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P Lu

Publications and source records attributed to P Lu.

At least 163 records · Page 9Linked to original sources
PubMed1994

[Analysis of the target epitopes recognized by two monoclonal antibodies directed to egg-associated fractions of Schistosoma japonicum].

Two IgM isotype monoclonal antibodies (McAb), 2H10 and 2H1, recognizing repetitive epitopes on Schistosoma egg-associated molecules were characterized and their specificities were identified in a two-site sandwich ELISA system. In consistent with the differences in immunological behaviour and specificity demonstrated with immunoelectrophoresis (IEP) and immunofluorescent antibody (IFA) techniques, absolutely negative reactions found in the heterologous detecting system with alternated capture and detecting McAbs of the two revealed a complete incompatibility giving evidences that epitopes on different molecules were recognized. Immunological liability of the target antigen SEA or SEA-TCA to the two McAbs were demonstrated on sodium periodate and trifluoroacetic acid treatment indicating the biochemical nature of these epitopes were glycosyolated molecules with apparently higher resistance to the oxidizing agent showing in 2H10 recognizing epitopes. By means of an ion-gradient Mono-Q FPLC system (Pharmacia), 2H10-reactive epitopes of SEA, being tested not so efficiently adsorbed by ConA-sepharose affinity column, was found successfully concentrated in the profile eluted with pH 8.0 PBS at 0.2-0.4 NaCl ionic strength. Repeated trials on SDS-PAGE and Western blotting analysis with the reactive fractions further showed a heterogeneity of molecular weight range as well as the non-transferable property of the CHO-reactive groups.

Animals↗
PubMed1994

[The comparative study of pattern visual evoked potential in the normal infants and adults].

This paper presents the pattern visual evoked potential (PVEP) results and comparative study between the two groups of 65 normal infants at the age of 3, 6, 9, 12 months and 18 normal adults. The different pattern spatial frequency stimulations (74', 25', 6') were given to the infants and adults. In the infant group, the latencies of P1 components of PVEP delayed significantly and amplitudes of PVEP decreased significantly between 74' and 6' spatial frequency with the spatial frequency of stimulation increasing. In the same spatial frequency stimulation, when 3 months infant group was compared with the 6, 9, 12 months infant and adult groups, the latencies of N1 and P1 waves showed significant difference, but no difference was found in the amplitudes of P1 wave. Our results might indicate that the visual function expressed by low, medium, and high spatial frequency in infants from 6 to 12 months could reach the level in adults, and in 3 months infants, not maturely develop.

Adult↗
PubMed1994

Cloning and expression of the cDNA for canine tumor necrosis factor-alpha in E. coli.

We have utilized the reverse transcription polymerase chain reaction (RT-PCR) to clone the protein coding region of canine tumor necrosis factor (TNF-alpha) cDNA. The gene displays 90% sequence homology to the corresponding human TNF-alpha cDNA. The predicted initial translation product is 233 amino acids and shows 88% homology to the human counterpart, and 92% homology with the human putative mature TNF-alpha protein. The canine TNF-alpha clone was used to engineer bacteria to express large amounts of the mature form of recombinant protein. A monoclonal antibody against human TNF-alpha cross-reacted with canine rTNF-alpha using Western blot and ELISA analysis. The purified canine rTNF-alpha had a cytotoxic effect on WEHI 164 clone 13 cells as well as increasing the cell surface expression of major histocompatibility class II antigens on canine kidney cortical cell line (MDCK) in vitro. The availability of canine rTNF-alpha will allow further studies on its role in immunoregulatory mechanisms in the canine transplantation model, both by itself and in conjunction with the already available canine specific recombinant interferon-gamma.

Amino Acid Sequence↗
PubMed1994

[Urinary luteinizing hormone surge and outcome of in vitro fertilization and embryo transfer].

