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Biomedical subjects

Y Luo

Publications and source records attributed to Y Luo.

At least 487 records · Page 27Linked to original sources

The unprocessed C-terminal dipeptide of recombinant beta-nerve growth factor determines three stable forms with distinct biological activities.

The processing of polypeptide neurotrophins in the nervous system is poorly understood. In this paper, we provide information on the effects of C-terminal processing of nerve growth factor. Three forms of recombinant mouse beta-nerve growth factor (rNGF) were produced and isolated from insect cells infected with a recombinant baculovirus. The three purified forms of rNGF exhibited distinct biological activities and differed in their abilities to compete with high affinity binding of mouse beta-nerve growth factor (mNGF). However, they were chemically and structurally indistinguishable from each other. All three forms of rNGF differed from mature mNGF from mouse submaxillary gland in that the C-terminal Arg-Gly dipeptide had not been proteolytically removed. Removal of the C-terminal dipeptide by gamma-NGF peptidase treatment converted the three forms into a single form identical with mature mNGF. The above results demonstrate that a single polypeptide of rNGF, due to the presence of a C-terminal dipeptide, exhibits three stable dimeric protein conformations with distinct biological activities. The apparent lack of gamma-NGF peptidase in the nervous system raises the possibility that the biologically significant form of NGF may differ from mature mNGF; such a difference may be of physiological relevance.

Animals↗

Neu differentiation factor: a transmembrane glycoprotein containing an EGF domain and an immunoglobulin homology unit.

We recently reported that a 44 kd glycoprotein secreted by transformed fibroblasts stimulates tyrosine phosphorylation of the product of the neu proto-oncogene and induces differentiation of mammary tumor cells to milk-producing, growth-arrested cells. A partial amino acid sequence of the protein, termed Neu differentiation factor (NDF), enabled cloning of the corresponding complementary DNA. The deduced structure of the precursor of NDF indicated that it is a transmembrane protein whose extracellular portion contains an EGF-like domain that probably functions as a receptor recognition site. In addition, the ectodomain contains one immunoglobulin homology unit. Despite the lack of a recognizable hydrophobic signal peptide at the N-terminus, a recombinant NDF, like the natural molecule, is released into the medium of transfected COS-7 cells in a biologically active form. Northern blot analysis indicated the existence of several NDF transcripts, the major ones being 1.8, 2.6, and 6.7 kb in size. Transformation by the ras oncogene dramatically elevated the expression of NDF in fibroblasts.

Amino Acid Sequence↗

Tolerance induction in T helper (Th1) cells by thymic macrophages.

Most macrophages in the peripheral tissues present Ag optimally to a variety of functionally distinct Th cells. Although thymic macrophages have been implicated in deleting autoreactive thymocytes, their role in influencing the functional capacities of mature T cells is not clear. We have established a normal untransformed macrophage cell line, named TMC, from the mouse thymus. The TMC line presents protein Ag to an IL-4-producing Th2 type Th clone after IFN-gamma treatment as evidence by T cell proliferation and the release of IL-3 and IL-4. However, these thymic macrophages are inefficient at stimulating a well characterized cytochrome C-specific IL-2-producing Th1 clone, A.E7. Ag presentation by TMC results in the production of IL-3 but not IL-2 production or proliferation of A.E7 cells. This selective Ag presentation defect to Th1 cells is corrected by the addition of live but not fixed allogeneic irradiated spleen cells, suggesting that the thymic macrophages lack the expression of costimulatory activity required for Th1 activation. This is further demonstrated by the failure of live thymic macrophages to provide costimulatory activity to A.E7 cells stimulated with fixed spleen cells plus the antigenic peptide 81-104. Exposure of A.E7 cells to paraformaldehyde-treated TMC in the presence of 81-104 peptide induces specific hyporesponsiveness, anergy. These data demonstrate that thymic macrophages can have a profound influence on the response of selected T cells to Ag. Furthermore, the nature of the T cell stimulus is also critical because Th1 and Th2 cells responded equally well to the T cell mitogen, Con A, and a bacterial superantigen presented by the thymic macrophages.

Animals↗

Investigation on the effect of selenium on T lymphocyte proliferation and its mechanisms.

