Uncoupling of transfer of the presequence and unfolding of the mature domain in precursor translocation across the mitochondrial outer membrane.

Translocation of mitochondrial precursor proteins across the mitochondrial outer membrane is facilitated by the translocase of the outer membrane (TOM) complex. By using site-specific photocrosslinking, we have mapped interactions between TOM proteins and a mitochondrial precursor protein arrested at two distinct stages, stage A (accumulated at 0 degrees C) and stage B (accumulated at 30 degrees C), in the translocation across the outer membrane at high resolution not achieved previously. Although the stage A and stage B intermediates were assigned previously to the forms bound to the cis site and the trans site of the TOM complex, respectively, the results of crosslinking indicate that the presequence of the intermediates at both stage A and stage B is already on the trans side of the outer membrane.

Novel proteoglycan linkage tetrasaccharides of human urinary soluble thrombomodulin, SO4-3GlcAbeta1-3Galbeta1-3(+/-Siaalpha2-6)Galbeta1-4Xyl.

O-linked sugar chains with xylose as a reducing end linked to human urinary soluble thrombomodulin were studied. Sugar chains were liberated by hydrazinolysis followed by N-acetylation and tagged with 2-aminopyridine. Two fractions containing pyridylaminated Xyl as a reducing end were collected.

Complete amino acid sequences and phosphorylation sites, determined by Edman degradation and mass spectrometry, of rat parotid destrin- and cofilin-like proteins.

Beta-adrenergic or cholinergic stimulation of the rat parotid gland was earlier shown to induce dephosphorylation of endogenous destrin- and cofilin-like proteins, which are phosphorylated in resting cells at Ser residues probably present near the N-terminals. The primary structures and phosphorylation sites were determined here. The rat destrin-like protein had a sequence 95% identical to the cDNA-derived sequence of porcine destrin.

cDNA cloning of a novel trypsin inhibitor with similarity to pathogenesis-related proteins, and its frequent expression in human brain cancer cells.

A novel trypsin inhibitor (P25TI) with an apparent molecular size of 25 kDa has previously been purified from the culture medium of human glioblastoma cells. In this study, the cDNA encoding P25TI was isolated by the polymerase chain reaction (PCR) screening system, and its complete amino acid sequence was determined. The cDNA consisted of 1440 nucleotides and encoded a sequence of 258 amino acids.

Development of a novel polyimide hollow fiber for an intravascular oxygenator.

The authors have synthesized a novel fluorinated polyimide to develop a membrane material for oxygenators and fabricated polyimide hollow fibers for use in an intravascular oxygenator. A dry/wet phase inversion process has been applied to a spinning process to prepare an asymmetric polyimide hollow fiber. The outer surface of the hollow fiber consists of an ultrathin, dense skin layer, with a calculated apparent thickness of approximately 60 nm.

Low-noise operation of fluoride-based erbium-doped fiber amplifiers with (4)I(11/2)-level pumping.

The low-noise operation of fluoride-based erbium-doped fiber amplifiers (EDFA's) is demonstrated with (4)I(11/2)-level pumping for what we believe to be the first time. A noise figure as low as 3.5 +/- 0.

Traumatic tricuspid regurgitation associated with congenital partial pericardial defect.

Traumatic tricuspid regurgitation (TR) and congenital pericardial defects are rare conditions, there being fewer than 200 recorded cases of each. Here we describe the interesting case of a patient with traumatic TR due to rupture of chordae tendineae in association with a congenital pericardial defect. We suggest that the association of traumatic TR and congenital pericardial defect has a high probability of occurrence.

Comparison between urea removal in hemodialysis and continuous ambulatory peritoneal dialysis using a mathematical kinetic model.

Urea removal by continuous ambulatory peritoneal dialysis (CAPD) was compared with that by hemodialysis (HD) using a mathematical kinetic model. The time-averaged urea-nitrogen concentration (TAC) in the blood of a patient on maintenance HD was calculated by the model. Clearance with HD giving a TAC value equal to the constant urea-nitrogen concentration in the blood of a patient on CAPD was obtained, leading to the relation of equivalent clearances between HD and CAPD on the assumption that the urea-generation rate of the patient on CAPD was equal to that of the patient on HD.

