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Biomedical subjects

W Yu

Publications and source records attributed to W Yu.

At least 397 records · Page 22Linked to original sources
PubMed1982

Penicillin-binding proteins in Bacillus subtilis mutants.

The penicillin-binding proteins (PBPs) from mutants of Bacillus subtilis were studied and related to morphology. In a previously described cloxacillin-resistant mutant of B. subtilis strain Porton, PBP 2a had an altered mobility by sodium dodecyl sulfate gel electrophoresis and was present in increased amounts. In addition, PBPs 1a and 1b were missing in this mutant. The only morphological change seen was a decrease in size of about 15%. Studies of two Triton-resistant morphological mutants of B. subtilis 168, Tr49 (small diameter) and Tr61 (helical form), revealed no change in the number of PBPs compared with that of the parent strain. However, PBPs 1a and 1b had an altered mobility in the mutant Tr49.

Bacillus subtilis↗
PubMed1980

Linear, uncross-linked peptidoglycan secreted by penicillin-treated Bacillus subtilis. Isolation and characterization as a substrate for penicillin-sensitive D-alanine carboxypeptidases.

Incubation of growing Bacillus subtilis with penicillin G led to the secretion of a peptidoglycan-related polymer and a nonglycan-bound pentapeptide into the culture medium. The secreted polymer was isolated and characterized as a linear cell wall glycan strand substituted predominantly by uncross-linked pentapeptide side chains. Polymer formation and secretion are most likely the result of continued synthesis and elongation of nascent glycan strands in the absence of subsequent processing by peptidoglycan transpeptidase or D-alanine carboxypeptidase enzymes. The nonglycan-bound pentapeptide, L-Ala-D-iso-Glu-meso-diaminopimelic acid-D-Ala-D-Ala, was probably formed by an N-acetylmuramyl-L-alanine amidase active on the peptide side chains of The uncross-linked polymer. The uncross-linked peptidoglycan polymer was shown to be a good substrate for penicillin-sensitive D-alanine carboxypeptidases purified from membranes of B. subtilis, Bacillus stearothermophilus, and Escherichia coli. D-alanine release was not, however, coupled to the cross-linking of peptide side chains, suggesting that these enzymes do not function as peptidoglycan transpeptidases in vivo. No transpeptidase or D-alanine carboxypeptidase activity was detected in mixtures of high molecular weight penicillin-binding proteins from B. subtilis, B. stearothermophilus, or Staphylococcus aureus. Possible reasons for the inability to demonstrate these activities are discussed. In addition, an N-acetylmuramyl-L-alanine amidase activity which copurifies with penicillin-binding proteins from B. subtilis, S. aureus, and E. coli was partially characterized.

Amino Acids↗
PubMed1980

Some aspects of chemical synthesis of oligoribonucleotides.

A simple synthesis of ribothymidine 3'-phosphate from uridine 3'-phosphate was described. Crystalline 5'-O-trityl- or 5'-O-monomethoxytrityl-derivatives of ribonucleotides are easily prepared and purified in relatively large batches through choices of appropriate solvents. Acylimidazoles together with a strong nitrogeneous base have been successfully used for the acylation of the hydroxyl and amino groups of the nucleoside and nucleotide, and the reaction mechanism was studied. The relative stability of different blocking groups of guanosine 3'-phosphate was compared. Oligoribonucleotide fragments including an octaribonucleotide of the T psi C- and D-loop of yeast alanine tRNA were synthesized. Quantitative analyses of the common ribo-nucleosides and -nucleotides were done by a modified RPC method.

Base Sequence↗
PubMed1977

Time-resolved fluorescence spectroscopy of spinach chloroplast.

Picosecond fluorescent kinetics and time-resolved spectra of spinach chloroplast were measured at room temperature and low temperatures. The measurement is conducted with 530 nm excitation at an average intensity of 2-10(14) photons/cm2, pluse and at a pulse separation of 6 ns for the 100 pulses used. The 685 nm fluorescent kinetics was found to decay with two components, a fast component with a 56 ps lifetime, and a slow component with a 220 ps lifetime. The 730 nm fluorescent kinetics at room temperature is a single exponential decay with a 100 ps lifetime. The 730 nm fluorescence lifetime was found to increase by a factor of 6 when the temperature was lowered from room temperature to 90 K, while the 685 and 695 nm fluorescent kinetics were unchanged. The time-resolved spectra data obtained within 10 ps after excitation is consistent with the kinetic data reported here. A two-level fluorescence scheme is proposed to explain the kinetics. The effect of excitation with high light intensity and multiple pulses is discussed.

