Biomedical subjects

Y Ai

Publications and source records attributed to Y Ai.

At least 19 recordsLinked to original sources

PubMed2001

Abnormal migration phenotype of mitogen-activated protein kinase-activated protein kinase 2-/- neutrophils in Zigmond chambers containing formyl-methionyl-leucyl-phenylalanine gradients.

Time-lapsed video microscopy and confocal imaging were used to study the migration of wild-type (WT) and mitogen-activated protein kinase-activated protein kinase 2 (MK2-/-) mouse neutrophils in Zigmond chambers containing fMLP gradients. Confocal images of polarized WT neutrophils showed an intracellular gradient of phospho-MK2 from the anterior to the posterior region of the neutrophils. Compared with WT neutrophils, MK2-/- neutrophils showed a partial loss of directionality but higher migration speed. Immunoblotting experiments showed a lower protein level of p38 mitogen-activated protein kinase and a loss of fMLP-induced extracellular signal-related kinase phosphorylation in MK2-/- neutrophils. These results suggest that MK2 plays an important role in the regulation of neutrophil migration and may also affect other signaling molecules.

Actins↗

PubMed2001

Leukocyte-specific gene 1 protein (LSP1) is involved in chemokine KC-activated cytoskeletal reorganization in murine neutrophils in vitro.

Leukocyte-specific gene 1 protein (LSP1) is a cytoskeletal-associated protein of leukocytes that in vitro cross-links F-actin into extensively branched bundles of mixed polarity. In this study, we examined chemotaxis and superoxide production in neutrophils prepared from wild-type (WT) and Lsp1 knockout mice. Compared to WT neutrophils, Lsp1-/- neutrophils showed impairment in both migration speed and chemotaxis direction during chemokine KC-directed chemotaxis. When examined by confocal microscopy, chemotaxing Lsp1-/- neutrophils showed abnormal morphologies. They had discontinuous primary actin-rich cortexes and large membrane protrusions. When stimulated by phorbol 12-myristate 13-acetate (PMA), Lsp1-/- peritoneal neutrophils produce more superoxide than WT. The data presented suggest that LSP1 plays important roles in the regulation of neutrophil morphology, motility, and superoxide production.

Actins↗

PubMed2000

A mouse model of galactose-induced cataracts.

Galactokinase (GK; EC 2.7.1.6) is the first enzyme in the metabolism of galactose. In humans, GK deficiency results in congenital cataracts due to an accumulation of galactitol within the lens. In an attempt to make a galactosemic animal model, we cloned the mouse GK gene (Glk1) and disrupted it by gene targeting. As expected, galactose was very poorly metabolized in GK-deficient mice. In addition, both galactose and galactitol accumulated in tissues of GK-deficient mice. Surprisingly, the GK-deficient animals did not form cataracts even when fed a high galactose diet. However, the introduction of a human aldose reductase transgene into a GK-deficient background resulted in cataract formation within the first postnatal day. This mouse represents the first mouse model for congenital galactosemic cataract.

Aldehyde Reductase↗

PubMed2000

P47(phox)-deficient NADPH oxidase defect in neutrophils of diabetic mouse strains, C57BL/6J-m db/db and db/+.

Deficiencies in neutrophil NADPH oxidase proteins have been demonstrated in humans with chronic granulomatous disease. However, no spontaneous mutation in murine NADPH oxidase has been reported. In this study we report that neutrophils from the diabetic mouse strains, C57BL/6J-m heterozygous lean (lepr(db/+)) and homozygous obese (lepr(db/db)) mice produced no superoxide on stimulation. An absence of intact p47(phox) but not other oxidase proteins was observed in both mouse strains through the use of immunoblotting. Molecular analysis by reverse transcriptase-polymerase chain reaction identified three abnormal p47phox mRNA transcripts. Sequencing of genomic DNA of p47(phox) revealed a point mutation at the -2 position of exon 8, which is consistent with aberrant splicing of the p47(phox) transcript. These results indicate that the C57BL/6J-m db/db and db/+ mice are the first spontaneously derived murine model of NADPH oxidase deficiency involving a p47(phox) mutation.

Animals↗

PubMed1999

MPTP-Induced pallidal lesions in rhesus monkeys.

