Search PubMed⌕ Search

Biomedical subjects

S C Li

Publications and source records attributed to S C Li.

At least 109 records · Page 6Linked to original sources
PubMed1994

Expression and specificity of human GM2 activator protein.

The cDNA encoding GM2 activator was expressed in the Escherichia coli/pT7-7 system. The yield of the GM2 activator with greater than 99% purity was about 3 mg per liter culture. The recombinant GM2 activator was found to be as active as that isolated from human kidney. The availability of the recombinant GM2 activator enabled us to critically examine the specificity of this activator protein. Our results show that the specificity of GM2 activator is not as strict as that reported previously. Although GM2 activator stimulates most efficiently the degradation of GM2 carried out by beta-N-acetylhexosaminidase A (Hex A), this activator also stimulates the following reactions: (a) conversion of GM2 to GA2 by clostridial sialidase; (b) hydrolysis of GalNAc from dipalmitoylphosphatidylethanolamine-II3NeuAcGgOse3 by Hex A; and (c) liberation of Gal from GM1 by beta-galactosidase at a high activator concentration. Thus, this activator does not differentiate between GM2 and dipalmitoylphosphatidylethanolamine-II3NeuAcGgOse3 or between Hex A and clostridial sialidase. The micellar forms of GD2 and GalNAc-GD1a were found to be more readily hydrolyzed by Hex A than GM2 in the absence of GM2 activator. Our results also show that saposin B can enhance the stimulatory activity of GM2 activator, but it cannot promote the stimulatory activity of sodium taurodeoxycholate. Taken together, our results suggest that the mechanism of action of GM2 activator is different from saposin B, and the action of GM2 activator is more than to solubilize lipid substrates. The effectiveness of GM2 activator in stimulating the hydrolysis of GM2 may be due to its ability to recognize the specific trisaccharide structure of the GM2 epitope, GalNAc beta 1-->4(NeuAc alpha 2-->3)Gal-, and to modify the GalNAc-NeuAc interaction in this structure.

Base Sequence↗
PubMed1994

A novel sialidase capable of cleaving 3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN).

We have examined the tissues of several species of fish and found that the liver of the loach (Misgurnus fossilis) contains a novel sialidase capable of efficiently hydrolyzing 3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN) from the 4-methylumbelliferyl alpha-ketoside of KDN, KDN alpha 2-->3Gal beta 1-->4GlcCer and KDN alpha 2-->6 N-acetylgalactosaminitol as well as Neu5Ac from the 4-methylumbelliferyl alpha-ketoside of Neu5Ac and GM3. The pH optimum for this enzyme was determined to be 4.6, and the Km using the 4-methylumbelliferyl alpha-ketoside of KDN and 4-methylumbelliferyl alpha-ketoside of Neu5Ac as substrates were 0.07 and 0.12 mM, respectively. The enzyme was stable in the pH range of 4 to 5 but very unstable above pH 6. This is the first report of a sialidase capable of efficiently cleaving glycosidically linked KDN.

Animals↗
PubMed1994

A measure of helical propensity for amino acids in membrane environments.

The frequent occurrence of beta-sheet promoting residues such as Ile, Val, and Thr in the alpha-helical transmembrane segments of most integral membrane proteins suggests that the helical propensities of these residues are altered in the hydrophobic environment of the lipid bilayer. Systematic studies of model peptides by circular dichroism models spectroscopy in various micellar/vesicular media allow the establishment of a ranking order of helical propensity for uncharged amino acids in the membrane environment. In contrast to their conformational preferences in water, the helical proclivity of amino acids in membranes is shown to be governed by their side chain hydrophobicity, and by the hydropathy of the local peptide segments in which the residues reside [corrected].

Amino Acid Sequence↗
PubMed1994

A measure of helical propensity for amino acids in membrane environments.