Chloroplasts↗
PubMed1975

Fluorescent kinetics of chlorophyll in photosystems I and II enriched fractions of spinach.

The fluorescent emission kinetics of spinach subchloroplast Photosystems I and II particles have been studied on a picosecond time scale. Using picosecond laser pulses and an optical Kerr gate, the fluorescent decay times are measured to be 60 plus or minus 10 ps, and 200 plus or minus 20 ps for Photosystems I and II, respectively. The quantum yields are calculated to be 0.004 for Photosystem I and 0.013 for Photosystem II. Theory of exciton energy transfer and trapping is applied for the determination of intermolecular potential energyin the photosystems.

Chlorophyll↗
PubMedDate not supplied

Early handling can attenuate adverse effects of fetal ethanol exposure.

The effects of early handling on physiological and hormonal responses of rats exposed to ethanol prenatally were studied. Male and female Sprague-Dawley rats from prenatal ethanol (E), pair-fed (PF), and ad lib-fed control (C) prenatal treatment groups were either handled (H) or nonhandled (NH) during the preweaning period and tested in adulthood. Early handling eliminated the deficit in preweaning weight gain observed in E compared to PF and C offspring. In adulthood, early handling eliminated the increased hypothermia observed in E and PF compared to C males following an ethanol challenge (2.0 g/kg, IP). In addition, H males displayed marginally less hypothermia overall than NH males. In contrast, handling accelerated the return to preinjection temperature in PF and C females but had no effect on E females. There were no significant differences among E, PF, and C rats in corticosterone (CORT) responses to ethanol challenge (1.5 g/kg, IP), but both males (marginally) and females in the H condition displayed higher CORT levels overall than NH rats. Early handling also eliminated the increased peak CORT response to restraint stress in E compared to C females, but did not affect the more prolonged elevation of CORT in E compared to PF and C females. There were no differences among E, PF, and C females in hippocampal type I and type II glucocorticoid receptor density or affinity. However, binding affinity of type II receptors was slightly but significantly increased in H compared to NH females. Together, these data indicate that early handling may modulate or attenuate some, but not all, of the adverse effects of fetal ethanol exposure on offspring growth and physiological responsiveness.

Animals↗
PubMedDate not supplied

Estimating the contributions of NMDA and non-NMDA currents to EPSPs in retinal ganglion cells.

Whole-cell recordings were obtained from retinal ganglion cells of the tiger salamander (Ambystoma tigrinum) in a superfused slice preparation to evaluate contributions of NMDA (N-methyl-D-aspartate) and KA/AMPA (kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid) receptors to excitatory postsynaptic potentials (EPSPs) of retinal ganglion cells. Synaptic activation of retinal ganglion cells was achieved through the use of a brief pressure pulse of hyperosmotic Ringer (Ringer + sucrose) delivered through a microelectrode visually placed in the inner plexiform layer while whole-cell recordings were obtained from adjacent cells in the ganglion cell layer. Separation of NMDA and KA/AMPA excitatory postsynaptic currents (EPSCs) was achieved through the application of the antagonists NBQX and D-AP7, while inhibitory currents were blocked by strychnine and picrotoxin. Simple addition of the two independent EPSCs showed, most often, that the sum of the KA/AMPA and NMDA currents was less than the control response, but in some cases the sum of the two currents exceeded the magnitude of the control response. Neither result was consistent with expectations based on voltage-clamp principles and the assumption that the two currents were independent; for this reason, we considered the possibility of nonlinear interactions between KA/AMPA and NMDA receptors. Computer simulations were carried out to evaluate the summation experiments. We used both an equivalent cylinder model and a more realistic, compartmental model of a ganglion cell constrained by a passive leakage conductance, a linear KA/AMPA synaptic current, and a nonlinear NMDA current based on the well-known, voltage-sensitive Mg2+ block. Computer simulation studies suggest that the hypo- and hyper-summation of NMDA and KA/AMPA currents, observed physiologically, can be accounted for by a failure to adequately space clamp the neuron. Clamp failure leads to enhanced NMDA currents as the ion channels are relieved of the Mg2+ block; their contribution is thus exaggerated depending on the magnitude of the conductance change and the spatial location of the synaptic input.