Dopamine neurons in the substantia nigra of the midbrain are the primary neuronal population affected by 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine (MPTP) toxicity, which produces the pathological and behavioral features of Parkinson's disease in nonhuman primates and man. We have identified another injury site in magnetic resonance imaging (MRI) brain scans in 13 of 37 rhesus monkeys taken 10-12 months after administration of this neurotoxin via the right carotid artery. Focal lesions, ranging in volume from 6.75 to 60 mm3 in the rostral globus pallidus region, were seen on the right side of the brain in these 13 animals in addition to the midbrain effects. While no significant differences were seen between globus pallidus lesioned and nonlesioned animals in the severity of MPTP-induced parkinsonian symptoms, the response to levodopa was muted in pallidal-lesioned animals. To confirm the role of neurotoxicity in producing the lesions, brain scans from an additional 12 monkeys were evaluated during the acute period following exposure to either MPTP (n = 6) or saline (n = 6). Focal lesions in the rostral globus pallidus were seen as early as 2-4 h following a carotid artery infusion in two of six MPTP recipients, but no evidence of injury was seen in saline recipients. The globus pallidus includes important components of the neural circuitry regulating motor functions. The present results indicate that in addition to midbrain dopamine neurons, a focal region of the rostral globus pallidus is selectively vulnerable to MPTP toxicity.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed1999

[A case of fetal valproate syndrome with intractable wheezing due to submucosal tumor below the vocal cord].

A full-term baby was born to an epileptic mother treated with two anti-epileptic drugs, sodium valproate and phenobarbital, throughout the pregnancy. She was given no information about the risk of teratogenesis of these drugs. At birth the patient was hypotonic and had clinical features specific for the fetal valproate syndrome. After a viral infection at three months of age, he had intractable and persistent wheezing. Suspecting the presence of congenital respiratory tract abnormality, we performed tracheobronchoscopy, which revealed a relatively big submucosal tumor on the left trachial wall below the vocal cord. Dyspnea and wheezing were remarkably improved by tracheolaryngotomy, but he died suddenly and unexplainedly at seven months of age.

Abnormalities, Drug-Induced↗

PubMed1998

The role of p38 MAP kinase in TGF-beta1-induced signal transduction in human neutrophils.

Transforming growth factor-beta 1 (TGF-beta 1) is the strongest chemoattractant yet described for human neutrophils. It activates neither phospholipase C nor phospholipase D. It does not induce rises in intracellular calcium, degranulation, or superoxide production. The signaling pathways utilized by TGF-beta 1 are largely unknown. This report demonstrates that TGF-beta 1 activates p38 MAP kinase. The kinase inhibitor SB203580 blocks the chemotactic responses as well as actin polymerization induced by TGF-beta 1. Potential cellular targets of the p38 MAP kinase pathway which could mediate these function are discussed.

Actins↗

PubMed1998

Construction and application of MCBL plate for facilitation of chromosome recombination in fungi.

A medium with camphor and benomyl MCBL plate was designed and constructed based on the proposed mechanism that d-camphor could induce the fusion of nuclear membrane while benomyl could induce the nondisjunctional recombination of chromosome in fungi. This so-called co-induction plate consisted of 0.1% d-camphor (W/V) and 0.5 microgram/L benomyl contained in Czapek's minimal medium. The precautions to be taken in the construction procedure of this plate was described in detail. One typical example of intergeneric fusion-cross, Aspergillus niger x Trichoderma reesei, was investigated, comparing the ratios of genotypes and phenotypes of fusant progenies produced by the co-induction of MCBL plate and by the step-by-step induction with camphor and benomyl separately. The results showed that the heterodiploid state was extremely transient and the recombinant haploid was hardly obtained when induced by the routine step-by-step method, whereas the ratios of apparent diplodization and nondisjunctional recombinant haplodization among all types of varied segregates on MCBL plate were greatly improved, compared with those on the routine plates containing single reagents, which indicated that the transient heterodiploid could be grasped and transformed further into nondisjunctional recombinant haploid when co-induced on the MCBL plate. The age of regenerated mycelium to be induced was found to have a dominant influence on the co-induction effects of MCBL plate. The mechanism of co-induction by MCBL plate and its promising applications were discussed.