Probing the environment along the protein import pathways in yeast mitochondria by site-specific photocrosslinking.

Artificially aminoacylated suppressor tRNAs were used to introduce photoreactive amino acids into model mitochondrial precursor proteins to probe the environment along the protein import pathway. Amino acids with benzophenone side chains of various lengths [DL-2-amino-3-(p-benzoylphenyl)propanoic acid (1) and DL-2-amino-5-(p-benzoylphenyl)pentanoic acid (2)] were incorporated at specific sites throughout the cytochrome b2-dihydrofolate reductase fusion proteins, pb2(220)-DHFR and pb2 delta 19(220)-DHFR, which were destined for the intermembrane space and the matrix in mitochondria, respectively. In vitro import of pb2(220)-DHFR and pb2 delta 19(220)-DHFR bearing 1 or 2 into isolated yeast mitochondria was arrested so that the N terminus reached the intermembrane space or the matrix, respectively, while the DHFR domain remained at the mitochondrial surface.

Enhancement of the sensitivity of a chemiluminescent immunoassay for estradiol based on hapten heterology.

Objective: To develop a highly sensitive chemiluminescent immunoassay (CLIA) that measures the serum estradiol (E2) levels in postmenopausal women.

Methods: The previously developed competitive CLIA for E2 consisted of an immobilized antigen and a labeled antibody with an N-functionalized acridinium ester that was modified to enhance its sensitivity. The modifications were changing the immobilized antigen from E2 to its analogues with its expected effect on hapten heterology and selecting optimal incubation conditions.

Isolation and characterization of the human chromosomal gene for prostacyclin-stimulating factor.

Prostacyclin-stimulating factor (PSF) is a protein which acts on vascular endothelial cells and stimulates the production of prostacyclin. Recently, we were able to purify PSF from the conditioned medium of cultured human diploid fibroblasts and clone PSF cDNA. In this study, we screened a human genomic library and isolated genomic clones to determine the structure of the human chromosomal PSF gene.

Molecular design of hybrid tumour necrosis factor-alpha. II: The molecular size of polyethylene glycol-modified tumour necrosis factor-alpha affects its anti-tumour potency.

To design hybrid tumour necrosis factor-alpha (TNF-alpha) applicable to systemic anti-tumour therapeutic use, we assessed the relationships among the molecular size of hybrid TNF-alpha, in vitro bioactivity and in vivo anti-tumour potency. Natural human TNF-alpha was covalently modified with polyethylene glycol (PEG) of various number-average molecular weights (Mn = 2000, 5000, 12,000). The in vitro bioactivity of PEG-modified TNF-alpha s decreased with an increase in the degree of PEG modification, irrespective of the molecular weight of PEG.

Molecular design of hybrid tumor necrosis factor-alpha III: polyethylene glycol-modified tumor necrosis factor-alpha has markedly enhanced antitumor potency due to longer plasma half-life and higher tumor accumulation.

We have reported that chemical modification of tumor necrosis factor-alpha (TNF-alpha) with polyethylene glycol (PEG) markedly increases its antitumor potency without any adverse side effects. MPEG-TNF-alpha, especially, in which 56% of the lysine amino groups of TNF-alpha are coupled with PEG, exhibits 100-fold more antitumor activity in vivo than native TNF-alpha in the Meth-A murine sarcoma model. In this study, we investigated the pharmacokinetics of PEG-modified TNF-alpha with various molecular sizes to clarify the mechanisms of the enhanced antitumor potency of MPEG-TNF-alpha.

Mass transfer of antibiotics adsorbed by human serum albumin in hemodialyzers.

The removal rate of a drug by a hemodialyzer is very important to determine the dosage of the drug to a patient without kidney function. Mass transfer of a drug in a hemodialyzer and a patient's body is more complicated than that of ordinary solutes because most drugs are adsorbed by serum albumin. In this paper, we reveal the adsorption characteristics of five clinical antibiotics onto human serum albumin (HSA) with new experimental methods and discuss the influence of adsorption characteristics on removal of antibiotics by hemodialyzers using mathematical models.

