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Biomedical subjects

M Imai

Publications and source records attributed to M Imai.

At least 451 records · Page 25Linked to original sources

Conditionally lethal nusAts mutation of Escherichia coli reduces transcription termination but does not affect antitermination of bacteriophage lambda.

Termination of transcription at bacteriophage lambda terminators as well as at the Escherichia coli trp a attenuator was examined in the conditionally lethal mutant (nusAts11) defective in the NusA protein of E. coli. Experiments using terminator-assay lambda vectors revealed that the efficiency of termination at both rho-dependent (lambda tL1) and rho-independent (lambda tL2 and trp a) terminators decreases in the mutant. The mutation does not block lambda phage growth at either permissive or nonpermissive temperatures, nor does it affect the lambda Q protein antitermination activity at the t6s terminator. These results indicate that NusA is required for transcription termination, and that lambda N and Q-mediated antitermination may not require the NusA protein function in the nusAts11 mutant.

Bacteriophage lambda↗

Lack of direct action of alpha-human atrial natriuretic polypeptide on the in vitro perfused segments of Henle's loop isolated from rabbit kidney.

We examined direct effects of alpha-human atrial natriuretic polypeptide (alpha-hANP) on water and NaCl transport across the segments of Henle's loop by the in vitro microperfusion technique. In the medullary and the cortical thick ascending limb, 10(-6) M alpha-hANP did not affect the transmural voltage (Vt) and the lumen to bath 36Cl flux whether it was added to the perfusate or to the bath. In the thin ascending limb, 10(-6) M alpha-hANP did not affect the lumen to bath 36Cl flux. The peptide may not affect permselectivity of the thin ascending limb, since it did not influence the diffusion potential generated in the presence of a NaCl gradient. In the descending limb, 10(-6) M alpha-hANP did not affect osmotic water permeability, whether it was added to the perfusate or to the bathing fluid. From these observations, we conclude that alpha-hANP does not show direct effects on water and NaCl transport in the segments of Henle's loop at least under our experimental conditions. Therefore, natriuresis by alpha-hANP may be caused either by an action on nephron segments other than Henle's loop or by an action on renal vasculatures.

Animals↗

Permselectivity for cations over anions in the upper portion of descending limbs of Henle's loop of long-loop nephron isolated from hamsters.

The permselectivity of the upper portion of long descending limb of Henle (LDLu) was investigated with electrophysiological methods in the isolated perfused tubule preparation of hamster kidney. The diffusion potential (Vt) was determined in three different protocols. In protocol 1, the tubules were initially perfused with modified Krebs Ringer's solution on both sides of the epithelium. Then the bath solution was exchanged consecutively with another solution in which 50 mmol/l NaCl replaced by 50 mmol/l KCl, RbCl, NH4Cl, CsCl, LiCl, NaBr, NaNO3, NaI, Na acetate or 75 mmol/l NaCl replaced by mannitol. The permeabilities for these ions relative to chloride were calculated by Goldman's constant field equation. The segment was found to be cation selective, with all cations being 5-9 times more permeable than all anions. The sequence of permeability was K+ greater than Rb+ greater than Li+ greater than NH+4 = Cs+ greater than or equal to Na+ much greater than Cl- greater than or equal to Br- greater than or equal to NO3- greater than or equal to I- greater than Acetate-. In protocol 2, pure 150 mM NaCl was used for the basal solution to avoid interference by other ions. The bathing solution was exchanged by other solutions which contained 150 mmol/l KCl, NH4Cl, CsCl, RbCl, LiCl, NaI, NaBr, NaNO3, Na acetate or 75 mmol/l NaCl with mannitol. Thus simple biionic substitution was performed. Again, the segment was found to be cation selective, with all cations being 4-10 times more permeable than all anions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Class distribution of immunoglobulin-containing plasma cells in the stroma of medullary carcinoma of breast.

A class distribution of plasma cells associated with the stroma in twenty-eight cases of medullary carcinoma of the breast was investigated by an unlabeled immunoperoxidase method. The stroma of the medullary carcinomas tested was found to contain predominantly IgG plasma cells except in two cases. Stroma of the other types of breast carcinoma, including ten cases of papillo-tubular carcinoma, five cases of scirrhous carcinoma, and six cases of medullary tubular carcinoma, contained predominantly IgG plasma cells, although few plasma cells were associated with carcinoma tissues in the latter group. Plasma cells associated with control specimens, including normal breast, fibroadenoma, cystic disease, and intraductal papilloma, were found to be predominantly of IgA type. Few carcinomatous epithelial cells contained secretory components in the cytoplasm, while a number of cells positive for secretory components were observed in acinar and ductular epithelia of normal breast tissues and in benign proliferative lesions of the breast. It is suggested that the lymphoid cells infiltrating the stroma of medullary carcinoma represent a sign of host immune response against the carcinoma cells which is related to the well-known favorable prognosis associated with this tumor.