The effects of selenium on T lymphocyte proliferation and its mechanisms have been investigated. The results showed that selenium is able to enhance lectin-stimulated T lymphocyte proliferation, to increase the production of interleukin 2 (IL-2) by lymphocytes and interleukin 1 (IL-1) by macrophages in the presence of lectin and to augment the response of T lymphoblasts to IL-2 and that of thymocytes to IL-1. The data presented suggest that selenium may be an important modulator for immune response. Selenium might enhance IL-2 production and response through the augmentation of IL-1 production and response, thereby promoting the enhancement of the proliferation and action of T lymphocytes and other immunocompetent cells. Based on these results, we may consider using selenium as an immunological enhancement agent to enhance or recover immune functions of the organism.

Animals↗

Effects of surgical trauma on interleukin 2 production and interleukin 2 receptor expression.

To elucidate the mechanism of immunologic abnormalities induced by surgical trauma, we measured interleukin 2 (IL-2) production and IL-2 receptor expression in peripheral blood mononuclear cells from 16 patients with cholelithiasis. Compared with preoperative levels, IL-2 production and IL-2 receptor expression were significantly reduced on postoperative day 4, and the reduction was consistent with the suppression of lymphocyte proliferative response to phytohemagglutinin.

Adult↗

Interleukin 2 production and its relationship with T lymphocyte subsets in patients with obstructive jaundice.

Sepsis remains a major risk in the high mortality and morbidity after surgery for obstructive jaundice. The reasons for the increased susceptibility to infection are unknown. This study examined interleukin 2 (IL-2) production and the lymphocyte response to PHA mitogen in 31 patients with obstructive jaundice. Among them, 18 patients were simultaneously investigated by enumeration of T lymphocyte subsets in peripheral blood with APAAP technique. The results showed that the patients had significantly decreased IL-2 production and lymphocyte response to PHA mitogen. The percentage of Leu 3a (helper/inducer T cell) in the patients was significantly lower than that in normal controls. Leu 3a/Leu 2a (suppressor/cytotoxic T cell) ratio was significantly lower in these patients. The reduction of IL-2 production correlated significantly with the suppression of lymphocyte proliferation but not with the percentage of Leu 3a cells. From these results, it may be suggested that the reduction of IL-2 production in the patients with obstructive jaundice is an important reason for the suppression of T lymphocyte proliferative response, not merely a reflection of the decrease of helper T cells.

Adult↗

Building protein backbones from C alpha coordinates.

An automatic procedure for building polyalanine backbones from guiding alpha-carbon positions is presented. Polyalanine backbones are built based on the geometric restraints of angle N-C alpha-C and the knowledge of main-chain dihedral angle distributions. A building module constructs a list of polyalanine backbones that follow exactly the C alpha trace. Then a selection module selects one backbone with the largest portion of phi-psi pairs in favoured regions. Several test cases on C alpha coordinates from X-ray refined structures give acceptable results. Less than 10% of the peptide planes are incorrectly built, and the result is not sensitive to random shift up to 0.5 A of C alpha coordinates.

Amino Acid Sequence↗

[The application of nasometer on the analysis of normal and cleft palate speech].

This paper reports the use of for assessment of speech by oronasal acoustic measures. Nasal energy during speech is reflected by nasolance value and graph pattern. The degree of velopharyngeal closure and the situation and movement of other speech organs can be demonstrated. The speech of 120 normal children and 60 preoperation cleft palate patients are studied. The scales of normal value and graph pattern are recorded. Comparing with normal speech, the scale of nasolance value and characteristics of graph pattern of cleft palate speech are obviously changed and may be assessed by this apparatus. The treatment and speech therapy plan can be made with the results obtained by this method as the basis.

Child↗

[Relationship among dosages of aminophylline, plasma levels of theophylline and variation of pulmonary arterial pressure].

The relationship among the dosages of aminophylline, plasma levels of theophylline and variations of mean pulmonary arterial pressure (mPAP) in 72 patients with COPD was investigated. The results showed that after a different loading dosage of aminophylline (6 mg/kg, 5 mg/kg and 4 mg/kg) was administered by intravenous injection, mPAP in the 6 mg/kg group was decreased more significantly (P less than 0.01) than that in the 4 mg/kg group. In the 6 mg/kg group, the decreased mPAP period sustained for 120 min, which was longer than that in the other 2 groups. The plasma levels of theophylline in the 6 mg/kg group of patients 30 to 120 min after loading dose injected were 115.54-79.04 mumol/L, which were higher than that in the others. Within the 120 min period of observation after the drug was administered no patients in any of these groups showed severe untoward effects. According to the results of this experiment, we suggest that the 6 mg/kg as a loading dose should be advised for the treatment of pulmonary hypertension in COPD. The optimum time to give the maintenance dosage should be set within 2 h after the loading dose. It is necessary to monitor the plasma levels of theophylline while aminophylline is administered, so that optimal therapeutic effects could be achieved without side effects.