Endogenous atrial natriuretic peptide inhibits endothelin-1 secretion in dogs with severe congestive heart failure.

Atrial natriuretic peptide (ANP) has been shown to counteract the response of endothelin-1 (ET-1), but whether endogenous ANP actually inhibits the systemic release of ET-1 in vivo has not yet been determined. We administered HS-142-1 (HS), a specific antagonist of the guanylate cyclase-coupled ANP receptor, to conscious dogs with severe congestive heart failure (CHF) produced by rapid right ventricular pacing (n = 5, for 22 days) at doses of 0.3, 1.

[Highly effective preoperative intraarterial infusion chemotherapy with CDDP for progressive uterine corpus cancer with a Sister Mary Joseph's nodule].

A 57-year-old female patient complained of atypical genital bleeding and a noxious emanation from her navel. A histological examination of the uterine body and the navel area confirmed a diagnosis of adenocarcinoma. We diagnosed it as IVb stage of uterine corpus cancer with a Sister Mary Joseph's nodule.

Differential contribution of metal complexation and dimerization to the chemotherapeutic potential of bicyclen-Zn2II complex against human immunodeficiency virus.

1,1'-(m-Xylenediyl)-bis(1,4,7,10-tetraazacyclododecane)-Z n2II complex (m-xylenediyl-bicyclin-Zn2II), a potent inhibitor of human immunodeficiency virus (HIV), was obtained from cyclin by a combination of dimerization and metal complexation. The ratio of median cytotoxic concentration against host cells (CC50) and median effective concentration against HIV cytopathogenicity (EC50), referred to as the selectivity index (SI), was regarded to be a measure of anti-HIV activity. These two chemical modifications contributed to a potent, in vitro anti-HIV activity of m-xylenediyl-bicyclin-Zn2II by respectively increasing the CC50 and decreasing the EC50 in comparison with those of cyclin.

Injection-incision--assisted snare resection of large sessile colorectal polyps.

Background: It can be difficult, even for experienced endoscopists, to completely remove large sessile colorectal polyps. We attempted to remove large sessile colorectal polyps without complication and residual tumors.

Method: Our new technique is characterized by submucosal pre-injection with a large volume of saline solution and then circumferential incision outside the lesion before resection using a special needle-tipped snare.

Competitive chemiluminescent immunoassay for estradiol using an N-functionalized acridinium ester.

We have compared three competitive chemiluminescent immunoassays (CLIA) for estradiol (E2) using an N-functionalized acridinium ester (AE). The assays were a standard competitive assay using immobilized antibody and directly labeled antigen (type A), an immobilized antibody and indirectly labeled antigen (type B), and an immobilized antigen and labeled antibody (type C). In an antibody-immobilized system, the assay using both AE- and E2-labeled thyroglobulin as a tracer (type B) was more sensitive than that using AE directly coupled with E2 (type A).

Flat adenomas and flat adenocarcinomas of the colorectal mucosa in Japanese and Swedish patients. Comparative histologic study.

Purpose: In recent years, flat adenomas of the colorectal mucosa have been intensively investigated by Japanese pathologists. Results of that work indicate that flat adenomas may antedate the development of colorectal carcinomas. Because of differences in the histologic definition of flat adenomas with severe dysplasia and with intramucosal carcinoma within the group, one single observer having both Western and Asian training in pathology reviewed the material.

Possible regulation of renin release by ANP in dogs with heart failure.

We investigated whether atrial natriuretic peptide (ANP) contributed to the regulation of renin release during the development of experimental heart failure in dogs. Significant increases of plasma ANP and plasma guanosine 3',5'-cyclic monophosphate (cGMP) levels were observed in dogs with mild heart failure induced by 6 days of rapid ventricular pacing. However, plasma renin activity (PRA) was not elevated, despite the significant decreases in renal plasma flow and mean arterial pressure.