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Biomedical subjects

M Yu

Publications and source records attributed to M Yu.

At least 397 records · Page 22Linked to original sources
PubMed1990

[Studies on ultraweak luminescence of bacteria].

Ultraweak luminescence of Escherichia coli, Bacillus subtilis and Brevibacterium ammoniagenes was measured with high sensitive single photon counting equipment (Made in China). The results obtained from ultraweak luminescence of as above three bacterial strains were as follows: spectral distribution curves, photon emission kinetic curves or emission intensity and its qualitative relationship between intensity and bacterial counts.

Bacillus subtilis↗
PubMed1989

Absence of Ig V region gene somatic hypermutation in advanced Burkitt's lymphoma.

Somatic hypermutation of rearranged Ig V region gene plays a major role in generating antibody diversity. Recently, V mutation has been established as a major mechanism of tumor escape from anti-Id immunotherapy. We cloned and sequenced the expressed Ig H and L chain V regions from a case of B acute lymphoblastic leukemia in order to evaluate B cell stages associated with V region mutation, and to determine which tumors would be better suited to Id directed immunotherapy. A consensus VH and V lambda sequence representing tumor at diagnosis was obtained by conventional cDNA cloning in lambda gt10 from a heterohybridoma. Primers which flanked both V regions were used in a modified polymerase chain reaction to generate multiple independent sequences from tumor cells harvested at relapse. In order to exclude mutations due to infidelity of the amplification procedure, single cDNA templates of known sequence were also amplified. The polymerase chain reaction proved to be an effective procedure to obtain multiple clones, but replication in M13 was associated with a low rate of base misincorporation. The results indicate that there is no evidence for biologically significant ongoing mutation in this t(8;14) B cell tumor when comparing sequences at diagnosis and relapse. Thus, V somatic mutation may be restricted to a discrete B cell stage whose malignant counterpart is follicular lymphoma.

Adolescent↗
PubMed1989

Rat liver ATP-sulfurylase: purification, kinetic characterization, and interaction with arsenate, selenate, phosphate, and other inorganic oxyanions.

ATP-sulfurylase (ATP:sulfate adenylyltransferase; EC 2.7.7.4), the first enzyme of the two-step sulfate activation sequence, was purified extensively from rat liver cytosol. The enzyme has a native molecular mass of 122 +/- 12 kDa and appears to be composed of identical 62 +/- 6-kDa subunits. At 30 degrees C and pH 8.0 (50 mM Tris-Cl buffer containing 5 mM excess Mg2+), the best preparations have "forward reaction" specific activities of about 20 and 2 units X mg protein-1 with MoO4(2-) and SO4(2-), respectively. The reverse (ATP synthesis) specific activity is about the same as the forward molybdolysis activity. The kinetic constants under the above conditions are as follows: KmA = 0.21 mM, Kia = 0.87 mM, KmB = 0.18 mM, KmQ = 0.65 microM, Kiq = 0.11 microM, and KmP = 5.0 microM where A = MgATP, B = SO4(2-), Q = APS, and P = total PPi at 5 mM Mg2+. PPi is a mixed-type inhibitor with respect to MgATP and SO4(2-). SeO4(2-) is an alternative inorganic substrate with a Vmax about 20% that of SO4(2-). The product, APSe, is unstable. But in the presence of a sufficient excess of APS kinase, APSe is completely converted to PAPSe. The rate constant for nonenzymatic PAPSe hydrolysis was determined from measurements of the final steady-state reaction rate in the presence of limiting initial SeO4(2-) and a large excess of MgATP, ATP sulfurylase, APS kinase, and the other coupling enzymes and their cosubstrates. The results yielded a k of 2.4 +/- 0.5 X 10(-3) sec-1 (t1/2 ca. 5 min). Phosphate is an effective buffer for enzyme purification and storage but inhibits catalytic activity, particularly at low substrate concentrations. In the presence of buffer levels of Pi, the MgATP reciprocal plot of the SO4(2-)-dependent reaction is concave-up. Inorganic monovalent oxyanions are dead end inhibitors competitive with SO4(2-) and apparently uncompetitive with respect to MgATP. The relative potencies are in the order ClO3- greater than ClO4- greater than FSO3- greater than NO3-. Thiosulfate is also competitive with SO4(2-) but noncompetitive with respect to MgATP. Several divalent oxyanions (MoO4(2-), WO4(2-), CrO4(2-), and HAsO4(2-] promote the enzyme-catalyzed cleavage of MgATP to AMP and MgPPi. The ratio Vmaxf/KmA ranged from 0.7 to 200 for various reactive inorganic substrates. The cumulative results suggest the random binding of MgATP and the inorganic substrate but the ordered release of MgPPi before APS.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine Phosphosulfate↗
PubMed1989

Attomole amino acid determination by capillary zone electrophoresis with thermooptical absorbance detection.

