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Biomedical subjects

J Y Song

Publications and source records attributed to J Y Song.

At least 55 records · Page 3Linked to original sources

Isolation of a root-specific cDNA encoding a ns-LTP-like protein from the roots of bean (Phaseolus vulgaris L.) seedlings.

A root-specific cDNA clone, PVR3, was isolated from a bean (Phaseolus vulgaris L.) root cDNA library by a differential screening procedure. The nucleotide sequence of PVR3 contains an open reading frame coding for an 11.14 kDa polypeptide of 102 amino acid residues; the first 25 amino acids correspond to the sequence characteristic of a signal peptide. Comparison of the deduced PVR3 polypeptide sequence with the polypeptide sequences of previously cloned genes indicates that PVR3 may encode a ns-LTP-like protein. Molecular modelling of the PVR3 protein predicts that it has a three-dimensional structure that is similar to the three-dimensional model determined from the maize ns-LTP. The PVR3 mRNA accumulated mainly in the roots of young seedlings. It can be detected at low levels in flowers, but it is not detected in other organs. Genomic Southern blot analysis indicates that the genomic DNA corresponding to PVR3 cDNA is encoded by a single gene or small gene family in the bean genome.

Amino Acid Sequence↗

Ultrastructural localization of activity of phosphatases by low temperature incubation of unfixed cryostat sections.

In the present study, we demonstrate the activity of several phosphatases ultrastructurally in long-term (up to 24 months) cold-stored (-80 degrees C) rat tissues. Phosphatase activity was histochemically studied with the use of unfixed cryostat sections in combination with low temperature (4 degrees C) incubation conditions in order to prevent inactivation of enzyme activity and to limit the loss of ultrastructure. 5'-Nucleotidase activity was observed at plasma membranes, mainly at bile canalicular membranes of hepatocytes in liver. Thiamine pyrophosphatase activity was detected not only in trans side cisternae but also in medial and cis side cisternae of Golgi complexes in the parotid gland. Glucose-6-phosphatase activity was localized in endoplasmic reticulum as well as at the outer membrane of the nuclear envelope. Acid phosphatase reaction product was found in lysosomes. Furthermore, the localization patterns of 5'-nucleotidase and thiamine pyrophosphatase activity were compared with those obtained after different fixation procedures such as immediate chemical fixation of tissues or fixation of tissues after freezing and thawing. The results showed similar localization patterns of these enzymes after the different pretreatments. However, with respect to the ultrastructural morphology, some damage was observed in unfixed material after incubation. It can be concluded that the procedure described here enables ultrastructural localization of activity of phosphatases in long-term cold-stored tissues. This procedure will be useful for a retrospective study on archival material when histochemical parameters are needed.

5'-Nucleotidase↗

Disturbed structural interactions between microfilaments and tight junctions in rat hepatocytes during extrahepatic cholestasis induced by common bile duct ligation.

Microfilaments in epithelial cells are important for the structural and functional integrity of tight junctions. In the present study, we examined the relationship between microfilaments and tight junctions in hepatocytes of rat liver following common bile duct ligation (CBDL) for up to 2 weeks. Actin filaments and tight junctions were studied by fluorescence microscopy using 7-nitrobenzene-2-oxa-1,3-diazole phallacidin (NBD-ph) and an anti-ZO-1 antibody, respectively. Double-stained sections were examined with confocal laser scanning microscopy (CLSM). Electron microscopy was applied for the assessment of structural alterations in microfilaments and in tight junctions with detergent-extraction and freeze-fracture preparations. Our results showed that F-actin was present at the entire plasma membrane of hepatocytes in control liver, whereas CBDL increased the amount of F-actin mainly at the bile canalicular and lateral plasma membranes. Simultaneously, the immunofluorescence of ZO-1 underwent striking changes, i.e., from a uniform to an irregular staining pattern with various fluorescence intensities. CLSM demonstrated a colocalization of ZO-1 and F-actin in control liver and its deterioration in CBDL liver. Electron microscopy showed marked alterations of microfilaments and tight junctions due to CBDL. It is concluded that actin filaments are intimately associated with tight junctions in normal hepatocytes. CBDL impairs this association by progressively diminishing the structural interaction between F-actin and ZO-1, which may in turn lead to functional disturbances of tight junctions.

Actin Cytoskeleton↗

Depot goserelin and danazol pre-treatment before rollerball endometrial ablation for menorrhagia.

