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Biomedical subjects

L Lin

Publications and source records attributed to L Lin.

At least 37 records · Page 2Linked to original sources
PubMed1996

Synthesis, chromatographic resolution, and anti-human immunodeficiency virus activity of (+/-)-calanolide A and its enantiomers.

The anti-HIV agent (+/-)-calanolide A (1) has been synthesized in a five-step approach starting with phloroglucinol [-->5-->6-->11-->18-->(+/-)-1], which includes Pechmann reaction, Friedel-Crafts acylation, chromenylation with 4,4-dimethoxy-2-methylbutan-2-ol, cyclization, and Luche reduction. Cyclization of chromene 11 to chromanone 18 was achieved by employing either acetaldehyde diethyl acetal or paraldehyde in the presence of trifluoroacetic acid and pyridine or PPTS. Luche reduction of chromanone 18 at lower temperature preferably yielded (+/-)-1. Reduction of chromone 12, synthesized by Kostanecki-Robinson reaction from chromene 11, failed to afford (+/-)-1. The synthetic (+/-)-1 has been chromatographically resolved into its optically active forms, (+)- and (-)-1. The anti-HIV activities for synthetic (+/-)-1, as well as resultant (+)- and (-)-1, have been determined. Only (+)-1 accounted for anti-HIV activity, which was similar to the data reported for the natural product, and (-)-1 was inactive.

Antiviral Agents↗
PubMed1996

Comparison of Osborne-Mendel and S5B/PL strains of rat: central effects of galanin, NPY, beta-casomorphin and CRH on intake of high-fat and low-fat diets.

The effects of central administration of galanin, neuropeptide Y (NPY), beta-casomorphin(1-7) and corticotropin releasing hormone (CRH) on intake of either a high-fat or low-fat diet have been compared in two strains of rat, the dietary fat-sensitive Osborne-Mendel (OM) rat and the dietary fat-resistant S5B/Pl rat. Injection of galanin (0.1, 0.3 nmoles) into the 3rd cerebral ventricle stimulated the intake of both a high-fat and a low-fat diet in OM rats in a dose dependent manner but the response was significantly smaller in rats fed the low-fat diet. In S5B/Pl rats, galanin had a small stimulatory effect on food intake but only at a high dose (2 nmole). Beta-casomorphin(1-7) (5 nmoles), an opioid-like peptide, increased the intake of the high-fat but not the low-fat diet in OM rats, whereas S5B/Pl rats fed either a high-fat or low-fat diet did not respond to beta-casomorphin(1-7). Both strains showed a similar stimulatory response to NPY (0.1, 0.5 nmoles) on the intake of the high-fat or low-fat diet, but the magnitude of the response was attenuated in S5B/Pl rats. In contrast, the anorectic effects of CRH (0.26 nmoles) on food deprived animals was similar in both strains for both diets. We speculate that the regulatory system controlling the intake of fat activated by galanin and beta-casomorphin(1-7) may be defective in S5B/Pl rats.

Animals↗
PubMed1996

HLA-B40, B18, B27, and B37 allele discrimination using group-specific amplification and SSCP method.

We developed a system for discriminating HLA-B40, B18, B27, and B37 alleles using a two-step PCR method followed by SSCP analysis. Fragments (0.8 kb) including exon 2, intron 2, and exon 3 were amplified in the first PCR. We used two sets of primers, one specific for HLA-B60-related alleles and the other specific for HLA-B61-related, B18, B27, and B37 alleles. No amplifications of other class I genes or pseudogenes were observed. In the second PCR, exon 2 and exon 3 were amplified separately, using diluents of the first PCR products as templates. HLA-B61-related, B18, B27, B37, and B60-related alleles were clearly discriminated in the SSCP analysis of the second PCR products. In a population study in which B61 alleles were analyzed, B*4003 was detected in two Japanese individuals in addition to two B61 alleles previously reported to occur in Japanese, B*4002 and B*4006. The relative frequencies of B*4002, B*4006, and B*4003 in Japanese were 58, 35, and 6%, respectively. The individuals having B*4003 are the first non-South Americans in whom this allele has been detected. The SSCP banding patterns of 18 HLA-B60-positive Japanese population samples were identical to those of a B*40012 sample for both exon 2 and exon 3. We also demonstrated that the B37 allele occurring in some Japanese is B*3701.

Alleles↗
PubMed1996

Allele typing of HLA-A10 group by nested-PCR-low ionic strength single stranded conformation polymorphism and a novel A26 allele (A26KY, A*2605).