Attomole quantities of 4-(dimethylamino)azobenzene-4'-sulfonyl chloride derivatized amino acids are separated by using capillary zone electrophoresis in a mixed acetonitrile/aqueous buffer system. Detection is performed with an on-column thermooptical absorbance detection technique based on a 130-mW argon ion pump laser. Detection limits for the concentration of analyte injected onto the column range from 5 x 10(-8) M for methionine to 5 x 10(-7) M for aspartic acid. Only 37 amol of methionine and 450 amol of aspartic acid are contained within the subnanoliter injection volume. It is interesting to note that these limits are a factor of 4 superior to the best fluorescence detection limit reported for chromatographic separation of amino acids. A subnanoliter sample of derivatized human urine was analyzed with this technique; quantities of amino acids contained within the sample are 3 orders of magnitude greater than the detection limit.

Amino Acids↗
PubMed1989

Fungicidal activity of cilofungin (LY121019) alone and in combination with anticapsin or other antifungal agents.

Cilofungin (LY121019) was shown to have potent fungicidal activity against clinical isolates of Candida albicans and Candida tropicalis but not Candida parapsilosis. Fungicidal activity was evident against both replicating and non-replicating Candida albicans and was progressive over the first 12 h of incubation. The combination of cilofungin (LY121019) with anticapsin but not with amphotericin B, ketoconazole or 5-fluorocytosine resulted in synergistic fungicidal activity. This compound warrants further investigation of its safety and efficacy in the treatment of Candida infections.

Alanine↗
PubMed1989

Heterogeneity of [3H]estradiol binding sites in the rat prostate: properties and distribution of type I and type II sites.

In order to assess the rat prostate as a target tissue for receptor-mediated estrogen action, we have studied the properties and distributions of estrogen binding sites in the dorsolateral (DLP) and ventral (VP) prostate. Saturation analyses over a wide range of [3H]estradiol ([3H]E2) concentrations (0.5-100 nM) revealed two distinct types of binding sites in the cytosol and nuclear fractions of DLP of intact rats. The high affinity (type I) estrogen binding sites saturated at 2-4 nM of [3H]E2 and had a capacity of 170 fmol/mg DNA in the cytosol and 400 fmol/mg DNA in the nuclei. DLP type I sites had ligand specificity similar to that described for the classical estrogen receptors (ERs) found in female target tissues. The moderate affinity (type II) estrogen binding sites saturated at 15-30 nM of [3H]E2 and had a capacity of 850 fmol/mg DNA in the cytosol and 1600 fmol/mg DNA in the nuclei. DLP type II sites shared some characteristics of the type II ERs described for the rat uterus; they were estrogen specific, heat labile, and sensitive to reducing agents such as dithiothreitol. Saturation analyses on VP cytosols and nuclear fractions revealed only high affinity sites but no moderate affinity sites in the tissue preparations. Our finding that prostatic type II estrogen binding sites are present exclusively in the DLP supports the concept that basic biological differences exist between the two major prostatic lobes of the rat. Furthermore, our findings may help elucidate the observed differences in susceptibility between these two lobes to the hormonal induction of proliferative prostatic lesions.

Animals↗
PubMed1989

Flow cytometric analysis of murine splenic cells after DNFB painting on Langerhans cell deficient skin.