OBJECTIVE: To compare the safety and efficacy of a GnRH agonist, depot goserelin, and danazol as preoperative treatments before rollerball endometrial ablation. METHODS: We performed an open, randomized study of women with clinically convincing histories of menorrhagia. Two treatment groups of equal size (goserelin acetate, 3.6 mg subcutaneous implant given monthly, n = 30; and danazol, 200 mg two times per day, n = 30) were treated for 2 months before undergoing endometrial ablation, which entailed using the rollerball with 80-100 watts of coagulating current. RESULTS: Measured menstrual blood loss decreased dramatically after ablation, with 74% of goserelin users and 62% of danazol users achieving complete amenorrhea by the end of the 6-month follow-up period. Among women who did not achieve complete amenorrhea, those in the danazol group were more likely to experience occasional episodes of moderate or heavy bleeding. Mean measured blood loss decreased from 94.8 to 1.1 mL at 3 months and 1.0 mL at 6 months after goserelin, and 97.9 to 15.0 mL and 7.4 mL after danazol. Menstrual pain also improved markedly. The median duration of surgery was 20 minutes (range 5-55) in both groups, median irrigation fluid deficit was 100 mL (range 0-800), and median operative blood loss was 20 mL (range 1-50). Endometrium was less than 2 mm thick in all goserelin and most danazol users. Side effects of goserelin and danazol therapy were all within the expected pattern for these drugs. CONCLUSION: Depot goserelin and danazol both provide adequate endometrial preparation before rollerball endometrial ablation for treatment of menorrhagia due to ovulatory dysfunctional uterine bleeding.

Combined Modality Therapy↗

Medium-term follow-up of women with menorrhagia treated by rollerball endometrial ablation.

OBJECTIVE: To assess the medium-term efficacy of rollerball endometrial ablation. METHODS: From January 1990 to December 1993, 142 women underwent rollerball endometrial ablation for menorrhagia, most with ovulatory dysfunctional uterine bleeding. Each was sent a detailed questionnaire 12-52 months later and then followed-up, with an overall 93% response rate. Thirty-two had uterine myomas (13 with hysteroscopic myoma resection) and 11 had a large endometrial polyp (four with both). Most of the women were pretreated with danazol. RESULTS: Thirty-two of 128 (25%) achieved long-term, complete amenorrhea and only three eventually resumed their periods after prolonged amenorrhea. Four women experienced late amenorrhea, which was probably due to menopause. The percentage of satisfactory outcomes decreased gradually with time (95.5, 85.5, and 75% for 13-24, 25-36 and more than 36 months respectively; P = .051); this was not statistically significant. One hundred twenty-one patients (85.2%) did not need further surgical treatment. The incidence of repeat ablation (8.5%) and hysterectomy (8.5%) increased gradually with time over the first 3 years but not thereafter, although total numbers are still small. Cyclic pelvic pain and premenstrual symptoms were much improved in both short and long term. Most women remained satisfied with the operation (84%) and would recommend it to others (89%). CONCLUSION: Rollerball endometrial ablation is a simple, effective, and acceptable procedure for the management of intractable menorrhagia due to ovulatory dysfunctional uterine bleeding. However, with the findings of slightly less satisfactory results with time, longer-term follow-up is still needed to establish the ultimate effectiveness of the procedure using different techniques.

Adult↗

Regulation of interleukin-4 on collagen gene expression by systemic sclerosis fibroblasts in culture.

This study was performed to evaluate the transcriptional regulation of interleukin-4 (IL-4) on collagen gene expression in systemic sclerosis (SSc) fibroblasts. The pro alpha 2(I) collagen promotor activity has been examined by transfection experiments and chloramphenicol acetyl transferase (CAT) assay. Maximal elevation of collagen synthesis was presented at a concentration of 5 ng/ml of IL-4. In the CAT assay, the percentage of acetylation was 7.0% +/- 2.0% in untreated controls and 12.5% +/- 3.5% in SSc fibroblasts at 5.0 ng/ml of IL-4. In normal skin (NS), 3.5% +/- 1.0%, and 10.2% +/- 2.5% were acetylated, respectively. The promoter activity was increased 1.8 +/- 0.7-fold and 2.9 +/- 1.3-fold in IL-4-treated SSc and NS, respectively, as compared to untreated groups. In Northern and dot-blot analysis, the level of types I and III collagen mRNA increased 1.1 +/- 0.3-fold and 1.3 +/- 0.3-fold, respectively, in IL-4-treated SSc fibroblasts compared to 3.0 +/- 0.4-fold and 3.0 +/- 0.3-fold, respectively, in NS fibroblasts. These data may indicate that IL-4 could be important in promoting biogenesis of collagen proteins by increased stability and transcription of the collagen mRNA. Also, transcriptional activation of collagen gene expression appears to have a less sensitive effect on SSc than on NS.