This study was based on 150 in vitro fertilization cycles carried out between June 1991 and July 1992. All the patients had blocked or dysfunctional fallopian tubes as the cause of infertility. Clomiphene citrate, human menopausal gonadotropin and hCG were used for controlled ovarian hyperstimulation. The results showed that an urinary luteinizing hormone (LH) surge (LH > 40 IU/L) occurred in 41 cycles (27.33%), a high LH level (LH = 40 IU/L) in 16 (10.67%), and normal (LH < 40 IU/L) LH level in 93 cycles, before hCG injection. The average number of oocytes recovered was not affected by the occurrence of a spontaneous LH surge, but the cleavage rate of the fertilized eggs was significantly lower than that in the cycles without LH surge and the rate of clinical pregnancy was also lower, although of no statistical significance. Our results demonstrated that the LH surge had an adverse effect on the egg quality and therefore would probably reduce the chances of pregnancy success.

Adult↗
PubMed1993

Origin of the asymmetrical contact between lac repressor and lac operator DNA.

The Escherichia coli lac operator DNA contains two sequence repeats related by a pseudo-dyad axis. Deviations from symmetry, in the central 21 bp sequence, occur at two pairs of symmetrically related sites (+15/+7, +13/+9) and at the central base-pair +11. Mutational analysis and DNA protection studies have suggested asymmetric interactions of lac repressor along this sequence. Previous biophysical studies on the lac repressor-operator system have typically employed symmetrized operator sequences to simplify analysis. As a result, it has remained difficult to assess the importance of the naturally occurring sequence deviations from symmetry. Here, 19F-NMR is used to determine if the wild-type E. coli lac operator DNA sequence itself specifies a pair of distinct half-site interactions with lac repressor DNA binding domains. To observe protein interactions simultaneously at operator half-sites using 19F-NMR, three pairs of naturally occurring, symmetry related thymine residues (at +6/+16, +8/+14 and +1/+21) were substituted pair-wise by 5-fluorodeoxyuridines (5-FdU). Two polypeptides corresponding to the N-terminal DNA binding domain of lac repressor "headpiece", residues 1 to 56 and 1 to 64, were employed to remove the steric constraints of subunit interaction in the wild-type tetramer. Spectral changes associated with headpiece binding to left side DNA sequences differ from those caused by binding to equivalent sequences on the right half-site. These results are similar to non-symmetric intact tetramer repressor interactions specified by the DNA sequence. Three mutant lac operator sequences with increased symmetry, bearing FdU substitutions were used to identify the relative importance of the three naturally asymmetric positions. Symmetrizing one pair of these sites alone or in addition to removing the central base-pair failed to produce identical NMR signal changes characteristic of symmetric headpiece-DNA complexes. However, symmetrizing both asymmetric pairs gave chemical shift changes expected from symmetric protein-DNA complexes. We propose that key interactions with the left side +9 (G.C) are altered at the symmetrically related right side +13 (A.T). The data show that the DNA sequence at +13 influences interactions three base-pairs away.

Base Sequence↗
PubMed1993

Bacteriophage T7 RNA polymerase. 19F-nuclear magnetic resonance observations at 5-fluorouracil-substituted promoter DNA and RNA transcript.

We have substituted 5-fluorodeoxyuridine (5-FdU) in place of thymidine in defined positions along synthetic bacteriophage T7 promoter DNA sequences. None of the fluoro-substitutions in the promoter DNA sequence reduced transcription yields with T7 RNA polymerase significantly. Substitutions on the coding template strand reduced transcription yields when placed at +3, but not at +4. 19F-n.m.r. spectra from transcription reactions and gel analysis of transcription products show that T7 RNA polymerase correctly and efficiently utilizes 5-FUTP as a RNA substrate analog. The fluorine atom provides a sensitive probe for monitoring the local environment, base sequence and solvent exposure at the DNA major groove through its 19F-n.m.r. resonance. Buffer dependencies of the fluorine chemical shift and digestion patterns with DNase I suggest that the T7 promoter base-pairs near the transcription start site are distorted with a more open minor groove and less solvent accessible major groove. Previous chemical footprinting data of promoter-polymerase complexes yield a picture that T7 RNA polymerase recognizes major groove features in the region from positions -7 to -11 and minor groove features on the same side of DNA flanking both sides of this region. Consistent with this, 19F-n.m.r. observations identify two additional positions, -8 and -17, involved in promoter recognition on this side of the DNA helix. On the other hand, our observations also implicate the opposite side of the DNA helix, primarily at positions -14 and -15, as major groove recognition sites for T7 RNA polymerase. In addition, n.m.r. spectra from 5-FdU-substituted base-pairs -2 and -3, suggest either additional interactions on the same side of the DNA helix as -14 and -15, or distortions in the DNA structure.

