Distribution of peripherin/rds mRNA in cone-dominant squirrel retina.
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Animals↗
Biomedical subjects
Y Nakashima
Publications and source records attributed to Y Nakashima.
Mucopolysaccharidosis IV A: assignment of the human N-acetylgalactosamine-6-sulfate sulfatase (GALNS) gene to chromosome 16q24.
Plasmid clones of three independent genomic fragments of the gene for human N-acetylgalactosamine-6-sulfate sulfatase (GALNS; EC 3.1.6.4) were utilized in a fluorescence in situ suppression hybridization study to assign the locus to chromosome 16q24. Enzyme assay for GALNS in a patient with del(16)(q22.1) confirmed this finding.
Blotting, Southern↗
The enhancement of catecholamine-induced Cl- current by cyclic GMP revealed using photolabile caged compounds in guinea-pig ventricular cells.
Whole cell currents were recorded in single myocytes dissociated from guinea-pig ventricles, and caged compounds were loaded intracellularly through the patch electrodes. Flash photolysis of caged cyclic GMP (cGMP) increased the amplitudes of both catecholamine-induced Cl- (ICl) and Ca2+ currents (ICa) which were pre-activated by submaximum doses of isoprenaline. Transient activation of ICl by photo-release of cyclic AMP (cAMP) showed a half decay time (t1/2) of 16.7 +/- 1.4 sec (mean +/- S.E.M., n = 14). This decay was markedly delayed by inhibiting phosphodiesterases using 3-isobutyl-1-methyl-xanthine (IBMX). The intracellular application of cGMP (10-50 microM) also prolonged the decay of the ICl response to caged cAMP (t1/2 = 38.0 +/- 7.1 sec, n = 12). These findings strongly support the hypothesis that cGMP facilitates the beta-adrenergic response of ionic currents through the inhibition of phosphodiesterase in mammalian cardiac myocytes.
Animals↗
Interleukin-5 induces tumor suppression by peritoneal exudate cells in mice.
The antitumor activity of peritoneal exudate cells (PEC) induced by murine interleukin-5 (mIL-5) was examined using Meth-A sarcoma cells transplanted into the peritoneal cavity of mice. Although in vitro treatment of Meth-A sarcoma cells with mIL-5 did not result in inhibition of their growth, treatment of mice intraperitoneally with mIL-5 (1 microgram/day) from day -5 to +5 (tumor cells were inoculated on day 0) led to a significant increase in survival or even rejection of tumor cells. This antitumor effect depended on the dose of mIL-5. Interestingly, there was identical therapeutic activity when the protocol of days -10 to -1 was used as opposed to -5 to +5. In addition, post-treatment with mIL-5 from day +1 to +10 was ineffective. This suggests that the therapeutic activity of IL-5 is largely prophylactic. Under the former condition, the number of PEC was found to increase over 50-fold when compared to levels in control mice. Moreover, the antitumor effect of mIL-5 was completely abolished by subcutaneous injection of anti-mIL-5 monoclonal antibodies. The treatment of mice injected intraperitoneally with human IL-2 also resulted in an increase in survival. Winn assay experiments using PEC recovered from mIL-5-treated mice (1 microgram/day, from day -10 to -1) revealed that these PEC could mediate antitumor activity against Meth-A sarcoma cells. Furthermore, when the cured mice were re-injected with Meth-A sarcoma cells or syngeneic MOPC104E cells, they could reject Meth-A sarcoma cells but not MOPC104E cells, indicating that immune memory had been generated. These results suggest that IL-5 augmented the PEC tumoricidal activity but we have no indication that the tumoricidal activity was mediated through a mIL-5-dependent mechanism.
Animals↗
Low serum apolipoprotein A-I level in patients with vasospastic angina.
The serum lipid and lipoprotein levels in 15 patients with vasospastic angina were compared with those in 33 patients with no angiographic coronary stenosis and no vasospastic angina after intracoronary acetylcholine infusion. The serum level of apolipoprotein A-I in patients with vasospastic angina (112 +/- 6 mg/dl) was significantly lower (p < 0.05) than that in patients without vasospasm (128 +/- 4 mg/dl). However, there were no differences between the two groups in the serum levels of cholesterol, triglycerides, high-density lipoprotein cholesterol, apolipoprotein A-II, and apolipoprotein B. Thus apolipoprotein A-I may play some role in the prevention of vasospastic angina.
Adult↗
Role of left ventricular regional nonuniformity in hypertensive diastolic dysfunction.
