[Enzyme inhibitors--current status and future scope].
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Animals↗
Biomedical subjects
R Kobayashi
Publications and source records attributed to R Kobayashi.
[Significance of the 14q32 translocations in childhood acute lymphoblastic leukemia].
To assess the frequency and significance of 14q32 translocation abnormalities in childhood acute lymphoblastic leukemia (ALL) and the differences between the clinical and cytogenetic features of patients with the 8; 14 translocation and those of patients with other 14q32 translocations, we analyzed our experience with 124 consecutive cases with completely banded karyotype. Eight cases (6.5%) with 14q32 translocation were identified :5 with the 8; 14 translocation and 3 with other 14q32 translocations. As compared with ALL children lacking 14q32 translocations, these 8 cases had a higher serum lactic dehydrogenase (LDH) level, more L3 (FAB classification), and a poorer outcome. On the other hand, in comparison with ALL patients with other 14q32 translocations, patients with the 8:14 translocation were likely to be younger (median age 4.5 years vs 10.4 years), to have a higher serum LDH level (median 5832 IU/l vs 504 IU/l), to have more L3 (3/5 vs 0/3), to have a higher induction failure rate (4/5 vs 1/3), and to have more partial duplication of the long arm of chromosome 1 (4/5 vs 0/3). These results helped clarify the characteristic features of ALL children with 14q32 translocations and showed that ALL children with the 8 ; 14 translocation have different clinical and cytogenetic findings from those of ALL children with other 14q32 translocations.
Adolescent↗
[Nonoliguric acute renal failure complicated with tumor lysis syndrome in chronic myelocytic leukemia in lymphoid crisis].
A case of nonoliguric acute renal failure complicated with tumor lysis syndrome is described. The patient is a 14-year-old boy who was diagnosed chronic myelocytic leukemia 17 months ago. On lymphoid crisis, he received vindesine-prednisolone therapy and acute renal failure occurred. Urine output was kept enough volume (2,500-4,000 ml/day), but blood urea nitrogen and serum creatinine levels rose and hyperkalemia, hyperphosphatemia and hypocalcemia were observed. Tumor lysis syndrome in patients with chronic myelocytic leukemia is rare, and acute renal failure with tumor lysis syndrome is oliguric or anuric in most patients. At therapy of lymphoproliferative disease, nonoliguric acute renal failure may occur. Physicians who treat patients with lymphoproliferative disease should pay attention to blood urea nitrogen and serum creatinine levels even if urine output is satisfactory.
Acute Kidney Injury↗
Three-step purification method and characterization of the bovine brain 90-kDa heat shock protein.
A protein that cross-reacted with antibody against the 90-kDa heat shock protein (HSP90) of a mouse lymphoma cell line was purified from bovine brain by three steps. Fifty milligrams of the 90-kDa protein was recovered from 350 g of the brain cortex. The sedimentation coefficient and Stokes radius of the purified protein were 6.0 s and 6.7 nm, respectively. The molecular weight was calculated to be 170,000. The molecule was composed of two identical 90-kDa subunits. A partial amino acid sequence (23 residues) of this protein was homologous (96%) to human HSP90 (the sequence of 174-196). These facts led to the identification of the 90-kDa brain protein with HSP90. In bovine tissues, the brain contained this protein at a remarkably high concentration. The brain HSP90 was separable from glucocorticoid receptor by heparin-agarose and DNA-cellulose columns. It is concluded that HSP90 is present in brain cytosol and mostly as free molecules. Immunohistochemical studies showed that the protein was localized in nerve excitable cells. It was not found in nuclei but in cytosol.
Amino Acid Sequence↗
Isolation and characterization of three distinct 34 kDa EDTA-extractable proteins from bovine lens.
EDTA-extractable protein (EEP) is known to be a major lens membrane protein with a molecular mass in the range 32 kDa to 38 kDa, and is also known to bind to the lens membrane and phospholipid-containing liposomes in a calcium-dependent manner. Recent results (Russell, P., Zelenka, P., Martensen, J., and Reid, T.W. (1977) Curr. Eye Res. 6, 533-538) on antibody cross-reactivity have demonstrated that a 34-35 kDa component of EEP is identical to calpactin I (lipocortin II). In this study, we have identified and purified three distinct 34 kDa components of EEP (designated as EEP-34A1, EEP-34A2 and EEP-34B) from bovine lens that inhibit phospholipase A2 activity. These proteins bind to phospholipid-containing liposome and F-actin in a calcium-dependent fashion. Two-dimensional electrophoresis demonstrates that the three proteins were distinct from one another. However, immunochemical studies and one-dimensional peptide mapping indicate that EEP-34A1 and EEP-34B are very similar. Our results also indicate that EEP-34A1 is very similar to calpactin II and that EEP-34A2 corresponds to calpactin I. The bovine lens 34-35 kDa component of EEP is a mixture of proteins rather than a single protein.
