Biomedical subjects

J H Lin

Publications and source records attributed to J H Lin.

At least 217 records · Page 12Linked to original sources

PubMed1995

Regiospecific intestinal absorption of the HIV protease inhibitor L-735,524 in beagle dogs.

PURPOSE: To evaluate regional intestinal absorption and the feasibility of sustained release dosage form development for an HIV protease inhibitor, L-735,524, METHODS: L-735,524 free base or sulfate salt was administered orally as suspension, solution or in solid dosage forms to fasted or fed Beagle dogs. Delayed-release dosage forms with "slow" or "fast" in vitro dissolution rates were evaluated in vivo to assess plasma concentration profiles. In addition, drug was administered directly into the jejunum or colon of animals, and drug concentrations determined in portal circulation to characterize absorption from these sites. RESULTS: L-735,524 sulfate was well absorbed orally form a solution or capsule formulation if fasted animals' stomachs were preacidified with citric acid solution. A free base suspension, delivered in divided doses to fed animals, was also well absorbed. Prototype extended release dosage forms of L-735,524 produced a reduction in peak plasma levels but failed to prolong absorption and extend plasma concentrations compared to an immediate release capsule. Administration of L-735,524 sulfate solution (pH < 3) as bolus solution or by infusion into the jejunum resulted in rapid but incomplete absorption compared to oral gavage. The free base suspension (pH 6.5) delivered into jejunal or colonic regions did not produce measurable systemic plasma concentrations. CONCLUSIONS: Extended release formulations did not prolong absorption of L-735,524 in dogs. Optimal L-735,524 absorption was dependent on solubility in an acidic environment in the duodenum.

Administration, Oral↗

PubMed1995

The effect of matrix phase morphology on the structure of Ag-Cu-Pd dispersed phase dental amalgam.

The effect of an Ag-Cu-Pd dispersant alloy on the structure of amalgams fabricated with two different particle types of low-copper amalgam matrices was investigated. In amalgams L and L (0), a low-copper lathe-cut amalgam Aristalloy was used as the matrix. In amalgams S and S(0), the matrix used was Spheralloy, a low-copper spherical amalgam. X-ray diffraction was used to analyse the relative content of the phases in the amalgams. The presence of Pd in the dispersant of amalgams L and S resulted in a decrease in the amount of eta' (Cu6Sn5) and an increase in the amount of unreacted particles when compared to the controls L(0) and S(0). Amalgams fabricated with a spherical alloy matrix (S and S(0)) showed a greater effect than amalgams fabricated with a lathe-cut matrix alloy (L and L(0)).

Chemical Phenomena↗

PubMed1995

Effects of ginseng on the blood chemistry profile of dexamethasone-treated male rats.

Ginseng, a panacea in the Orient, has been widely investigated in the last two decades and found to possess a wide range of pharmacological activities including anti-fatigue properties, a transient regulatory action on metabolism and blood pressure, and an increase in the hypothalamo-pituitary-adrenocortical activities. However, a panoramic clinical chemistry study including adrenal and thyroid functions has never been done before. Two experiments with the same design but different concentrations of dexamethasone were performed in this study. The results obtained from the two experiments indicated that ginseng administration at this regime did not influence the blood chemistry profiles in normal rats, but significantly decreased AST and ALT levels from those in dexamethasone-treated ones. It implies that ginseng has a liver-protective effect. Meanwhile, ginseng therapy restores the adrenal and thyroid functions of rats inhibited by dexamethasone treatment.

Animals↗

PubMed1995

Uroplakins, specific membrane proteins of urothelial umbrella cells, as histological markers of metastatic transitional cell carcinomas.

