Biomedical subjects
Publications and source records attributed to S Hara.
Blood cysts of the heart in adults are extremely rare, and their origin is usually from the valvular apparatus. However, a case was encountered of a blood cyst originating from the right atrial septum of a 59-year-old woman. Furthermore the cyst contained calcium stone. The cyst with the stone was Successfully excised. Neither situation has been reported previously in the literature.
The positions of the reactive site and the disulfide bridges in fungal protease inhibitor F (FPI-F) from silkworm (Bombyx mori), which has a unique amino acid sequence and inhibitory specificity, were investigated. At pH 3.0, subtilisin BPN', which is one of target proteases of this inhibitor, specifically cleaved the peptide bond of the inhibitor at Thr(29)-Val(30). The cleaved bond was regenerated by subtilisin BPN' at pH 8.0. These results indicate that the Thr(29)-Val(30) bond of the inhibitor is the reactive site. The locations of disulfide bridges were determined to be Cys(3)-Cys(35), Cys(14)-Cys(27), Cys(18)-Cys(55), and Cys(37)-Cys(49). Based on the positions of the reactive site and the disulfide bridges, FPI-F is considered to be a member of a new family of serine protease inhibitors. We propose the designation Bombyx family for the new inhibitor family of which FPI-F is a member.
Fungal protease inhibitor F (FPI-F) from silkworm hemolymph is a novel serine protease inhibitor of the Bombyx family. The cDNA of FPI-F was introduced into a baculovirus vector and a recombinant virus was isolated and plaque-purified. The protease inhibitory activities increased in the culture medium of insect cells and in the hemolymph of silkworms infected with the recombinant virus. Judged from the behavior on ion-exchange and reversed-phase chromatographies, amino acid compositions, amino-terminal sequences, and CD spectra, the recombinant FPI-F was identical with native FPI-F. Infection with the recombinant virus caused inhibition of larval development of the silkworm. However, the degree of the effect was different in two strains, Shinryukaku and Taiheichoan, indicating that the selection of the strain of silkworm is important in using the baculovirus expression system.
The complete amino acid sequence of a white kidney bean (Phaseolus vulgaris) alpha-amylase inhibitor (PHA-I), which is composed of two kinds of glycopolypeptide subunits, alpha and beta, was established by conventional methods. The polypeptide molecular weight of PHA-I determined by the light-scattering technique, considered together with the sequence molecular weights revealed for the subunits, indicated that PHA-I has the subunit stoichiometry of (alpha beta)2 complex. Inhibition test of PHA-I with increasing amounts of porcine pancreatic alpha-amylase (PPA) suggested that an inactive 2:1 complex is formed between PPA and PHA-I. In fact, two complexes differing from each other in the molar ratio of PPA to PHA-I were separated by gel filtration, and molecular weight estimation by the light-scattering technique confirmed that they are complexes of PHA-I with one or two PPA molecules. The binding of PPA to PHA-I appeared to follow simple binomial statistics, suggesting that two binding sites on PHA-I are independent and of high affinity for PPA.
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A patient with short bowel syndrome (SBS) and renal failure developed a disturbance of consciousness with hyperammonemia. Abnormally low concentrations of ornithine, citrulline, and arginine were observed on the plasma aminogram. These results suggested that the activities of amino acid synthetase localized in the small intestinal flora were lost. The small intestine is required for arginine synthesis; thus, infusion limited to the essential amino acids to SBS patients will cause a deficiency of the urea cycle intermediates, ornithine, citrulline, and arginine and may lead to hyperammonemia. In addition, the renal insufficiency may have caused decreased excretion of ammonia. In this patient, supplemental arginine improved the symptoms.
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We could prepare the microsomal fraction of mouse liver, without using an ultracentrifuge but with a low speed centrifuge. The procedure includes 1) lyophilization of post-mitochondrial fraction (9,000 x g supernatant) of mouse liver, 2) powdering of the lyophilized sample, 3) the addition of 1.15 per cent potassium chloride solution or distilled water, which afforded microsomal aggregates, 4) sedimentation of microsomal fraction by low-speed centrifugation (20,000 x g, 20 min). The sedimented microsomal fraction showed normal contents of cytochrome P-450 and cytochrome b5, and gave a normal pattern on SDS polyacrylamide gel electrophoresis and normal electron microscopic feature. This method should be convenient for rapid and large-scale preparation of microsomes, especially for the preparation of cytochrome b5 and cytochrome P-450.
