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Biomedical subjects

Y Dai

Publications and source records attributed to Y Dai.

At least 217 records · Page 12Linked to original sources

Selective extraction of outer-membrane proteins from membrane complexes of Pseudomonas maltophila by chloroform-methanol.

An organic phase partitioning method is described for the selective purification of outer-membrane proteins (OMPs) from the total membrane complex of the opportunistic human and sheep pathogen, Pseudomonas maltophila. SDS-PAGE analysis confirmed that OMPs purified by chloroform-methanol treatment of the total membrane complex were not only identical to OMPs extracted from outer-membrane vesicles separated by sucrose gradient density centrifugation, but also possessed little or non-detectable levels of inner-membrane contaminants. Further analysis by enzyme linked immunosorbent assay (ELISA) and immunoblotting established that OMPs extracted by organic phase partitioning with chloroform-methanol retained antigenicity and serological activity indistinguishable from OMPs that were present in outer-membrane vesicles resolved by isopycnic sucrose density centrifugation of sarkosyl-treated membrane complexes.

Animals↗

Karyophilic properties of Semliki Forest virus nucleocapsid protein.

Semliki Forest virus capsid (C) protein molecules (Mr, 33,000) can be introduced efficiently into the cytoplasm of various target cells by electroporation, liposome, and erythrocyte ghost-mediated delivery (M. Elgizoli, Y. Dai, C. Kempf, H. Koblet, and M.R. Michel, J. Virol. 63:2921-2928, 1989). Here, we show that the transferred C protein molecules partition rapidly from the cytosolic compartment into the nucleus. Transport of the C protein molecules into the nucleus was reversibly arrested by metabolic inhibitors, indicating that the transfer process is energy dependent. Fractionation of isolated nuclei revealed that the delivered C protein preferentially associates with the nucleoli. This finding was confirmed by morphological studies, showing that in an in vitro system containing ATP isolated nuclei rapidly accumulated rhodamine-labeled C protein in their nucleoli. Furthermore, in this assay system, the lectin wheat germ agglutinin prevented transfer of C protein through nuclear pores. These results are in agreement with our observation that nucleoli contain measurable amounts of newly synthesized C protein as early as 5 h after infection of cells with SFV. Thereafter, nucleolar-associated C protein increased progressively during the course of infection.

Adenosine Triphosphate↗

ADPRT-mediated decrease of cellular NAD content and the detection of chemically induced DNA damage--development of a new short-term screening test for mutagens.

It was found that the DNA-damaging agents N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), methyl-methanesulphonate (MMS) and 4-nitroquinoline-N-oxide (4NQO) could stimulate ADP-ribosyl transferase (ADPRT) activity and reduce the cellular NAD content in a dose-dependent way. The reduction of NAD after DNA damage could be partially or completely prevented by ADPRT inhibitors, 3-aminobenzamide or nicotinamide, which showed no influence on reduction of NAD induced by metabolic blocking agents. Therefore, a simple and specific method to detect DNA-damaging mutagens by measuring ADPRT-mediated decrease of cellular NAD content was explored. Using beta-naphthoflavone, a mixed function oxygenase inducer, together with induced or uninduced human amnion FL cells, it was found that aflatoxin B1, benzo(a)pyrene, 2-acetylaminofluorene, 9,10-dimethylanthracene and ethylcarbamate could induce the ADPRT-mediated decrease of cellular NAD content, while 4-acetylaminofluorene, anthracene, isopropyl-N-(3-chlorophenyl)-carbamate, beta-propiolactone, gamma-butyrolactone, cyclophosphamide and safrol could not. The results indicate that this is a cheap and specific method to detect DNA damage caused by chemical carcinogens/mutagens with a specificity approaching that of the unscheduled DNA synthesis assay.

Amnion↗

Semliki Forest virus capsid protein acts as a pleiotropic regulator of host cellular protein synthesis.

The Semliki Forest virus capsid (C) protein was introduced into various target cells by electroporation-, liposome-, and erythrocyte-ghost-mediated delivery. Data are presented which show that the incorporated C protein is biologically active and, at low concentrations (10(3) to 10(4) molecules per cell), markedly induces host cellular protein synthesis (average value, up to 90%). On the other hand, high concentrations (10(5) to 10(6) molecules per cell) led to a significant inhibition (average value, up to 60%). The cellular response to C protein was found to be identical in P3X63Ag8 suspension cells, CV-1 cells, and GpBind4 cells. Following electroporation-mediated delivery of C-protein molecules, both induction and repression of cellular protein synthesis were immediate, whereas with liposome-mediated delivery these events were delayed by about 1 h. Maximum stimulation and repression occurred between 0 and 1 h after delivery of C protein and decreased thereafter to reach control values at about 4 h. The analysis of the proteins synthesized suggests that low amounts of microinjected C protein are responsible for the induction of classes with specific Mrs, whereas high amounts lead to an inhibition of overall protein synthesis.