Uroplakins (UPs) Ia, Ib, II, and III, transmembrane proteins constituting the asymmetrical unit membrane of urothelial umbrella cells, are the first specific urothelial differentiation markers described. We investigated the presence and localization patterns of UPs in various human carcinomas by applying immunohistochemistry (avidin-biotin-peroxidase complex method), using rabbit antibodies against UPs II and III, to paraffin sections. Positive reactions for UP III (sometimes very focal) were noted in 14 of the 16 papillary noninvasive transitional cell carcinomas (TCCs) (88%), 29 of the 55 invasive TCCs (53%), and 23 of the 35 TCC metastases (66%). Different localization patterns of UPs could be distinguished, including superficial membrane staining like that found in normal umbrella cells (in papillary carcinoma), luminal (microluminal) membrane staining (in papillary and invasive carcinoma), and, against expectations, peripheral membrane staining (in invasive carcinoma). Non-TCC carcinomas of various origins (n = 177) were consistently negative for UPs. The presence of UPs in metastatic TCCs represents a prime example of even advanced tumor progression being compatible with the (focal) expression of highly specialized differentiation repertoires. Although of only medium-grade sensitivity, UPs do seem to be highly specific urothelial lineage markers, thus operating up interesting histodiagnostic possibilities in cases of carcinoma metastases of uncertain origin.

Amino Acid Sequence↗

PubMed1995

pH-dependent oral absorption of L-735,524, a potent HIV protease inhibitor, in rats and dogs.

L-735,524, a potent and specific inhibitor of human immunodeficiency virus protease, is currently under investigation for the treatment of acquired immunodeficiency syndrome. The aqueous solubility of L-735,524 was pH-dependent, > 100 mg/ml at pH below 3.5 and 0.03 mg/ml at pH 6. When L-735,524 was given orally as a suspension in 0.5% methocel (pH 6.5) at 10 mg/kg, the bioavailability was approximately 16% for both dogs and rats. When the same dose of the drug was administered in 0.05 M citric acid (pH 2.5), the bioavailability increased 4.5-fold in dogs (72%), but only slightly in rats (24%). The pH- and species-dependent differences in bioavailability observed in rats and dogs may be because of differences in the rate of gastric acid secretion and in the magnitude of hepatic first-pass effect. Gastric acid secretion is poor in dogs but substantial in rats. When L-735,524 was administered in 0.5% methocel, a large portion of the drug in dogs, but not in rats, remained undissolved, resulting in poor absorption in dogs. On the other hand, when L-735,524 was administered in citric acid, most of the drug would be in solution allowing for better absorption in dogs. The hypothesis of pH-dependent absorption was further supported by the findings that absorption was significantly increased in dogs after feeding, but substantially decreased in rats after pretreatment with famotidine, a potent H2-receptor antagonist. L-735,524 underwent an extensive first-pass metabolism in rats, but not in dogs.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1995

Time- and dose-dependent pharmacokinetics of L-754,394, an HIV protease inhibitor, in rats, dogs and monkeys.

L-754,394 is a potent and specific inhibitor of the HIV-1 encoded protease that is essential for the maturation of the infectious virus. The drug exhibited dose-dependent kinetics in all species studied (rat, dog and monkey); the apparent clearance decreased when the dose was increased. However, the dose-dependency cannot be explained by Michaelis-Menten kinetics. L-754,394 in plasma declined log-linearly with time, but with an apparent half-life that increased with dose. The apparent terminal half-life of L-754,394 in rats increased from 20 min at 0.5 mg/kg i.v. to 118 min at 10 mg/kg i.v. Furthermore, L-754,394 exhibited time-dependent pharmacokinetics. After chronic i.v. doses for 7 days (1 mg/kg/dose/day), the apparent clearance of L-754,394 in rats decreased from 87 ml/min/kg after the first dose to 25 ml/min/kg after the last dose. Similar results were observed in dogs and monkeys. In vitro spectral studies indicated that approximately 40 to 60% of the content of cytochrome P-450 was inactivated when L-754,394 (10 microM) was incubated with rat, dog and monkey liver microsomes in the presence of NADPH. Little or no inactivation of cytochrome P-450 was observed when either NADPH or L-754,394 was omitted. In addition, L-754,394 selectively inhibited CYP 2C11-dependent testosterone 2 alpha- and 16 alpha-hydroxylase activity and CYP 3A1/2-dependent testosterone 6 beta-hydroxylase activity, but not CYP 2D1/2-dependent bufuralol 1'-hydroxylase activity nor CYP 1A2-dependent phenacetin O-deethylase activity in rat liver microsomes.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1995

[Application and evaluation of multiple exposure photography and motion analysis system in gait measurement].