Adenocarcinoma↗

Nude mice bearing human primary hepatocellular carcinoma that produces hepatitis B surface, core, and e antigens, as well as deoxyribonucleic acid polymerase.

A primary hepatocellular carcinoma, from a patient carrying hepatitis B virus, was transplanted to athymic nude mice, and maintained through eight passages involving 39 mice. Hepatitis B surface and e antigens were detected in the circulation of tumor-bearing mice. Hepatitis B surface, core, and e antigens were demonstrated in tumor cells. Hepatitis B core particles were visualized in the tumor extract by immune electron microscopy, and they exhibited an activity of deoxyribonucleic acid polymerase. In the tumor tissue, deoxyribonucleic acid of hepatitis B virus was present both in integrated and extrachromosomal forms. Nude mice carrying the tumor would provide opportunities for studying the replication of hepatitis B virus and the expression of its various proteins, and also for evaluating the efficacy of virustatic drugs in interrupting the persistent infection.

Aged↗

Large hepatitis B surface antigen polypeptides of Dane particles with the receptor for polymerized human serum albumin.

Large hepatitis B surface antigen polypeptides with apparent molecular sizes of 39,000 and 43,000 daltons (P39 and P43) were liberated from a purified preparation of Dane particles of subtype adr. They were tested for reactivity with monoclonal antibodies raised against three synthetic oligopeptides representing fundamental sequences of the pre-S region in deoxyribonucleic acid of hepatitis B virus (subtype adr), as well as with monoclonal antibody against the major surface antigen polypeptide (P22) coded for by the S gene. Both P39 and its glycosylated form P43 bound to all four monoclonal antibodies, thereby indicating that they were coded for by the sequence of 1200 nucleotides, from the second ATG codon in the pre-S region to the stop codon of the S gene. Both P39 and P43 bound to polymerized human and chimpanzee albumins, but not to polymerized albumin from species without susceptibility to hepatitis B virus. Due to their presence in Dane particles and the expression of a polyalbumin receptor, the immune responses against P39 and P43 may have significance in infection with hepatitis B virus and its immunoprophylaxis.

Antibodies, Monoclonal↗

Enhanced myocardial protection by systemic deep hypothermia in children undergoing total correction of tetralogy of Fallot.

The effectiveness of systemic deep hypothermia for myocardial protection was evaluated retrospectively in 36 consecutive children who underwent total correction of tetralogy of Fallot in the four-year period 1980 to 1984. Moderate hypothermia combined with potassium-induced cold cardioplegia and topical cardiac cooling was employed in 16 patients (Group A), and deep hypothermia together with cold cardioplegia and topical cooling was used in 20 patients (Group B). A higher incidence of spontaneous defibrillation, a higher postoperative right ventricular cardiac index, a significant decrease in the maximal requirement of isoproterenol hydrochloride, a significant increase in the mean urinary output, and much better operative results were obtained in Group B compared with Group A. Postmortem histopathological examination of the heart in 3 patients in Group A disclosed various degrees of hypoxic change in the myocardium, which were more pronounced in the right ventricle than in the left ventricle. It is concluded that the myocardial protection obtained with cold cardioplegia and topical cooling under moderate hypothermia may well be insufficient for repair of tetralogy of Fallot, a condition characterized by increased non-coronary blood flow to the myocardium and abundant collateral bronchial flow. However, when combined with systemic deep hypothermia, such myocardial protection is quite safe and effective.

Child↗

Modulation by verapamil of hormonal action on the Henle's loop of mice.

