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Biomedical subjects

X Lin

Publications and source records attributed to X Lin.

At least 379 records · Page 21Linked to original sources
PubMed1994

Differences in the metabolism of 18:2n-6 and 18:3n-6 by the liver and kidney may explain the anti-hypertensive effect of 18:3n-6.

The present study examined the in vitro and in vivo metabolism of 18:2n-6 and 18:3n-6 by kidney and liver in the male adult spontaneously hypertensive (SHR) and normotensive (WKY) rats. In liver and kidney slices incubated for 1 h with either [1-14C]18:2n-6 or [1-14C]18:3n-6 (60 microM), substantial amounts of radioactivity were incorporated into triacylglycerol and phospholipid fractions. Approximately 15% of the radiolabeled 18:2n-6 was converted into 18:3n-6 in liver slices but no conversion was found in kidney slices. When incubated with radiolabeled 18:3n-6, over 40% of the radioactivity was metabolized mainly to 20:4n-6 in liver slices, but evenly to 20:3n-6 and 20:4n-6 in kidney slices. There were no differences between the results from SHR and those from WKY. In WKY rats given an oral bolus of radiolabeled 18:3n-6, most of the radioactivity was recovered in the liver and significantly less in the kidney. In both tissues, the radioactivity was associated initially only with 18:3n-6 and later with its elongation product, 20:3n-6. These findings indicated that the kidney, although unable to metabolize 18:2n-6, could metabolize 18:3n-6 taken up from the circulation. The effectiveness of 18:3n-6, compared to 18:2n-6, as an anti-hypertensive agent may result from the provision of a post-delta 6-desaturation metabolite which can be directly converted to blood pressure-regulating eicosanoids in the kidney.

Animals↗
PubMed1994

Use of k0-NAA in the life sciences.

The applicability and usefulness of k0-based reactor neutron activation analysis (NAA) in the life sciences is evaluated from the following examples: 1. Instrumental NAA of NIST SRM 1633a coal fly ash, as a quality assessment; 2. Radiochemical NAA of Versieck's reference human serum, and--herewith associated--the development of practical correction procedures for neutron-induced reaction interferences and of improved methods to evaluate the detection efficiency and the correction for true coincidence; and 3. Determination of the lanthanides in plant leaves and lichens near a Portuguese coal-fired power station, which led to the introduction of the Westcott formalism and to the use of a low-energy photon detector. As concluded, k0-based NAA is at present capable of tackling a large variety of analytical problems when it comes to the multielement determination in environmental and biological matrices.

Biology↗
PubMed1994

Isolation of a novel cDNA encoding a putative membrane receptor with high homology to the cloned mu, delta, and kappa opioid receptors.

A rat brain cDNA library was screened for clones homologous to the recently cloned mouse delta-opioid receptor (DOR-1). Among the clones isolated was Hyp 8-1, a clone with a unique nucleotide sequence capable of encoding a putative protein which is 57-58% identical to the amino acid sequences of the cloned delta, mu and kappa opioid receptors, indicating a close relationship of Hyp 8-1 with the opioid receptor family. Several cDNAs representing possible splice variants of Hyp 8-1 were also isolated. Binding studies of COS-7 cells transfected with clone Hyp 8-1 failed to demonstrate specific binding with several 3H-opioid ligands. In situ hybridization studies indicate that the mRNA for Hyp 8-1 is distributed discretely throughout the rat brain, in an overall pattern which is different from that of several other G-protein-coupled seven transmembrane receptors. Thus, it is likely that the Hyp 8-1 cDNA encodes a novel peptide receptor.

Amino Acid Sequence↗
PubMed1994

Engineering aspartic proteases to probe structure and function relationships.

Recently, protein engineering has been used to interconvert homodimeric and homologous single-chain aspartic proteases, with some success. The independent folding of the domains of these proteases has also permitted the engineering of domain-rearranged protease zymogens and the use of individual domains as probes for structural denaturation. In addition, site-directed mutagenesis has provided insights into the catalytic mechanism and specificity of this family of proteases.

