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Y Lu

Publications and source records attributed to Y Lu.

At least 919 records · Page 51Linked to original sources
PubMed1992

The reverse DNA sequencing using Bst DNA polymerase.

The reverse DNA sequencing (RDS) [1] is a rapid method used to check the DNA sequences by sequencing them from the opposite orientation. Because the RDS is basically a double stranded sequencing, the quality of the sequence patterns so obtained generally is not as good as those obtained by the single stranded sequencing, and extra bands and higher background are produced more frequently. This paper shows that the RDS could now generate as good sequence patterns as those obtained by sequencing on the single stranded DNA template if Bst DNA polymerase instead of the conventional enzymes, such as the Klenow enzyme, was used in the RDS. Bst DNA polymerase is heat stable (optimum reaction temperature 65 degrees C) and has recently been successfully used in the conventional DNA sequencing. The RDS has recently been further simplified to meet the need of large DNA sequencing projects such as the human genome project. The combination of the simplified RDS and the use of Bst polymerase should be expected to facilitate greatly the work on sequence confirmation and correction.

Base Sequence↗
PubMed1992

[Chemical constituents of Eupolyphaga sinensis Walker].

Five compounds were isolated from the n-hexane extract and n-butanol of extract Eupolyphaga sinensis. These compounds were identified as octacosanol, beta-sitosterol, batyl alcohol, 2,4-pyrimidinedione and allantoin.

Allantoin↗
PubMed1992

Comparative ultrastructure of the tip of ventricular papillary muscle.

Electron microscopic studies of the tips of left ventricular papillary muscles from seven human, two monkey, three sheep, and two chicken hearts were done to elucidate the fine structure of myotendinous junctions. The human specimens were from normal hearts obtained 3 to 9 hours postmortem from persons aged 7 months to 30 years (mean, 13.3 years). We found no significant ultrastructural differences between the human hearts and those of monkey, sheep, and chicken. Myocardial fibers were elongated and thinner (tapered) in the tips of papillary muscles. In addition to usual working myocardial cells, the distal end of narrowing muscle fibers also contained small pale cardiocytes containing fewer myofibrils and smaller mitochondria. These cells were similar to P cells or transitional cells in the conduction system. Nerve axons and Schwann cells were commonly seen in the interstitium, usually in association with capillaries. Fibroblasts and axon varicosities were occasionally seen extremely close to the cardiocytes. These specialized myocardial cells associated with rich neural tissue in the papillary muscle tip possibly function as foci of local automaticity. This histologic organization may also represent neurosensory function responding to and monitoring local pressure changes, efferent adrenergic or cholinergic neural activity, or both.

Adolescent↗
PubMed1991

Retinal [125I]iodomelatonin binding sites in the tree shrew (Tupaiidae).

The characteristics of melatonin-binding sites labelled by [125I]iodomelatonin in membrane preparations from the tree shrew retina were determined. Specific binding of [125I]iodomelatonin to the membrane preparations of tree shrew retina was rapid, stable, saturable, and reversible. Among the indoles tested only 6-chloromelatonin, melatonin and N-acetylserotonin had significant affinities to the [125I]iodomelatonin binding site. Scatchard analysis of the membrane preparations revealed a dissociation constant (Kd) of 51.0 +/- 16 pM and a total number of binding sites (Bmax) of 1.97 +/- 0.6 fmol/mg protein.

Animals↗
PubMed1991

Binding characteristics and regional distribution of [125I]iodomelatonin binding sites in the brain of the human fetus.

The binding and pharmacological characteristics of [125I]iodomelatonin binding sites in human fetal brain membrane preparations were determined. Membrane preparations of the whole brain revealed a equilibrium binding constant (Kd) of 17.5 pmol/l with a total number of binding sites (Bmax) at 0.8 fmol/mg protein. Among the various brain regions studied, [125I]iodomelatonin binding was highest in the hypothalamus, and lowest in the mid-brain, pons-medulla, and cerebral cortex. The Kd of the hypothalamus was calculated to be 26.1 pmol/l and the Bmax 5.4 fmol/mg protein. Only 6-chloromelatonin, melatonin and N-acetylserotonin had significant inhibition in the binding. Our results suggest that melatonin receptors are present in the human brain and that melatonin may exert a central action.

Binding, Competitive↗
PubMed1991

[125I]iodomelatonin binding sites in spleens of birds and mammals.

The specific binding of [125I]iodomelatonin to duck spleen membrane preparations was studied in detail. These binding sites were stable, saturable, reversible and of high affinity. Scatchard analysis of the binding revealed a equilibrium binding constant (Kd) of 73.1 +/- 5.4 pM and a total number of binding sites (Bmax) of 3.64 +/- 1.38 fmol/mg protein. Studies on the relative binding capacities of [125I]iodomelatonin to the spleen in different species showed the following order: duck greater than chicken greater than guinea pig greater than pigeon greater than mouse. No binding site was detected in the rat spleen. The presence of [125I]iodomelatonin binding sites in the spleen of birds and mammals suggested a direct action of pineal melatonin on the immune system.