Animals↗

[Anti-inflammatory effect of fructus Ligustri Lucidi].

Fructus Ligustri Lucidi (FLL) inhibits the edema of hind Paw induced by injection of 1% carrageenin 0.1 ml, fresh egg 0.1 ml or 2.5% formaldehyde 0.1 ml in rats, and suppresses the increased vascular permeability induced by 0.7% acetic acid in mice and the proliferation of granuloma induced by cottonpellet in rats. The swelling of ear induced by xylene in mice and the content of prostaglandin E in inflammatory tissue of rats are also decreased by FLL. FLL increase the weight of adrenal in rats, but has no marked effect on the weight of thymus of rats.

Adrenal Glands↗

[Effects of oleanolic acid on immune system and type I allergic reaction].

Oleanolic acid (OLA)50, 100 mg/kg sc antagonised the inhibitory effect of cortisone on the weights of spleen and thymus, decreased the clearance rate of charcoal particles in mice. The production of serum specific antibody hemolysin was not clearly affected by OLA. The administration of OLA 100 mg/kg enhanced the development of serum antibody immunoglobulin G in mice and showed marked inhibition on the hemolytic activity of the total complement by classical pathway in guinea pigs. In addition, the homologous passive cutaneous anaphylaxis in mice or rats and the degranulation of mast cells of calvarial periosteum in rats were significantly prevented by OLA 50, 100 mg/kg. The increased capillary permeability in the rat skin caused by intradermic injection of histamine was also reduced by OLA 100 mg/kg. These results indicate that OLA inhibited the type I allergic reaction.

Animals↗

Effect of graded low-dose noradrenaline infusions on noradrenaline clearance in normals and essential hypertensives.

1. In normal subjects and essential hypertensives, plasma noradrenaline (NA) levels, and NA clearance (3H-NA constant infusion), were measured during graded infusion of NA. 2. Plasma levels of NA were similar in both groups, with parallel increases to levels approximately three times basal, but still in the upper normal range. 3. Clearance did not increase in either group with increasing plasma NA levels. There was no correlation between plasma NA achieved and NA clearance, which was similar in hypertensives and normotensives. 5. Insofar as plasma clearance reflects tissue clearance, these studies provide no support for the contention that altered NA metabolism might predispose to elevated NA levels in essential hypertension.

Adult↗

Three-dimensional microscopy data exploration by interactive volume visualization.

This paper presents a new volume visualization approach for three-dimensional (3-D) interactive microscopy data exploration. Because of their unique image characteristics, 3-D microscopy data are often not able to be visualized effectively by conventional volume visualization techniques. In our approach, microscopy visualization is carried out in an interactive data exploration environment, based on a combination of interactive volume rendering techniques and image-based transfer function design methods. Interactive volume rendering is achieved by using two-dimensional (2-D) texture mapping in a Shear-Warp volume rendering algorithm. Image processing techniques are employed and integrated into the rendering pipeline for the definition and searching of appropriate transfer functions that best reflect the user's visualization intentions. These techniques have been implemented successfully in a prototype visualization system on low-end and middle-range SGI desktop workstations. Since only 2-D texture mapping is required, the system can also be easily ported to PC platforms.

Actins↗

Simplified techniques in rat heterotopic small bowel transplantation.

AIM: Establish a simplified heterotopic small bowel transplantation (SBT) in the rat. METHODS: Ninety pairs of male Wistar rats were used as donors and recipients. The whole small intestine with a vascular pedicle composed of superior mesenteric artery (SMA) and portal vein (PV) was harvested as the graft. Revascularization was accomplished by end-to-side anastomosis between donor SMA and recipient infrarenal aorta and cuffed end-to-end anastomosis between donor PV and left renal vein of recipient. The distal end of graft was exteriorized to form an enterostoma. RESULTS: Average time of an operation was 130 minutes and the mean warm ischemia time of grafts was 30 minutes. The technical success rate of this model was 100% and 7-day survival was 95.6% (86/90). CONCLUSION: This simplified technique was effective and practical to improve the outcome of rat heterotopic SBT.

Animals↗

Effects of ulinastatin on postoperative systemic inflammatory response of recipients of rat small bowel transplantation.

AIM: We sought to evaluate the effects of ulinastatin on postoperative systemic inflammatory responses of recipients of rat small bowel transplantations (SBT). METHODS: Twenty-four recipients of rat heterotopic SBT were randomly divided into a control group and a treated group. Ulinastatin (50,000 U/kg(-1)/d(-1)) was injected intravenously 30 minutes before graft revascularization. Measured variables included plasma concentrations of tumor necrosis factor (TNF), interleukin (IL)-6, and C-reactive protein (CRP) on postoperative days 1 and 3. RESULTS: Administration of ulinastatin attenuated the postoperative increases in plasma concentrations of TNF, IL-6, and CRP. CONCLUSION: Ulinastatin attenuated the postoperative systemic inflammatory response of rat recipients of SBT.