HLA-A26 is one of the most polymorphic HLA-A locus antigens among the Japanese population. Four HLA-A26 subtypes have so far been defined: A*2601-2604 [1]. We developed a means of typing alleles of the HLA-A10 group by nested PCR low ionic strength single-stranded conformation polymorphism (NPCR-LIS-SSCP) that is simple and cost effective. We used it to type 200 DNA samples from unrelated Japanese individuals who were serologically HLA-A26 positive. We found a novel A26 allele that had been suggested by PCR-SSO. Sequence analysis of A26KY (officially assigned A*2605, Accession No. D50068) revealed that the allele differs from A*2601 by a single nucleotide substitution at position 299, which leads to an amino acid substitution Ala-->Glu at position 76 in the alpha helix loop of the alpha 1 domain. From our results, A*2605 is likely to originate from A*2601 by a single point mutation. HLA-A*2601 showed the highest frequency (61.9%), followed by A*2603 (19.5%), A*2602 (17.6%), A*2604 (0.5%), and A*2605 (0.5%) in Japanese.

Alleles↗
PubMed1996

Visualization of D1 dopamine receptors on living nucleus accumbens neurons and their colocalization with D2 receptors.

To examine the substrate for dopamine (DA) synaptic action in the nucleus accumbens (nAcc), we visualized the cellular and subcellular distribution of DA receptors on postnatal nAcc neurons in culture using fluoroprobe derivatives of DA receptor ligands. Previously, we have shown that rhodamine-N-(p-aminophenethyl)-spiperone (NAPS) (10 nM), a derivative of the D2 antagonist spiperone, labels D2-like receptors on living nAcc neurons. We now show that rhodamine-Sch-23390 (30 nM), a derivative of the D1 antagonist, labels D1-like receptors. Putative specific membrane labeling reached a plateau after about 20 min. Labeling was stereospecific, as it was unaffected by competition with (-)-butaclamol, but blocked with (+)-butaclamol. We found that 52 +/- 7% of nAcc medium-sized neurons showed D1 labeling, which extended onto the dendrites. Labeling was also seen on presynaptic terminals, often abutting D1-positive and D1-negative cell bodies, consistent with a presynaptic modulatory role for D1 receptors. Larger neurons, which may be GABAergic or cholinergic interneurons, were also labeled. By sequential labeling first with rhodamine-Sch-23390 and then rhodamine-NAPS, we found that 38 +/- 6% of medium-sized neurons express both D1- and D2-like receptors, indicating that D1-D2 interactions may occur at the level of single postsynaptic neurons.

Animals↗
PubMed1996

Modular organization of the PDZ domains in the human discs-large protein suggests a mechanism for coupling PDZ domain-binding proteins to ATP and the membrane cytoskeleton.

The human homologue (hDIg) of the Drosophila discs-large tumor suppressor (DIg) is a multidomain protein consisting of a carboxyl-terminal guanylate kinase-like domain, an SH3 domain, and three slightly divergent copies of the PDZ (DHR/GLGF) domain. Here have examined the structural organization of the three PDZ domains of hDIg using a combination of protease digestion and in vitro binding measurements. Our results show that the PDZ domains are organized into two conformationally stable modules one (PDZ, consisting of PDZ domains 1 and 2, and the other (PDZ) corresponding to the third PDZ domain. Using amino acid sequencing and mass spectrometry, we determined the boundaries of the PDZ domains after digestion with endoproteinase Asp-N, trypsin, and alpha-chymotrypsin. The purified PDZ1+2, but not the PDZ3 domain, contains a high affinity binding site for the cytoplasmic domain of Shaker-type K+ channels. Similarly, we demonstrate that the PDZ1+2 domain can also specifically bind to ATP. Furthermore, we provide evidence for an in vivo interaction between hDIg and protein 4.1 and show that the hDIg protein contains a single high affinity protein 4.1-binding site that is not located within the PDZ domains. The results suggest a mechanism by which PDZ domain-binding proteins may be coupled to ATP and the membrane cytoskeleton via hDlg.

Adenosine Triphosphate↗
PubMed1996

Detection of p53 gene abnormality by sequence analysis of archival paraffin tissue. A comparison with fresh-frozen specimens.

This parallel study was designed to compare the sensitivity and specificity of detection of point mutations in fresh-frozen and formalin-fixed, paraffin-embedded breast cancer tissue. Sequence analysis of exon 5 of p53 gene was performed on polymerase chain reaction-amplified DNA from 25 infiltrating ductal carcinomas of the breast. Four tumor showed mutations with identical base substitutions in their respective codons of both frozen and paraffin-embedded specimens. We conclude that subtle genetic alterations can be detected in archival paraffin tissue with an accuracy comparable to that of fresh-frozen histologic samples.

Breast Neoplasms↗
PubMed1996

Further molecular diversity in the HLA-B15 group.