Aspartic Acid Endopeptidases↗
PubMed1994

Emulsification and fatty acid chain length affect the kinetics of [14C]-medium-chain triacylglycerol utilization by neonatal piglets.

In vivo oxidation kinetics of medium-chain triglycerides (MCT) composed of [1-14C]hexanoate (tri-6:0) or [1-14C]octanoate (tri-8:0) were compared after being fed to day-old piglets (1.4 kg; n = 4/treatment). Animals were fitted with arterial and oro-gastric catheters and placed into respiration chambers held at 35 degrees C. The radiolabeled MCT were fed in equimolar amounts (6.5 mmol/kg0.75), in either nonemulsified form or as 30% (v/v) emulsions, using polyoxyethylene sorbitan monooleate as an emulsifier. Expired CO2 was quantified and specific radioactivity (Bq/mumol) determined at 20-min intervals over 24 h, and serial blood samples were drawn for measurement of plasma hexanoate and octanoate by HPLC. The rate [mumol/(h.kg0.75)] and extent (mmol/kg0.75) of MCT oxidative utilization (i.e., composite of digestion, absorption and oxidation) were calculated from the kinetics of 14CO2 expiration. Emulsification increased both the rate and extent of MCT oxidative utilization by > 20% regardless of fatty acid chain length (P < 0.05). Maximal MCT oxidation rates occurred at 3.5 h after feeding and were sufficient to meet up to 48% of the piglets' energy expenditure. The maximal rate of tri-6:0 oxidative utilization exceeded that of tri-8:0 by 37% (P < 0.05), but the extent of utilization was not affected by fatty acid chain length. Plasma medium-chain fatty acid concentrations were in agreement with these observations, being higher in pigs fed emulsified MCT than in pigs fed nonemulsified MCT (P < 0.05) and being higher in pigs fed tri-6:0 than in animals fed tri-8:0 (P < 0.05). Collectively, these data demonstrate that newborn piglets can effectively digest, absorb and oxidize MCT, that utilization is improved with emulsification, and that tri-6:0 is utilized more rapidly than tri-8:0.

Analysis of Variance↗
PubMed1994

DQB1*0602 and DQA1*0102 (DQ1) are better markers than DR2 for narcolepsy in Caucasian and black Americans.

In the present study, we tested 19 Caucasian and 28 Black American narcoleptics for the presence of the human leucocyte antigen (HLA) DQB1*0602 and DQA1*0102 (DQ1) genes using a specific polymerase chain reaction (PCR)-oligotyping technique. A similar technique was also used to identify DRB1*1501 and DRB1*1503 (DR2). Results indicate that all but one Caucasian patient (previously identified) were DRB1*1501 (DR2) and DQB1*0602/DQA1*102 (DQ1) positive. In Black Americans, however, DRB1*1501 (DR2) was a poor marker for narcolepsy. Only 75% of patients were DR2 positive, most of them being DRB1*1503, but not DRB1*1501 positive. DQB1*0602 was found in all but one Black narcoleptic patient. The clinical and polygraphic results for this patient were typical, thus confirming the existence of a rare, but genuine form of DQB1*0602 negative narcolepsy. These results demonstrate that DQB1*0602/DQA1*0102 is the best marker for narcolepsy across all ethnic groups.

Black People↗
PubMed1994

Transformation of human osteoblasts to anchorage-independent growth by insoluble nickel particles.