The frequent occurrence of beta-sheet promoting residues such as Ile, Val, and Thr in the alpha-helical transmembrane segments of most integral membrane proteins suggests that the helical propensities of these residues are altered in the hydrophobic environment of the lipid bilayer. Systematic studies of model peptides by circular dichroism spectroscopy in various micellar/vesicular media allow the establishment of a ranking order of helical propensity for uncharged amino acids in the membrane environment. In contrast to their conformational preferences in water, the helical proclivity of amino acids in membranes is shown to be governed by their side chain hydrophobicity, and by the hydropathy of the local peptide segments in which the residues reside.

Amino Acid Sequence↗
PubMed1994

Expression of I mu-C gamma hybrid germline transcripts subsequent to immunoglobulin heavy chain class switching.

Germline CH transcripts initiate from a non-coding I exon and terminate downstream of the associated CH exons. Ig heavy chain class switch recombination from the VDJ-C mu gene to particular downstream CH genes appears to be regulated by a process that involves mitogen and/or cytokine induction of germline CH transcripts from the downstream genes. We have examined the expression of germline C mu transcripts (I mu-C mu transcripts) in splenic B cells and pre-B cell lines after cytokine and mitogen stimulation. In contrast to the expression of the germline transcripts from downstream CH genes, expression of germline C mu transcripts was constitutive and unaffected by mitogen and cytokine treatment. After a primary switch recombination event, the germline I mu promoter, which is now associated with a downstream CH gene, continues to be active--leading to the generation of a novel germline transcript consisting of the I mu exon spliced to the CH exons of the switched CH gene. We discuss the potential role of the expression of hybrid I mu-containing transcripts in the class switch process. We also describe a novel and sensitive assay, based on the detection of the hybrid I mu-containing transcripts, that allows detection of class switch recombination events even in heterogeneous populations of cells.

Animals↗
PubMed1994

Absence of toxicity in patients with malignant otitis externa following long-term treatment with high dosage tobramycin.

An in-vitro model of aminoglycoside dosing was used to demonstrate that the bactericidal activity of tobramycin against Pseudomonas aeruginosa is directly related to the peak concentration of the drug. In addition, six patients who were being treated for malignant otitis externa with long-term, high dosage tobramycin were monitored for oto- and nephro-toxicity; the only adverse effects were transient increases in the serum creatinine concentration in two patients.

Aged↗
PubMed1994

Tuberculous arthritis in children. Diagnosis by needle biopsy of the synovium.

In two children ages 8 months and 4 years, tuberculous synovitis of the knee was suggested by pathologic examination of synovial tissue obtained through needle biopsy. Culture of the biopsy material was positive for Mycobacterium tuberculosis in one case. In this child, the tuberculin test was negative, in the absence of anergy, at the time the child was first evaluated, and the primary lung lesion was not identified by the radiologist. Tuberculous synovitis has not previously been recognized as part of primary tuberculosis during the early weeks when the tuberculin skin test may be negative. Magnetic resonance imaging (MRI) is a sensitive modality for demonstrating joint fluid, synovial hypertrophy, and associated osteomyelitis, if present. With the increasing frequency of cases of tuberculosis in the population, greater awareness of the risk of tuberculous arthritis in childhood is important in order to recognize this unusual presentation. If negative early on, the tuberculin skin test should be repeated after 6 weeks of arthritis. A needle biopsy of the synovium is helpful and appropriate in all children with monoarticular arthritis who have a positive tuberculin skin test.

Biopsy, Needle↗
PubMed1994

Hysteroscopic tubal catheterization and hydrotubation for treatment of infertile women with tubal obstruction.

We reported the efficacy of interval hysteroscopic tubal catherization and hydrotubation for three months in 54 infertile women with tubal obstruction (102 fallopian tubes). By comparison of HSGs before and after treatment, 86 of 102 tubes (84%) showed the improvement of tubal patency (P < 0.01). 56 tubes were patent (54.9%). This method is effective for partially patent tubes (87.5%), intramural tubal obstruction (62.5%), and the obstruction at the distal portion of the fallopian tube (13.3). One -12 month follow-up after treatment, 12 of 35 women whose tubes became completely patent achieved intrauterine pregnancies.