In order to further clarify the diversity of the HLA-B15 antigens and the correspondence of serological types with alleles in Asians, we screened various B15 serological splits by means of a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method. Subsequently, the genes encoding various B15 variants were sequenced. Two novel alleles, B*1528 and B*1529, were identified: the nucleotide sequence of the former contained a single-base substitution at position 263 in exon 2 as compared to that of the B*1501 allele, which results in an amino acid change at position 64 in the alpha 1 domain, and the nucleotide sequence of the latter differs from that of B*1518 by a single-base substitution at position 272 of exon 2 which results in an amino acid change at position 67 of the alpha 1 domain. One new allele, B*1521, described recently in Australian Aborigines was also identified in Asians in the present study. Moreover, the results of sequencing demonstrated that Asian HLA-B62, B70, and B77 antigens are encoded by B*1501, B*1518, and B*1513, respectively. Two splits of B75 antigens, B75V (TS-1) and B15N, which have been proposed to exist in the Japanese population were encoded by B*1511 and B*1502, respectively. Most of the B15 alleles detected in the present study showed positive associations with other locus antigens. Especially, B*1502 was strongly associated with Cw8, while B*1521 was strongly associated with A34 and Cw6.

Alleles↗
PubMed1996

Temporal variation in nephrotoxicity of low doses of isepamicin in rats.

The temporal variation in the nephrotoxicity of low doses of isepamicin was studied in male Sprague-Dawley rats treated with a single daily intraperitoneal injection of saline (NaCl, 0.9%) or isepamicin (80 mg/kg of body weight) at either 0800, 1400, 2000, or 0200 h for 4 and 10 days. On day 10, the cellular regeneration (incorporation of [3H] thymidine into DNA of renal cortex) and cortical accumulation of isepamicin were significantly higher in animals treated at 1400 h than at 0200 h (P < 0.01). Immunogold labeling studies showed that isepamicin was essentially localized in the lysosomes of proximal tubular cells in all treated groups, but the density of the gold particles over the lysosomes was higher in animals treated at 1400 than at 0200 h. The results of the present study show that the renal toxicity of isepamicin was maximal at 1400 h (midlight period) and minimal at 0200 h (middark period).

Animals↗
PubMed1996

A glycine-rich region in NF-kappaB p105 functions as a processing signal for the generation of the p50 subunit.

Transcription factor NF-kappaB is generally considered to be a heterodimer with two subunits, p50 and p65. The p50 subunit has been suggested to be generated from its precursor, p105, via the ubiquitin-proteasome pathway. During processing, the C-terminal portion of p105 is rapidly degraded whereas the N-terminal portion (p50) is left intact. We report here that a 23-amino-acid, glycine-rich region (GRR) in p105 functions as a processing signal for the generation of p50. A GRR-dependent endoproteolytic cleavage downstream of the GRR releases p50 from p105, and this cleavage does not require any specific downstream sequences. p50 can be generated from chimeric precursor p105N-GRR-IkappaBalpha, while the C-terminal portion (IkappaBalpha) can also be recovered, suggesting that p105 processing includes two steps: a GRR-dependent endoproteolytic cleavage and the subsequent degradation of the C-terminal portion. We have also demonstrated that the GRR can direct a similar processing event when it is inserted into a protein unrelated to the NF-kappaB family and that it is therefore an independent signal for processing.

Amino Acid Sequence↗
PubMed1996

Continuous measurement of 13C16O2 production from [13C]pyruvate by intact liver mitochondria: effect of HCO3-.

We have measured continuously the production of mass 45 CO2(13C16O2) from 13C-labeled pyruvate in a guinea pig liver mitochondrial suspension and simultaneously the O2 consumption at 37 degrees C and pH 7.4. The reactions took place in a closed 3-ml volume, stirred, thermoregulated chamber separated from the ion source of a mass spectrometer by a gas-permeable membrane that permitted recording the mass peaks of any gas dissolved in the reaction mixture with a response time as fast as 3 s. If the pyruvate was labeled on C-2, no 13C16O2 was formed, even after 1 h, indicating that C-2 and C-3 were not metabolized in the citric acid cycle. We found that production of 13C16O2 was five times greater in the presence of 25 mM HCO3- than in its absence. A probable mechanism of this CO2/HCO3- effect is carboxylation of pyruvate to oxaloacetate, which would react with acetyl CoA to form citrate and with NADH to form malate, thus removing two major inhibitors of pyruvate dehydrogenase. We conclude that CO2/HCO3- has a potent and hitherto unappreciated regulatory effect on liver pyruvate dehydrogenase.

Animals↗
PubMed1996

Rate of uptake of CO by hemoglobin in pig erythrocytes as a function of PO2.