Bacteriophage T7↗
PubMed1993

A primary intestinal helminthic infection rapidly induces a gut-associated elevation of Th2-associated cytokines and IL-3.

The immune response that is characteristic of parasitic helminth infections includes components associated with immediate-type hypersensitivity: elevated serum IgE, eosinophilia, and intestinal mast cell hyperplasia. In infection with the parasitic nematode, Heligmosomoides polygyrus, IL-4 mediates protective immunity, suggesting the presence of a host-protective Th2 response. In this investigation, we examined early stages of immune responsiveness to H. polygyrus infection to determine whether and at what stage a specific Th2-like pattern first appears. Using a quantitative reverse transcriptase-polymerase chain reaction assay, we analyzed changes in IL-2, IFN-gamma, IL-3, IL-4, IL-5, IL-6, IL-9, and IL-10 gene expression in the spleen, mesenteric lymph node, and Peyer's patch at various time points after infection. Our results demonstrate a highly specific and reproducible pattern of cytokine gene expression that remains localized to the enteric region. By 6 h after infection, IL-5 and IL-9 mRNA were elevated in the Peyer's patch and IL-3 was elevated by 12 to 24 h after infection. IL-4 RNA became elevated by 4 to 6 days after infection, but little change was observed in IFN-gamma, IL-2, or IL-10 mRNA levels. The early increases in IL-3, IL-5, and IL-9 gene expression after infection were probably T cell-independent, inasmuch as they were observed in Peyer's patches of congenitally athymic mice and anti-CD4, anti-CD8 mAb-treated conventional mice. However, treatment with these mAb considerably decreased cytokine gene expression 6 days after infection, and 8 days after infection, increased IL-4 gene expression in mesenteric lymph node cells was restricted to the CD4+ population. Thus, H. polygyrus infection induces cytokine gene expression that is restricted to some Th2-associated cytokines, is initiated by a T-independent response, and culminates in a T-dependent response.

Animals↗
PubMed1993

Taurine in toad brain and blood under different conditions of osmolality: an immunohistochemical study.

The concentrations of taurine in blood and brain regions of the toad Bufo boreas have been measured. Most of these values are considerably lower than those found in mammals. Using an antibody prepared against conjugated taurine, the distribution of taurine in three brain regions of the toad has been visualized. The possible osmoregulatory functions of taurine have been investigated by making toads hyper- or hypo-osmotic in vivo. Induction of hypoosmolality is accompanied by a massive taurine tide in blood plasma, but has no immediate effects upon the taurine concentrations in the brain areas studied. However, histochemical visualization indicates a marked redistribution of taurine between cellular components and extracellular space of brain tissues. This may indicate that taurine has an osmoregulatory function in brain tissue under hypo-osmotic conditions. Hyperosmolality results in no elevation of the taurine concentration in blood plasma of toads, but rather in a very gradual decline of total plasma taurine content over a prolonged time period. Histochemical studies reveal little change in frontal cortex after 1 hour but deeper staining of many neurons in optic lobe accompanied by greater staining in the extracellular fluid. By 3 hours there is a depletion of taurine from all compartments of cerebral cortex tissues. No evidence of any prolonged direct osmoregulatory role for taurine is indicated under hyperosmotic conditions. A possible indirect osmoregulatory function of taurine is discussed.

Animals↗
PubMed1993

Visualization of taurine, GABA and glutamate in developing feline cerebellum by immunohistochemistry.