Nickel compounds are well established by epidemiologic studies as human carcinogens. Although the carcinogenicity of nickel compounds has been studied in experimental animals and in a variety of cultured mammalian cells, there are only sporadic reports of nickel-induced transformation of human cells. In attempts to study the mechanisms of nickel-induced carcinogenesis in human cells, an immortalized human osteoblastic cell line (HOS) that could not grow in soft agar or form tumors in athymic nude mouse was repeatedly treated with a water-soluble nickel compound (NiCl2) or a less water-soluble nickel compound crystalline (NiS). After three rounds of NiS treatment, there was an increase in anchorage-independent (AI) colony formation. This was not found in untreated or NiCl2-treated cells. Ten AI colonies obtained from NiS-treated cells were isolated. All of these clones showed changes in cell morphology, including the appearance of uniform polygon shape, growth in multilayers, and heavy staining with Giemsa. Most of these clones were retested for their ability to grow in soft agar and showed growth efficiencies of 5 to 50%. It has been shown by other investigators that aggregate growth is well correlated with tumorigenic potential in viral or chemical transformants of HOS cells. Four of seven tested NiS-transformed clones were able to form large aggregates compared to their untransformed counterparts, and continued to proliferate in aggregate form when they were plated on 0.9% agar. Current investigations focus on the molecular and genetic changes induced by nickel compounds in these human cells.

Carcinogens↗
PubMed1994

[Construction of human glioma cDNA library].

Total RNA from a human glioma was extracted by acid guanidinium-thiocyanate-chloroform one step method, and poly(A+)RNA was separated by affinity chromatography on oligo (dT) cellulose. The cDNA was synthesized by using poly (A+) RNA as template, the length of cDNA being 0.2-5 kb. The glioma cDNA inserts could be cloned into lambda gt11 vector, the resulting recombinant DNA was packaged in vitro. The cDNA library of 1.12 x 10(5) pfu/ml was obtained, and the cloning efficiency was 4.8 x 10(3)/ng cDNA. This library might contribute an important basis to study of the structure and function of human glioma genes.

Brain Neoplasms↗
PubMed1994

NMDA receptor activation in differentiating cerebellar cell cultures regulates the expression of a new POU gene, Cns-1.

POU/homeobox genes encode transcription regulatory proteins that are important in defining cellular phenotypes. Expression of these genes may be critical for to the regulation of CNS cellular differentiation. We have identified a cDNA corresponding to a new member of the POU/homeobox gene family. Expression of RNA encoded by this new gene occurs predominantly in the CNS. Thus, this new gene was designated Cns-1. Cns-1 transcripts are expressed in differentiating cells cultured from the early postnatal cerebellum. Treatment of these cultured cells with NMDA results in an increase in the level of Cns-1 RNA. This increase is blocked by simultaneous treatment with the specific NMDA receptor antagonist amino-5-phosphonovaleric acid. Continued activation of the NMDA receptor allows maintenance of this new steady state level of Cns-1 mRNA for at least 5 d in these cultured cells. A transcription runoff assay suggests that this increase in the level of RNA is due, at least in part, to an increase in transcription from the Cns-1 gene. The NMDA-induced increase in Cns-1 mRNA was reduced by pretreatment with calcium chelators EGTA or 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) tetrakis(acetoxymethyl). These studies suggest that specific activation of the NMDA receptor in cultures of differentiating cerebellar cells increases Cns-1 gene expression and that calcium entry through the NMDA channel may be required for this response. This change in Cns-1 expression may modify phenotypic characteristics of these cultured cells.

Amino Acid Sequence↗
PubMed1994

[Hepatic ultrasonographic imaging and serum amino acid levels in cured schistosomiasis cases].

Ultrasonography (B mode) of liver and serum amino-acid levels were studied in 15 cases of schistosomiasis japonica who had been cured for more than 5-15 years. Typical B-ultrasonic images of hepatic fibrosis due to schistosomiasis were found in all patients which could be classified into three types, namely, the spotty type (I), the tortoise-shell type (II) and the net-patchy type (III) according to the patterns of echogenic bands. Moderate to marked echogenic thickening of portal vein wall, and dilation of portal and splenic veins were revealed in patients with type II and III images. Free amino acids in sera were determined by using HPLC-AAAS. The result revealed that the concentration of 8 nonessential and 3 essential amino acids (threonine, valine, tryptophan) were significantly decreased in the 15 patients. The ratio of aromatic to aliphatic amino acid was markedly elevated in 3 patients of type III. The serum amino acid imbalance was attributed to the severe hepatic damage and impaired liver function.

Adult↗
PubMed1993

Recombinant canditropsin, an extracellular aspartic protease from yeast Candida tropicalis. Escherichia coli expression, purification, zymogen activation, and enzymic properties.