Adult↗

A comparison of mechanisms underlying disturbances of bleeding caused by spontaneous dysfunctional uterine bleeding or hormonal contraception.

Recent research has not been able to demonstrate close endometrial morphological correlations with specific abnormalities of menstrual bleeding, but has pointed to an increasing number of molecular mechanisms that may be involved in the occurrence of certain forms of abnormal uterine bleeding. Ovulatory and anovulatory dysfunctional uterine bleeding (DUB) and progestogen-related breakthrough bleeding (BTB) are three conditions with quite different clinical characteristics. It is also probable that the local endometrial molecular mechanisms associated with these three menstrual disturbances are quite different. Ovulatory DUB is associated with a series of vascular and haemostatic disturbances that all appear to contribute to increased loss of blood and tissue fluid at menstruation. Anovulatory DUB is associated with obvious disturbances of endometrial histology, vascular morphology and fragility, with variable and increased blood flow. Progestogen-related BTB is associated with a multitude of morphological and functional endometrial changes that appear to relate predominantly to a patchy capillary origin for the bleeding. Many molecular and cellular changes have been observed in all three conditions. It is not yet known whether there is a single, but different, underlying mechanism responsible for these multiple abnormalities in each of the three clinical situations.

Anovulation↗

Effects of high dose progestogens on white cells and necrosis in human endometrium.

Endometrium was collected from 20 high-dose progestogen-treated patients and examined for leukocyte populations by immunohistochemistry and phloxine-tartrazine staining. A labelled streptavidin-biotin-alkaline phosphatase technique was used with antibodies against leukocyte common antigen (LCA), T cells (CD3), neutrophils (NP57), macrophages] (CD68, KP1) and B cells (CD20). The numbers of LCA (1070 +/- 117/mm2), CD3 (459 +/- 60/mm2), CD68 (129 +/- 21/mm2) positive cells and endometrial granulated lymphocytes (EGL) (236 +/- 41/mm2) were significantly higher than those in the control group (P < 0.001). Of these, EGL increased most (6.7 times). NP57 positive (NP57+) neutrophils were present in five out of 20 progestogen-treated samples and NP57 negative (NP57-) neutrophils in another six out of 20; while a neutrophil was only identified in one control tissue (P = 0.002). Three progestogen-exposed endometrial samples had either focal or extensive necrosis, and many NP57+ and NP57- neutrophils were present in the necrotic areas. EGL, neutrophils and macrophages are known to release a number of cytolytic and cell toxic molecules which may play a role in the initiation or acceleration of progestational endometrial necrosis.

Antigens, CD↗

Redistribution of Ca2+, Mg2+-ATPase activity in relation to alterations of the cytoskeleton and tight junctions in hepatocytes of cholestatic rat liver.

Ca2+, Mg2+-ATPase is a membrane-bound enzyme localized at the bile canalicular membranes of hepatocytes. Cytoskeleton and tight junctions are important for maintenance of the polar distribution of plasma membrane proteins. In order to understand the mechanisms involved in the redistribution of Ca2+, Mg2+-ATPase due to cholestasis, the relationship between Ca2+, Mg2+-ATPase, microfilaments and tight junctions was examined. Cholestasis was induced in rat liver by common bile duct ligation (CBDL) for 2 weeks. Localization of Ca2+, Mg2+-ATPase activity was studied at the light and electron microscopic level. Double-staining of the enzyme and F-actin was performed using phase-contrast and fluorescence microscopy of 7-nitrobenzene-2-oxa-1,3-diazole phallacidin (NBD-ph), respectively. Immunofluorescence microscopy of ZO-1 was applied for the observation of tight junctions. Furthermore, cytoskeleton and junctional complexes were investigated electron microscopically in saponin-extracted tissues. The results showed that CBDL induced redistribution of Ca2+, Mg2+-ATPase activity from the apical to the entire plasma membrane of hepatocytes, which seemed to occur independently of F-actin. F-actin was present at all membrane domains of hepatocytes in control liver, whereas CBDL increased the amounts of F-actin mainly at the bile canalicular membranes. An inverse distribution pattern of Ca2+, Mg2+-ATPase activity and F-actin was found in epithelial cells of bile ducts in control and cholestatic livers. Marked alterations in microfilaments were observed at the bile canaliculi, which were defined as hypertrophy and atrophy and were in association with changes in tight junctions. Structural impairment of the tight junctions was proven by disordered immunofluorescence of ZO-1. It is concluded that changes in the distribution of Ca2+, Mg2+-ATPase and F-actin due to CBDL are independent of each other. CBDL-induced disorders of microfilaments are related to impairment of structural integrity of tight junctions that is suggested to be responsible for the redistribution of Ca2+, Mg2+-ATPase in hepatocytes.

