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J K Li

Publications and source records attributed to J K Li.

At least 73 records · Page 4Linked to original sources
PubMed1993

Glycosylation of the major outer capsid protein of bluetongue viruses.

The major outer capsid protein, VP5, of five US bluetongue viruses, was found to be glycosylated. Enzymatic removal of the carbohydrate moiety did not affect the electrophoretic mobility of VP5 in SDS-PAGE, indicating the presence of only short and possibly unbranched oligosaccharide chains. The surface accessibilities and immunogenic specificities of two conformational-dependent antigenic epitopes on VP5 of BTV were not affected by deglycosylation. Selective binding of those lectins which have well-defined sugar specificities suggests that the potential short N-linked carbohydrate chain present on BTV VP5 might be composed of sialic acid alpha(2-6)-N-acetylgalactosamine-beta(1-3)-galactose-beta(1-4)-N- acetylglucosamine-beta(1-2)-mannose-N-acetylglucosamine-Asn. In accordance with the standard nomenclature, BTV-VP5 should now be termed GP5 due to the presence of the carbohydrate moiety.

Antibodies, Monoclonal↗
PubMed1993

Repeated reflection of waves in the systemic arterial system.

Traditional analysis of pulse-wave propagation and reflection in the arterial system treats measured pressure and flow waves as the sum of a single forward wave (traveling away from the heart) and a single backward wave (traveling toward the heart). The purpose of this study was to develop a more general wave reflection theory that allows repeated reflection of these waves. The arterial system was modeled as a uniform viscoelastic tube terminating in a complex load with reflections occurring at the tube load interface and the heart tube interface. The resulting framework considers the forward wave to be the sum of an initial wave plus a series of antegrade waves. Similarly, the backward wave is the sum of a series of retrograde waves. This repeated reflection theory contains within it the traditional forward/backward wave reflection analysis as a special case. In addition, the individual antegrade and retrograde waves, at the tube entrance, are shown to be independent of the tube length. Aortic pressure and flow data, from dog experiments, were used to illustrate the phenomenon of repeated reflections. Alteration of the arterial system loading conditions, brought about through pharmacological intervention, affected the number and morphology of repeated waves. These results are compared with those found in traditional forward/backward reflection analysis.

Animals↗
PubMed1992

Evolutionary analyses of five US bluetongue viruses using the cognate S2 genes.

Full-length cDNA copies of S2 genes (segment 8), coding for non-structural protein 2 (NS2) of bluetongue virus serotypes 2, 11, 13 and 17 were selectively synthesized by a modified polymerase chain reaction (Clamp-R) and cloned into the PstI site of the pUC19 plasmid. Each of these S2 cognate genes was 1125 nucleotides in length with an initiation and a termination codon at nucleotides 20-22 and 1082-1084, respectively, resulting in a long open reading frame capable of coding for a protein of 354 amino acids. The deduced amino acid sequence of NS2 protein had a high concentration of lysine and contained a relatively low number of tryptophan and histidine residues. There was a highly conserved hydrophilic region at the carboxyl termini of predicted NS2 proteins in all five BTV serotypes, even though the amino acid sequence in this region in BTV-2 was more variable than in the other four serotypes. There was significant sequence homology of the cognate S2 genes at both the nucleotide and the amino acid levels. Phylogenetic analyses using the S2 gene sequences indicated that BTV-10, -11, -13 and -17 were more closely related and BTV-2 was the most distantly related serotype among the five US bluetongue viruses.

Amino Acid Sequence↗
PubMed1992

Production and characterization of recombinant lignin peroxidase isozyme H2 from Phanerochaete chrysosporium using recombinant baculovirus.

Recombinant Phanerochaete chrysosporium lignin peroxidase isozyme H2 (pI 4.4) was produced in insect cells infected with a genetically engineered baculovirus containing a copy of the cDNA clone lambda ML-6. The recombinant enzyme was purified to near homogeneity and is capable of oxidizing veratryl alcohol, iodide, and, to a lesser extent, guaiacol. The Km of the recombinant enzyme for veratryl alcohol and H2O2 is similar to that of the fungal enzyme. The guaiacol oxidation activity or any other activity is not dependent upon Mn2+. The purified recombinant peroxidase is glycosylated with N-linked carbohydrate(s). The recombinant lignin peroxidase eluted from an anion exchange resin similar to that of native isozyme H1 rather than H2. However, the pI of the recombinant enzymes is different from both H1 and H2 isozymes. Further characterization of native isozymes H1 and H2 from the fungal cultures revealed identical N-terminus residues. This indicates that isozymes H1 and H2 differ in post-translational modification.

