[Retinal-binding protein and photopigment systems].
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Biomedical subjects
Publications and source records attributed to T Hara.
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We present our most recent technique of endocapsular phacoemulsification and aspiration (ECPEA) and report the results achieved using it in 50 consecutive eyes of 47 patients. Using a bent 0.9-mm Fragmatome tip, we removed relatively hard cataracts through 1.5-mm anterior capsular openings. With an average follow-up period of 12 months, the success rate was 72%. Causes of failure were anterior capsular openings enlarged more than 2.6 mm in nine eyes, and posterior capsular rupture in five eyes. After the incisions were extended to 6 mm, 11-mm posterior chamber lenses were inserted into the capsular bags. In the successful 36 eyes, postoperative visual acuity of better than 20/20 was achieved in 20 eyes (56%), and better than 20/40 vision in 31 (86%). Average corneal endothelial cell loss was 3.7%. Ample anterior capsular transparency was retained in 26 eyes (72%).
Pyrexia which occurs at late stage after prosthetic heart valve replacement must be suspected to be caused by prosthetic valve endocarditis (PVE). However, since confirmatory diagnosis is difficult, we are concerned about the treatment for so many cases. We encountered two cases who were suspected PVE at late stage after prosthetic aortic valve replacement and diagnosed miliary tuberculosis at autopsy. Here, we present a report of the two cases.
The bacteria in 37 stones obtained from 37 patients with urinary stone diseases, that is, 11 renal stones (containing 2 staghorn calculi), 21 ureteral stones, 4 bladder stones and 1 urethral stone, were studied, according to the Nemoy & Stamey's method. The stones were collected by partial nephrectomy (1 case), nephrolithotomy (1 case), pyelolithotomy (1 case), percutaneous nephrolithotripsy (PNL) (6 cases), 12 ureterolithotomies, transurethral ureterolithotripsy (2 cases), cystolithotripsies (4 cases) and spontaneous deliveries (10 cases). According to a stone analysis by infrared spectrophotometer revealed 30 were noninfection stones (81.1%) and 7 infection stones (18.9%). Of these 37 stones, 5 stones (13.5% of 11 stones) had bacteria within them. These stones consisted of 4 infection stones (57.1% of all infection stones) and 1 noninfection stone (3.3% of all noninfection stones). Out of 4 patients having bacteria within their stones, urine culture of whom were carried out before stones were collected, only one patient (25%) had the same species of bacterium (E. coli) both within the stone and in urine. The E. coli within the stone and that in urine, however, showed quite different reactions to some antibiotics. The fact that all organisms within stones cannot be detected by urine culture before collecting stones of the patients in our present study, suggests that patients undergoing endourological surgery, such as extracorporeal shock wave lithotripsy and PNL, may have a risk of complications, such as severe urinary tract infection and urosepsis, if the possibility of the presence of organisms within stones is not taken into consideration.
1. A system to train nurses to be first and solo assistants of microsurgery has been practiced in our hospital for the past 13 years. 2. Under our unique weekly rotation system, nurses are educated to be all-around nurses. Microsurgical assistance is only one of approximately eight sections of the rotation. 3. Under this system, nurses can become extremely efficient solo assistants after 3 years.
The tumor promoter PMA has been shown to induce the expression of a 28-kDa/32-kDa early activation Ag, termed EA 1, on resting T cells. Under nonreducing conditions, EA 1 was detected by SDS-PAGE as a diffuse band in the 60-kDa region. In this study, this diffuse band was resolved into 56-kDa and 60-kDa bands. Endoglycosidase F treatment of EA 1 resulted in the appearance of a single band with a Mr of 48 kDa. Upon reduction, the 48-kDa band was shown to be composed of 24-kDa peptides. Diagonal gel electrophoresis showed that the major band of EA 1 was composed of a series of disulfide-linked homodimers with subunits of the same 24-kDa core protein that were differentially glycosylated. This analysis also revealed in a minor population of the EA 1 molecules, the presence of proteins of different Mr associated with the core protein. The signal requirements for the induction of EA 1 were investigated. The putative cellular action of PMA is the activation of protein kinase C (PKC). To further investigate the role of PKC activation in the expression of EA 1, the synthetic diacylglycerol, 1,2-sn-dioctanoylglycerol (diOG) was examined for its ability to substitute for PMA. DiOG induced EA 1 expression in a dose dependent manner. H-7, a relatively selective inhibitor of PKC, blocked diOG and PMA induced EA 1 expression. HA1004, a selective inhibitor of cAMP- and cGMP-dependent protein kinases, had no effect. In kinetic studies, EA 1 expression was seen as early as 1 h in diOG- and PMA-activated T cells. However, diOG did not completely mimic PMA-induced EA 1 expression. By 18 h, diOG-induced EA 1 expression was markedly reduced, whereas PMA-induced EA 1 expression was persistent. The role of calcium in EA 1 expression was investigated. mAb against CD3 potentiated diOG-induced EA 1 expression. This potentiation appeared to correlate with the ability of the anti-CD3 mAb to induce rises in intracellular calcium. Addition of EGTA to the media blocked the potentiation of diOG induced EA 1 expression by these mAb. The role of calcium in EA 1 expression was further demonstrated by the ability of ionomycin to potentiate EA 1 expression. These results demonstrate that PKC activation is the primary pathway for the induction of EA 1. However, calcium-dependent pathways appear to have a secondary role.