This study was initiated to obtain data on the rate of carbon monoxide (CO) uptake (theta CO) by hemoglobin in pig erythrocytes to derive, in a later study, the pulmonary capillary blood volume (Qc) in pigs from the Roughton-Forster relationship. Blood from five different female pigs was used. The theta CO, the milliliters of CO taken up by 1 ml of whole blood per minute per Torr CO tension, was determined on each blood sample with a continuous-flow rapid-mixing apparatus and double-beam spectrophotometry at 37 degrees C and pH 7.4 at four or five different PO2 values. Because the individual regression lines of theta CO vs. PO2 were not significantly different, a common regression equation was calculated: 1/theta CO = 0.0084 PO2 + 0.63. The slope of this regression line is significantly steeper than the reported slopes of regression lines for human and dog erythrocytes measured under the same conditions. Our results revealed that calculation of Qc in pigs by using theta CO values for human or dog erythrocytes would result in an underestimation of 51 and 50%, respectively.

Animals↗
PubMed1996

Differentiation of rat preadipocytes is accompanied by expression of thyrotropin receptors.

To investigate the regulation of expression of the TSH receptor (TSHR) in extrathyroidal tissues, the level of TSHR messenger RNA (mRNA) and TSH-dependent signal transduction were determined in isolated rat adipocytes and cultured preadipocytes. The epididymal, sc, and perirenal, but not the interscapular brown adipose tissues, possessed TSHR mRNA and increased cAMP responses to TSH and were thus used as the source of preadipocytes. Morphological analysis revealed that the combination of insulin and T3 most effectively caused the differentiation of rat preadipocytes. These differentiated preadipocytes exhibited increased cAMP production in response to TSH. The addition of FCS to the culture medium inhibited the differentiation of rat preadipocytes as well as TSH-stimulated production of cAMP. The stimulation of differentiation was associated with an increased expression of TSHR mRNA levels, whereas the inhibition of differentiation was associated with a decreased expression of TSHR mRNA, as detected by Northern blot analysis. The results indicate that the expression and function of the TSHR in cultured rat preadipocytes are closely related to cellular differentiation. Cultured rat preadipocytes appear to provide a useful system for studying the mechanism of extrathyroidal expression of TSHR.

Adipocytes↗
PubMed1996

Day-night treatment difference of tobramycin serum and intrarenal drug distribution and nephrotoxicity in rats: effects of fasting.

The effects of short-term food deprivation on the serum and renal distribution and nephrotoxicity of tobramycin were studied in female Sprague-Dawley rats maintained on a 14-h light/10-h dark cycle (light on: 06:00). For the distribution study, a single injection of tobramycin (40 mg/kg, i.p.) was administered at 14:00 or 02:00 to normally fed animals or to animals fasted for 12 h before tobramycin injection; these treatment times correspond to the peak and trough of tobramycin nephrotoxicity as previously determined in other studies. The serum and cortical levels of tobramycin were significantly higher 60, 120, and 240 min after the injection in fasted animals treated at 02:00 compared with normally fed animals treated at the same time (p < 0.05). In animals injected at 14:00, similar levels of tobramycin were measured in both fasted and fed rats. In the nephrotoxicity study, female Sprague-Dawley rats were fasted for 12 h before and 24 h after the timed single injection of tobramycin (150 mg/kg, i.p.). The 24-h urinary excretion of beta-galactosidase was significantly higher in fasted animals treated at 02:00 than in fed rats treated at the same time of day. Seventy-two hours following tobramycin injection, serum creatinine levels and cortical levels of tobramycin were significantly higher in fasted rats treated at 14:00 than at 02:00 and in fed rats treated at 14:00. These data suggest that a short period of food deprivation modulates the temporal variations of tobramycin nephrotoxicity.

Animals↗
PubMed1996

Effect of herba artemisiae scopariae on cytogenetic damage induced by aflatoxin B1.

It was found that the per mill rate of micronucleus of marrow cells in mice induced by AFB1 decreased with the increasing dosage of Herba Artemisiae Scopariae, and that there were dosage-effect relations in the lowering of percentage of chromosomal aberration induced by AFB1 and in the inhibition effect on some sister chromatid exchanges per cell.

Aflatoxin B1↗
PubMed1996

[Quality of semen Hydnocarpi anthelminiticae introduced to various places].

The result shows that the Semen Hydnocarpi Anthelminticae introduced in Guangdong, Hainan and Yunnan Provinces is similar to the control medicinal materials in fatty oil content, index of refraction, effective component content and trace element content. But the saponification value of the introduced products is higher. The quality of the Semen Hydnocarpi Anthelminticae introduced to the above places is up to the level of the control medicinal materials.

China↗