Aspergillus niger↗

PubMed1997

LSP1 is the major substrate for mitogen-activated protein kinase-activated protein kinase 2 in human neutrophils.

In intact cells, mitogen-activated protein kinase-activated protein (MAPKAP) kinase 2 is rapidly activated by various cytokines, stresses, and chemotactic factors. The small heat shock protein p27 has been shown to be a substrate for MAPKAP kinase 2. Recently, we identified a novel substrate, designated p60, for MAPKAP kinase 2 in human neutrophils (Zu, Y.-L., Ai, Y., Gilchrist, A., Labadia, M. E., Sha'afi, R. I., and Huang, C.-K. (1996) Blood 87, 5287-5296). To further understand the signaling pathway of MAPKAP kinase 2, we have purified p60 from a heat-treated neutrophil lysate by DEAE-cellulose chromatography and SDS-polyacrylamide gel electrophoresis. Microsequencing of five peptides derived from purified p60 indicates that p60 is lymphocyte-specific protein 1 (LSP1). Furthermore antibodies specific for human and mouse LSP1 react with human and mouse p60. The sequence of human LSP1 indicates two serine residues at positions 204 and 252 as potential phosphorylation sites. The amino acid sequences surrounding these two sites are in agreement with the consensus sequence (Xaa-Xaa-Hyd-Xaa-Arg-Xaa-Xaa-Ser-Xaa-Xaa) for phosphorylation by MAPKAP kinase 2. Both serine residues in human LSP1 and the corresponding conserved serine residues in mouse LSP1 are in the basic C-terminal F-actin binding domain. Various fusion proteins of wild type and truncated mouse LSP1 with glutathione S-transferase were tested for their capacity to be phosphorylated by MAPKAP kinase 2. The results indicate that LSP1 is a substrate for MAPKAP kinase 2 in vitro and that the phosphorylation sites are located in the basic C-terminal domain of LSP1. Because both the small heat shock proteins and LSP1 are F-actin binding proteins, these results suggest a role for MAPKAP kinase 2 in the regulation of cytoskeletal structure or function.

Amino Acid Sequence↗

PubMed1997

High expression and activation of MAP kinase-activated protein kinase 2 in cardiac muscle cells.

Recently, three mammalian mitogen-activated protein (MAP) kinases, ERK, SAPK/JNK, and p38/HOG-1 have been identified, each with apparently unique signal transduction pathways. The p38 MAP kinase mediates an intracellular stress-activated signaling pathway by regulating down-stream molecules, such as MAP kinase-activated protein (MAPKAP) kinase 2. To study the tissue specificity of MAPKAP kinase 2, mRNA blots containing multiple human tissues were hybridized with a specific oligonucleotide probe corresponding to human MAPKAP kinase 2. The Northern blot analysis revealed that two mRNA species of MAPKAP kinase 2, with sizes of 4.8 and 3.3 kb, were expressed in high levels in both human heart and skeletal muscle tissues. To better understand how MAPKAP kinase 2 is regulated in myocardium, cultured rat cardiac myoblast (H9c2) cells were stimulated with heat shock, H2O2-induced oxidative stress, or phorbol ester (PMA). Enzymatic activity of cellular MAPKAP kinase 2 in the cell lysates was evaluated using an in vitro kinase assay. Exposure of H9c2 cells to heat shock or oxidative stress induced a transient increase of cellular MAPKAP kinase 2 activity, which reached its peak level within 5 min. In contrast, stimulation of H9c2 cells with PMA, a potential myocardial hypertrophic factor, induced a sustained increase of cellular MAPKAP kinase 2 activity that was detectable for over 1 h. In addition, in vitro protein phosphorylation analysis with recombinant MAPKAP kinase 2 showed that small heat shock protein (hsp25) served as a major substrate molecule for the kinase in H9c2 cells and the protein phosphorylation of cellular hsp25 was stimulated by H2O2-induced oxidative stress or PMA treatment in intact H9c2 cells. Moreover, exposure of H9c2 cells to H2O2-induced oxidative stress or PMA rapidly activated cellular p38 MAP kinase as detected by the induced protein phosphorylation of the kinase. Taken together, these results strongly suggest that MAPKAP kinase 2 may be involved in stress-activated signal transduction in myocardium.

Animals↗

PubMed1997

[Scalp expansion in treatment of scalp defect with skull exposure].