A method for determining 17-hydroxyprogesterone in plasma by isotope dilution-mass spectrometry is described. For the internal standard 17-hydroxy [2H4]progesterone is used. Extraction of plasma is followed by conversion into the 3,20-dienol,17-tristrimethylsilyl ether derivative and analysis by capillary gas chromatography-mass spectrometry with selected-ion monitoring, at a resolution of 6000. The lower limit of quantitation was 1 pg, judged from a criterion of a signal-to-noise ratio of 10. The precision and accuracy of the method were satisfactory.
The binding of methotrexate (MTX) to IgG in conjugates was examined by studies on a direct MTX conjugate with a monoclonal antibody (aMM46) to mouse mammary tumor MM46 cells and corresponding irrelevant antibody and normal gamma-globulin conjugates, all prepared by the active ester method with MTX N-succinimidyl ester (MTX-OSu). The binding was examined in terms of effects on the potency and selectivity of the cytotoxic activity of the aMM46 conjugate. The results obtained supported the following conclusions: (a) MTX-OSu reacts not only with the amino group of IgG to give an amide bond, but also with another group(s) to give a less stable bond(s) such as an ester bond; (b) in contrast to the amide bond-linked MTX, which is taken up by the cells by endocytic internalization, a substantial portion of the MTX linked by an ester or other less stable bond(s) is released from the conjugates extracellularly and enters the cells by the MTX active transport system, as revealed by the inhibitory effect of thiamine pyrophosphate on the cytotoxicity; (c) this MTX linked by a less stable bond(s) that causes nonspecific cytotoxicity can be removed by treatment with hydroxylamine; (d) the overall cytotoxicity of aMM46-MTX decreased on removal of this less stably linked MTX, suggesting that the lysosomal degradation of the conjugate carrying amide bond-linked MTX to liberate MTX derivatives of low molecular weight is insufficient; (e) the liberation of low-molecular-weight substances in the lysosomes is probably more important for efficient entry of active substances into the cytosol, than for inhibition of the activity of dihydrofolate reductase, because after hydroxylamine treatment, the amide bond-linked MTX showed greater decrease in drug cytotoxicity than in inhibitory activity against dihydrofolate reductase; (f) in combination with hydroxylamine treatment, insertion between MTX and IgG of a linkage capable of ready cleavage in lysosomes deserves exploitation as a method for making potent conjugates with less nonspecific activity.
Thirty-one children with leukemia or lymphoma treated with multi-drug chemotherapy for more than 2 years were reviewed to identify and characterize the occurrence of febrile episodes related to vincristine (VCR) injection. In nine of the 31 children, more than two febrile episodes apparently attributable to VCR during maintenance therapy were identified. No such episodes were found to have occurred during induction therapy. The median age at diagnosis of these nine children was 3.2 +/- 1.6 years, significantly lower than the 6.8 +/- 4 years of the other 22 children (P less than 0.01). No significant difference was observed between these two groups in laboratory data obtained before the VCR injections, and the drugs used in combination with VCR were apparently unrelated to the incidence of febrile episodes. All of these febrile episodes began within 24 hours after VCR injection. Peak levels of 38 degrees C to 39 degrees C occurred 6 to 24 hours after onset. The episodes also were accompanied by mild, general fatigue and a loss of appetite. They ranged in duration from half of a day to 4 days, but those persisting more than 3 days occurred in three of the five children whose regimens did not include corticosteroid. The results thus suggest that fever is an immediate reaction to the toxicity of VCR in young children, and that the duration of fever can be shortened by combining VCR with corticosteroid.