One week after a single painting of 50 microliters of 0.5% DNFB in acetone-olive oil on the tail skin of C3H mice, the spleens were removed from the animals. A single cell suspension was prepared from the spleens, and flow cytometric analysis was performed for L3T4 and Lyt2 positive cells, as well as for the IJk expression of the Lyt2+ cells. Results showed that the percentages and absolute numbers of Lyt2+ L3T4- cells, Lyt2- L3T4+ cells, Lyt2+ IJk+ cells, and Lyt2+ IJk- cells in the spleens of the DNFB-treated mice were not significantly different from those in the non-treated mice. However, in the treated mice, the IJk expression of Lyt2+ cells intensified. These results indicate that, following a single painting of DNFB onto Langerhans cell-deficient skin, the numbers of Lyt2+ cells do not change significantly, but do change functionally.

Animals↗
PubMed1989

Transcription of the folC gene encoding folylpolyglutamate synthetase-dihydrofolate synthetase in Escherichia coli.

The folC gene of Escherichia coli is cotranscribed with an upstream gene from two promoters located in the noncoding region 5' to the coding sequence of the upstream gene. Virtually all of the expression of the folC gene product, folylpolyglutamate synthetase-dihydrofolate synthetase, is therefore due to the upstream gene promoters. No promoter activity was found in the coding sequence of the upstream gene or in the 72-base-pair noncoding region between the two genes. It is shown that a third gene, which may overlap the coding sequence of the folC gene by 8 base pairs at the 3' end, nevertheless, has an promoter independent from that of the upstream gene-folC operon. These results contrast with those presented by Nonet et al. (M. L. Nonet, C. C. Marvel, and D. Tolan, J. Biol. Chem., 262:12209-12217, 1987), who concluded that folC was cotranscribed with the gene at its 3' end and the gene upstream to folC was cotranscribed with the gene(s) further upstream. A stable stem-loop structure resembling a rho-independent terminator is present within the noncoding region between the upstream gene and the folC gene. Folypolyglutamate synthetase expression is 6- to 15-fold lower than that of the upstream gene product, suggesting that the stem-loop terminates some of the transcription from the upstream gene promoter. We found by deletion mutagenesis and cloning sequences containing the stem-loop structure into a termination reporter plasmid that this stem-loop does not act as an effective terminator of transcription. We also found that the stem-loop does not protect the upstream gene message from degradation, since expression of the upstream gene product in maxicell experiments is the same whether the stem-loop structure is present or deleted.

DNA Mutational Analysis↗
PubMed1989

[Expression of human interferon beta gene controlled by SV40 late promoter in mouse myeloma SP2/0 cells].

The HindII fragment of human interferon beta (IFN-beta) gene was inserted downstream from SV40 late promoter in pSV2-dhfr and cotransfered with pSV2-gpt into the mouse myeloma SP2/0 cells which were hypoxanthine guanine phosphoribosyl transferase (HGPRT) deficient. After the selection in HAT (Hypoxanthine Aminopterine Thymidine) medium containing myciphenolic acid and xanthine, the efficient constitutive expression of IFN-beta could be detected in the supernatant of the survive cells.

Animals↗
PubMed1989

[Antigen presenting cells and T lymphocytes in the induction phase of contact hypersensitivity].

When a sensitizing substance that induces contact hypersensitivity, fluorescein isothiocyanate (FITC), was painted on abdominal skin of mice, FITC+ cells appeared in the inguinal lymph node after 24 hours. The FITC+ cell in the lymph node was relatively large in size, and it did not appear to be a T lymphocyte. When FITC was painted on either murine tail skin or skin pre-treated by tape stripping, the number of FITC+ cells in the inguinal lymph node was significantly less than that in the positive control. In the mesenteric lymph nodes, which have a different lymph flow from that of the skin regional lymph node, FITC+ cells did not increase in number, and the few FITC+ cells were not significantly different in number among above-mentioned experimental systems. In the inguinal lymph nodes on the 4th day after painting of picryl chloride (PCl) on the abdominal skin of mice, L3T4+ cells, which expressed an interleukin 2 receptor (IL-2R), increased in number. On the other hand, when PCl was painted on either tail skin or skin treated by tape stripping, L3T4+ IL-2R+ cells did not increase in the skin regional lymph nodes. The number of L3T4+ IL-2R+ cells in the mesenteric lymph nodes did not increase in any of the experimental systems mentioned above. These results suggest some relationship between antigen presenting cells and T lymphocytes, as well as one between the skin and the regional lymph nodes, in an induction phase of sensitization in contact hypersensitivity.

Animals↗
PubMed1989

[Constitutive expression of human fibroblast interferon gene controlled by SV40 early promoter in CHO cells].