Actins↗
Purification, biological properties and partial sequence analysis of 67-kDa calcimedin and its 34-kDa fragment from chicken gizzard.
Calcimedin is a group of proteins, originally isolated from chicken gizzard, which are able to bind to several hydrophobic matrices in the presence of Ca2+. Although the molecular properties have been partially discovered, the physiological functions of calcimedins have not yet been clearly defined. In this study, we describe the isolation and characterization of 67-kDa calcimedin and its 34-kDa fragment from chicken gizzard. Both structural and functional studies establish that 67-kDa calcimedin is a member of the calpactin/lipocortin family: it displays phospholipase A2 inhibitory activity, Ca2(+)-dependent F-actin binding and phospholipid binding activity similar to those of calpactins (lipocortins). By comparing the sequence of 67-kDa calcimedin with the predicted sequence of 67-kDa calelectrin, we concluded that the primary structure of these 67-kDa proteins is highly conserved. In particular, the sequences GLGTDEGAIIXVLTQR and EGAGTDESTLIEIMATR conform with the annexin consensus sequence which is characteristic of the calpactin/lipocortin family. A 34-kDa fragment of 67-kDa calcimedin was also purified and their relatedness has been confirmed by antibody cross-reactivity. The sequence data further support that the 34-kDa fragment is derived from the C-terminal portion of 67-kDa calcimedin by limited proteolysis. The 34-kDa fragment, which contains the annexin consensus sequence, preserves the phospholipase A2 inhibitory activity, and binds F-actin and phospholipids.
Actins↗
Identification of the 32 kDa components of bovine lens EDTA-extractable protein as endonexins I and II.
EDTA-extractable protein (EEP) is a mixture of major lens membrane proteins with molecular masses ranging from 32 kDa to 40 kDa. These bind to the lens membrane in a Ca2(+)-dependent manner. In the present study we have identified and purified two distinct 32 kDa components of EEP (designated as EEP 32-1 and EEP 32-2) from bovine lens that inhibit phospholipase A2 activity. Both EEP 32-1 and EEP 32-2 bind to phospholipid-containing liposomes and actin filaments in a Ca2(+)-dependent fashion. Immunochemical studies and two-dimensional electrophoreses demonstrate that the two proteins are distinct from one another. Both EEP 32-1 and EEP 32-2 are clearly different from calpactin (lipocortin) or its proteolytic fragments because they did not react with anti-[human placenta calpactin (lipocortin)] antibody. Our results also indicate that EEP 32-1 is very similar to endonexin I and that EEP 32-2 corresponds to endonexin II.
Actins↗
Purification, characterization, and partial sequence analysis of 32-kDa calcimedin from chicken gizzard.
Calcimedin is a group of proteins which has a binding ability to several hydrophobic matrices or cellular membrane fractions in the presence of Ca2+. Although the molecular properties were partially clarified, the physiological functions of calcimedins have not been clearly defined. In this study, we describe the isolation and characterization of 32-kDa calcimedin from chicken gizzard. Both structural and functional studies establish that 32-kDa calcimedin is a member of the calpactin/lipocortin family. The 32-kDa calcimedin displays phospholipase A2 inhibitory activity, Ca2(+)-dependent F-actin binding activity, and phospholipid binding activity similar to those of calpactins/lipocortins. Antiendonexin II antibody recognized 32-kDa calcimedin. However, antibodies against calpactin I (lipocortin II), calpactin II (lipocortin I), 35-kDa calcimedin, and 67-kDa calcimedin did not cross-react with 32-kDa calcimedin. One-dimensional peptide maps of the 32-kDa calcimedin and the 35-kDa calcimedin are different, confirming that they are distinct proteins. By comparing the sequence of 32-kDa calcimedin with the predicted sequence of endonexin II, we concluded that the primary structure of the 32-kDa protein is highly conserved. In particular, the sequences AMKGMGTDDEXEIXL, GMGTDEEEIL, VLTEILASR, and ILTSR conform to the endonexin consensus sequence, which is characteristic of the calpactin/lipocortin family.
Actins↗
In situ hybridization of muscle mitochondrial mRNA in mitochondrial myopathies.