The mAb Tm 1 was obtained from a fusion of SP2/O tumor cells with spleen cells from CF1 mouse immunized with T cells modulated by an IgM anti-CD3 mAb.mAb Tm 1 reacted with IgM anti-CD3 modulated T cells (66.6%) but not with unmodulated T cells (4.4%). Tm 1 was not expressed on T cells modulated with either IgG2a or IgG1 anti-CD3 mAb. Immunoprecipitation from 125I-labeled CD3-modulated T cells showed that Tm 1 Ag is a single polypeptide of 33 kDa under reducing and nonreducing conditions. Kinetic studies revealed that Tm 1 was detectable on T cells 10 min after incubation and maximally expressed after 4 h of incubation with IgM anti-CD3 mAb. CD3 expression was markedly modulated by this anti-CD3 mAb after the same period of incubation. Studies with cycloheximide revealed that Tm 1 expression on T cells does not require new protein synthesis. Tm 1 expression persisted long after CD3-reexpression 24 h later. Tm 1 was present on a small fraction of circulating T cells, B cells, and monocytes and absent from granulocytes, platelets, E, and thymocytes. Tm 1 was not expressed on T cells after various activation stimuli but was expressed on B cells upon activation. Additional studies indicate that IgM mAb against other T cell differentiation Ag and IgM mAb against B cell Ag also lead to the expression of Tm 1 on these cells. Thus, modulation of surface Ag by IgM mAb externalizes this cytoplasmic Ag. However, one exception has been noted. Purified mAb Tm 1 was not mitogenic and was unable to block either the T cell proliferation induced by 12-O-tetradecanoyl phorbol-13-acetate plus anti-CD3 mAb and other T cell stimuli, or the B cell proliferation induced by B cell mitogens. The role of Tm 1 on lymphocyte function remains to be determined.
We studied the morphology and removal of lens epithelial cells in the following ways. In 49 senile cataractous eyes with a mean (+/- SD) age of 69 +/- 9 years, mean (+/- SD) cell density was 3574 +/- 512 cells/mm2 and mean (+/- SD) cell size was 281 +/- 48 micron 2. With age there was a significant decrease in cell density and an increase in cell size. Immediately after each of the following procedures was performed on the central anterior capsule during routine cataract surgery, the capsule was removed and observed. Procedures used were (1) aspiration, (2) aspiration with ultrasound, (3) instant single cryopexy application, (4) double cryopexy, and (5) double cryopexy followed by aspiration with ultrasound. Each procedure could destroy or remove lens epithelial cells completely. The procedure that we recommend is single cryopexy application followed by aspiration with ultrasound.
11C-methylated inulin, supposedly useful for imaging of brain edema and pulmonary edema, was prepared using cyclotron produced 11CO2. The synthesis consists of the production of 11C-methyl iodide and its coupling with inulin alkoxide sodium in dimethylsulfoxide as solvent. 11C labeled inulin was purified by alcohol precipitation. The radiochemical yield of pure 11C-inulin was 34% of 11CO2 30 min after the end of bombardment. The blood clearance and body distribution of 11C was observed in rabbits after i.v. injection of 11C-inulin. The blood clearance curve was composed of a sum of three exponential functions. The gamma camera image showed that the 11C activity in blood moved quickly to kidneys and urine and a small dose of radioactivity remained persistently in edematous tissues, i.e. the edematous lung tissues produced by oleic acid treatment.
Positron emission tomography (PET) using 11C-inulin was demonstrated to be applicable to the clinical measurement of blood brain barrier permeability and cerebral interstitial fluid volume. Kinetic data were analyzed by application of a two compartment model, in which blood plasma and interstitial fluid spaces constitute the compartments. The blood activity contribution was subtracted from the PET count with the aid of the 11CO inhalation technique. The values we estimated in a human brain were in agreement with the reported values obtained for animal brains by the use of 14C-inulin.
Thrombocytopenia was observed in 10 (2.0%) of 486 children with Kawasaki disease. In nine of the ten, the minimal platelet count of 94,000 +/- 38,000 (SD)/mm3 was seen on day 6.8 +/- 2.2 (SD) of illness and the platelet counts were elevated to the normal level in 1-2 weeks. Thrombocytopenia in the nine appeared to be caused via coagulation-mediated platelet consumption, while the remaining child was diagnosed as having idiopathic thrombocytopenic purpura. One of the two who had severe coagulation-mediated thrombocytopenia of less than 50,000/mm3 developed coronary aneurysms persisting over 1 year.