Accurate techniques to evaluate and quantitate gait patterns have been available in clinical applications for the last 20 years. In this study, by employing multiple exposure photography, the walking velocity, stride length, step cadence and joint angle of the lower limbs were recorded for normal young men during the gait cycle. Data were compared with results from the motion analysis system and related researches. The results showed that multiple exposure photography can also provide quantitative analysis of several important time-distance parameters in the gait cycle.

Adult↗

PubMed1995

Metabolites of L-735,524, a potent HIV-1 protease inhibitor, in human urine.

L-735,524, N-[2(R)-hydroxy-1(S)-indanyl]-5-(2(S)-(1,1- dimethylethylaminocarbonyl)-4-[(pyridin-3-yl)methyl]piperazin++ +-1-yl)-4(S)- hydroxy-2(R)-phenylmethylpentanamide, is a potent and specific inhibitor of the human immunodeficiency virus type 1 protease and is undergoing clinical evaluation. In an initial clinical study, noninfected male volunteers were administered single, 1000 mg oral doses of nonlabeled compound. Urine samples were collected over a period of 48 hr. Metabolic profile of the urine was determined by HPLC-UV comparison with that from a human liver slice incubation of radiolabeled L-735,524. Seven significant metabolites were isolated from pooled human urine, and were characterized by NMR, MS, and/or chromatographic comparisons with authentic standards. The major metabolic pathways were identified as: a) glucuronidation at the pyridine nitrogen to yield a quaternized ammonium conjugate, b) pyridine N-oxidation, c) para-hydroxylation of the phenylmethyl group, d) 3'-hydroxylation of the indan, and e) N-depyridomethylation. A minor product was identified as 2',3'-trans-dihydroxyindan analog. Urinary excretion of L-735,524 and its metabolites represented a minor pathway of elimination. The intact parent compound seemed to be the major component in the urine, whereas the level of each metabolite was relatively low.

Chromatography, High Pressure Liquid↗

PubMed1995

Potent and selective inactivation of human liver microsomal cytochrome P-450 isoforms by L-754,394, an investigational human immune deficiency virus protease inhibitor.

L-754,394, N-[2(R)-hydroxy-1(S)-indanyl]-5-[2(S)-(1,1-dimethylethylaminocarbonyl )-4- [(furo[2,3-b]pyridin-5-yl)methyl]piperazin-1-yl]-4(S)-hydroxy-2(R) - phenylmethylpentanamide, is a potent and specific inhibitor of the human immune deficiency virus (HIV) protease. The drug selectively inhibited human liver microsomal CYP 3A4-dependent testosterone 6 beta-hydroxylase and CYP 2D6-dependent bufuralol 1'-hydroxylase activities in a time- and concentration-dependent manner in the presence of an NADPH-generating system. L-754,394 was found to be a very potent inactivator of CYP 3A4. Thus, for testosterone 6 beta-hydroxylase, the inactivation kinetic constants, Kl and kinact, were 7.5 microM and 1.62 min-1, respectively, and the partition ratio (moles product formed per moles enzyme inactivated) was approximately 1.35. To a lesser extent, L-754,394 also was an inactivator of CYP 2D6, for which the corresponding values for Kl, kinact and partition ratio were 32 microM, 0.18 min-1 and 40, respectively. CYP 3A4 inactivation was reduced markedly by ketoconazole, a selective CYP 3A4 inhibitor. Similarly, CYP 2D6 inactivation also was prevented by quinidine, a specific competitive inhibitor of this isoform. However, exogenously added nucleophiles (GSH, semicarbazide and N-acetylcysteine) failed to protect against P-450 inactivation. These results suggest that the inactivation process likely is mediated by a reactive metabolite of L-754,394 that alkylates, and thereby destroys, the enzyme. Furthermore, this electrophilic intermediate may not be released into the medium before the inactivation event.

Cytochrome P-450 CYP2D6↗

PubMed1995

A study of different postures on isometric lifting strength in normal college students.