Animals↗
PubMed1991

Regulation of glycan processing by Golgi enzymes from red kidney bean (Phaseolus vulgaris) seedlings.

The effect of a protein matrix on the processing of glycoprotein glycans by Golgi enzymes from plant seedlings has been determined with an artificial glycoprotein system, comparing the processing rates of glycan-(biotinyl)Asn (or glycan-(biotinamidohexanoyl)Asn) substrates either free or bound to avidin. An analysis of the pooled glycoproteins from the seedlings suggested that the most common glycan structure is a complex one (GlcNAc-Man3GlycNAc2-protein), and consistent with this processing end-product, mannosidases I and II and GlcNAc transferases I and II were all found to be present in the seedling Golgi membrane preparations. The effect of the avidin matrix either in a proximal (biotinyl substrates) or distal (N-(biotinamido)hexonoyl substrates) association with the appropriate glycan substrate for these four enzymes was assessed from the direct comparison of the apparent first-order rate constants for the free and avidin-bound substrate-product conversions. All four plant enzymes were inhibited by the association of the glycan substrates with avidin, but the inhibition was much less pronounced than that observed with the corresponding enzymes from rat liver and hen oviduct. The rate effect shows a progression from 3- to 10-fold rate decreases in the proximal complexes and 2- to 3-fold in the distal complexes in going from the first (mannosidase I) to the fourth (GlcNAc transferase II) enzyme; with the mammalian and avian enzymes the largest effects were for the first ones and much larger absolute rate effects were observed. The results suggest that the nature of the processing enzymes in terms of this response to the avidin glycan substrates may differ in different organisms.

Carbohydrate Sequence↗
PubMed1991

Identification of aecA mutations in Bacillus subtilis as nucleotide substitutions in the untranslated leader region of the aspartokinase II operon.

Recent genetic mapping of the aspartokinase II (lysC) operon of Bacillus subtilis [M. Petricek. L. Rutberg & L. Hederstedt (1989) FEMS Microbiology Letters 61, 85-88; N.Y. Chen. J. J. Zhang & H. Paulus (1989) Journal of General Microbiology 135, 2931-2940] has shown its chromosomal location to be close to the aecA locus, the mutation of which leads to highly increased levels of aspartokinase II. In order to examine the relationship between lysC and aecA, we have cloned the control regions of the lysC operon from several independent aecA mutants and determined their nucleotide sequences. The nucleotide sequences of the aecA mutants differed from the wild-type sequence by the substitution of one or two nucleotides at two widely separated sites in the transcribed leader region of the lysC operon. To confirm that the observed nucleotide changes are indeed responsible for the AecA phenotype and not simply the reflection of sequence polymorphisms in different B. subtilis strains, we introduced the same nucleotide substitutions as those observed in the aecA strains into the leader region of the wild-type lysC operon by oligonucleotide-directed mutagenesis. The expression of the mutagenized genes was analysed after transcriptional or translational fusion to lacZ in a single-copy integration vector. The levels of beta-galactosidase were greatly elevated by the nucleotide substitutions, with similar increases observed in transcriptional and translational fusions. The high level of expression of beta-galactosidase in the lysC'-lac'Z strains with nucleotide substitutions corresponding to the aecA mutations was resistant to repression by L-lysine but was completely abolished by the inactivation of the lysC promoter.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Proopiomelanocortin gene promoter elements required for constitutive and glucocorticoid-repressed transcription.

The POMC gene is expressed predominantly in the anterior pituitary. The high level of POMC transcription in this tissue is modulated by peptide hormones and repressed by glucocorticoids. In this present study we have investigated promoter elements required for the high basal transcription and glucocorticoid repression using transient transfection and in vitro transcription assays. We first determined that the region between -77 to -51 of the promoter, which has previously been shown to harbor a glucocorticoid receptor-binding site, is required for high basal expression both in vivo and in vitro. This promoter domain is also required for glucocorticoid repression of transcription in vivo. Two site-directed mutants within this area both decreased basal transcription, but were fully repressed by glucocorticoids, implying that the -77 to -51 region is a complex regulatory region harboring separable basal and glucocorticoid-repressible elements. Electrophoretic mobility shift and exonuclease III footprinting analysis revealed the existence of two factors that bind in this region. We also examined the effect of broad promoter deletions on basal expression and glucocorticoid repression. These experiments revealed that the region between -480 and -320 is also required for glucocorticoid repression. Taken together, the data suggest a model in which high basal transcription is generated by direct interaction of factors binding between -480 to -320 and -77 to -51. Glucocorticoid repression could occur by direct receptor disruption of these interactions.

Animals↗
PubMed1991

Regulation of testicular P-450 cholesterol side-chain cleavage and P-450 C17-20 lyase/C17 hydroxylase enzymes in the neonatal and adult rat.

