and experimental animals, the microcirculatory alterations in particular. This study was designed to develop a new sinus vein thrombosis model and to further elucidate pathophysiological events such as the relationship between local and regional cerebral blood flow and haemoglobin oxygen saturation (HbSO2), changes of the microvasculature, leukocyte behaviour and brain tissue damage. METHODS: In a first experimental series, animals were divided into two groups which resulted from different procedures of inducing SVT. In the SSS middle occlusion group (SMO group), SVT was induced by the ligation of the superior sagittal sinus right in the middle between the bregma and the confluence sinum. In the SSS posterior occlusion group (SPO group) the ligation was performed close to the confluence sinum. Regional cerebral blood flow (rCBF) was assessed at 36 identical locations by laser Doppler flowmetry together with regional haemoglobin oxygen saturation (HbSO2). In a second series of experiments SVT was induced by ligation of the SSS close to the confluence sinum (SVT group) to study effects on the cortical microcirculation. A sham operation was performed in six animals (sham group). In both groups, an intravital microscopic double tracing technique was utilised for evaluating microvessel structures and leukocyte behaviour. The images were recorded on videotape for evaluating alterations of microvessel (venules, arterioles and capillaries) diameters and numbers of leukocyte rollers and stickers by a digital video analyser. Animals were sacrificed for histological evaluation after 5 days. RESULTS: The posterior sinus ligation caused a significant decrease of rCBF and HbSO2 and brain tissue damage which was not seen in the SMO group. Alteration of rCBF and HbSO2 were positively correlated with infarct size in the SPO group only, where venous infarction was easily reproduced. Therefore, it is suggested that this model is suitable for studying SVT in Mongolian gerbils. Intravital microscopy of the cortical microcirculation revealed no significant changes of vessels diameter in the sham group, whereas a significant dilation of veins and capillaries was seen in the SVT group. Numbers of leukocyte rollers and stickers were positively correlated with infarct size. CONCLUSION: Microcirculatory alterations and brain tissue damage from SVT in the Mongolian gerbil depend on the SSS occlusion site. The newly established mongolian gerbil sinus-vein thrombosis model has advantages compared to previously reported sinus-vein thrombosis models such as easy handling, easy technique, highly reproducibility, and good observation of microcirculatory event. The model allows for studies of cerebral low-flow conditions such as expected to occur in an ischaemic penumbra zone.

Animals↗

PubMed2001

Kalopanaxsaponin A is a basic saponin structure for the anti-tumor activity of hederagenin monodesmosides.

Hederagenin, delta-hederin [hederagenin alpha-L-arabinoside], kalopanax-saponin A [hederagenin 3-O-alpha-L-rhamnosyl(1-->2)-alpha-L- arabinoside], kalopanaxsaponin I [hederagenin 3-O-beta-D-xylosyl(1-->3)-alpha-L- rhamnosyl(1-->2)-alpha-L-arabinoside], and sapindoside C [hederagenin 3-O-beta-D-glucosyl(1-->4)-beta-D-xylsyl (1-->3)-alpha-L-rhamnosyl(1-->2)-alpha-L-arabinoside] were isolated from stem bark of Kalopanax pictus Nakai (Araliaceae). Among glycosides of hederagenin, disaccharide (kalopanaxsaponin A, commonly also called alpha-hederin), trisaccharide (kalopanaxsaponin I), and tetrasaccharide (sapindoside C) showed significant cytotoxicity on several types of tumor cells, while hederagenin itself exhibited only weak cytotoxicity and its monosaccharide (delta-hederin) was non-cytotoxic. From these results, it suggests that the arabinosyl moiety at C-3 blocks the activity of hederagenin and the position of the second sugar for glycoside linkage is also important for cytotoxicity. In the in vivo experiments, kalopanaxsaponin A (15 mg/kg, i.p.) apparently increased the life span of mice bearing Colon 26 and 3LL Lewis lung carcinoma, as well as cisplatin (3 mg/kg, i.p.). These results indicated that kalopanaxsaponin A has potential anti-tumor applications.

Animals↗

PubMed2001

Comparative biokinetics of tritium in rats during continuous ingestion of tritiated water and tritium-labeled food.

PURPOSE: The biokinetics of tritium during continuous ingestion of tritiated water and tritiated wheat were investigated to estimate the radiation dose rates at the end of two modes of chronic exposure. MATERIALS AND METHODS: Wistar strain male rats continuously ingested tritiated water as drinking water and tritiated wheat as food for 14 weeks. Urine and tissue samples were obtained and total tritium in the fresh wet samples and organically bound tritium (OBT) in the freeze-dried samples were determined. RESULTS: The biokinetics of tritium was different between the two modes of exposure. The concentration of total tritium in the tissues exposed to tritiated water attained a steady-state condition by 2-3 weeks. The steady-state condition in the case of exposure to tritiated wheat was not observed for 10 weeks after the start of exposure in the majority of tissues. The relatively efficient and prolonged OBT formation during chronic exposure to tritiated wheat resulted in relatively high incorporation and retention of tritium in the tissues compared with those for exposure to the same activity of tritiated water. CONCLUSION: Radiation dose rates estimated at the end of continuous ingestion showed that tritiated wheat gave higher dose rates than tritiated water by a factor of 1.3 to 4.5, but the factors were within 2.0 in the majority of tissues except for small intestine and adipose tissue.