The localization of taurine, GABA and glutamate in developing feline cerebellum was performed using antibodies raised against the amino acids conjugated to bovine serum albumin with glutaraldehyde. Distinct patterns of immunostaining were observed for each of the amino acids. Taurine-like immunoreactivity reached a peak at 4 weeks after birth, as did GABA-like immunoreactivity, whereas glutamate-like immunoreactivity was greatest in the mature cerebellum. Purkinje cells are all taurine-positive in cerebellum from neonatal animals, whereas in the mature cerebellum they appear to contain only GABA and glutamate, with virtually no taurine, in contrast to observations reported with rodent cerebellum. Ultrastructural studies and immunogold labelling visualized by electron microscopy show that the band of taurine-like immunoreactivity observed in newborn feline cerebellum is localized within dendrites, axons and glial processes. Granule cells migrating through this region also show prominent taurine-like immunoreactivity.

Animals↗
PubMed1993

Differential effects of IFN-gamma on kidney cell expression of MHC class II molecules, kidney cell associated molecules and their stimulatory capacity in mixed lymphocyte kidney cell culture.

Mixed cell co-cultures of lymphocytes responding to kidney cells (MLKC), islets of Langerhans (MLIC) and mixed lymphocyte culture (MLC), were used to clarify mechanisms in allogeneic and autoimmune (tissue-associated) antigen presentation. Fresh kidney cortical tubular cells (KC), Madin-Darby canine kidney (MDCK), and dog kidney (6247) (DK) cell lines were used in MLKC reactions, and islets of Langerhans were used in the MLIC as putative antigen presenting cells (APC). The stimulating cells were treated with purified or recombinant dog interferon-gamma (IFN-gamma), and the detection of class II MHC molecule expression was assessed by a moneclonal antibody (mAb) (B1F6). Transcription of MHC class II mRNA and IFN-gamma mRNA was measured by semiquantitative polymerase chain reaction, and detection of a kidney cell (tissue-associated) antigen molecule was assessed by the mAb I1F6 that recognizes 72 and 150 kDa tubular cell protein(s) (KT1). The MDCK cell line constitutively expressed low levels of MHC class II molecules and KT1. The steady-state level of the MHC class II mRNA transcription was virtually unaltered by treatment with IFN-gamma (400 units) for 48 hours; however, the MHC cell surface protein expression was enhanced. The KC and DK cell lines constitutively expressed KT1, but not MHC class II molecules; these cells required a minimum of 4000 units, and a 62-hour incubation with IFN-gamma was needed to upregulate both surface MHC class II molecules and the transcription of corresponding specific mRNA. In the MLKC reaction both the MDCK and DK cell lines, as well as fresh KC cells, could serve as lymphocyte activators. This could be amplified by exogenous IFN-gamma. The removal of APC from the responding T cell population did not reduce the IFN-gamma effect. This indicates that IFN-gamma treatment allows for the expression of all of the co-stimulating factors and/or adhesion molecules necessary for these cells to serve as (surrogate) APC (direct as opposed to indirect antigen presentation). The requirements for purified IFN-gamma to increase this amplification was greater in the MLKC reactions with kidney cells than in the MLC reactions. The mAbs anti-IFN-gamma and I1F6 differed in their ability to inhibit lymphocyte proliferation depending on the different cell types involved. The I1F6 inhibited the MDCK and DK cell-driven MLKC (in the absence of exogenous IFN-gamma).(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1993

Production and characterization of recombinant canine interferon-gamma from Escherichia coli.

We have used the recently cloned cDNA for canine interferon-gamma (IFN-gamma) to engineer bacteria to produce large amounts of the recombinant cytokine. The resulting protein can be recognized by monoclonal and polyclonal antibodies largely species specific for canine IFN-gamma. The purified recombinant IFN-gamma (rIFN-gamma) also had biological activity in vitro in three assay systems: (i) vesicular stomatitis virus plaque inhibition, (ii) class II major histocompatibility complex antigen upregulation on canine kidney parenchymal cells, and (iii) amplification of in vitro tissue-associated lymphoproliferation, all known to be effected by native IFN-gamma (nIFN-gamma). The availability of large amounts of active canine rIFN-gamma will be an important tool in studies of the role of this cytokine in the widely used experimental canine organ transplant model and also will be of diagnostic and therapeutic veterinary interest.