To determine whether a mitochondrial mRNA deficiency exists in mitochondrial myopathies, muscle biopsies from a patient with chronic progressive external ophthalmoplegia (CPEO) and a patient with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) were studied using in situ hybridization. Histochemistry and immunohistochemistry were performed along with hybridization. Hybridization reactions were widely distributed over the sarcoplasm of all muscle fibers in the patient with MELAS. In the patient with CPEO, 80% of the fibers showed a marked decrease in density of autoradiographic grains. This marked decrease corresponded to the histochemical and immunohistochemical findings of a very weak staining of cytochrome c oxidase (CCO). The isotope-labeled cDNA probe used in in situ hybridization in this study complements a part of subunit I of CCO and a part of subunit II of complex I in the mitochondrial gene. Our results suggest a defect in the mRNA in this CPEO patient.
Adult↗
Simple and rapid method for purification of vinculin from bovine aorta.
A new rapid purification procedure has been developed for mammalian smooth muscle vinculin. Bovine aorta vinculin has been purified by a two-step procedure employing hydrophobic chromatography on phenyl-Sepharose and gel filtration. This procedure should be particularly useful for purifying vinculin from tissues in which it is present in a relatively low concentration. In addition, the new purification protocol provides highly pure vinculin free from active contaminants which reduce the low-shear viscosity of F-actin solution.
Amino Acids↗
Isolation and some properties of bovine brain 100 kDa heat shock protein.
1. The 100 kDa protein was purified from bovine brains. 2. The antibody against the 100 kDa brain protein was prepared and was monospecific to the antigen. 3. The antibody cross-reacted with HeLa cell HSP100 (100 kDa heat shock protein). 4. The physicochemical, immunochemical properties and a partially amino acid sequence indicated that the 100 kDa protein was HSP100. 5. Peptide mapping using Staphylococcus aureus V8 protease showed a core peptide with 10 kDa molecular mass common to both HSP100 and HSP90. 6. The amino acid sequence of the 10 kDa fragment of the 100 kDa protein showed a high homology with that of human HSP90 (38-60); the difference was only two of 23 amino acid residues determined.
Amino Acid Sequence↗
The distribution of serotonin immunoreactivity in the rat locus ceruleus after intraventricular injections of either 5,6- or 5,7-dihydroxytryptamine with special reference to serotonin synthesis.
The localization of serotonin-immunoreactivity (5-HT-IR) in the locus ceruleus (LC) of rats was studied by the peroxidase-anti-peroxidase method using a purified antibody obtained from a rabbit. Antibody production was performed according to the method of Grota and Brown (1974). The antibody was applied to serial cryostat sections with alternate counterstaining by cresyl violet, after intraventricular injections of 5,6-dihydroxytryptamine or 5,7-dihydroxytryptamine prior to treatment with pargyline and a precursor of 5-HT. The majority of LC neurons were immunopositive, and more than half of all LC neurons clearly showed 5-HT-IR. Although core cells were the most predominant, all types of neurons were immunopositive, and randomly scattered throughout the LC. The uptake inhibitor, Lilly 110140, administered in sufficient amounts prior to an injection of pargyline, did not reduce 5-HT-IR within the LC. The results suggest that LC neurons receive 5-hydroxytryptophan (5-HTP) through an afferent vascular-neuronal channel and/or by diffusion from blood capillaries much more than 5-HT itself. We consider from these results that all types of LC neurons throughout the nucleus are masked 5-HT cells, and that the majority of LC neurons utilize blood-borne 5-HTP as an immediate precursor for intraneuronal 5-HT synthesis.
5,6-Dihydroxytryptamine↗
Detection and isolation of a 30 kDa abnormal protein in avian dystrophic muscle.
We have studied the protein composition of the pectoralis superficialis muscle of genetically dystrophic (New Hampshire line 413) and normal control (line 412) chickens by one- and two-dimensional gel electrophoresis. A protein, referred to hereafter as the 30 kDa abnormal protein, was specifically detected in the affected muscle. It was purified to homogeneity, and its molecular properties were studied. It is a monomer with a molecular mass of approximately 30 kDa and an isoelectric point of about pI 8.4. We have screened by Western blotting a variety of muscles from line 412 and line 413 chickens for the presence of the 30 kDa protein. While the pattern of total protein is very similar in all cases, the 30 kDa protein was not detected in the pectoralis superficialis muscle of line 412 chickens. However, the immunoreactive bands were detected in the sartorius muscle and the tensor fasciae latae muscle from dystrophic and normal chickens. Interestingly, the immunoreactive bands of normal skeletal muscles are smaller in molecular weight than those of dystrophic skeletal muscles. To determine the early time sequence of the appearance of the abnormal protein, we studied muscles from embryos and post-hatched chickens at various ages. The abnormal protein was detected in dystrophic muscles as early as 15 days ex ovo and occurred throughout development up to six months ex ovo. Although the implication of the dystrophy-associated appearance of the 30 kDa protein in the affected muscle is not clear at present, it would be of particular interest to elucidate the biochemical functions of the 30 kDa protein in the affected muscle (pectoralis superficialis muscle) of genetically dystrophic chicken.