The purpose of this study was to describe norms of isometric lifting strength for normal college students and to investigate the effects of gender, body weight, height, and upper and lower limb length on lifting strength. Three types of lifting (i.e., arm, back, and leg lifting) were measured in a sample of 104 college students aged between 18 and 26 years. A Force Evaluation and Testing System (FET 5000) was used for strength measurements with three standard lifting positions. The average of three trials for each lifting strength test was used as the subject's test score. The results showed that the highest mean lifting strength was recorded for leg lifting in both males and females. Both groups also had the lowest score in arm lifting. Of the total sample, there was a difference of strength of nearly three- times between leg and arm lifting. Males were stronger than females in all lifting patterns. In addition, lifting strengths were significantly affected by such variables as sex, weight and height. Body weight was an effective predictor of arm lifting strength, and height an effective predictor of leg lifting strength. Finally, we found that strength in one position had positive relations with strength in other positions. These findings and the establishment of data base can in future provide therapists with an objective evaluation regarding lifting strength of individuals for clinical use.

Adolescent↗

PubMed1995

Species similarities and differences in pharmacokinetics.

One of the fundamental challenges drug metabolism scientists face in drug discovery and development is the extrapolation of metabolic data and risk assessment from animals to humans. Although it is generally believed that data from animal studies can be reasonably extrapolated to humans with the application of appropriate pharmacokinetic principles, there are certainly some limitations. The purpose of this review is to show some species similarities and differences in absorption, distribution, metabolism, and excretion, with an attempt to address the questions of whether animal data can be extrapolated and what the limitations are. The underlying mechanisms responsible for these similarities and differences are discussed and examples given for illustration. In addition, the concepts of allometric and physiological models for extrapolation of kinetic data are briefly highlighted. In conclusion, although pharmacokinetics has been advanced greatly in recent years, it is not yet possible to predict all of the pharmacokinetic parameters of a drug in humans from animal studies. Nevertheless, under certain well-defined conditions, it may be possible to make reasonably good predictions. For example, the intrinsic absorption of a given drug across the wall of the gastrointestinal tract is probably similar among species, because the nature of the biomembrane of epithelial cells is similar in mammals and because the absorption process is basically an interaction between the drug and the biomembrane. However, other factors, such as pH-dependent solubility and first-pass metabolism that affect the absorption, may differ from species to species resulting in species differences in absorption. Predicting the renal excretion of drugs in humans has been relatively successful using the glomerular filtration rate ratio between humans and animals. Similarity, if the elimination of a drug is mainly by the liver and the rate of elimination is limited by hepatic blood flow, one could predict the clearance of the drug in humans by the hepatic blood flow. However, biochemical parameters, such as protein binding and drug metabolism, are less predictable. These parameters vary considerably among species.

Absorption↗

PubMed1994

Modulation of glycosaminoglycan addition in naturally expressed and recombinant human thrombomodulin.

The two major glycoforms of full-length human thrombomodulin (TM), one with (TM(CS+)) and one without (TM(CS-)) chondroitin sulfate (CS) were analyzed on Western blots of primary and transformed cells and in cells expressing recombinant TM. TM on the surface of Chinese hamster ovary and COS-7 cells is solely TM(CS-). Primary arterial endothelial cells (HAEC and HPAEC) express a greater fraction of TM with CS attached than venous cells (HUVEC). Human lung carcinoma cells (A549) express more TM(CS+) than primary cells and recombinant TM on human melanoma cells (CHL-1) occurs in two very high molecular weight forms of TM(CS+). We explored this variation in TM(CS+) with soluble recombinant TM in several cell lines and analyzed the ambiguous CS addition site in human TM by site-directed mutagenesis. Mutation of Ser474 to Ala blocks CS addition in Chinese hamster ovary and COS-7 cells but not CHL-1 cells which add CS to Ser472 and Ser474. Structure of the O-link domain affects partitioning into TM(CS+) since substituting with the decorin CS addition sequence, substituting all Ser and Thr except Ser474 with Ala, and deleting around the potential beta-turn all increase the ratio of TM(CS+) to TM(CS-). A combination of the decorin substitution and deletion of the remaining O-link domain yields the most TM(CS+).

Amino Acid Sequence↗

PubMed1994

Expression efficiency of the human thrombomodulin-encoding gene in various vector and host systems.