Nine patients underwent full thickness chest wall resection and reconstruction in our department between January 1981 and December 1994. There were chest wall recurrence of breast cancer in 5 cases, primary chest wall chondrosarcoma in 2, primary chest wall malignant fibrous histiocytoma in 1 and metastatic sternal renal cell carcinoma in 1. Seven of 9 cases underwent partial sternal resection. Sizes of chest wall defects were from 10 x 7 cm to 15 x 14 cm. In eight cases of 9, chest wall reconstruction was by double Marlex mesh repairs and various flaps (major pectoral muscle in 3, major pectoral myocutaneous flap in 2, latissimus dorsi myocutaneous flap in 1 and pedicled omentum in 1). The last case underwent repair with rectus abdominis myocutaneous flap without mesh. There was no operative death. Postoperative complications occurred in 4 patients: partial skin necrosis in 2, skin dehiscence in 1 and respiratory failure in 1. Eight cases are alive now from 9 months to 14 years and 8 months after chest wall reconstruction. One patient with metastatic renal cell carcinoma died of recurrence 3 years after operation. Full thickness chest wall resection and reconstruction is a safe operation and may provide a long-term survival.
Diabetic nephropathy can be regarded mainly as a type of microangiopathy, but is a disease that may also include aspects of macroangiopathy. This is especially true of renal disease in non-insulin dependent diabetes mellitus (NIDDM), which is characterized not only by diabetic glomerulosclerosis, but also by atherosclerosis. We performed morphological studies on the kidney, using computed tomography (CT), focusing on such points as: (1) abdominal aortic calcifications at the level of kidney, (2) calcifications in the renal artery, and (3) wedge-shaped defects on the renal surface. We noted that these findings became more prominent in NIDDM patients during end-stage renal failure than during normal renal function, and were significantly more common in those two NIDDM groups than in age-matched nondiabetic patients without hypertension, hyperlipidemia or gout. NIDDM patients exhibited these features more frequently than IDDM patients.
We have experienced a case of bilateral partial anomalous pulmonary venous return without atrial septal defect. Right upper pulmonary vein returned to the superior vena cava and left upper pulmonary vein returned to the left brachiocephalic vein. This type of bilateral partial anomalous pulmonary venous return was extremely rare, and only 4 such cases have been reported in the Japanese literature. The patient, aged 3 years and 11 months, was successfully operated on, using a direct anastomosis between the right atrial appendage and the superior vena cava. This procedure is considered to be advantageous because both this right atriotomy line and suture line avoid injury of the sinus node and the sinuatrial node artery and no foreign material is used. A decrease in postoperative arrhythmia can also be expected from this procedure.
We investigated the intraarterial concentration of CDDP following continuous hyperthermic pleural perfusion (CHPP, 43 degrees C for 30 to 60 minutes) with 100 mg CDDP diluted in distilled water or saline in 6 lung cancers (2 pleural dissemination, 1 malignant pleural effusion, 3 positive wash cytology), 2 metastatic lung cancer (from breast cancer, 1 rhabdomyosarcoma with pleural dissemination), and 1 metastatic pleural tumor (adenocarcinoma, unknown origin). The level of CDDP was 9.16 to 24.1 micrograms/ml (average 16.1) in perfusion fluid, 0.3 after 5 minutes, 0.41 to 0.85 after 60 minutes and 0.15 to 0.27 (average 0.22) after 24 hours. There were no severe side effects nor any difference in effects by dilution fluid between distilled water and saline. Only 1 patient suffered pleural effusion after this treatment. The results suggested that CHPP with CDDP diluted by saline was effective for prevention and treatment of pleural dissemination.
It has been recognized that glucose intolerance is a common finding in patients with chronic renal failure (CRF). Insulin resistance and impaired insulin secretion contribute to the pathogenesis of glucose intolerance. The resistance to insulin action is commonly observed in chronic renal failure and mainly due to impaired glucose uptake by muscle. In contrast, insulin secretion in response to hyperglycemia is variable: increased, decreased and normal. Recent studies reported that high PTH levels impaired insulin secretion from pancreatic islets. In this review, we mention the mechanism of glucose intolerance in chronic renal failure.