HindII fragment encoding human fibroblast interferon (IFN beta) gene coding sequence was fused at 60 bp downstream from the RNA start site of SV40 early gene to be a constitutive expression plasmid pSVE beta. This recombinant plasmid was transfected into the dihydrofolate reductase (dhfr)-deficient Chinese hamster ovary (CHO) cells together with a selectable dhfr gene. About half of transformants continuously secreted IFN beta into the supernatant without inducement. One of the subclone transformants constitutively produced up to 852U IFN beta/2 X 10(6) cells/ml. 48hr in common medium.

Animals↗
PubMed1989

[Course and prognosis of multiple sclerosis. Report of 90 cases].

90 cases of multiple scleroses were studied in connexion with the course and prognosis of the disease. The course of this disease may be classified into 4 type: benign, remitting progressive, and chronic progressive types. Relapse took place in 75% of the patients. The frequency of relapse was on the average 0.45 time per year. Severe disability occurred mostly during the first five years. The present study suggested that the factors indicating relatively poor prognosis would be the onset at the age over 40 years, chronic insidious onset, early severe impairment of the pyramidal tract function and multiple involvement of the different systems in the CNS.

Adolescent↗
PubMed1988

High-affinity binding of [3H]estradiol-17 beta by an estrogen receptor in the liver of the turtle.

Specific [3H]estradiol-17 beta ([3H]E2) binding activity (EBA) with characteristics of an estrogen receptor (ER) was demonstrated in cytosols and nuclear extracts of the female turtle, Chrysemys picta. Three different receptor assays (dextran-coated charcoal assay, hydroxylapatite batch procedure, and DNA-cellulose chromatography) were evaluated in terms of their applicability in analyzing large numbers of samples. For the measurement of cytosolic EBA, the hydroxylapatite batch procedure was found to be the most reliable assay. On the other hand, the dextran-coated charcoal assay was found to be the most appropriate method for the measurement of nuclear EBA. Turtle hepatic EBA binds [3H]E2 with high affinity (cytosolic, 17.4 +/- 2.8 X 10(9) M-1; nuclear, 17.7 +/- 1.9 X 10(9) M-1), limited capacity (cytosolic, 133.7 +/- 4.6 fmol/g tissue; nuclear, 81.1 +/- 9.0 fmol/g tissue), and strict steroid specificity. The EBA bound natural estrogens (E2, estrone, estriol) as well as the nonsteroidal estrogen, diethylstilbestrol, but exhibited little affinity for androgens, progesterone, or corticosterone. The turtle hepatic EBA resembled mammalian and avian ERs in terms of binding characteristics; however, unlike mammalian and avian ERs it was shown to be heat-labile. Incubation at 30 degrees caused rapid loss of [3H]E2 binding activity in both cytosolic and nuclear fractions. The exchange between [3H]E2 and the endogenously bound estrogen was slow at 4 and 15 degrees, but the exchange process was facilitated in the presence of the chaotropic salt, NaSCN. Establishment of quantitation methods for both cytosolic and nuclear forms of EBA will enable future investigation of the mechanism and regulation of estrogen action in the liver of this turtle species.

Animals↗
PubMed1988

Influence of in vitro susceptibility testing conditions on the anti-candidal activity of LY121019.

LY121019 is a new antifungal antimicrobic that is structurally similar to the lipopeptide agents echinocandin B and aculeacin A. Because of the importance of in vitro test conditions on the activity of other antifungal agents, we studied the effects of inoculum size, time and temperature of incubation, pH, and medium composition on the in vitro activity of LY121019 against Candida albicans, Candida tropicalis, and Candida parapsilosis. LY121019 was highly active against Candida albicans and Candida tropicalis and inactive against Candida parapsilosis. The in vitro activity of LY121019 is marked by a paradoxical dose-response with isolates of Candida albicans and Candida tropicalis and is influenced by choice of inoculum size, time and temperature of incubation, medium composition, and pH. We recommend the use of an inoculum size of less than 10(5) organisms/ml, a defined medium buffered to a pH of 7.0, and incubation at 30 degrees C for 24 hr for future in vitro studies of LY121019.

Amphotericin B↗
PubMed1988

Calcium entry into the inositol 1,4,5-trisphosphate-releasable calcium pool is mediated by a GTP-regulatory mechanism.