Expression systems were developed for evaluating recombinant human thrombomodulin (TM) production in different host cell lines by investigating the performance of five mammalian expression vectors. The expression vectors were constructed so that they contain multiple monocistronic gene cassettes which include a gene encoding a dominant selectable marker, HyR (hygromycin B phosphotransferase), under the regulation of the thymidine kinase promoter, the target gene which encodes a truncated human re-TM under the regulation of various promoters, an amplifiable gene (Dhfr) encoding murine dihydrofolate reductase under the regulation of either the SV40 early or late promoter along with the SV40 enhancer and the SV40 ori. We tested the performance of the five expression vectors in human embryonic kidney cells (HEK293), baby hamster kidney cells (BHK), human melanoma cells (CHL-1) and Dhfr- Chinese hamster ovary cells (CHO/Dhfr-). We found that the efficiency of DNA uptake, transient expression and stable expression of the different expression vectors were all cell-line dependent. However, the myeloproliferative sarcoma virus (MPSV) LTR promoter consistently showed higher expression levels in all cell lines, particularly in HEK293 cells. These results were confirmed by the distribution curves of the level of expression of individual clones. Furthermore, by amplifying Dhfr in transfected CHO/Dhfr- cells with 100 nM methotrexate, we achieved a 20-fold increase in re-TM production using the SV40 late promoter to control murine Dhfr expression. Our data from DNA and mRNA analysis reveal that pMPSV-TM has a high transcription efficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Identification and biological characterization of an epidermal growth factor-related protein: cripto-1.

The human and mouse cripto-1 (CR-1) genes can code for proteins related in structure to epidermal growth factor (EGF). A specific 36-kDa immunoreactive protein was detected by Western blot analysis in human cell lines that express CR-1 mRNA but not in cell lines that fail to express this transcript. Immunoprecipitation of GEO colon carcinoma or mouse embryonal carcinoma cells detected 27-29-kDa and 24-kDa proteins, respectively. Cell lysates and conditioned medium that were prepared from several CHO clones and were expressing either a recombinant human or mouse CR-1 cDNA contained immunospecific 27-29-kDa and 24-kDa proteins, respectively. Monensin or tunicamycin treatment resulted in a shift of the 27-29-kDa human CR-1 protein to 24 kDa and 20 kDa, respectively. The 20-kDa protein was also observed after digestion of the 27-29-kDa human CR-1 protein with N-glycosidase F. Using two CR-1 synthetic refolded peptides that correspond to the EGF-like domain of the human CR-1 sequence or conditioned medium obtained from human CR-1 expressing CHO cells, growth stimulatory activity could be detected on non-transformed human mammary epithelial cells and on two human breast cancer cell lines. EGF receptor-blocking antibody did not inhibit the growth stimulatory action of the CR-1 protein. Likewise, the CR-1 refolded peptides or conditioned medium from the human CR-1-expressing CHO cells failed to inhibit the binding of 125I-EGF in an EGF-radioreceptor assay. These data demonstrate that the CR-1 is a glycoprotein that can function as a growth factor through an EGF receptor-independent pathway.

Amino Acid Sequence↗

PubMed1994

Mammalian uroplakins. A group of highly conserved urothelial differentiation-related membrane proteins.

The asymmetric unit membrane (AUM) forms the apical plaques of mammalian urothelium and is believed to play a role in strengthening the urothelial apical surface thus preventing the cells from rupturing during bladder distention. We have shown previously that purified bovine AUMs contain four major integral membrane proteins: the uroplakins Ia (27 kDa), Ib (28 kDa), II (15 kDa), and III (47 kDa). This contradicts some previous reports indicating that some of these proteins are absent in AUMs of several species. Using an improved procedure, we isolated AUMs from, in addition to cattle, eight mammalian species (human, monkey, sheep, pig, dog, rabbit, rat, and mouse). The AUMs of these species appear morphologically similar bearing crystalline patches of 12-nm protein particles with a center-to-center spacing of 16.5 nm. Using antibodies raised against synthetic oligopeptides or individual bovine uroplakins, we established by immunoblotting that the four uroplakins are present in AUMs of all these species. The DNA-deduced amino acid sequences of bovine and mouse uroplakin II revealed 83% identity. These results indicate that uroplakins Ia, Ib, II, and III are the major protein components of probably all mammalian urothelial plaques, and that the sequence and three-dimensional structure of uroplakin molecules are highly conserved during mammalian evolution.