Adult↗
PubMed1993

Peptide environment specifies conformation. Helicity of hydrophobic segments compared in aqueous, organic, and membrane environments.

Transmembrane segments in integral membrane proteins exist characteristically as helices in lipid bilayers, yet are often rich in residues considered helix-destabilizing (Val, Ile, Gly) in soluble proteins. We propose that helicity of a transmembrane segment is likely to be affected by factors other than the "intrinsic" helical propensities of its component amino acids. This hypothesis is tested by comparing the conformation(s) in aqueous, organic, membrane-mimetic (micellar), and membrane (bilayer) environments of designed model peptides with systematically altered helical propensity and/or segmental hydrophobicity. Peptides of prototypic sequence NH2-(Ser-Lys)2-Ala5-Leu6-Ala7-Ala8-Leu9-Ala10-++ +Trp11-Ala12-Leu13-Ala14- (Lys-Ser)3-OH were synthesized, which incorporate a hydrophobic core "guest" segment (residues 5-14) into a water-soluble hydrophilic host matrix. Related peptides featured substitution of Leu6,9,13-->Gly, Leu6,9,13-->Ala, and Ala7,10,14-->Gly. Circular dichroism spectra revealed that algorithms for soluble proteins correctly predicted peptide helical proclivities in aqueous solutions, but peptide helicity in organic (trifluoroethanol) solvents, membrane-mimetic SDS micelles, and negatively charged lipid bilayer vesicles, was found to be governed almost exclusively by the segmental hydrophobicity of the peptide mid-hydrophobic core segment. In related Trp fluorescence studies, peptide-membrane association was similarly correlated with extent of hydrophobic interaction.

Amino Acid Sequence↗
PubMed1993

Replacement of germ-line epsilon promoter by gene targeting alters control of immunoglobulin heavy chain class switching.

Recent work has shown that the ability of cytokines to direct immunoglobulin heavy chain class-switch recombination to particular heavy chain constant (C) region (CH) genes correlates with the induction of specific germ-line CH transcripts. To test the role of germ-line transcripts in class switching, we have used homologous recombination to mutate the immunoglobulin heavy chain locus of the 18.81A20 murine pre-B-cell line. In the parent cell line, the combination of interleukin-4 (IL-4) and lipopolysaccharide (LPS) induces germ-line epsilon locus transcription prior to class switching to epsilon. The heavy chain locus of the mutated cell line contains the immunoglobulin heavy chain enhancer and variable region gene promoter in place of the LPS/IL-4-responsive germ-line epsilon promoter. The mutant cell line constitutively transcribes the epsilon locus in the absence of IL-4. Strikingly, the mutant cell line also switches to epsilon in the absence of IL-4. This result demonstrates that, at least in the 18.81A20 cell line, germ-line epsilon transcription plays a direct role in class switching to the epsilon locus. In addition, the ability to change the pattern of class switching by altering transcriptional activity indicates that transcription of germ-line CH is mechanistically important in regulation of class switching.

Abelson murine leukemia virus↗
PubMed1993

Bactericidal effect of gentamicin peak concentration provides a rationale for administration of bolus doses.

A study was made of the influence of peak gentamicin concentration (Cmax) independent of the area under the antibiotic concentration-time curve (AUC) on the bactericidal effect of gentamicin against Escherichia coli NCTC 10418. Bacteria in the lag phase were exposed to in-vitro gentamicin concentration-time profiles which modelled complete profiles determined in vivo in patients after an intravenous bolus (1 min) or infusion (30 min) regimen. An additional in-vitro profile reflected antibiotic removal 30 min after bolus dosing. Bacterial viable cell counts and gentamicin concentrations were measured before and during antibiotic exposure. Both Cmax and AUC were shown to be factors determining antibacterial activity; however Cmax independently represented some 35% of total exposure effect. These findings indicate grounds for preferring bolus intravenous gentamicin dosing with appropriate attention to potential toxicity.