OBJECTIVES: This study investigated 1) the role of left ventricular diastolic nonuniformity in hypertensive left ventricular diastolic dysfunction, and 2) the effects of a calcium channel antagonist on diastolic nonuniformity in hypertensive and normotensive subjects. BACKGROUND: Augmented left ventricular diastolic nonuniformity contributes to diastolic dysfunction in hypertrophic cardiomyopathy. Impaired left ventricular diastolic function with preserved systolic function has been recognized in hypertension. Therefore, abnormal ventricular regional nonuniformity might also be involved in hypertensive diastolic dysfunction in a milder form of hypertrophy. METHODS: Thirteen patients with established hypertension underwent radionuclide ventriculography before and after nifedipine administration. Indexes of left ventricular function were derived by computer analysis of the time-activity curve. After a computer subdivided the left ventricle into four regions, a time-activity curve of each region was constructed to determine an index of left ventricular diastolic nonuniformity. This index was calculated as the sum of the absolute values of time difference between global and regional peak filling in the septal, the apical and the lateral region. Ten normotensive subjects were studied for comparison. Echocardiography was performed in both groups. RESULTS: The two groups were matched for age, gender, heart rate, echocardiographic dimensions and systolic function. In the hypertensive group, left ventricular diastolic filling indexes were impaired, with a negative correlation between peak filling rate and the diastolic nonuniformity index. Although the change in ejection fraction after nifedipine administration was similar in the two groups, the increase in peak filling rate was larger in the hypertensive patients. The diastolic nonuniformity index decreased after nifedipine in the hypertensive but not in the control group. This decrease correlated with improved peak filling rate in the hypertensive group. CONCLUSIONS: In hypertensive patients with preserved systolic function, left ventricular diastolic nonuniformity increases, causing early diastolic dysfunction. Decreased diastolic nonuniformity after pharmacologic intervention contributes to lessened ventricular filling dysfunction, regardless of changes in loading conditions in hypertension. Thus, diastolic nonuniformity is an important determinant of left ventricular filling abnormality and might be a target of pharmacologic intervention in hypertensive patients.
Adult↗
Does transesophageal echocardiography improve postoperative outcome in patients undergoing coronary artery bypass surgery?
The incidence and characteristics of ischemic episodes as detected by transesophageal echocardiography (TEE) and their relationship to postoperative myocardial infarction (MI) and adverse clinical outcome were studied in patients undergoing coronary artery bypass grafting (CABG). Seventeen of 50 patients had 21 TEE-defined ischemic episodes: 4 patients (8%) had 4 ischemic episodes in the pre-cardiopulmonary bypass (CPB) period, and 15 patients (30%) had 17 ischemic episodes in the post-CPB period, whereas 19 patients had 20 ECG ischemic episodes: 3 patients (6%) had 3 ischemic episodes in the pre-CPB period and 17 (34%) had 17 ischemic episodes in the post-CPB period. In 14 patients, ischemic episodes were detected by both TEE and ECG. Clinicians were made aware of the TEE data and appropriate treatments were undertaken during ischemic episodes: of the 15 patients with TEE-defined post-CPB ischemia, 4 had an additional saphenous vein graft placed, 8 had an intra-aortic balloon pump (IABP) inserted, 3 were given sublingual nifedipine, and 13 received nitroglycerin. These treatments resulted in improvement in regional wall motion abnormalities (RWMA) by the end of surgery in 11 of the 15 patients (73%), including the 4 with postoperative MI and 2 who died with cardiogenic shock. The authors conclude that: (1) significantly more patients had TEE-defined ischemic episodes in the post-CPB period (30%) than in the pre-CPB period (8%); (2) a poor graft and/or inadequate myocardial protection were strongly suggestive of post-CPB ischemia, which was significantly related to adverse outcome; and (3) TEE was a useful tool enabling detection of problem areas at an early stage and timely and appropriate treatment to support and sustain patients.
Adult↗
The diagnostic use of low molecular weight keratin expression in sebaceous carcinoma.