Amino Acid Sequence↗

PubMed1994

Column-switching high-pressure liquid chromatographic method for the determination of a new carbapenem antibiotic, L-739,428, in rat and monkey plasma.

A column-switching HPLC method for determination of a new carbapenem antibiotic, L-739,428 (I), was developed that allowed direct injection of rat and monkey plasma. Following dilution with a pH 6.5 buffer, samples were injected without further cleanup into an extraction column dry-packed with 10-microns particle size Maxsil C18 reversed-phase adsorbent. Endogenous plasma components were washed to waste for 6 min with a weak mobile phase of 0.025 M sodium phosphate, 0.005 M hexanesulfonate (pH 6.5). Compound I, retained on the extraction column, was then backflushed with a mobile phase which consisted of a mixture of the preceding buffer-ion pair solution and acetonitrile (96:4, v/v) into a Partisphere C18 analytical column and detected by ultraviolet absorption at 299 nm. Chromatographic retention time was 11.5 min. Stability of I in plasma was maximized by use of a refrigerated autosampler which maintained plasma at 5 degrees C until analyzed. The limit of quantification was 0.1 microgram/ml using the equivalent of 75 microliters plasma.

Animals↗

PubMed1994

Precursor sequence, processing, and urothelium-specific expression of a major 15-kDa protein subunit of asymmetric unit membrane.

The asymmetric unit membrane (AUM) is a highly specialized biomembrane elaborated by terminally differentiated urothelial cells. It contains quasi-crystalline arrays of 12-nm protein particles each of which is composed of six dumbbell-shaped subdomains. In this paper we describe the precursor sequence, processing and in vitro membrane insertion properties of bovine uroplakin II (UPII), a 15-kDa major protein component of AUM. The cDNA-deduced amino acid sequence revealed that UPII is synthesized as a precursor protein containing a cleavable signal peptide of approximately 26 amino acids, a long pro-sequence of approximately 59 residues harboring three potential N-glycosylation sites, and the mature polypeptide of 100 residues. In vitro translation of UPII mRNA demonstrated that UPII is indeed first synthesized as a 19-kDa precursor, which loses its signal peptide upon insertion into added microsomes; this process is accompanied by the acquisition of high mannose-type oligosaccharides giving rise to a 28-kDa precursor which is completely protected from the digestion by exogenous proteases. These results, together with the presence of a stretch of 25 hydrophobic amino acids at the C terminus, suggest that UPII protein is anchored to the lipid bilayer via its C-terminal membrane-spanning domain with its major N-terminal domain exposed luminally. The formation of the 15-kDa mature UPII requires the removal of the pro-sequence by a furin-like endoprotease. Since only mature UPII devoid of this pro-sequence can interact with 27-kDa uroplakin I, the proteolytic processing of UPII precursor may play an important role in regulating the assembly of AUM. Finally, we showed that genomic sequences cross-hybridizing with bovine UPII cDNA are present in many mammals suggesting that UPII performs a highly conserved function in the terminally differentiated cells of mammalian urinary bladder epithelium.

Amino Acid Sequence↗

PubMed1994

Splenic lymphoma with circulating villous lymphocytes: report of seven cases and review of the literature.

Splenic lymphoma with villous lymphocytes (SLVL) is a relatively new entity with only a few reports published. We report seven cases of SLVL with detailed clinicopathologic and comprehensive immunophenotypic studies to further characterize this lymphoma, which is frequently confused with hairy cell leukemia and other low-grade B-cell lymphoid neoplasms. The diagnostic criteria we used include 1) prominent splenomegaly, 2) insignificant or no lymphadenopathy, 3) lymphocytosis without leukopenia, 4) presence of circulating villous lymphocytes, 5) characteristic cytologic and histologic features, and 6) specific phenotypic and cytochemical findings. Our studies show that SLVL does not represent a pure entity but rather a morphologically heterogeneous group of low-grade lymphomas with various cytologic and histologic features. Although immunophenotyping is helpful in differential diagnosis, multiparameter studies are necessary to confirm the diagnosis. In our series, only two patients died of SLVL, who probably developed transformation to a higher-grade lymphoma.

Aged↗