Administration, Oral↗

PubMed2001

Cloning and functional expression of an E box-binding protein from rat granulosa cells.

Ovarian granulosa cells undergo cell growth and cytodifferentiation during follicular maturation. In a number of tissues, the gene expression that is responsible for the cytodifferentiation is largely dependent on E box(es) located upstream of the responsible genes. In this study, we report on the cloning of cDNA(s) encoding E box (5'-CACGTG-3')-binding protein from a rat granulosa cell cDNA library using a yeast one-hybrid system. When multiple E box sequences were used as target, we obtained a positive clone that encodes the rat homologue of upstream stimulatory factor 2 (USF2). An analysis of the nucleotide sequence and its deduced amino acid sequence reveals that rat USF2 protein consists of 346 amino acid residues and belongs to the basic helix-loop-helix/leucine zipper protein family. Northern blot analysis shows that rat USF2 mRNA exists as multiple forms between 1.6 and 2.2 kilobases. The size of the cloned insert was identical to that of the transcript of maximal length. Electrophoretic mobility shift assays showed that in vitro-translated rat USF2 specifically binds to the E box. In addition, cotransfection experiments with luciferase-reporter constructs in HepG2 cells reveal that the overexpression of rat USF2 leads to an increase of luciferase activity in the E box sequence-dependent manner. Thus, we report molecular cloning, expression, and functional characterization of full-length rat USF2 cDNA.

Amino Acid Sequence↗

PubMed2001

Elevated levels of free tissue factor pathway inhibitor antigen in cases of disseminated intravascular coagulation caused by various underlying diseases.

Tissue factor pathway inhibitor (TFPI) is primarily synthesized by vascular endothelial cells and is found in vivo in association with endothelial cells, lipoproteins, or in free form. Free TFPI is the most potent and important type, because it is released from endothelial cells following an injection of heparin, or as a result of pathological stimuli. In order to study the role of TFPI in disease, the concentration of free form TFPI was measured in the plasma of 114 patients suffering from disseminated intravascular coagulation (DIC), as the result of several underlying diseases. Plasma antigen levels of free TFPI were significantly higher even in those patients not exhibiting DIC than in normal healthy subjects. These levels were even higher among patients exhibiting DIC, especially those with acute promyelocytic leukemia or cancer, receiving continuous heparin drip infusions. A significant correlation was observed between the plasma antigen levels of free form TFPI and those of fibrin/fibrinogen degradation products, and free form TFPI and plasmin inhibitor complex (r = 0.428, P < 0.0001 and r = 0.329, P < 0.0001, respectively) among 114 DIC patients. There were no significant differences between the plasma levels of free TFPI in DIC patients with or without multiple organ failure. It has been suggested that the plasma levels of free TFPI are closely related to the levels of fibrinolysis occurring in DIC patients, although further study is required to clarify the degree to which TFPI is expressed by endothelial cells during DIC.

Case-Control Studies↗

PubMed2001

Quantification of limited augmentation of myocardial (99m)Tc-tetrofosmin uptake at exercise in stable coronary artery disease.

We evaluated the diagnostic accuracy of a new method for quantitative analysis of myocardial perfusion at exercise using (99m)Tc-tetrofosmin tomographic imaging. (99m)Tc-tetrofosmin imaging of an exercise-rest sequence was performed in 30 patients with coronary artery disease (CAD) and eight age-matched control subjects. The exercise-induced myocardial count increase was calculated as the relative value (per cent increase ratio, %IR) to that at rest by correcting for physical decay for (99m)Tc and injected doses and by a subtraction technique. Exercise and rest (99m)Tc-tetrofosmin images were assessed visually and quantitatively using the per cent peak count and %IR of the myocardial count at exercise. Segments with significant coronary stenosis (diameter stenosis=75% or more) showed a significantly lower %IR than did those without significant coronary stenosis in the CAD patients (37+/-19% vs 63+/-21%, P<0.05). The diagnostic efficacy of visual analysis for detecting coronary stenosis was as follows: sensitivity, 58.1% and specificity, 81.4%. When %IR=37% was used for detecting significant coronary stenosis, sensitivity and specificity increased to 74.2% and 93.2%, respectively. Furthermore, the quantitative analysis significantly (P=0.04) improved the overall diagnostic accuracy from 73.3% to 86.7% compared to that of visual assessment. Thus, augmentation of myocardial (99m)Tc-tetrofosmin uptake at exercise is blunted in the myocardium with significant coronary stenosis. The calculation of myocardial count increase at exercise relative to that at rest can improve the diagnostic value of (99m)Tc-tetrofosmin SPECT imaging and may contribute to more accurate quantification of myocardial ischaemia and impaired tracer uptake in coronary artery disease.