Actins↗

The effect of high-dose medium- and long- term progestogen exposure on endometrial vessels.

A total of 19 paraffin-embedded endometrial tissue blocks were obtained from high-dose progestogen-exposed patients. A labelled streptavidin-biotin-alkaline phosphatase method was used with antibodies against von Willebrand factor (vWF) and CD34. The density of CD34 and vWF positive (CD34+ and vWF+) vessels in progestogen-exposed endometria (103 +/- 9.6/mm2 and 106 +/- 8.7/mm2) was significantly lower than in endometria from women with normal cycles (169 +/- 9.3/mm2 and 136 +/- 8.0/mm2) (P < 0.05). In women with normal menstrual cycles the concentration of CD34+ vessels was significantly higher than the number of vWF+ vessels (P = 0.0001). By comparison, the concentration of CD34+ vessels was similar to the concentration of vWF+ vessels in progestogen-exposed endometria. The ratios of vascular density as determined by vWF+ and CD34+ staining the control and progestogen groups were 0.81 and 1.05 respectively (P = 0.0001). Dilated venules were seen in the progestogen group. This study has demonstrated firstly that CD34 antibody detected the endothelial cells in a higher proportion of small endometrial vessels than vWF, and secondly that high-dose progestogen exposure significantly decreased the density of microvessels and increased the number of dilated venules in endometrium.

Antigens, CD↗

Effects of progestogens on human endometrium.

The effect of progestogens on endometrium depends on the dosage, duration of exposure, the type of progestogen, and the presence or absence of estrogen. Mechanisms of progestogen action in endometrium are mainly expressed through the binding of hormone to specific nuclear receptors. Exogenous progestogens seem to influence the hypothalamic-pituitary-ovarian-endometrial axis differently in different individuals. Endogenous hormones resulting from ovarian secretion have effects on the endometrium independent of, and in combination with, exogenous progestogens. Endometrial morphological changes with progestogens vary from suppression of endometrial glandular growth, through stromal decidualization and leukocytic infiltration to glandular atrophy and stromal focal necrosis. In a minority of cases resulting from very prolonged treatment connective tissue fibers increase to some degree and may be accompanied by endometrial fibrosis and calcification. Clinical and histological data have demonstrated that all these changes, including fibrosis and calcification, return to normal in a short period after discontinuing treatment. Endometrial changes during progestogen therapy are often accompanied by leukocyte infiltration, especially when necrosis occurs. White blood cells constitute an important component of normal endometrium. The number and the type of leukocytes change during the normal menstrual cycle apparently related to circulating ovarian hormonal changes. Exogenous progestogens also influence white blood cells, by increasing total numbers and certain specific cell types. Changes in endometrial white blood cell function as a consequence of exogenous progestogens are unclear, but it is possible that the increase of leukocyte infiltration resulting from exogenous progestogens plays an important role in evoking progestational endometrial necrosis.

Cell Movement↗

Ventricular tachycardia and exercise related syncope in children with structurally normal hearts: emphasis on repolarisation abnormality.

OBJECTIVE: To emphasize the importance of ventricular tachycardia associated with repolarisation abnormality in syncope associated with exercise. DESIGN: Retrospective analysis of data on children presenting with syncope between 1985 and 1993. PATIENTS: 5 apparently normal children with recurrent exercise related syncope associated with electrocardiographically abnormal TU complexes. RESULTS: 3 children were diagnosed as having an intermediate form of the long QT syndrome and catecholamine sensitive ventricular tachycardia because the abnormal TU complexes were associated with polymorphic ventricular tachycardia that was not typical of torsades de pointes. Tachycardia was induced by exercise in all patients and by isoprenaline in the one patient who was tested. One patient also had sinus node dysfunction. One child had incessant salvos of polymorphic ventricular arrhythmias and intermittent abnormal TU complexes suggestive of repolarisation abnormalities. The other had typical congenital long QT syndrome. Treatment was effective in three patients; two patients took a beta blocker alone and one took a beta blocker and low doses of amiodarone. One patient died suddenly, death being associated with sinus node dysfunction. In one patient with incessant ventricular arrhythmias treatment with a beta blocker, amiodarone, or Ic drugs was ineffective and always associated with proarrhythmia or syncope. He was not given further treatment and was asymptomatic despite having mild cardiomegaly. CONCLUSIONS: Ventricular tachycardia associated with repolarisation abnormality was an important cause of exercise related syncope in apparently normal children. TU complex abnormalities can be identified by repeated electrocardiography. beta Blockers are effective in preventing recurrent episodes. The role of amiodarone in this type of ventricular tachycardia needs further evaluation.