Adult↗

Optimal low dosage of acetylsalicylic acid (ASA) for the prevention and treatment of ischemic cerebrovascular disease in geriatric patients.

A series of 108 geriatric patients with ischemic cerebrovascular disease were treated with low dose aspirin (acetylsalicylic acid, ASA). Daily doses of 100 mg, 50 mg and 25 mg were administered to three groups of 36 patients. Changes in platelet aggregation responses were dynamically observed in 64 (22 normal subjects, 42 patients). Monitoring of 22 normal subjects revealed inhibition of platelet aggregation at a dose of 300 mg or 100 mg which could last as long as 7 days. This suggests that 100 mg or less could be clinically effective. Satisfactory inhibitory effects on platelet aggregation were observed in all three groups, with 14 patients in each group given daily doses of 100 mg, 50 mg and 25 mg during four weeks' observation. The most effective inhibition was obtained in the 50 mg group. Therefore, the authors recommend 50 mg/d as the optimal dosage for low dose aspirin therapy in geriatric patients.

Aged↗

Potential use of luminol-dependent chemiluminescence for estimation of free radicals produced in hepatic microsomes and reconstituted cytochrome P-450 systems.

The evaluation of luminol-dependent chemiluminescence (CL) for determination of free radicals generated during the oxidative reactions of microsomal enzymes in microsomes and reconstituted systems has been studied. The results indicated that the CL not only depended on NADPH induction but also on intact cytochrome P-450 enzymes. The light emission signal was strongly scavenged by SOD with inhibition rate of 93%. A concentration of 24 mumol.L-1 cadmium acetate and 2.5 mumol.L-1 mercuric chloride also inhibited the signal significantly as compared with the control. The study of CL in detecting free radicals formed by biotransformation of 44 xenobiotics suggested that some of the compounds known as free radicals producing agents by bioactivation increased the light emission in cytochrome P-450 enzyme system, such as carbon tetrachloride, chloroform, tetrachloroethylene etc., the others forming toxic metabolites by biotransformation induced the CL in some cases, such as carbon disulfide, benzene, methyldursban, methylparathion etc. or decreased the light emission, such as parathion, aniline and polychlorinated byphenyls etc., and still others whose toxic effects have no relationship with their biotransformation had no influence or an inhibitory effect on the CL, such as aminopyrine, TOCP, and acrylamide. CL in the cytochrome P-450 enzyme system remained unchanged during the biotransformation of carcinogens such as diethylnitrosamine, dipropylnitrosamine and benzo(a)pyrene as compared with the control. However, 3-methylcholanthrene and dimethylnitrosamine had an enhancing effect on the CL, while 2-acetylaminofluorene and 2-aminoanthracene showed an inhibition to the signals.

Acetates↗

Discrete functional stages of vaccinia virus early transcription during a single round of RNA synthesis in vitro.

We have developed a system for analysis of discrete steps in vaccinia virus early mRNA synthesis during a single round of transcription in vitro. A synthetic early promoter is used to direct transcription by vaccinia RNA polymerase of a G-less cassette in linear duplex DNA. Omission of GTP from transcription reactions leads to the formation of ternary elongation complexes paused stably at the end of the G-less cassette. These complexes can be induced to elongate by provision of GTP. While initiation of transcription is sensitive to low concentrations of salt and Sarkosyl, elongation is relatively resistant to these agents. Termination can be studied in a single synthetic cycle by forming transcription complexes paused just proximal to the termination signal TTTTTNT that can subsequently elongate and terminate. By selectively incorporating the termination-inhibiting analog BrUMP into proximal and distal portions of the nascent transcript, we localize the termination signal within or near the sequence UUUUUNU in the nascent RNA. We show that access of the vaccinia termination factor (VTF/capping enzyme) to the transcriptional apparatus can occur subsequent to initiation and synthesis of a 390-nucleotide nascent RNA. Termination is more sensitive to inhibition by salt and Sarkosyl than in elongation. This sensitivity is not reversed by preincubation of VTF with the transcription complex. Finally, we confirm the identity of VTF and vaccinia mRNA capping enzyme by demonstration of VTF activity associated with capping enzyme expressed in Escherichia coli.