Animals↗

Association and linkage analysis of candidate chromosomal regions in multiple sclerosis: indication of disease genes in 12q23 and 7ptr-15.

Four recent genome-wide screen studies in multiple sclerosis (MS) identified a number of candidate regions for susceptibility genes in addition to the HLA complex in 6p21. However, none of these regions provided formally significant evidence for genome-wide linkage. We have investigated such regions in 46 Swedish multiplex MS families, 28 singleton families, 190 sporadic MS patients and 148 normal controls by parametric and nonparametric linkage and association analysis. One microsatellite marker, in 12q23, provided evidence for association in addition to suggestive transmission distortion and slightly positive linkage. In addition, a marker in 7ptr-15 showed a significant transmission distortion as well as a highly significant score in affected pedigree member analysis, but not quite significant deviations in association analysis. One of three markers in 5p, a region implicated in all four previous studies, showed a weakly positive lod score, but no other evidence of importance. Markers in 2p23, 5q11-13, 6q25, 7q21-22, 11q21-23, 13q33-34, 16p13.2, 18p11.32-23, Xp21.3 provided little or no evidence of importance for MS. In summary, these data support the importance of genome-wide screens in the identification of new candidate loci in polygenic disorders.

Alleles↗

Changes in activity of energy dissipating mechanisms in wheat flag leaves during senescence.

Excitation energy dissipation, including the xanthophyll cycle, during senescence in wheat flag leaves grown in the field was investigated at midday and in the morning. With progress of senescence, photosynthesis (Pn) and actual PSII photochemical efficiency (PhiPSII) decreased markedly at midday. The decrease in extent of Pn was greater than that of PhiPSII. However, there was no significant decline in Pn and PhiPSII observed in the morning, except in leaves 60 days after anthesis. The kinetics of xanthophyll cycle activity, thermal dissipation (NPQ), and q (f) observed at midday during senescence exhibited two distinct phases. The first phase was characterized by an increase of xanthophyll cycle activity, NPQ, and q (f) during the first 45 days after anthesis. The second phase took place 45 days after anthesis, characterized by a dramatic decline in the above parameters. However, the qI, observed both at midday and in the morning, always increased along with senescence. A larger proportion of NPQ insensitive to DTT (an inhibitor of the de-epoxidation of V to Z) was also observed in severely senescent leaves. In the morning, only severely senescent leaves showed higher xanthophyll cycle activity, NPQ, q (f), and qI. It was demonstrated that, at the beginning of senescence or under low light, wheat leaves were able to dissipate excess light energy via NPQ, depending on the xanthophyll cycle. However, the xanthophyll cycle was insufficient to protect leaves against photodamage under high light, when leaves became severely senescent. The ratio of (Fj - Fo)/(Fp - Fo) increased gradually during the first 45 days after anthesis, but dramatically increased 45 days after anthesis. We propose that another photoprotection mechanism might exist around reaction centres, activated in severely senescent leaves to protect leaves from photodamage.

Aging↗

Oocyte maturation and embryonic failure.

Embryonic development is readily compromised by imperfections introduced during the process of oocyte maturation. We discuss the nature and causes of these imperfections, particularly in oocytes exposed to inappropriate hormonal regimes in vivo or to culture systems designed to induce the maturation of oocytes in vitro. The acquisition of developmental competence involves the synthesis and storage of a wide range of molecules during oocyte growth followed by the reprogramming and ordered utilization of these stored products during maturation, fertilization and early embryogenesis. The regulatory signals for these molecular changes are produced by the follicle cells in response to circulating levels of gonadotrophins; we report that some ovarian stimulation protocols distort these signals thereby disrupting molecular reprogramming of the oocyte and reducing subsequent developmental competence. The aspiration of immature oocytes from antral follicles followed by their maturation in vitro is a potential alternative to hormonal stimulation of patients in IVF treatment. Although relatively successful in a variety of animals, the production of fully viable human embryos by in-vitro maturation is still unsatisfactory despite the use of a wide variety of culture protocols. Our data suggests that the key to maturation and embryo viability in vitro resides in the follicle cell compartment rather than the oocyte. Because of rapid luteinization changes, follicle cells in culture probably fail to provide the maturing oocyte with the necessary ordered set of instructive signals and nutrients needed for the acquisition of developmental competence. Although much remains to be discovered about the nature, concentration and transmission of signals, nevertheless it is already clear that different steroids, matrix metalloproteinases and growth factors are involved in conferring viability on the maturing oocyte. Major improvements in the yield of viable embryos from in-vitro matured oocytes can be anticipated from a systematic analysis of somatic signals from the pre-ovulatory follicle.

Animals↗