Amino Acid Sequence↗
PubMed1993

Brucella abortus induces a novel cytokine gene expression pattern characterized by elevated IL-10 and IFN-gamma in CD4+ T cells.

Immunization of BALB/c mice with killed Brucella abortus (BA) has previously been shown to increase serum IgG2a levels and long-term T cell clones from these mice secrete Th1-associated cytokines: IFN-gamma and IL-2 but not IL-4 or IL-5. We analyzed cytokine gene expression following primary immunization with BA to determine when CD4+ T cells first express cytokine genes and whether specific hypothesized cytokine patterns (e.g. Th precursor, Th0) could be identified prior to a Th1-like pattern. Our results demonstrated a highly consistent and novel pattern of Th1/Th2 cytokine gene expression characterized by elevated IL-10 and IFN-gamma in CD4+ T cells which rapidly manifests itself and is sustained for at least 10 days after immunization. No elevation in IL-2 cytokine gene expression was observed and treatment of BA-immunized mice with blocking anti-IL-2 antibodies had no effect on the cytokine gene expression pattern, although treatment with anti-IFN antibodies resulted in increased IL-4, IL-5, and IL-9 cytokine gene expression, in the absence of any change in IFN-gamma or IL-10 as early as 4 days after immunization. These results suggest that a whole pathogen may trigger sufficient costimulatory signals to rapidly induce effector T cells in the absence of elevated IL-2 and that IL-10 is specifically elevated in certain Th1-like responses.

Animals↗
PubMed1993

Immunohistochemical localization of taurine in rat renal tissue: studies in experimental disease states.

Taurine, a sulfur amino acid, is present in abundant amounts in cells throughout the body. The kidney regulates taurine balance by modulating proximal tubule reabsorption in response to fluctuations in dietary intake of this nutrient. There is no information about the localization of taurine within the kidney in normal and diseased renal parenchyma. Therefore, using an antibody to a taurine-glutaraldehyde-BSA conjugate, we examined the distribution of taurine in renal tissue. Normal rats, those with streptozocin diabetes, puromycin aminonucleoside nephropathy, bilateral ureteral ligation, and 5/6 nephrectomy were studied. In normal animals, taurine was found primarily in medullary tubules, with minimal staining of proximal tubules and glomeruli. There was increased taurine staining of all structures, especially medullary tubules, in rats with streptozocin diabetes and puromycin aminonucleoside nephropathy. These changes were more pronounced in diabetic rats and were unrelated to renal medullary osmolality. The distribution of taurine within the kidney was unchanged in the models of acute and chronic renal failure. Alterations in the immunohistochemical localization of taurine correlated with the beneficial effect of this amino acid to preserve renal function in the rats with chronic diabetes and puromycin aminonucleoside nephropathy. These results suggest that taurine is preferentially localized in the medullary regions of the kidney, where it exerts a protective effect against renal injury in select disease states.

Animals↗
PubMed1993

[Using myocardial perfusion tomography to estimate infarct size and to determine prognostic value in patients with acute myocardial infarction].

Single-photon emission computerized tomography with Tc-99m-MIBI was used to measure the infarct size in 41 patients with acute myocardial infarction (AMI), among them 11 patients (accounting for 27%) were dead or recurred with perfusion defect > 31% in 6 cases including 3 deaths and 2 recurrences. The perfusion defect was between 11%-30% in 12 patients, including one death and two recurrences. The perfusion defect was 10% in 23 patients, including 3 with recurrence. During the period from 6 months to 2 years following-up, the total death and patients with recurrence were 11/41 (27%) cases. The results showed that the prognosis of patients with AMI was closely related to the size of myocardial perfusion defect, and those with perfusion defect > 31% had a poorer prognosis.

Adult↗
PubMed1993

Taurine distribution in different cat muscles as visualized by immunohistochemistry: changes with stimulus state.