Sebaceous carcinoma is an infrequent skin tumor and its histological features sometimes closely resemble those of squamous cell carcinoma (SCC) and basal cell epithelioma (BCE), which often leads to a misdiagnosis. In the present immunohistochemical study, however, sebaceous carcinoma exhibited quite a different expression of keratins from SCC and BCE. We immunohistochemically examined 26 excised specimens of sebaceous carcinoma, 10 of SCC and 12 of BCE of the eyelids, using two monoclonal antibodies against high molecular weight keratins, 34 beta B4 (68kd) and 34 beta E12 (56kd, 56.5kd, 58kd), and two monoclonal antibodies against low molecular weight keratins, 35 beta H11 (54kd) and CAM5.2 (39kd, 43kd, 50kd). The cases of sebaceous carcinoma were positive with 34 beta B4 (23%), 34 beta E12 (54%), 35 beta H11 (81%) and CAM5.2 (73%). Of the four anti-keratin antibodies used in this study, 35 beta H11 was negative in all cases of SCC or BCE. These findings indicate that when sebaceous carcinoma is suspected, but no fat staining appropriate materials are available, a monoclonal antibody against low molecular weight keratin, 35 beta H11 (54kd), can be a useful tool to immunohistochemically rule out both SCC and BCE.
Adenocarcinoma, Sebaceous↗
Differential biological significance of tissue-type and urokinase-type plasminogen activator in human breast cancer.
Plasminogen activator (PA) is a serine protease existing in two forms known as tissue-type (t-PA) and urokinase-type (u-PA). To examine whether PA is related to the postoperative clinical course of human breast cancer, total PA activity, t-PA activity, u-PA activity, and immunoreactive t-PA were determined in tissue extracts from 144 breast cancer specimens. The patients were initially divided into four groups according to the postoperative clinical course: Group I (83 patients who are disease-free), Group II (20 patients whose first metastases were found only in bone), Group III (19 patients whose first metastases were found in both bone and lung), and Group IV (22 patients whose first metastases were found only in lung). Total PA activity was significantly lower in Groups, II, III and IV than in Group I. Both t-PA activity and t-PA antigen levels were also significantly lower in Groups II, III and IV than in Group I, while no significant difference was found in u-PA activity among these groups, indicating that low activity of total PA in Groups II, III and IV was due to a decrease in t-PA but not in u-PA. In the multivariate analyses, t-PA activity was found to be an independent prognostic factor for relapse-free survival. When four groups of patients were further analysed in terms of nodal status, both t-PA activity and antigen levels were markedly decreased in the node-negative Group II compared with the node-negative Groups III and IV or with the node-positive Groups II, III and IV. Of additional interest, u-PA activity was significantly higher in node-positive patients than in node-negative patients with any group. The clinico-pathologic analyses of the patients in this series showed that node involvement and lymphatic invasion were more frequently positive in Groups III and IV than in Groups I and II. When 144 breast cancers were categorised in terms of combinations of oestrogen receptor (ER) and progesterone receptor (PgR) status, breast cancers which were positive for both receptors were found to contain the highest t-PA activity and antigen. This study provides provocative evidence suggesting a possible differential significance of t-PA and u-PA expression in human breast cancer.
Bone Neoplasms↗
Breast cancer prognosis is poor when total plasminogen activator activity is low.
Plasminogen activator (PA) is a serine protease which exists in two forms: tissue-type (t-PA) and urokinase-type (u-PA). The total PA activity was measured in tumour extracts of 235 breast cancer patients who were followed for a median of 8.5 years after surgery. Patients were initially divided into three groups with low (< 60 units mg-1 protein), intermediate (60-300 unit mg-1 protein), or high (> 300 unit mg-1 protein) total PA activity in tumour extracts. The PA activity was not significantly associated with the recognised prognostic factors of age, menstrual status, tumour size, lymph node involvement, histologic type, grade of anaplasia, and/or vessel involvement. A significant association was found between total PA activity and the oestrogen receptor (ER) or progesterone receptor (PgR) status. Among receptor-positive tumours, a significantly greater proportion of patients had high PA activity in their tumour extracts. Breast cancer patients with low total PA activity had a significantly shorter disease-free and overall survival rate when compared to those with intermediate or high PA activity. In univariate and multivariate analyses, total PA activity (< 60 unit mg-1 vs > or = 60 unit mg-1 protein) was found to be a significant prognostic factor for disease-free and overall survival of about the same import as lymph node involvement. Furthermore, the combination of total PA activity and nodal status could be even more precise in predicting survival times and probabilities in individual patients. This retrospective study demonstrates the total PA activity is a valuable prognostic factor in determining prognosis in human breast cancer.
Breast Neoplasms↗
Expression of secreted immunoglobulin heavy chain genes and immunoglobulin-secreting cells in human lymphocytes.