We repaired the scalp defect and skull exposure with scalp expansion. The technique consisted of two stages. First, expander was implaned an then N. S injected to its enough volume. The size of the expander was estimated by the formula that 1 cm2 needs 4 ml expansive volume. The incision was made at normal skin. Second flap was transferred. The expander was removed and the external skull which was necrotic severed. There were five patient with eight flaps at all, followed up for 6 months to 1 year. The hair grew. The results were satisfactory. This method not only treated the disease, but also avoided alopecia.

Adolescent↗

PubMed1997

Hybridization dominance of kinetics in recombinant ATH-1376 obtained via protoplast fusion between Aspergillus niger and Trichoderma reesei.

Comparisons of the kinetics of mycelium growth, cellulase biosynthesis, and the degradation of filter paper to accumulate reducing-sugar by the filtrates of cultures were carried out among the recombinant strain ATH-1376 and its two parents, Aspergillus niger AMS11 and Trichoderma reesei QM9414. The results showed that both the specific mycelium growth rate and the cellulase biosynthesis rate of the recombinant were dramatically dominant over those of the two parents. In addition, the negative correlation between the specific mycelium growth rate and the cellulase biosynthesis rate of the recombinant ATH-1376 was much lower than those of its parents. In terms of the amount of reducing-sugar accumulated from the hydrolysis of filter paper by culture filtrates, there were great differences among the three different treatments, i.e., fermentation filtrate of single parental strain, mixture of the fermentation filtrates from two parental strains with different ratios (v:v), and filtrate from the mixed culture of the two parental strains. Out of these, the second approach, particularly with the ratio of 1:1, was best for the accumulation of reducing sugar. Within various tested periods of enzymic hydrolysis, the amounts of reducing-sugar produced by the recombinant were 1.19 to 2.26 times as much as the maximum amounts produced in parallel by the mixture of filtrates (1:1) from separate fermentations of the two parental strains. These results suggested that constructing the engineered strains with hybridization dominance of these two typical genera of far-heredity could be effective to overcome the great deficiencies of routine mixculture, single-strain fermentation, or double fed-batch fermentations.

Aspergillus niger↗

PubMed1996

Activation of MAP kinase-activated protein kinase 2 in human neutrophils after phorbol ester or fMLP peptide stimulation.

In response to extracellular stimulation, one of the earliest events in human neutrophils is protein phosphorylation, which mediates signal transduction and leads to the regulation of cellular functions. Mitogen-activated protein (MAP) kinases are rapidly activated by a variety of mitogens, cytokines, and stresses. The activated MAP kinases in turn regulate their substrate molecules by phosphorylation. MAP kinase-activated protein (MAPKAP) kinase 2, a Ser/Thr kinase, has been shown to be phosphorylated by p38 MAP kinase both in vivo and in vitro. Phosphorylation of the Thr-334 site of MAPKAP kinase 2 results in a conformational change with subsequent activation of the enzyme. To better define the role of MAPKAP kinase 2 in the activation of human neutrophils, its enzymatic activity was measured after stimulation by either a phorbol ester (phorbol myristate acetate [PMA]), a potent protein kinase C activator, or the tripeptide fMLP, which is a chemotactic factor. The in vitro kinase assays indicate that both PMA and fMLP stimulated a transient increase in the enzymatic activity of cellular MAPKAP kinase 2. The induced kinase activation was concentration-dependent and reached a maximum at 5 minutes for PMA and 1 minute for fMLP. To identify potential substrate molecules for MAPKAP kinase 2, a highly active kinase mutant was generated by mutating the MAP kinase phosphorylation site in the C-terminal region. The replacement of threonine 334 with alanine resulted in a marked augmentation of catalytic activity. Analysis of in vitro protein phosphorylation in the presence of the active kinase indicates that a 60-kD cytosolic protein (p60) was markedly phosphorylated and served as the major substrate for MAPKAP kinase 2 in human neutrophils. Based on the MAPKAP kinase 2 phosphorylation site of Hsp27, a competitive inhibitory peptide was synthesized. This competitive inhibitory peptide specifically inhibited MAPKAP kinase 2 enzymatic activity, as well as the in vitro and in vivo kinase-induced p60 phosphorylation. To assess the contribution of MAPKAP kinase 2 in neutrophil function, the oxidative burst response after manipulation of endogenous kinase activity was measured. Intracellular delivery of the competitive inhibitory peptide into human neutrophils reduced both PMA- and fMLP-stimulated superoxide anion production. Thus, the results strongly suggest that MAPKAP kinase 2 is involved in the activation of human neutrophils.