Intracellular Ca2+ release activated by inositol 1,4,5-trisphosphate (InsP3) plays a pivotal role in Ca2+ signaling in cells. A controlling mechanism for InsP3-induced Ca2+ movements is suggested by results showing that the InsP3-releasable Ca2+ pool is directly modified by a specific and sensitive GTP-regulated Ca2+-translocating process. By using saponin-permeabilized N1E-115 neuroblastoma cells or DDT1MF-2 smooth muscle-derived cells, InsP3 releases 30-50% of Ca2+ accumulated through intracellular high-affinity ATP-dependent Ca2+-pumping activity. Oxalate-promoted Ca2+ uptake is reversed by InsP3, indicating oxalate permeability of the InsP3-releasable pool, which is consistent with this compartment being the endoplasmic reticulum. GTP (10 microM) activates release of 50-70% of accumulated Ca2+ from cells. In the presence of 5-10 mM oxalate, GTP induces a biphasic Ca2+ flux response; initially (1-2 min) GTP induces rapid Ca2+ release followed thereafter by a profound increase in Ca2+ uptake. Thus, GTP-activated Ca2+ influx and efflux compete for Ca2+ access to the oxalate-permeable Ca2+ pool. The nonadditive effects of InsP3 and GTP suggest that InsP3 releases Ca2+ from a subcompartment of the GTP-releasable pool. Most significantly, InsP3 is observed to block the GTP-activated uptake phase in the presence of oxalate, indicating that GTP induces Ca2+ entry into the pool from which InsP3 activates release. Hence, the results provide direct evidence that loading of Ca2+ into the InsP3-sensitive Ca2+ pool is controlled by a GTP-regulated Ca2+-translocating mechanism. Such a process could be significant in regulating the extent and duration of the InsP3-induced Ca2+ signal, a crucial step in the inositol phospholipid signaling pathway.

Animals↗
PubMed1988

UV photoelectron spectroscopy and ab initio characterization of valence orbital structures and conformations of neutral phosphate esters.

The HeI UV photoelectron spectrum of trimethyl phosphate (TMP) has been measured and interpreted with the aid of SCF molecular orbital calculations carried out with STO-3G, STO-3G* and 4-31G basis functions. The photoelectron spectrum of TMP is more accurately reproduced by results from 4-31G calculations than by results from STO-3G or STO-3G* calculations. However, all three basis sets yield results which predict the same assignment of the photoelectron spectrum. Results at the 4-31G level indicate that whether calculations are based on crystallographic bond angles and bond lengths or on STO-3G optimized geometries has little effect on the energetic ordering of the upper occupied orbitals. The energetic ordering of orbitals is also found to be only weakly dependent upon the torsional angle phi, describing rotation of ester groups about P-O bonds and upon the torsional angle psi, describing rotation of methyl groups about C-O bonds. For trimethyl phosphate, with C3 symmetry, the vertical ionization potentials of the upper occupied orbitals are 10.81 eV (8e), 11.4 eV (9a), 11.93 eV (7e), 12.6-12.9 eV (8a and 6e), 14.4 eV (7a) and 15.0-16.0 eV (5e and 6a). Calculations at the 4-31G level indicate that many of the highest occupied orbitals in neutral dimethyl phosphate and methyl phosphate have energies and electron distributions similar to orbitals in TMP. For TMP, a search for optimized values of phi and psi has been carried out at the STO-3G*level. In agreement with previous NMR studies and with classical potential calculations, the STO-3G* results indicate that both the gauche (phi = 53.1 degrees) and anticlinal (phi = 141.9 degrees) conformations are thermally accessible. Also in agreement with the classical potential calculations, the STO-3G* results predict that in the all gauche conformation energy is minimized when the methyl groups assume a staggered geometry (psi = 60 degrees to 80 degrees) and that an energy maximum occurs for an eclipsed geometry (phi = 0 degrees to 20 degrees). A study of the dependence of optimized values of O-P-O ester bond angles on the torsional angles, phi, was carried out at the STO-3G, STO-3G* and 4-31G levels. The results demonstrate that for C3 symmetry, the coupling of O-P-O angles to phi is influence by repulsive steric interactions.

Chemical Phenomena↗