Animals↗
The 30 kDa abnormal protein in avian dystrophic muscle is indistinguishable from carbonic anhydrase III by physicochemical, immunological, and enzymological criteria.
In the accompanying paper, we described the existence, molecular characterization, and ontogeny of a 30 kDa abnormal protein in chicken dystrophic muscles. In this study, we have purified chicken carbonic anhydrase III and the 30 kDa protein and directly compared them. In terms of its enzymological features, the 30 kDa protein is a typical carbonic anhydrase III. Like carbonic anhydrases, it contains one mole zinc per mole of protein. The protein selectively cross-reacted with a chicken carbonic anhydrase III antibody. Antibody to the 30 kDa protein cross-reacted with chicken skeletal muscle carbonic anhydrase III. Moreover, the distribution of the abnormal protein is exactly identical to that of carbonic anhydrase III; however, there is a possibility that the 30 kDa protein is a variant of carbonic anhydrase III. Slight differences were found in antigenicities and in the apparent molecular weights of the two proteins. We have compared the two proteins by 125I-labeled two-dimensional peptide mapping. Tryptic maps have shown that the two proteins are highly homologous. Combined, these results strongly indicate that the 30 kDa protein and carbonic anhydrase III are similar, if not identical.
Animals↗
Phosphorylated and dephosphorylated types of non-activated glucocorticoid receptor.
We purified glucocorticoid receptors quickly but very partially using DEAE-resin. [3H]-Triamcinolone acetonide-labeled and non-activated receptors in the quickly purified fraction were found to be separated into two fractions (P-2 and P-3) by hydroxyapatite column chromatography. The P-2 receptor was the main component, and the ratio of P-2/P-3 was around 2. The molecular weights of the two receptors were calculated to be the same, 242,000: Rs = 6.2 nm and s20,w = 9.0. Treatment of the receptor with catalytic subunits of phosphoprotein phosphatase 2A1 reduced the P-2/P-3 ratio from 2 to 0.5, while treatment with catalytic subunits of cAMP-dependent protein kinase and ATP increased it to 2.5. The isolated P-3 receptor could be converted into the P-2 type by the kinase treatment. Tungstate, a phosphatase inhibitor, stabilized the P-2 receptor, and the P-2/P-3 ratio was larger than 3 when the DEAE-fraction was prepared in the presence of tungstate. However, the tungstate effect was not very strong, and the P-2 type tended to change into the P-3. [3H]-Triamcinolone acetonide-labeled and non-activated receptors were purified very highly by using an affinity gel; the procedure required more than 10 h. Only the P-3 form was observed in the preparation of highly purified receptors. Hormone-free receptors were affected by neither the phosphatase nor the kinase. The results indicate that the hormone binding makes the receptor sensitive to phosphatase. The reversibly dephosphorylated receptor is more stable than the non-dephosphorylated one, and can be activated.
Animals↗
[Determination of cadmium, cobalt, copper, nickel and lead in urine by inductively coupled argon plasma emission spectrometry after solvent extraction].
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Argon↗
Transient paraplegia caused by acute aortic dissection--case report.
A case of acute aortic dissection (AAD) presenting as sudden, transient paraplegia and severe back pain is reported. The patient was a 66-year-old male with a 10-year-history of hypertension. The pain characteristically migrated from the back to the neck and then returned to the back. He showed complete transverse myelopathy at the level of the 9th thoracic cord. Computed tomography disclosed internal displacement of aortic intimal calcifications, without abnormalities in the spinal canal, and myelography showed no spinal canal block or stenosis. Electrocardiography and chest x-ray indicated nonspecific changes of high amplitudes and mild cardiomegaly, respectively. Together, these findings suggested acute aortic dissection with spinal cord ischemia. The initial systolic blood pressure of 220 mmHg was lowered with medication, and the pain was controlled with morphine. He recovered fully and was discharged 80 days after the onset of symptoms, with no neurological deficits. AAD carries a very poor prognosis unless treated immediately. Therefore, it is very important to promptly differentiate this disorder from spinal vascular conditions that also produce back pain and paraparesis.
Aged↗
[A study of ultrasonic scaling in combination with povidone-iodine solution. (1)].
The effects of ultrasonic scaling in combination with povidone-iodine solution were examined clinically. The results obtained were as follows: 1. Three days after instrumentation, significant improvement was found in all experimental groups. Thereafter the improvement was at the same level and backing in each group. 2. Clinical findings, with exception of plaque index, showed improvement with ultrasonic scaling in combination with povidone-iodine solution in comparison with ultrasonic scaling alone and root planing.
Combined Modality Therapy↗