2-Amino-5-phosphonovalerate↗
PubMedDate not supplied

Cytoplasmic lipid bodies in eosinophils: central roles in eicosanoid generation.

BACKGROUND: Lipid bodies are lipid-rich cytoplasmic inclusions which form in diverse cell types, including eosinophils. Lipid body numbers increase in vivo in leukocytes participating in inflammatory processes. Our interest in lipid bodies relates to the roles that these structures play in arachidonate metabolism by eosinophils and other leukocytes involved in inflammation. METHODS: Specific agonists, platelet-activating factor (PAF), two cis-unsaturated fatty acids (arachidonic and oleic acids) and a diglyceride (1-oleyl-2-acetyl-glycerol (OAG)), were used to stimulate lipid body formation in human eosinophils. Lipid bodies were enumerated and eosinophils were stimulated with submaximal calcium ionophore to generate leukotriene C4 (LTC4), which was quantitated by immunoassay. RESULTS: Lipid body formation was rapidly inducible in eosinophils by specific intracellular signaling pathways. PAF, the two cis-unsaturated fatty acids and OAG each stimulated lipid body formation in eosinophils. Increased numbers of lipid bodies correlated quantitatively with the 'priming' response of eosinophils to form enhanced amounts of the 5-lipoxygenase-derived eicosanoid, LTC4. CONCLUSION: Lipid bodies in eosinophils function as intracellular domains that are both depots of esterified arachidonate and sites at which regulated enymatic events relevant to arachidonate metabolism can occur. In conjunction with our findings that key eicosanoid-forming enzzymes, including cyclooxygenase, 5- and 15-lipoxygenase and LTC4 synthase, are localized at lipid bodies in eosinophils, the finding that induction of lipid body formation correlated quantitatively with enhanced LTC4 production indicate that lipid bodies are structurally distinct, inducible, non-nuclear sites for enhanced synthesis of paracrine eicosanoid mediators of inflammation.

Cells, Cultured↗
PubMedDate not supplied

Recyclable islet isografts: a biological insulin pump to facilitate surgery in diabetic rats.

A method is described to facilitate extensive abdominal surgery in diabetic rats using temporary renal subcapsular islet isografts. The procedure is performed in three stages--each separated by 2 wk. After each rat is made diabetic (Step 1), an isogenic islet transplant is performed beneath the left renal capsule (Step 2). Step 3 is the operative procedure, followed by a left nephrectomy. Finally, it is shown how the islet isograft can be excised from the nephrectomy specimen and "recycled" into another diabetic rat (Step 2). Using this three step process, it is possible to decrease the mortality rate following orthotropic liver transplantation in diabetic rats from 100% to about 30%. This procedure can also be used to facilitate other extensive surgical procedures in diabetic rats.

Animals↗
PubMedDate not supplied

Perigastric lymph nodes with metastasis in gastric cancer.

BACKGROUND/AIMS: We analyzed the significance of metastasis to the subdivided perigastric lymph node stations according to the distance from the primary gastric cancer, and the appropriateness of the recent change in the Union Internacional Contra la Cancrum (UICC) tumor node metastasis (TNM) system. METHODOLOGY: Gastrectomy was performed in 753 patients with gastric cancer. The perigastric lymph nodes were divided into 6 stations according to the Japanese classification. These were subdivided into 2 categories according to the distance from the primary tumor: -1, nodes within 3 cm of the edge of the tumor; and -2, nodes more than 3 cm from the edge of the tumor. Survival rates were calculated with the Kaplan-Meier method, and the difference between each group was evaluated by the log-rank method. RESULTS: The frequency of metastasis to the subdivided perigastric lymph node stations, numbered 1-1 to 6-2, varied between 10.0% and 41.1%. The 5-year survival rate of the patients with positive 6-1 lymph node was higher than that of the patients with positive 6-2 lymph node (31.5% and 17.5%, P = 0.0032). There were no statistically significant differences in survival between subgroups of patients who had metastatic lymph node in the other 5 stations. The frequency of metastasis to other regional lymph nodes in patients with N2 perigastric lymph nodes was higher than that in patients with N1 perigastric lymph nodes. CONCLUSIONS: Subdivision of the perigastric lymph nodes had little advantage. Elimination of the old system of classifying perigastric lymph nodes according to distance from the tumor is appropriate.

Female↗