The distribution of endogenous taurine in the cardiac pectinate muscle, the intestinal wall smooth muscle and in nine striated muscles of the cat was studied by immunohistochemistry. It was found that taurine-like immunoreactivity (TLI) is distributed homogeneously in the cardiac and smooth muscles selected where it is present in every single fibre. In contrast, TLI appeared unevenly distributed in eight out of nine skeletal muscles, the soleus being atypical. Under resting conditions, TLI is present in only a portion of the fibre population, the proportion being specific for each muscle. Under nerve stimulation the TLI negative/positive ratio increased, suggesting release activity, while an opposite trend occurred after denervation. Glutamate-like immunoreactivity (GLI), although present in every fibre, showed a stronger reaction in those fibres positive to taurine, suggesting parallel movements of both amino acids. Results are discussed in terms of two functional possibilities.

Animals↗
PubMed1992

Perforin expression in human peripheral blood mononuclear cells. Definition of an IL-2-independent pathway of perforin induction in CD8+ T cells.

Perforin gene expression upon in vitro stimulation was studied at the mRNA level in normal human PBMC and in subpopulations. Freshly isolated PBMC express low levels of perforin mRNA. Increased perforin expression is rapidly induced by the calcium ionophore A23187 and by rIL-2. Phorbolesters (PMA), by comparison, are poor inducers of perforin RNA. Perforin induction by Ca-ionophore, unlike granzyme 2 and IL-2 induction, did not synergize with phorbolesters in PBMC or in purified T cells. Instead, perforin mRNA induction by A23187 in purified T cells requires the presence of adherent cells. Ca-ionophore plus adherent cell-induced perforin occurred in CD8+ T cells and was abolished by depletion of CD8+ T cells but not by depletion of CD4+ T cells. Adherent cells alone did not express perforin under any condition. Perforin mRNA induction by both A23187 and by rIL-2 is independent of de novo protein synthesis. The half-life of perforin mRNA induced by either stimulus is approximately 100 min. Cyclosporin A completely abrogates perforin induction by A23187 but only slightly inhibits the effect of rIL-2 on perforin mRNA expression. These data show that A23187 activates perforin gene expression in CD8+ cells by an IL-2-independent pathway and that the molecular mechanism of perforin expression may be different from the one induced by IL-2. Granzyme 2 (human leukocyte protease-HLP, homologous to murine granzyme B) mRNA expression was studied in comparison to perforin. Granzyme 2 in contrast to perforin responds to the synergistic action of phorbolester and Ca-ionophore in PBMC. In addition, the kinetics of the induction of granzyme and perforin mRNA, by various signals are different. Our data suggest that situations in vivo may exist that allow perforin expression in CD8+ cells in the absence of cytokines by a combination of Ca signals and accessory receptor ligation. The same signals may not be sufficient for granzyme 2 expression in any T cell subpopulation.

Base Sequence↗
PubMed1992

Transcriptional enhancer related DNA sequences: anomalous 1H NMR NOE crosspeaks.

A dynamic heterogeneity which correlates with the function of the operator DNA in the lactose operon of E. coli. was previously observed (1) as a local minimum in the thymine imino proton T1 centered at a GTG/C-CAC sequence. Since this triplet occurs frequently in DNA regulatory regions, it was proposed that these sequences may be part of a structural element for specific protein interaction. We examine here three additional biologically significant 17 base pair duplexes containing GTG/CAC triplets: (1) a sequence from the mouse heavy chain immunoglobulin enhancer, (2) a sequence from the critical core of the Simian Virus 40 (SV40) enhancer, and (3) a sequence from pBR322 plasmid used as control for experiments with the SV40 DNA sequences. The 1H NMR resonance assignment for nearly all the nonexchangeable protons for both eukaryotic enhancer duplexes with the exception of the H5'/H5" protons was accomplished to use for structural analysis of these duplexes. The data presented show several NOE's associated with the GTG/CAC triplets which suggest structural variation from uniform B-DNA. In addition, anomalous broad crosspeaks for the fixed thymine methyl to its own H6 proton in combination with the imino proton kinetics associated with these triplets reinforces the original observation of a sequence dependent dynamic variation.

Animals↗