A novel antibacterial peptide that shows antibacterial activity against Staphylococcus aureus was isolated from the hemolymph of the silkworm, Bombyx mori. The novel peptide consisted of 42 amino acids and was highly basic. This peptide indicated no significant similarity with other antibacterial peptides. The peptide showed antibacterial activity against several Gram-negative and -positive bacteria and had a higher activity against Gram-positive bacteria than cecropin B1, a major antibacterial peptide of B. mori. The novel peptide was inducible by bacterial injection. These results suggest that the peptide is responsible for the antibacterial activity in B. mori against Gram-positive bacteria. The effects of the peptide on bacterial and liposomal membranes showed that a target of the peptide is the bacterial cytoplasmic membrane. The results also suggest that the N-terminal portion of the peptide, containing a predicted alpha-helix, is responsible for an increase in the membrane permeability. We propose the name "moricin" for this novel antibacterial peptide isolated from B. mori.
To define the local effects of prostacyclin (PGI2) on the growth of vascular smooth muscle cells (VSMC), we transfected VSMC with an expression vector harboring the cDNA for PGI2 synthase (PGIS), which catalyzes the rearrangement of prostaglandin H2 to PGI2. Transfection of the human PGIS cDNA into rat VSMC did not affect DNA synthesis under serum-free basal conditions, but it increased PGI2 synthesis and decreased DNA synthesis under serum-stimulated conditions (in the presence of 1 or 5% fetal calf serum). These results demonstrated that locally synthesized PGI2 can exert autocrine and/or paracrine inhibitory effects on VSMC growth. It was also suggested that in vivo transfer of PGIS gene may be useful for the gene therapy for vascular disease such as neointimal hyperplasia.
The procedure of aspiration biopsy cytology by fine needle aspiration (ABC) is as option in establishing definitive diagnoses for breast cancers. In this series, a needle size of 21G was considered most suitable for ABC as well as flow cytometric DNA analysis. Histograms from fresh samples aspirated by fine needle clearly delineated a sharp peak in G&sup0;G(1) phases and also a better CV was obtained than with paraffin-embedded preparations. In addition, fresh samples gave more reliable DI and suggested the value of measuring nuclear DNA contents. It is believed that the prognoses of breast cancers are closely associated with DNA ploidy patterns. In this sense, flow cytometric DNA analysis of fresh samples of ABC is regarded as important in clinical use.
There exist two distinct isozymes of prostaglandin-endoperoxide synthase (PES). PES-2 mRNA is synergistically induced by lipopolysaccharide (LPS) and 12-O-tetradecanoylphorbol-13-acetate (TPA) in bovine arterial endothelial cells. On the other hand, PES-1 mRNA is constitutively expressed under these conditions. Therefore, the promoter activities of the human genes for PES-1 and -2 in bovine arterial endothelial cells were examined. The 5'-flanking region of the human PES-2 gene (nucleotides -327 to +59) showed promoter activity inducible by LPS and TPA using transient transfection analysis, whereas that of the PES-1 gene (nucleotides -1010 to +69) showed constitutive promoter activity. Destruction of both consensus sequences for the nuclear factor responsible for the interleukin-6 expression (NF-IL6) site (nucleotides -132 to -124) and the cyclic AMP response element (CRE) (nucleotides -59 to -53) of the human PES-2 gene markedly reduced the promoter activity (25%) of the PES-2 gene after combined treatment with LPS and TPA, although single destruction of the NF-IL6 site or the CRE slightly reduced the promoter activity (60 or 90%, respectively). Moreover, cotransfection experiments showed that a trans-acting factor, CCAAT enhancer binding protein delta (C/EBP delta), which binds to both the NF-IL6 site and the CRE, increased the promoter activity of the PES-2 gene mainly through the CRE. C/EBP delta mRNA was rapidly induced by LPS. Collectively, these results suggest that transcription of the PES-2 gene in vascular endothelial cells is regulated through combination of the NF-IL6 site and the CRE and that C/EBP delta functions as one of the trans-acting factors.
A cDNA encoding lebocin, a novel member of insect antibacterial peptides, was isolated from the fat body cDNA library of Bombyx mori larvae immunized with Escherichia coli. The cDNA was 844 bp long and had an open reading frame (ORF) containing a probable signal peptide (16 amino acids), a putative prosegment (104 amino acids) and a mature peptide (32 amino acids) followed by 27 additional amino acids at its carboxyl-terminus. Northern blot analysis showed that lebocin gene expression was inducible by bacterial injection, occurred tissue-specifically in the fat bodies and continued at least for 48 h post-infection. These results suggest that lebocin has a unique precursor structure and shows typical gene expression pattern as insect antibacterial peptide.