Amino Acid Sequence↗
PubMed1992

Iontophoretically enhanced transdermal delivery of an ACE inhibitor in induced hypertensive rabbits: preliminary report.

Both conventional direct current (DC) and pulsed-mode DC constant-current iontophoresis were used to investigate enhanced transdermal delivery of the angiotensin-converting enzyme (ACE) inhibitor captopril to rabbits with acutely induced hypertension. Passive transdermal captopril administration and pulsed DC constant-current iontophoresis of the vehicle were studied as control experimentation. Mean arterial pressure (MAP) was not significantly (p > 0.05) altered following passive transdermal delivery of captopril (n = 4) or after iontophoretic delivery of the vehicle alone (n = 4). Pressure reduction was evident within 10 minutes of iontophoretic enhancement of transdermal captopril delivery. DC mode constant-current (n = 4) iontophoretic transdermal captopril administration caused MAP to fall by 21% from a mean hypertensive level of 66 +/- 5 mmHg to a mean post-treatment level of 52 +/- 6 mmHg (p < 0.05) within 60 minutes. Pulsed DC mode constant-current (n = 4) iontophoresis of captopril caused mean MAP to fall on average by 27% from 62 +/- 6 to 45 +/- 5 mmHg (p < 0.05), also within 60 minutes. This paper provides the first report on the enhanced efficiency during iontophoretic delivery of an ACE inhibitor. We have concluded that both modes of constant-current iontophoresis of captopril offer a safe and effective means of pressure reduction in rabbits with induced hypertension and that there is no significant difference in efficacy between the two forms of enhanced delivery. These results have potential applications for enhanced transdermal delivery of ACE inhibitors in humans.

Administration, Cutaneous↗
PubMed1992

Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses.

Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.

Amino Acid Sequence↗
PubMed1992

Sequence conservation among the cognate nonstructural NS3/3A protein genes of six bluetongue viruses.

Full-length cDNA copies of segment 10 genes of bluetongue virus serotypes 2, 11, 13 and 17 were synthesized by the Clamp-R method and inserted into the plasmid pUC19. The complete nucleotide sequences of these four cognate genes were sequenced and determined to be 822 nucleotides in length, smallest of the 10 genes in the bluetongue virion. These four cognate gene segments contained two in-phase and overlapping open reading frames capable of coding for two non-structural proteins of 229 and 216 amino acids with net charges of +4.5 and +5.5, respectively, at neutral pH. Comparative analyses of the predicted amino acid sequences of bluetongue virus serotypes 1, 2, 10, 11, 13 and 17 revealed (i) a high degree of sequence homology and conservation, (ii) a single conserved tryptophan located at residue 159, (iii) the presence of two conserved cysteines at residues 137 and 181 and two potential N-linked glycosylation sites at residues 63-65 and 150-152, (iv) a cluster of six prolines within a 15-amino acid region near the amino terminus, and (v) the longest 3' noncoding sequence of 113 bases among the 10 bluetongue viral genes. Phylogenetic analyses indicated that BTV-10 and -11 are very closely related and BTV-2 is the distantly related serotype of the five US bluetongue virus serotypes.

Amino Acid Sequence↗
PubMed1992

Complete genomic sequences of the GP5 protein gene of bluetongue virus serotype 11 and 17.

The complete nucleotide sequences of full-length copies of genomic segment 6 or M3 of US bluetongue virus serotype 11 and 17 consisted of 1638 nucleotides. The plus-strand contained an open reading frame for a protein of 526 amino acids which was equivalent to about 59,000 Da, similar to the molecular weight of GP5 as determined by SDS-PAGE analysis. This long open reading frame was flanked by a 5' non-coding region of 29 nucleotides and a 3' non-coding region of 28 bases. When the predicted amino acid sequences of GP5 of BTV-11 and -17 were aligned and compared with those of BTV-2, -10, -13, -1AU and -1SA, four major highly conserved domains interrupted by several variable regions were detected. The potential significance of these discrete domains is discussed. Evolutionary and phylogenetic characteristics of these US BTV serotypes were consistent with our finding concerning BTV-1AU and -1SA.

Amino Acid Sequence↗
PubMed1992

Comparative sequence analyses of the cognate structural protein VP6 genes of five US bluetongue viruses.