In order to know the role of cytosolic calcium in the modulation of the hormone action on sodium chloride transport across the thick ascending limbs of Henle's loop, we examined whether verapamil, a blocker of cellular calcium entry, can modulate the effects of arginine vasopressin (AVP) or glucagon in stimulating transepithelial voltage (Vt) and cyclic AMP generation in the mouse medullary thick ascending limb (MAL). The pretreatment of the renal tubule with 5 X 10(-5)M verapamil reduced the Vt stimulated with 200 microU/mliter AVP from 1.7 +/- 0.3 mV to 0.4 +/- 0.4 mV (N = 7, P less than 0.05). The changes in Vt were well correlated with those of unidirectional Cl flux from the lumen to the bath. However, verapamil did not influence the Vt stimulated with 10(-3) M dibutyryl cyclic AMP. The pretreatment of the MAL with 10(-5) M verapamil also inhibited the cyclic AMP generation in the MAL from 72.1 +/- 17.9 to 50.6 +/- 13.6 fmoles/mm/7 min (N = 7, P less than 0.05) as well as in the medullary collecting tubule from 147.6 +/- 46.6 to 121.2 +/- 41.6 fmoles/mm/7 min (N = 4, P less than 0.05). The effect of verapamil in inhibiting the AVP-stimulated cAMP was dose-dependent: the cAMP generation was inhibited by 28.9 +/- 6.8 and 61.1 +/- 9.3% with 10(-5) M and 10(-4) M verapamil, respectively. When verapamil was added to the medium simultaneously with AVP, the generation of cyclic AMP was unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nucleotide sequence of a cloned hepatitis B virus genome, subtype ayr: comparison with genomes of the other three subtypes.

The entire nucleotide sequence of genomic DNA was determined for hepatitis B virus (HBV) of subtype ayr, which had been derived from the blood of a Japanese asymptomatic carrier. The genome was 3215 nucleotides long, and differed in DNA sequence by 10% from that of subtypes adw or ayw, but by only 2% from that of subtype adr. Amino acid sequences coded for by the S, C, P and X genes, as well as by the pre-S region, closely resembled those of subtype adr, indicating that the evolution of HBV/ayr from HBV/adr was more recent than the differentiation of the other three subtypes. In the product of the S gene, the mutually exclusive subtypic determinants of the surface antigen, d and y, were associated with variation of amino acid residues at only the 68th and 122nd positions from the N terminus, in contrast to the variation at as many as seven positions for the other set of subtypic determinants, w and r. Sequences representing high local hydrophilicity in the product of the S gene were involved in subtypic variation, although such sequences in the pre-S region were shared by HBV genomes of the various subtypes. In particular, a hydrophilic sequence of 19 amino acid residues, coded for by the pre-S(2) region and implicated in the presumed hepatotropism of HBV, was possessed in common by HBV/adr, HBV/ayr and HBV/ayw, and differed in HBV/adw by only one residue at the 9th position. This amino acid sequence appears to be a promising candidate for a synthetic peptide vaccine.

Amino Acid Sequence↗

Integration of region X of hepatitis B virus genome in human primary hepatocellular carcinomas propagated in nude mice.

Tissues of human primary hepatocellular carcinoma (PHC) from six patients infected with hepatitis B virus (HBV) were propagated in nude mice, as well as a strain of hepatitis B surface antigen-positive PHC (PLC/PRF/5). Integration of viral DNA into chromosomal DNA of tumour cells was evaluated by the capacity to hybridize with radiolabelled DNA probes, each representing fundamental parts of the HBV genome, that is S and C genes and regions pre-S and X. All PHC cells possessed region X integrated in their chromosomes. However, integration of the S gene, C gene and region pre-S was found in only six of the seven PHCs. Based on these findings, the integration of region X seems to be most closely associated with carcinogenesis in HBV infection.

Animals↗

Autogenous regulation of the gene for transcription termination factor rho in Escherichia coli: localization and function of its attenuators.

We present evidence that the expression of rho is regulated by rho-dependent attenuation of transcription. Gene fusion analysis with nested series of deletions of rho indicated that the transcription of rho is attenuated in a rho-dependent manner in the leader region and that neither a read-through transcription from the upstream gene, trxA, nor a modulation of transcription initiation of the rho promoter is involved in the self-control of rho. S1 mapping and Northern hybridization analyses localized at least six transcription attenuation or termination sites in the region ranging from the 3' end of the trxA structural gene to the middle of the rho structural gene. Among them, the most upstream site overlapping the rho promoter sequence was assigned to the terminator for the trxA gene, and the second and third sites, mapping about 80 and 50 nucleotides upstream from the start codon of rho, were suggested to function as the major attenuation sites for regulation of the rho expression. Further, the start points of the trxA and rho RNAs were determined in an in vitro transcription system to be located 111 nucleotides (U) and 255 nucleotides (G) upstream from their respective start codons. These results necessitate revisions of previous predictions on the sites of transcriptional signals in the trxA and rho genes (S. Brown, B. Albrechtsen, S. Pedersen, and P. Klemm, J. Mol. Biol. 162:283-298, 1982; C.-J. Lim, D. Geraghty, and J. A. Fuchs, J. Bacteriol. 163:311-316, 1985; B.J. Wallace and S.R. Kushner, Gene 32:399-408, 1984).