Dose-Response Relationship, Drug↗
PubMed1993

Kill kinetics and regrowth patterns of Escherichia coli exposed to gentamicin concentration-time profiles simulating in vivo bolus and infusion dosing.

The relative influence of peak concentration (Cmax) versus the area under the antibiotic concentration-time curve (AUC) on the bactericidal effect of gentamicin against Escherichia coli NCTC 10418 was studied. Bacteria in the lag phase were exposed to an in vitro gentamicin concentration series which mirrored the concentrations determined in patients after 80-mg intravenous bolus (1 min) and 80-mg intravenous infusion (30 min) doses. Bacterial viable cell counts and gentamicin concentrations were measured before and during antibiotic exposure. Both the Cmax and AUC were shown to be factors determining antibacterial activity; however, the Cmax was an independent determinant of effect. These findings indicate that bolus intravenous dosing with gentamicin could maximize bactericidal activity. Increased efficacy could result at any given daily antibiotic dose if delivered via bolus with long intervals (12 to 24 h) between doses if appropriate precautions to avoid toxicity are taken.

Colony Count, Microbial↗
PubMed1993

[Effects of purified VLDL injection on experimental hyperlipidaemia].

Forty-two neonate NZW rabbits were randomly allocated to two experimental groups (II and III) and a control group (I). Groups II and III were intraperitoneally injected with purified VLDL 0.5mg and 1.0mg, respectively within 12 hours after their birth, and then were repeated with same doses twice on the 7th and 15th day, respectively. From the 18th day of their life, all the rabbits were fed with high cholesterol forage to establish atherosclerosis models. Serum levels of TG, TCh, and HDL-C were determined in the 17th, 30th, and 40th week of their life. Serum levels of TG in Groups II and III were much lower than those in Group I in the 17th week. Serum levels of TG and ratio of LDL-C to HDL-C were significantly lower and of HDL-C much higher in Groups II and III than those in Group I in the 30th week. The results determined in the 40th week were the same as above, and serum levels of TCh in Groups II and III were significantly lower than those in Group I. There Were no significant differences in serum levels of LDL-C and TCh among the three groups. The results suggested peritoneal injection of purified VLDL into rabbits in their earliest stage of life had effects on the formation of hyperlipidaemia, and could Prevent them from atherosclerosis. Its mechanism was briefly discussed in the paper.

Animals↗
PubMed1993

[Selective salpingography and tubal cannulation through hysteroscopy].

In this paper the procedure of selective salpingography and tubal cannulation through hysteroscopy for diagnosis and treatment of fallopian tube obstruction at the interstitial portion was introduced. We combined hysteroscopic tubal cannulation with selective salpingography under fluoroscopic quidance in 28 infertile women with 49 obstructed interstitial portion of fallopian tubes diagnosed previously. After the procedures 27 tubes (55.1%) became patent in 16 cases. In 16 women with patent tubes followed up over 6 months after the procedures 5 intrauterine pregnancies occurred. This procedure is a safe and simple diagnostic method to identify fallopian tube obstruction at the interstitial portion and may alos serve as a therapeutic approach in some of these patients.

Adult↗
PubMed1993

The endoscopic diagnosis of iliac colonic fistula and review of literature.

Although the efficacy of upper gastrointestinal endoscopy in the diagnosis of a graft-enteric fistula is defined, the clinical utility of colonoscopy in this problem is not well described. We report a case of a 65-yr-old white male with gastrointestinal bleeding and a suspected graft-enteric fistula which was diagnosed by an immediate preoperative colonoscopic examination. The colonoscopy performed under general anesthesia in the operating room not only documented the presence and location of the graft colonic fistula but also illustrated the necessity for a more radical surgical resection before any planned revascularization. The current literature on the various methods of diagnosis and management of graft-enteric fistula was reviewed.

Aged↗
PubMed1992

Glycine and beta-branched residues support and modulate peptide helicity in membrane environments.