The numbers of immunoglobulin-secreting cells in peripheral blood mononuclear cells and the expression of mRNA for secreted type of immunoglobulin heavy chains were investigated in healthy children, compared with the percentages of surface immunoglobulin-bearing cells and the expression of mRNA for membrane-bound type of immunoglobulin heavy chains, respectively. Although a difference between expression of mu s mRNA and mu m mRNA was unclear, mu mRNA was well transcribed. The expression of gamma s mRNA or alpha s mRNA was markedly higher than that of gamma m mRNA or alpha m mRNA. However, although the detection methods could be of different sensitivities, the percentage of IgM-, IgG-, or IgA-secreting cells was markedly low, compared with the percentage of surface IgM-, IgG-, or IgA-bearing cells, respectively. Therefore, additional regulation of the pattern of the immunoglobulin gene expression may be exerted at the translational and post-translational stages.
Antibody-Producing Cells↗
A fixative suitable for in situ hybridization histochemistry.
We compared the morphology and stability of hybridization signals between paraffin sections of rat retina fixed with commonly used 4% paraformaldehyde/PBS and those fixed with a fixative containing glutaraldehyde in in situ hybridization histochemistry, using a digoxigenin-labeled RNA probe complementary for beta-galactoside alpha 2,6-sialyltransferase mRNA. Retinal detachment was frequently observed in the sections fixed with 4% paraformaldehyde-PBS, whereas the morphology was satisfactorily preserved in those fixed with either 0.5% glutaraldehyde, 4% paraformaldehyde-PBS, or 2.5% glutaraldehyde-PBS. Without glutaraldehyde, it was difficult to determine the most appropriate length of proteinase K digestion of tissue sections for facilitating probe penetration, since the optimal time for definite hybridization was variable among the retinal cells in heterogeneous layers. By addition of glutaraldehyde to paraformaldehyde or with glutaraldehyde alone, it was easy to establish the appropriate time for the unmasking procedure, since intense mRNA signals were constant throughout the retina by proteinase K digestion for more than 30-40 min. Using a fixative that causes stronger cross-linking (e.g., glutaraldehyde) is recommended to improve not only the morphology but also the stability of hybridization signals in in situ hybridization histochemistry with paraffin embedding and digoxigenin-labeled RNA probes.
Animals↗
Amino-acid sequence of the alpha D- and beta-polypeptide chains of the Japanese quail hemoglobin.
Two hemoglobin components are recognized in the erythrocytes of the adult Japanese quail: a major (Q-II) and a minor (Q-I) component. We have determined the amino-acid sequence of the alpha D-globin of the minor component and the beta-globin which is common to both components by conventional protein sequence analysis. The sequences of both the alpha D- and the beta-globins showed close homology with those of their counterpart constituents in other avian hemoglobins. Proline at position 119 in the alpha D-globin which is known to be critical for the altitude respiration by the alpha 1 beta 1 interface is conserved in the Japanese quail hemoglobin.
Amino Acid Sequence↗
Pathological extent of interventricular septal infarction in patients with acute anteroseptal myocardial infarction with and without right bundle branch block.
The purpose of the present study is to elucidate the difference in extent of myocardial infarction (MI) between those cases with and those without right bundle branch block (RBBB) occurring during the course of acute MI. We examined postmortem hearts from 20 patients with acute anteroseptal MI; 10 with (group A) and 10 without RBBB (group B). The extent of MI was studied pathologically in the interventricular septum (IVS) and reconstructed. The longitudinal extent of MI did not distinctly differ between groups A and B. In 5 hearts of group B, the anterior limit of the MI extended as high, or as close to the cardiac base, as in group A hearts. Transmural MI was relatively common and seen more frequently in group A than in group B. However, the MI did not always extend evenly to the left and right ventricular sides in the IVS. Left-sided predominance in extent of the MI was more frequently observed in group B than in group A. Right-sided predominance in the extent of septal MI was relatively rare and was seen in 5 cases at the portion where the right bundle branch came down. Four of these were group A patients. Thus, involvement of the right bundle branch might partly depend on the unevenness of the mural extent of the MI as well as the height of extension of the MI in the IVS.
Aged↗
Evaluation of the effects of different cardioplegic techniques on left ventricular function during coronary artery bypass surgery using transesophageal echocardiography.