Adult Leydig cells respond to LH or hCG with an initial stimulation of testosterone secretion followed by LH receptor down-regulation and blockade of androgen biosynthesis. In contrast, fetal Leydig cells respond with increased LH receptor number and enhanced steroidogenesis. In this study, the molecular mechanisms of high-dose hCG treatment on steroidogenesis in adult and neonatal testes (containing predominantly the fetal generation of Leydig cells) were examined using two recombinant DNA clones specific for enzymes of the rat steroidogenic pathway (P-450 cholesterol side-chain cleavage enzyme, P-450scc and P-450 17 alpha-hydroxylase/C17-20 lyase, P-450c17). We treated adult (60 days of age) and neonatal (2 days of age) rats with a single high dose of hCG (600 IU/kg), sc. The high dose of hCG caused neonatal testicular P450scc and P450c17 mRNA levels to increase, and stimulated adult testicular P450scc mRNA levels, but caused a decrease in adult P450c17 mRNA levels. These studies suggest that high doses of hCG regulate testosterone production differently in adult and fetal Leydig cells at a pretranslational level of the P450c17 enzyme, while mRNA for P450scc is stimulated in both the adult and fetal Leydig cell.

Aging↗
PubMed1991

[A study of textile noise influence on maternal function and embryo-growth].

Ninety hundred seventy-eight female textile workers exposed to noise (greater than or equal to 85 dB (A)) were recruited as the subjects of observation group, and 402 nonexposed female workers were chosen as the controls (less than 75 dB (A)). According to noise level, the observation group was subdivided into three groups: group 1, 100-104 dB (A), 416 workers; group 2, 95-99 dB, (A), 310 workers; group 3, 85-94 dB (A), 252 workers. Non-conditional logistic regression model was used to exclude the effect of mixed factors, such as age and working life. The result showed that noise-exposure could have some adverse effect on reproductive function of women and embryo growth. The workers exposed to noise had significantly increased incidence rates for irregular menstrual cycle, dysmenorrhea, lumbago, hypertension in pregnancy, threatened abortion, spontaneous abortion, premature labour, dystocia and low birth weight of newborns. When the noise intensity heightened, the sorts of abnormal function of reproduction increased and the risk of incidence raised too. These demonstrated a dose-response relationship between the noise intensity and incidence rate. The noise-working life had significant effects on the incidence of hypertension in pregnancy and prematurity. Although the age had some influence, it was not statistically significant. In terms of the maternal function and embryo growth, the factors in order of risk are: noise intensity greater than working life greater than age.

Abortion, Spontaneous↗
PubMed1991

[Correlation between neonatal Apgar scores and the results of maternal and umbilical cord blood gas analysis].

The correlation between the Apgar scores of 162 neonatal 1 minute after birth and the result of maternal arterial and umbilical cord blood gas analysis was studied. The results were as follows: (1) With decreasing neonatal Apgar score, the value of the pH, PO2, HCO3, TCO2 and actual base excess (ABE) of the maternal arterial and umbilical cord blood also decreased markedly, while the PCO2, except that of the maternal arterial blood, increased obviously, and mixed acid base imbalance and hypoxemia also significantly increased (P less than 0.01). (2) Of the neonates with umbilical arterial blood pH less than or equal to 7.2, the proportion of neonates with Apgar score less than or equal to 3 amounted to 100%; that with on Apgar score of 4-7, was 77.7%; And that with on Apgar score of 8-10, was 56.31%. The difference was highly significant statistically (P less than 0.01).

Adult↗
PubMed1990

Expression of chimeric human transferrin genes in vitro.

Transferrin (TF), a major plasma protein, binds and transports ferric iron. Evidence exists for unique roles for TF in brain in oligodendrocyte differentiation, myelination and neuronal development. In this study, 5' flanking regions of the TF gene important in regulating gene expression were identified by transfected cell studies and a comparison of 5' flanking sequences of the human TF and TF receptor genes. Human glioma cell lines HTB-16 and HTB-17 were shown to synthesize TF identical in size and immunological reaction to TF synthesized by liver. The expression of a series of human chimeric TF genes in glioma cells was compared with hepatoma and HeLa cells. A difference in transient expression was observed in hepatoma and glioma cells transfected with TF chimeric genes containing 3.9 kb of the 5' region; hepatoma cells demonstrated significantly more expression than did glioma cells, suggesting that a DNA region present in the 3.9-kb construct is important either in liver-specific expression or in repression of brain expression, or in both. Smaller constructs containing less than or equal to 0.622 kb of the 5' regulatory region of the TF gene failed to demonstrate cell-specific expression; they were expressed in HeLa cells, a line that does not synthesize TF. High levels of expression of 0.15-kb TF constructs were also observed in hepatoma and glioma cell lines, but not in transgenic mice. Possible explanations of differences observed in expression of shorter TF constructs in vitro and in vivo are discussed.

Animals↗