The significance of primary tumor resection in gastric and colorectal cancer patients with liver metastasis (H(+)) was evaluated in terms of operative mortality and survival rate by dividing the materials [293 gastric cancer and 80 colorectal cancer patients (53 colon and 27 rectum) with synchronous liver metastasis] into the following groups: Firstly, with or without peritoneal dissemination (P), secondly, with or without resection of the primary tumor and thirdly, with or without postoperative adjuvant chemotherapy. The following results were obtained: (1) The direct operative death rate of primary tumor resection, excluding death from other causes, showed an absence of statistically significant differences between the P0H(+) and P(+)H(+) gastric and colorectal cancer patients. (2) There was no significance in the prognosis between the primary tumor resection + postoperative chemotherapy group and the non-resectable group in the P(+)H(+) gastric and colorectal cancer patients, revealing no prognostic value of the primary tumor. (3) In the P0H(+) gastric and colorectal cancer patients, the primary tumor resection + postoperative chemotherapy group was significantly more favorable in prognosis than was the primary tumor resection alone group or the non-resectable group, showing the value of primary tumor resection.
SDS polyacrylamide gel electrophoresis and immunoblotting were used to examine soluble proteins from the eyes of six species of cephalopods i.e. Lolliguncula brevis, Sepia officinalis, Octopus maya, Octopus bimaculoides, Rossia pacifica and Loligo opalescens. All species had a protein ("IRBP") with molecular weight virtually identical with vertebrate interstitial retinol-binding protein (IRBP) averaging 132,400 +/- 700 (n = 6). "IRBP" reacted on nitrocellulose blot transfers with rabbit antibovine IRBP and rabbit antifrog IRBP antibodies. Unlike vertebrate IRBP, cephalopod "IRBP" (from L. brevis) did not bind exogenous retinol or concanavalin A. The N-terminal amino acid appeared to be blocked in samples electroeluted from SDS gels. The antifrog IRBP antibodies also reacted with a series of proteins with molecular weights between 46,000 and 47,000, identified as retinal-binding protein (RALBP) with anti-RALBP antibodies. Anti-IRBP also reacted with pure RALBP prepared from Todarodes pacificus. Occasionally, anti-RALBP antibodies were seen to react weakly with "IRBP" in some cephalopods. We conclude that RALBP, cephalopod "IRBP" and vertebrate IRBP share a common but distant ancestry, and that a protein resembling IRBP appeared before the vertebrates diverged from the invertebrates. Both RALBP and IRBP appear to have analogous functions in shuttling retinoids between rhodopsin and the corresponding isomerizing system, retinochrome in the cephalopods and retinol isomerase in the vertebrates. The function of cephalopod "IRBP" is unknown.
The effects of benzodiazepines (BDZs) and GABA system on tonic and clonic component of postdecapitation convulsion (PDC) were studied in mice. Mice decapitated at the occipito-cervical junction, exerted biphasic convulsions, i.e., initially tonic and subsequently clonic convulsions. BDZs such as diazepam or clonazepam increased tonic and clonic components of PDC. These effects were not antagonized by Ro 15-1788, a benzodiazepine receptor antagonist. The increased tonic component was antagonized by the GABA receptor antagonists, bicuculline and picrotoxin, whereas the clonic component was augmented by them. Aminooxyacetic acid, which increases the endogenous GABA content by inhibiting the GABA-transaminase, increased the tonic component significantly; this increase was antagonized by both bicuculline and picrotoxin. Muscimol, a GABA agonist, however did not affect the tonic components but rather augmented the clonic component. Bicuculline and picrotoxin did not antagonize this effect of muscimol. These results indicate that endogenous GABA may play a crucial role in mediating the tonic component of PDC and the facilitation of this component by BDZs may also be due to the activation of GABA in the spinal cord. Furthermore, the mechanisms of the tonic component may be different from that of the clonic component.
Flow cytometric quantitative analysis of cytochrome b on outer surface membrane and of oxidative product formation in polymorphonuclear leukocytes (PMNLs) from patients with chronic granulomatous disease was carried out with the use of monoclonal antibody against cytochrome b of human neutrophils and 2', 7'-dichlorofluorescin diacetate. Cytochrome b was present on the outer surface membrane of PMNLs in normal individuals, and its absence on the outer surface membrane was of value in the diagnosis and classification of chronic granulomatous disease. This study has shown that a major type of chronic granulomatous disease is one of the monoclonal antibody-defined surface membrane diseases.