Transmembrane (TM) segments of integral membrane proteins are putatively alpha-helical in conformation once inserted into the membrane, yet consist of primary sequences rich in residues known in soluble proteins as helix-breakers (Gly) and beta-sheet promoters (Ile, Val, Thr). To examine the specific 2 degrees structure propensities of such residues in membrane environments, we have designed and synthesized a series of 20-residue peptides with 'guest' hydrophobic segments--expected to provide three turns of incipient alpha-helix content--embedded in 'host' hydrophilic (Lys-Ser) matrices. Circular dichroism (CD) spectra of the model peptides in water showed that significant helical content was observed only for peptides with high Ala content; others behaved as 'random coils'. However, in the membrane-mimetic environment of sodium dodecylsulfate (SDS) micelles, it was found that Gly can be accommodated as readily as Ala, and Ile or Val as readily as Leu, in hydrophobic alpha-helices. Further subtleties of structural preferences could be observed in electrically-neutral lyso-phosphatidylcholine (LPC) micelles, where helical propensity decreased in the order Ala-Leu-rich > Gly-Leu-rich > Gly-Ile(Val)-rich hydrophobic segments. The results conjure a role of environment-dependent helix-modulation for Gly, Ile, and Val residues--and suggest that these residues may provide, in part, the structural basis for conformational transitions within or adjacent to membrane domains, such as those accompanying membrane insertion and/or required for transport or signalling functions.

Amino Acid Sequence↗
PubMed1992

Prospective audit of an aminoglycoside consultative service in a general hospital.

OBJECTIVE: To investigate the impact of the introduction of a consultative service on the use, efficiency of dosing and clinical toxicology of the aminoglycoside antibiotics, gentamicin and tobramycin, in a general hospital. METHODS: Two audits were conducted six months and 18 months after the introduction of the consultative service. The audits reviewed the use of drug assay services, the adequacy of drug administration (as measured by serum antibiotic concentrations), indications for prescription, adverse outcomes (by noting markers of nephrotoxicity) and the antibiotic sensitivity of Gram-negative pathogens. The results were compared with the results of an audit conducted before the consultative service was instituted. RESULTS: There was a significant (P < 0.001 by chi 2 test) increase in the use of assays, with drug assays performed in 67% (first audit) and 77% (second audit) of aminoglycoside courses compared with 48.2% in the pre-intervention audit. Sample timing was greatly improved, with more than 70% of the samples collected at the appropriate times. Assay wastage in terms of uninterpretable assay results decreased significantly (P < 0.001) from 42.9% of total assays to 6.3% at the first audit and 3.8% at the second audit. The percentage of assay results in the desirable range increased significantly (P < 0.001) from 39.1% to 71.9% (first audit) and 75.4% (second audit). Pharmacokinetic recommendations were made in 39.1% and 64% of all aminoglycoside courses during the first and second audits respectively, with clinician acceptance of dosage recommendations at 83.1% and 82.8% respectively. For aminoglycoside courses prescribed for therapeutic reasons, 97.9% (first audit, n = 325) and 98.6% (second audit, n = 280) of indications for use were judged as clinically appropriate. The incidence of suspected aminoglycoside-induced nephrotoxicity was reduced from 8.9% of patients to 1.6% (first audit, P < 0.001) and 2.4% (second audit). Bacterial sensitivity audits showed that the great majority of clinical isolates of target organisms (n = 3523, Year 1 and n = 3385, Year 2) were sensitive to gentamicin (92.2% and 91.5% respectively) and tobramycin (98.1% and 98.8% respectively); these aminoglycosides exceeded all alternative agents in effectiveness, including first and third generation cephalosporins. CONCLUSIONS: The overall results indicate that introduction of the consultative service had a positive impact on the effective use of aminoglycosides, with a marked decrease in clinical toxicity. These influences were shown to persist for at least 18 months. The availability of reliable predictive techniques to reduce toxicity allows active promotion of aminoglycosides as the agents of choice on grounds of efficacy and economy.

Blood Specimen Collection↗