Adult↗

PubMed2001

Octreotide for severe acute bleeding from portal hypertensive colopathy: a case report.

A 56-year-old man developed severe lower gastrointestinal bleeding. He was classified as Child-Pugh grade C. Colonoscopy revealed multiple angiodysplasia-like lesions and mucosal friability throughout the entire colon (portal hypertensive colopathy, PHC). Haemostasis was immediately achieved with octreotide treatment, although melaena recurred after discontinuation of the infusion. Propranolol treatment before discontinuation of octreotide infusin prevented the recurrence of bleeding from PHC. Octreotide is a safe and effective treatment for severe acute bleeding from PHC, especially if the patient is not a candidate for transjugular intrahepatic portosystemic shunt (TIPS) or treatment with a beta-blocker due to the severity of liver disease or haemodynamic instability. However, a sufficient reduction of portal pressure by propranolol or other medical treatment may be needed in order to discontinue octreotide infusion without the recurrence of bleeding.

Acute Disease↗

PubMed2001

Enhancement of in vivo antitumor activity of a novel antimitotic 1-phenylpropenone derivative, AM-132, by tumor necrosis factor-alpha or interleukin-6.

TK5048 and its derivatives, AM-132, AM-138, and AM-97, are recently developed antimitotic (AM) compounds. These 1-phenylpropenone derivatives induce cell cycle arrest at the G2 / M phase of the cell cycle. TK5048 inhibited tubulin polymerization in human lung cancer PC-14 cells in a concentration-dependent manner. In a polymerization assay using bovine brain tubulin, AM-132 and AM-138 were quite strong, AM-97 was moderately strong, and TK5048 was a relatively weak inhibitor of tubulin polymerization. A murine leukemia cell line resistant to a sulfonamide antimitotic agent, E7010, which binds to colchicine-binding sites on tubulin, was cross-resistant to the in vitro growth-inhibitory effect of AM compounds. Inhibition of tubulin polymerization is therefore one of the mechanisms of action of these AM compounds against tumor cells. To profile the antitumor effect of AM compounds, the in vivo antitumor effect of AM-132 was evaluated against cytokine-secreting Lewis lung carcinoma (LLC). Tumor-bearing mice were treated with intravenous AM-132 using three different treatment schedules. LLC tumors expressing tumor necrosis factor-alpha (TNF-alpha), granulocyte macrophage colony-stimulating factor (GM-CSF), or interleukin (IL)-6 were very sensitive to AM-132. In particular, LLC tumors expressing IL-6 were markedly reduced by AM-132 treatment, and showed coloring of the tumor surface and unusual hemorrhagic necrosis. These results suggest a combined effect of AM-132 and cytokines on the blood supply to tumors.

Animals↗

PubMed2001

Solution structures of the inactivation gate particle peptides of rat brain type-IIA and human heart sodium channels in SDS micelles.

The Ile-Phe-Met (IFM) motif located in the Ill-IV linker of voltage-gated sodium channels has been identified as a major component of the fast inactivation gate. If Gln was substituted for Phe, the role in the gate was disrupted completely. If Ile was replaced by Gln inactivation became slightly incomplete and if the Thr, which is adjacent to the IFM motif (-IFMT-), was replaced by Met, inactivation became much more incomplete than in the I/Q mutation, but not as vigorous as in the F/Q mutation. Previously, we studied the structures of the inactivation gate-related peptide (K1480-K1496 in rat brain type-IIA, MP-3A) and its F1489/Q substituted one (MP-4A) in SDS micelles and found that the conformational change of the IFM hydrophobic cluster due to the F/Q substitution may be a reason for disrupting the gate. In this study, in order to obtain supporting evidence for this view and also to further knowledge of the effect of I/Q and T/M mutations on the structure of the IFM cluster, we studied the structures of 11488Q [MP(rb)-3QFMT] and T1491M [MP(rb)-31FMM] substituted peptides. The fragment peptide K1477-K1493 [MP(hh)-3A] and its T1488M substituted peptide [MP(hh)-3IFMM] in the human heart sodium channel were also studied. It was found that the backbone structures around the IMF motif of MP-3A, MP(hh)-3A and MP(rb)-3QFMT resemble one another in such a manner that the residues Ile(Gln) and Thr are brought so close together that they form a unique type of lid to occlude the pore. In contrast, the residues between Ile and M1491 of MP(rb)-3IFMM or M1488 of MP(hh)-3IFMM were fairly far apart from each other. We conclude that Thr plays an important role in forming a structure of the IFM hydrophobic cluster for inactivation.

Amino Acid Motifs↗