Chromosome Deletion↗

Left renal vein hypertension in patients with left renal bleeding of unknown origin.

The pullback pressure from the left renal vein (LRV) to the inferior vena cava was studied in 16 patients with left renal bleeding of unknown origin (group A), 15 patients with hematuria of miscellaneous laterality (group B), and nine control subjects (group C). The mean pressure gradients were 5.0 +/- 2.0 mm Hg (mean +/- SD) in group A, 1.9 +/- 1.0 mm Hg in group B, and 1.1 +/- 0.9 mm Hg in group C. The mean pressure gradient of group A was significantly higher than that of groups B and C (P less than .001). From the results of control studies, we regarded pressure gradients greater than or equal to 3.0 mm Hg as indicative of LRV hypertension. With this criterion, 88% (14 of 16) of the patients in group A had LRV hypertension, which was considered a cause of hematuria. Analysis of the results of renal angiography in the 31 patients revealed that opacification of collateral pathways of the LRV was a significant angiographic finding in LRV hypertension.

Adolescent↗

Diluting segment in avian kidney. I. Characterization of transepithelial voltages.

Renal tubules from the Japanese quail, Coturnix coturnix, were perfused in vitro to characterize the transepithelial voltage (Vt). The thick limb (TL) of the mammalian-type (MT) nephron showed Vt positive in the lumen (+9.1 +/- 0.7 mV, n = 35). The Vt decreased with increases in hydrostatic perfusion pressure. Furosemide (lumen), Na cyanide (bath), and ouabain (bath) reversibly reduced Vt. Removal of Cl or Na from the perfusate and the bath decreased Vt of the TL from +10.3 +/- 3.0 to -0.3 +/- 0.3 (n = 5) and from +7.4 +/- 1.5 to +1.2 +/- 0.2 mV (n = 8), respectively. The distal tubule of the reptilian-type (RT) nephron showed two types of Vt: a lumen-negative Vt in the late segment and a lumen-positive Vt in the early segment that is in close contact with the parent glomerulus. Both voltages were reversibly reduced by ouabain and Na cyanide. These results suggest that in quail the TL of the MT nephron resembles the thick ascending limb of the mammalian kidney with both Na and Cl required for generation of luminal positivity and the distal tubule of the RT nephron appears functionally heterogeneous.

Animals↗

Localization of binding sites for alpha-rat atrial natriuretic polypeptide in rat kidney.

To determine the intrarenal localization of receptors for atrial natriuretic polypeptide (ANP) in the rat, we performed autoradiography and binding assay by using 125I-labeled alpha-rat ANP (alpha-rANP). Autoradiography at the slice and microscopic level in the kidney and binding assay in isolated glomeruli demonstrated that receptors in the renal cortex were distributed mainly in glomeruli. Although dense silver grains were distributed diffusely both in inner medulla and outer stripe of outer medulla, a marked displacement of the grains was observed only in the inner medulla. Autoradiography at the microscopic level also showed that silver grains were distributed in the renal artery, renal pelvis, and inner medullary collecting tubule (IMCT) prepared by the microdissection method, but not in the arcuate artery, interlobular artery, and afferent or efferent arterioles. Specific binding was demonstrated in the isolated glomeruli and the preparation was rich in fragments of IMCT. Apparent binding affinity (Kd) and receptor density (R) for 125I-labeled alpha-rANP in isolated glomeruli and IMCT were Kd = 3.2 and 21 X 10(-9) M, and R = 320 and 420 fmol/mg protein, respectively. These observations suggest that alpha-rANP has a physiological action on glomeruli and possibly on the inner medullary collecting tubules in addition to the renal artery.

Animals↗

Intrarenal localization of receptors for alpha-rat atrial natriuretic polypeptide: an autoradiographic study with [125I]-labeled ligand injected in vivo into the rat aorta.

We examined intrarenal localization of receptors for alpha-rat atrial natriuretic polypeptide (alpha-rANP) by injecting [125I]-labeled ligand in vivo into the rat aorta. We found that the receptors for alpha-rANP are distributed also on the vasa recta of the outer and inner medulla in addition to the previously reported sites, i.e., the renal arteries, renal pelvis, glomeruli, and inner medullary tissues including collecting tubules. In the vascular bundle of the outer medulla, the majority of grains was preferentially localized on the arterial vasa recta. The electron microscopic autoradiography of the glomerulus showed that the binding sites were mainly localized on the foot process of the podocyte. Since alpha-rANP injected into the aorta under physiological conditions was bound to the glomerulus and vasa recta in the kidney, the effect of ANP on these binding sites may be important in the mechanism of natriuresis.

Animals↗