Fifty consecutive patients undergoing coronary artery bypass grafting surgery were studied to evaluate the effect of intermittent anterograde cold cardioplegia (IACCH) and intermittent combined anterograde-retrograde cold cardioplegia (IRCCN) on left ventricular function using transesophageal echocardiography. Global function did not significantly change in both groups, but significantly more inotropes were required in IACCH. Newly developed abnormalities of regional wall motion after cardiopulmonary bypass, which were indicative of ischemic myocardium, were detected in the segments supplied by the right coronary artery in both groups (IACCH, 20%; IRCCN, 16%), by the left circumflex coronary artery only in IACCH (12%) and by the left anterior descending coronary artery only in IRCCN (12%). It is concluded that although global evaluation of left ventricular function did not show any significant change after IACCH and IRCCN under routine management, analysis of abnormalities of regional wall motion provided specific information. In both groups, complete protection of the myocardium was not achieved, and the characteristics of poorly protected areas were dependent on the difference in the two methods. Myocardium supplied by the right coronary artery seemed to be particularly vulnerable, and a special effort to protect these segments is mandatory for a successful outcome.
Aged↗
Effect of a high linoleate and a high alpha-linolenate diet on general behavior and drug sensitivity in mice.
Semi-purified diets supplemented with either a high linoleate (n-6) (safflower) oil or a high alpha-linolenate (n-3) (perilla) oil were fed to mouse mothers and their offspring through 6 weeks of age. The proportions of n-3 and n-6 highly unsaturated fatty acids in brain phospholipids reflected the n-3/n-6 balance of the diets while no difference was found in phospholipid compositions or cholesterol/phospholipid ratios. In the elevated plus maze task, the total number of entries into the open- and enclosed-arms was smaller and the time spent in the dark enclosed arms tended to be longer in the perilla group than the safflower group. The time required to reach a safe platform in Morris's water maze test was less in the perilla group, but no significant difference was observed in the entries into the arms darkened with a movable cover in Y-maze dark-preference task. The safflower group was more sensitive to pentobarbital; the anesthesia onset time was less and the anesthetic time was longer than in the perilla group. Increased locomotion induced by scopolamine injection was less in the safflower group as compared with the perilla group. These results indicate that in mice the dietary alpha-linolenate/linoleate balance affects the n-3/n-6 ratio of brain phospholipid acyl chains and that this is accompanied by general behavioral changes as well as changes in sensitivities to drugs known to affect behavior.
Analgesia↗
Demonstration of peripherin/rds mRNA in normal and light-damaged rat retinas by in situ hybridization histochemistry.
Cellular expression of the mRNA for peripherin/rds was studied in rat retinas by in situ hybridization histochemistry with antisense and sense probes prepared from polymerase chain reaction-amplified cDNA of the bovine peripherin/rds. Predominant mRNA signals were detected in the inner segments and distal region of the outer nuclear layer, but not in other layers of the retina including retinal pigment epithelium or in the choroid, giving evidence that the gene product, peripherin/rds, is synthesized specifically in the inner segment ribosomes to form outer segment disc membranes of photoreceptor cells. The in situ hybridization technique was applied to explore how mRNA for peripherin/rds is affected by light-induced retinal damage in rats. Following 3 days of continuous exposure to low-intensity fluorescent light, the mRNA expression in adult rats was found to be defective, although photoreceptor cells were still observed by light microscopy. After longer exposure to continuous light, the mRNA expression was severely damaged or undetectable, together with loss of photoreceptor cells.
Animals↗
Role of uremic and endothelial factors in the development of beta 2-microglobulin amyloidosis.
To clarify the importance of uremic and endothelial factors in the development of beta 2-microglobulin (beta 2m) amyloidosis, in vitro evaluations were carried out using cultured synoviocytes and synovial tissues. Synoviocytes of patients with chronic renal failure were cultured with media containing uremic or normal serum. In cultures with uremic serum, greater proliferation of synoviocytes and stronger expression of CD68, beta 2m, interleukin 1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) on synoviocytes were observed, compared with those containing normal serum. Uremic serum stimulated greater production of IL-1 beta and TNF alpha than normal serum (P < 0.05 for IL-1 beta, P < 0.01 for TNF alpha). Addition of supernatant of endothelial cell culture to medium containing uremic serum significantly accelerated production. After a three month culture of uremic synovial tissue, embedded in 3% type 1 collagen gel with uremic serum, beta 2m-positive 10 nm fibrils were recognized around the synoviocytes. In culture with uremic serum plus supernatant of endothelial cell culture, positive Congo red staining was noted. These findings indicate that uremic serum plays a significant role in the development of synovial beta 2m amyloidosis and that endothelial cell-derived factors contribute to the formation of beta 2m amyloid.
Amyloidosis↗