A cDNA fragment which encodes the zymogen of canditropsin, the extracellular aspartic protease from the yeast Candida tropicalis (Togni,G., Sanglard, D., Falchetto, R., and Monod, M. (1991) FEBS Lett. 286, 181-185) was cloned into a T7 expression vector for the synthesis of the recombinant zymogen in Escherichia coli. Recombinant canditropsinogen (Ctg), which was expressed as inclusion bodies in the cytosol of E. coli, was refolded by dialysis from an 8 M urea solution and purified to homogeneity using chromatographies on Sephacryl S-300 and on MonoQ columns. The purified Ctg was converted into canditropsin by either acid activation or trypsin conversion. The specificity of the resulting recombinant canditropsin toward polypeptide substrates is significantly different from other aspartic proteases. Canditropsin hydrolyzes oxidized insulin B chain between Ala-Leu and many other minor cleavage sites. Canditropsin also hydrolyzes keratin and collagen, which are components of connective tissues known to be hydrolyzed by canditropsin during Candida infections. Canditropsin was strongly inhibited by the universal aspartic protease inhibitor pepstatin (Ki = 1.75 x 10(-8) M) and inactivated by two aspartic protease inactivators, DAN and EPNP. Canditropsin is weakly inhibited by leupeptin and antipain, with an apparent Ki of 1.74 x 10(-4)M and 1.5 x 10(-5) M, respectively.

Amino Acid Sequence↗
PubMed1993

Conformational instability of the N- and C-terminal lobes of porcine pepsin in neutral and alkaline solutions.

Pepsin contains, in a single chain, two conformationally homologous lobes that are thought to have been evolutionarily derived by gene duplication and fusion. We have demonstrated that the individual recombinant lobes are capable of independent folding and reconstitution into a two-chain pepsin or a two-chain pepsinogen (Lin, X., et al., 1992, J. Biol. Chem. 267, 17257-17263). Pepsin spontaneously inactivates in neutral or alkaline solutions. We have shown in this study that the enzymic activity of the alkaline-inactivated pepsin was regenerated by the addition of the recombinant N-terminal lobe but not by the C-terminal lobe. These results indicate that alkaline inactivation of pepsin is due to a selective denaturation of its N-terminal lobe. A complex between recombinant N-terminal lobe of pepsinogen and alkaline-denatured pepsin has been isolated. This complex is structurally similar to a two-chain pepsinogen, but it contains an extension of a denatured pepsin N-terminal lobe. Acidification of the complex is accompanied by a cleavage in the pro region and proteolysis of the denatured N-terminal lobe. The structural components that are responsible for the alkaline instability of the N-terminal lobe are likely to be carboxyl groups with abnormally high pKa values. The electrostatic potentials of 23 net carboxyl groups in the N-terminal domain (as compared to 19 in the C-terminal domain) of pepsin were calculated based on the energetics of interacting charges in the tertiary structure of the domain. The groups most probably causing the alkaline denaturation are Asp11, Asp159, Glu4, Glu13, and Asp118.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Effect of salt-loading and spontaneous hypertension on in vitro metabolism of [1-14C]linoleic and [2-14C]dihomo-gamma-linolenic acids.