The S3 segment (the small segment 3), encoding the structural protein, VP6, from the five United States (US) prototype bluetongue virus (BTV) serotypes were amplified by the Clamp-R method and cloned as full-length entities. The complete nucleotide sequence of each cognate gene segment was determined. Each cognate S3 segment of BTV-10, 11, 13 and 17 was 1049 nucleotides long and contained an open reading frame (ORF) capable of encoding a 325-amino acid protein. However, the S3 segment of BTV-2, which also contained 1049 nucleotides, had a longer 5'-non-coding region of 99-nucleotide and contained an ORF capable only of encoding a 301-amino acid protein. Comparative analyses of the predicted amino acid sequences of S3 segments of BTV-2, 10, 11, 13 and 17 revealed that VP6 was unusually high in glycine and contained few aromatic amino acids, but a high concentration of charged amino acids, which is a characteristic of a hydrophilic protein. Phylogenetic analyses indicated that BTV-11, 13 and 17 were more closely related than the other two US BTV serotypes. BTV-2 was the most distantly related.

Amino Acid Sequence↗
PubMed1992

Temporal relationship between left ventricular and arterial system elastances.

Arterial compliance is an important component of ventricular afterload. Although its pressure dependence has been recognized, its temporal relationship to ventricular elastance (Elv(t)) has not been established. We investigated this in five open chest anesthetized dogs where simultaneous aortic pressure and flow and left ventricular pressure were measured. Elv(t) was derived using an elastance-resistance model of the left ventricle assuming an ejection fraction of 0.50 and a dead volume (Vd) of 3.0 mL. The nonlinear pressure-dependent compliance (C(P)) of the arterial system was incorporated in a three-element Windkessel model and determined by accurate prediction of aortic pressure from aortic flow. The resulting arterial elastance (Eas(t)) was computed as Eas(t) = 1/C(P). Results show that Eas(t) reaches a minimum value at or near the start of ventricular ejection and attains its peak value at or near the same time maximum LV elastance (Emax) is reached, at end-systole. Finally, numerical simulation of the model demonstrates its ability to adequately reproduce measured pressure and flow. Thus, the arterial system, in terms of elastance, is dynamically and temporally coupled to the left-ventricle during ejection.

Animals↗
PubMed1992

Cardiac adaptation of sarcomere dynamics to arterial load: a model of hypertrophy.

In the past, the dynamics of the left ventricle were studied by its response to altered venous and arterial load for a given heart. This led researchers to propose the concept of an arterioventricular match or optimal point of function. The model of this paper reverses that idea by fixing preload and afterload while computing cardiac function due to altered left ventricular size or shape, resulting from modification of the number of parallel and series sarcounits. A mathematical model of physiological hypertrophy is introduced. Series and parallel arrangements of sarcounits constitute a cylindrical model of the left ventricle. Filling occurs from a venous reservoir with constant pressure through a valve, while ejection takes place into a three-element model of the systemic arterial system through another valve. It is found that the dynamics of the myofibrils can be matched to those of the left ventricle by choosing a ventricular shape that results in a minimum in myocardial O2 consumption (MVO2) for any constant ventricular load. A unique solution for the size of the ventricle results if the rate of MVO2 is specified. The model is able to predict correctly hypertrophy due to hypoxia and due to pressure (concentric) and volume (eccentric) overloads.

Adaptation, Physiological↗
PubMed1991

Diastolic mechanics and the origin of the third heart sound.

The third heart sound (S3) is observed for various hemodynamic conditions in both the normal and diseased heart. A theory is proposed in which myocardial viscoelasticity is primarily responsible for S3. A mathematical model is developed based on the mechanical aspects of diastolic function: nonlinear elasticity, viscoelasticity, and pressure generation. The model is provided as an electrical analogy of the left ventricle and circulatory system. S3 is predicted for the normal heart and the heart with dilated cardiomyopathy. An elevation of S3 intensity is indicated for cardiomyopathy, as is often observed in the clinic. S3 is produced experimentally by volume loading of the open-chest canine preparation and mathematically by imposing the conditions of volume loading on the model. Consistency of theory and experiment imply that it is valid to attribute S3 to myocardial viscoelasticity. The animal whose heart possessed the largest constant of viscoelasticity produced the greatest level of S3, in both cases. Nonlinear ventricular compliance is not found to be an essential requirement for sound generation, although increased compliance led to an increase in sound. S3 is predicted to change in response to venous return, ventricular stiffness, contractility, heart rate, and duration of contraction, as observed by others. In general, the coupling of these quantities to S3 is explained in terms of an excitation of viscous properties of the ventricle.

Animals↗
PubMed1991

Heterologous expression and characterization of an active lignin peroxidase from Phanerochaete chrysosporium using recombinant baculovirus.