Amino Acid Sequence↗

PubMed1996

Research on soft-tissue expander permeability to metronidazole and procaine.

This in vitro study was designed to determine if in fact silicone expanders are readily permeable to metronidazole and procaine. The expanders were filled with 0.2% metronidazole or 2% procaine through the filler valves and then immersed wholly in normal saline. At several intervals over 120 hours, a certain amount of the surrounding saline was sampled and the drug levels subsequently determined. In this study, the silicone expanders were indeed readily permeable to the drugs, as measured with an ultraviolet spectrophotometer. A consistent diffusion curve was demonstrated. The rate of diffusion of a drug is inversely proportional to its molecular weight; i.e., the smaller the given drug's molecular weight, the greater is its ability to permeate an expander. In view of this, in the course of expansion, 0.2% metronidazole could be used in clinical expansion instead of normal saline, and a certain amount of antibiotic would diffuse out of the expander to prevent and control the infection.

Humans↗

PubMed1996

Fine structure of the human galactokinase GALK1 gene.

Defects in the human GALK1 gene result in galactokinase deficiency and cataract formation. We have isolated this gene and established its structural organization. The gene contains 8 exons and spans approximately 7.3 kb of genomic DNA. The GALK1 promoter was localized and found to have many features in common with other housekeeping genes, including high GC content, several copies of the binding site for the Sp1 transcription factor, and the absence of TATA-box and CCAAT-box motifs typically present in eukaryotic Pol II promoters. Analysis by 5'-RACE PCR indicates that the GALK1 mRNA is heterogeneous at the 5' terminus, with transcription sites occurring at many locations between 21 and 61 bp upstream of the ATG start site of the coding region. In vitro translation experiments of the GALK1 cDNA indicate that the protein is cytosolic and not associated with the endoplasmic reticulum membrane.

Amino Acid Sequence↗

PubMed1996

An attempt on using the method of R-Q double-factor analysis to identify and group fusants.

A subsequent numeric taxonomy method for identifying and grouping the fusants was explored on the basis of characterization of the protein profiles of the fusants. By these two means, several typical excellent candidates of recombinants could be searched out quickly. Among the fusants from a definite fusion-cross, the different sister-strains were regarded as the samples of observation (n = N), the positions of the all bands of protein profiles as the objects (p = P), and the photometer-scanning area of the specific band as the experimental value (X) (zero was taken when the specific band of a certain strain was absent). The genetic multirelationships among the inter- and intra-sister-strains in terms of the positions and contents of the protein bands after fusion recombination occurred from this definite fusion-cross could therefore be determined on the same orientational factor-plate by using the computer program of the R-Q double-factor method to analyze this data matrix (Xnxp). These sister-strains could then be identified and grouped from the deduced heredity relationship between the fusants and parents.

Aspergillus niger↗

PubMed1996

[The application of expanded delto-pectoral flaps in the faciocervical region].

The delto-pectoral flap has been used to repair burned scar in the faciocervical region for some years. However, its limited size restricts its application. Furthermore, direct transfer of the flap may result in a swelling and inexpressive face and the donor site needs skin grafting. To avoid the above disadvantages, we have tried pre-expansion of the flap. In this article, the authors report the experiences in the application of the method to eighteen cases, including the surgical procedure, the applied anatomy, and typical cases as well. Also included in the article are the comparison between various therapies to the burned scar of the face, the key points for successful pre-expansion of the delto-pectoral flap, accurately maintaining the desired position. The method has been proved to have many advantages and can be widely applied.

Adolescent↗

PubMed1996

[Endoscopic control during facelifting].

The endoscopic techniques were just recently applied to aesthetic surgery. The paper reports 22 cases of video endoscopic facelift that have been performed since April 1994. The operative results were satisfactory. Clinical experience with forehead plasty, resection of corrugator and procerus muscle and other cosmetic procedures using endoscopic technique is presented.

Adult↗