Adrenergic beta-Antagonists↗

Effects of x-irradiation on survival and extracellular matrix gene expression of cultured keloid fibroblasts.

Survival and extracellular matrix gene expression were studied by viable cell count assay and Northern transfer analysis to compare the sensitivity of normal skin and keloid fibroblasts towards x-irradiation. As the dosage of radiation increased, the numbers of viable cells in irradiated groups were remarkably decreased exponentially, with no significant difference between normal and keloid cell lines. By Northern blot analysis, there was no change in size of the mRNAs for pro alpha 1(I) collagen, fibronectin and beta-actin. By slot-blot hybridization, pro alpha 1(I) collagen mRNA levels in x-irradiated fibroblasts were markedly decreased compared with non-irradiated controls. The amounts of fibronectin and beta-actin mRNAs were also decreased. This study suggests that both normal skin and keloid fibroblasts are sensitive to x-irradiation, and that extracellular matrix gene expression is also affected by such exposure.

Actins↗

Decreased expression of collagen and fibronectin genes in striae distensae tissue.

Striae distensae are characterized by a thinning of connective tissue stroma to produce linear, atrophic-appearing skin. Excessive adrenocortical activity, genetic factors and inherited defects of connective tissues, etc. are important causative factors in the formation of striae distensae, but the basic aetiology is not known. Total RNA was extracted from skin biopsies of five patients with striae distensae. The expression of genes coding for types I and III procollagen, elastin, fibronectin and beta-actin were studied and compared with those of four sex- and age-matched healthy individuals. The percentages of types I and III procollagen mRNA were 9.9 +/- 2.9% (mean +/- s.d.) and 10.6 +/- 1.6%, respectively, of the corresponding controls. The value for fibronectin mRNA in striae distensae was 7.3 +/- 1.8% of the control. The steady-state ratio fibronectin/type I procollagen mRNAs was 0.12 +/- 0.01 in striae distensae and 0.18 +/- 0.01 in the control. These observations suggest that expression of collagens, elastin and fibronectin genes are apparently decreased, and that there is a marked alteration of fibroblast metabolism, in striae distensae.

Adolescent↗

Anxiety and depression in patients with abdominal bloating.

Eighteen patients with a syndrome of abdominal bloating and discomfort were examined to explore the relationship between their symptoms and their emotional problems. They were compared with 33 patients with Crohn's disease and 38 normal, healthy volunteers. Using the Hospital Anxiety and Depression Scale, patients with bloating were found to resemble patients with Crohn's disease. Both groups showed increased anxiety and depression. After controlling for age, sex, education, occupation, personality variables and childhood experience, there was a trend towards more anxiety in the bloating group compared with normal subjects and a significant difference for depression. These characteristics appear to be related to the effects of the illness or to selection, but not to personality or childhood experience. Although psychiatric problems are common among patients with abdominal bloating and pain who stay in touch with a clinic, they are not the primary cause of the disorder.

Abdominal Pain↗

Stage-specific expression of plasmodial proteins containing an antigenic marker of the intraerythrocytic cisternae.

A monoclonal antibody, LWLI, recognized 3 proteins of 45, 50 and 102 kDa in Plasmodium falciparum-infected erythrocytes. The 45- and 50-kDa proteins were parasite-encoded and displayed markedly different peptide maps, indicating that they were distinct plasmodial polypeptides with a common antigenic epitope rather than differentially processed forms of a primary translational product. The 45-kDa protein was present throughout intraerythrocytic growth, while the 50-kDa molecule was not detected earlier than 11 h in the life cycle. The 102-kDa protein was only expressed in trophozoite- and schizont-infected red cells: its structural relationship to the 45- and 50-kDa proteins, if any, remains undefined. By indirect immunofluorescence and immunoelectron microscopy, LWLI bound to flattened intraerythrocytic cisternae exported into the erythrocyte cytoplasm. The results support the theory that proteins recognized by the antibody were concentrated in these compartments and their common antigenic epitope may serve as a marker for the cisternae. Stage-specific expression of LWLI reactive proteins implicates developmental regulation of cisternal functions during asexual parasite development.

Animals↗