The cDNA clone lambda ML-1 encoding one of the extracellular lignin peroxidases from the white rot fungus, Phanerochaete chrysosporium, was heterologously expressed in an active form using a recombinant baculovirus system. The glycosylated extracellular form of the recombinant protein contained the ferriprotoporphyrin IX moiety and was capable of oxidizing both iodide and the model lignin compound, veratryl alcohol. In comparative peroxidase assays using guaiacol and Mn(II), the recombinant lignin peroxidase did not appear to be Mn(II) dependent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that the heterologously expressed peroxidase had an apparent molecular weight similar to that of the native fungal isozyme H8. The elution profile of the active recombinant enzyme derived by ion-exchange chromatography and immunoblot analysis using an anti-H8 monoclonal antibody provided further evidence that the lambda ML-1 DNA encodes the lignin peroxidase H8.

Baculoviridae↗
PubMed1991

Bluetongue virus evolution: sequence analyses of the genomic S1 segments and major core protein VP7.

The S1 segments, encoding the group-specific antigen, VP7, from the five United States prototype BTV serotypes were cloned as full-length entities. The nucleotide and deduced amino acid sequences of segment S1 of BTV-2 were determined and compared with BTV-10, -11, -13, and -17, completing the sequencing of this cognate gene segment from all five US BTV serotypes. Each segment is 1156 bp long and contains an open reading frame encoding the 349-amino acid VP7 protein. Most (greater than 94%) of the amino acids of VP7 among the serotypes are conserved, including the location (position 255) of a single lysine residue. Secondary structure analyses of VP7 predict a putative eight-stranded beta-barrel between amino acid positions 150 and 250, a structure similar to that observed in ssRNA viruses. The S1 genes are flanked by conserved 5' and 3' noncoding regions. Stem-loop structures are predicted at the 3' end of each gene (nucleotide positions 1058-1097). The S1 segments of BTV-2, -10, -11, and -17 have greater than 93% of the nucleotides conserved, while less than 80% of their bases are identical with BTV-13. Analyses of nucleotide mismatches in each codon position of the VP7 open reading frame, transition frequencies, and evolutionary distances show that of the five, BTV-13 is the most distantly related and that BTV-10 and -17 are the most closely related serotypes. Evolutionary distance calculations of segment L2 from BTV-10, -11, and -17 concur with these observations. Comparison of this relationship with hybridization data of segment M3, which codes for VP5, suggests that BTV-17 has evolved by a combination of genetic drift and genomic reassortment. The data also indicate that the five US BTV serotypes are derived from two distinct gene pools. Evolution distances were used to estimate an evolution rate of 2.2 x 10(-3) nucleotide substitution/site/year for BTV segment S1. This rate is similar to the genes of retroviruses and implies an absence of RNA polymerase proofreading activity for dsRNA viruses.

Amino Acid Sequence↗
PubMed1991

Similar pressure pulse propagation and reflection characteristics in aortas of mammals.

Similar pressure and flow waveforms recorded in mammalian aortas suggest that pulse transmission characteristics may also be similar. We examined the validity of this hypothesis, utilizing allometric equations of pertinent hemodynamic parameters and a model of the arterial system. Results show that both the reflection coefficient and the propagation constant times the aortic length are essentially invariant across the mammalian species investigated. Resolved forward and reflected propagating waves are also similar. These findings suggest that the arterial system in these mammals indeed functions in a similar manner.

Animals↗
PubMed1990

Concurrent compliance reduction and increased peripheral resistance in the manifestation of isolated systolic hypertension.

The hemodynamic mechanisms responsible for producing isolated systolic hypertension (ISH) in the elderly are generally attributed to a decrease in arterial compliance. However, no consistent theoretical or experimental model has been proposed for the production of ISH. This problem was investigated with the use of computer simulation of the modified Windkessel model, an often-used tool in the study of arterioventricular function. Aortic pressure (Pa(t] and aortic flow (Qa(t] data were used to obtain the model parameters: peripheral resistance (Rs), arterial compliance (C) and characteristic impedance of the proximal aorta (Zo). Using Qa(t) as the input to the model, the effects of altered vascular properties on Pa(t) were studied by changing these model parameters. Graded reductions of C (25, 50 and 75%) alone increased systolic pressure (Ps), but also decreased diastolic pressure (Pd) to values below those found in ISH. On the other hand, an increase in Rs of 25% along with a 50 to 75% increase in C resulted in percent changes in Ps and Pd that would result in ISH from a normal pressure level. These results were consistent for a wide range of pressures. Decreased arterial compliance alone is not always responsible for the production of ISH. Rather, isolated systolic hypertension is usually the result of greatly reduced arterial compliance along with a smaller but significant increase in peripheral resistance.

Aged↗