The present study compared the effect of spontaneous hypertension and salt-loading on in vitro metabolism of 18:2n-6 (linoleic acid) and 20:3n-6 (dihomo-gamma-linolenic acid). Ten weanling spontaneously hypertensive rats (SHR) and 10 normotensive Wistar-Kyoto rats (WKY) maintained on a rodent lab chow were given tap water with (n = 5) or without (n = 5) addition of 1% NaCl for 4 weeks. Thereafter, animals were killed and liver microsomes were prepared. Aliquots of microsomes suspended in the phosphate-sucrose buffer containing MgCl2, ATP, CoA, and NADPH were incubated with 0.3 microCi of [1-14C]-18:2n-6 or [2-14C]-20:3n-6 at 37 degrees C for 15 min. The activity of delta 6- and delta 5-desaturases, and the distribution of radioactivity in different lipid fractions and in phospholipid fatty acids were determined. Results showed that both spontaneous hypertension and salt-loading suppressed the desaturation of radiolabeled 18:2n-6 and of 20:3n-6. Incubation of microsomes with [1-14C]-18:2n-6 resulted in 29% of radioactivity being associated with phospholipid fraction, of which 3% was associated with 18:3n-6. Incubation with radiolabeled 20:3n-6 resulted in 30% of the radioactivity being incorporated into phospholipids, of which 50% was associated with 20:4n-6 (arachidonic acid). Salt-loading suppressed the incorporation of radiolabeled fatty acids into phospholipids, more so in SHR than in WKY. Thus, salt-loading not only suppressed the desaturation of 18:2n-6 and 20:3n-6, but also interfered with the acylation of n-6 fatty acids into the phospholipid fraction.

Animals↗
PubMed1993

Superoxide production by neutrophils from diabetics and normal subjects in response to glucose and galactose.

Glucose added to the medium was found to enhance superoxide production by isolated circulating neutrophils from both diabetic and normal subjects, but quantitatively the enhancement decreased from 4 to 50 mmole/liter. Galactose up to 50 mmole/liter had no effect on superoxide production in cells from the control subjects, but appeared to depress it in those from diabetics. No correlations were found between indices of the degree of hyperglycemia (plasma glucose and hemoglobin A1c) and the magnitude of the respiratory burst in cells from diabetics. When the isolated cells from normal and diabetic subjects were restored to a medium containing glucose at the original concentration in plasma at phlebotomy, the rate of superoxide production was approximately doubled in every case and there was no significant difference between diabetic and normal cells. Preincubation of cells for 1 hr in the presence of 0-50 mmole/liter glucose or galactose prior to activation had no significantly depressant effect on the respiratory burst except at 50 mmole/liter glucose in diabetic cells. It is concluded that circulating neutrophils from the diabetic population under the conditions studied are just as competent as control cells in their ability to sustain superoxide production over a wide range of energy availability.

Adult↗
PubMed1993

An epidemiological model for HIV/AIDS with proportional recruitment.

A model for HIV transmission is formulated for a homosexual population of varying size, with recruitment into the susceptible class proportional to the active population size and with stages of progression to AIDS. Analysis of this model includes identifying the threshold that determines whether the disease dies out or proportions remain endemic and establishing criteria that determine whether the population size decays asymptotically exponentially to zero or grows asymptotically exponentially to infinity. In an analogous heterosexual model, the conservation of heterosexual contacts is shown to imply that this two-sex model reduces to the one-sex model.

Female↗
PubMed1993

Intracellular diversion of glycoprotein GP160 of human immunodeficiency virus to lysosomes as a strategy of AIDS gene therapy.

A potential gene therapy strategy against human immunodeficiency virus (HIV-1) is to disrupt the intracellular transport of viral proteins. We report here the binding and transporting of HIV-1 glycoprotein gp160 to lysosomes as a result of the expression of fusion genes consisting of soluble CD4 and lysosome targeting domains. The effective lysosome targeting domain tested includes a lysosomal protease zymogen, procathepsin D, and the COOH-terminal domains of three lysosome membrane proteins: lamp-1, lamp-2, and lysosomal acid phosphatase. We demonstrated that cell fusion (syncytium), caused by the transport of gp160 to the surface of HeLa-CD4+ cells, was completely abolished by the expression of these fusion genes. The lysosomal localization of gp160 in HeLa cells coexpressing CD4-fusion genes was also established. From pulse-chase experiments, we observed that gp160 and the fusion proteins were degraded, as expected of lysosomal activities. Additionally, T lymphoblastoid cells transiently and permanently expressing these fusion genes strongly retarded the propagation of human immunodeficiency virus type 1. Thus, these fusion genes can deprive HIV of newly synthesized envelope protein gp160 for the assembly of new virions and are potentially useful in gene therapy against AIDS.

Acid Phosphatase↗