Base Sequence↗

Splice site requirement for the efficient accumulation of polyoma virus late mRNAs.

Polyoma virus late nuclear primary transcripts are giant and heterogeneous, containing tandem repeats of the late strand of the circular viral genome. Late pre-mRNA processing involves the splicing of noncoding 'leader' exons to each other (removing genome-length introns), with the joining of the last leader to a coding 'body' exon. We have constructed a number of mutants blocked only in leader-leader splicing, or blocked in both leader-leader and leader-body splicing. We examined the accumulation of both nuclear and cytoplasmic late-strand RNAs in NIH3T3 cells. Consistent with our previous results, mutants lacking the 3' splice site of the late leader (leader-leader splicing blocked) showed a 10-20 fold defect in late RNA accumulation. Mutants which lacked the leader 5' splice site (leader-body splicing blocked) had a more profound defect, exhibiting virtually no late-strand cytoplasmic or nuclear RNA. This result was unexpected as a substantial proportion of wild type late cytoplasmic messages are unspliced. A mutant with no intron, but having functional 3' and 5' splice sites bordering the leader exon, is capable of producing large amounts of unspliced late mRNA. This demonstrates that an excisable intron is not a requirement for late mRNA accumulation. The accumulation of polyoma late mRNAs requires the presence of leader exons bordered by functional 3' and 5' splice sites, whether or not these sites are used during pre-mRNA processing.

Base Sequence↗

Effect of arginine on immune function in rats with obstructive jaundice.

In this study we used an experimental model produced by bile duct ligation (BDL) in rats to demonstrate relation between immune function and susceptibility to infection associated with obstructive jaundice and immunoregulatory effect of arginine in BDL rats. Our experimental results showed that thymus weight, content of lymphocytes in thymus and responsiveness to Con A in vitro were dramatically reduced in BDL rats as compared to rats with sham operation and that the impairment of lymphocyte function was significantly related to the susceptibility to infection. Arginine, as a T cell stimulator, markedly improved immune function and decreased susceptibility to infection of BDL rats.

Animals↗

Antitermination of vaccinia virus early transcription: possible role of RNA secondary structure.

Transcription of vaccinia early genes by the viral RNA polymerase terminates downstream of a signal sequence TTTTTNT in the nontemplate DNA strand. Signal recognition occurs at the level of the sequence UUUUUNU in nascent RNA and depends on a virus-encoded termination factor (VTF). The presence of TTTTTNT elements within protein encoding regions of some early genes requires that these 5' proximal signals be ignored in order to achieve early expression of the full-sized proteins. In the case of the A18R gene, which contains a proximal terminator that is not utilized in vivo (Pacha et al., J. Virol. 64, 3853-3863 (1990)), the TTTTTNT sequence can be folded into a potential hairpin structure such that UUUUUNU would be part of a duplex stem in the nascent RNA. We find that the A18R putative hairpin is unable to promote factor-dependent termination in a purified in vitro transcription system. Sequence manipulations that abrogate the potential to form an RNA hairpin restore the activity of the TTTTTNT motif. The in vitro studies suggest that antitermination at the proximal site of the A18R gene may be mediated by secondary structure in the nascent RNA, and that early termination involves recognition by VTF and/or RNA polymerase of the UUUUUNU sequence in single-stranded form.

Base Sequence↗

Changes in cutaneous microcirculation, hemorrheology and platelet aggregation function in dermatomyositis.

Marked disturbance of microcirculation was observed in 85 cases of dermatomyositis: decrease in number and irregular pattern of the capillaries, many being tortuous and dilated and having a slow and granular blood stream, as well as an increase of vascular permeability. The changes were more marked in cases with visceral disorders, or in active disease; however, there was no correlation between these changes and the duration of illness. Blood hyperviscosity was found in an hemorrheological study. The cause is considered to be due to high plasma viscosity, high fibrinogen levels, and increase of aggregation of erythrocytes and platelets. Modulation of microcirculation is advisable in the treatment of dermatomyositis.

Adolescent↗