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Biomedical subjects

Y L Lin

Publications and source records attributed to Y L Lin.

At least 127 records · Page 7Linked to original sources

Cottontail rabbit papillomavirus L1 protein-based vaccines: protection is achieved only with a full-length, nondenatured product.

Papillomas induced by the cottontail rabbit papillomavirus (CRPV) progress at a high frequency to carcinomas. In this regard, CRPV and its tumors can serve as an animal model for highly oncogenic human papillomaviruses. We have previously shown that immunization with major structural protein L1 elicits neutralizing antibodies and protects rabbits from papilloma development (Y.-L. Lin, L.A. Borenstein, R. Selvakumar, R. Ahmed, and F.O. Wettstein, Virology 187:612-619, 1992). In this study, we demonstrated that vaccination with the TrpE-L1 fusion protein not only protected rabbits from papilloma development but also prevented latent infection. This was indicated by the failure to amplify CRPV sequences by polymerase chain reaction in biopsies from infection sites of immunized animals. Furthermore, we showed that TrpE-L1 immunization protected rabbits from papilloma formation induced by virus but not from that induced by viral DNA. To explore the possibility of developing vaccines based on L1 subfragments, we mapped the linear L1 epitopes recognized by TrpE-L1-immunized rabbits and by virus-infected rabbits resistant to superinfection. Sera from papilloma-bearing rabbits reacted with one major epitope located at the carboxy-terminal end of L1, between amino acids (aa) 480 and 505. A second epitope, and in some animals a third one, was located in the amino-terminal region, between aa 78 and 101, as well as between aa 37 and 62. Sera from TrpE-L1-immunized animals recognized only one major epitope, located between aa 6 and 37. Immunization of rabbits with L1 subfragment fusion proteins led to seroconversion, but no neutralizing antibodies were produced and the animals were not protected against papilloma formation. The data indicate that a successful papillomavirus vaccine must be based on immunization with full-length native L1 and that further simplification to smaller peptides containing major linear epitopes is not feasible.

Animals↗

[Effects of bromocriptine microinjection into VTA on the DOPAC level in nucleus accumbens].

The experiment was performed in sodium pentobarbital anesthetized male SD rats. In vivo differential pulse voltammetry (DPV) was used to monitor a metabolite of dopamine-3,4-dihydroxyphenylacetic acid (DOPAC) level in the ipsilateral nucleus accumbens following microinjection of bromocriptine (2.5 micrograms/0.5 microliters, n = 5) into left ventral tegmental area (VTA). The results indicated that after microinjection the amplitude of DOPAC peak increased gradually and reached to 128% and 143% compared to the control at the 50th and 80th min respectively (P < 0.01). The duration of the effect was about two hours. One hundred and eighty min after injection, the amplitude of DOPAC decreased to 111% of control (P > 0.05). SCH23390 (2 nmol/0.5 microliters, n = 4), a selective D1 antagonist was given into left VTA 40 min prior to microinjection of bromocriptine. In this group, the above-mentioned effect of bromocriptine was blocked, the amplitude of DOPAC peak showed no increase. The results suggest that the microinjection of bromocriptine into VTA induces an increase of activity of mesolimbic dopaminergic neurons probably mediated by D1 receptor.

3,4-Dihydroxyphenylacetic Acid↗

[Uses of monomolecular lipid film for control of udo aphid (Toxoptera odinae (Van der Goot))].

The biological character of udo aphid was observed and monomolecular lipid film was used to control it. The results showed that at 100 and 200 times the lipid film, the hatch rate of over-winter eggs was lower than the control by 30.76% and 12.15% respectively, and at 50, 100 and 200 times the lipid film the mortality of young nymph aphids was 100% and those of old nymph aphids and adult aphids were 8.31%, 32.62% and 3.31% respectively.

Animals↗

Activation of human platelets by the rabbit anticardiolipin antibodies.

Affinity purified anticardiolipin antibodies (ACLA) raised in rabbits showed cross-reactivities with various negatively charged phospholipids as shown by both the solid phase enzyme-linked immunosorbent assay (ELISA) and inhibition studies. In ELISA, ACLA showed strong cross-reactivity to both sphingomyelin (SM) and phosphatidylethanolamine (PE), but the inhibition studies showed that ACLA failed to bind the aqueous suspensions of SM, PE, and PE/PC (1:1). ACLA bound to resting gel-filtered human platelets (GFP) as shown by both inhibition study and flow cytofluorometric analysis. Western blotting procedure showed that ACLA strongly cross-reacted to an 80-Kd plasma membrane protein. ACLA activated platelet response in a concentration-dependent manner. At less than 10 micrograms/mL, ACLA induced both platelet shape change to spiculate irregular forms as shown by scanning electron microscopy and the phosphorylation of 20-Kd protein. ACLA at more than 10 micrograms/mL caused platelet aggregation and secretion. The aggregation was inhibited by EDTA; aspirin; antimycin A plus 2-deoxyglucose; PGE1; and the F(ab')2 fragment of ACLA. It was not inhibited by monoclonal antibody to Fc receptor (MoAb FcR2). The biochemical events of ACLA-induced platelet response involved the elevation of (1) thromboxane A2 formation, (2) cytosolic free calcium ion concentration ([Ca2+]i), and (3) 47-Kd protein phosphorylation. In addition, the subaggregatory concentration of ACLA showed synergistic platelet activation with that concentration of thrombin, collagen, and epinephrine. The study showed the mechanism involved in ACLA-induced platelet responses.

Animals↗

Effective vaccination against papilloma development by immunization with L1 or L2 structural protein of cottontail rabbit papillomavirus.

Immunization of rabbits with either L1, the major structural protein, or L2, a minor structural protein of cottontail rabbit papillomavirus (CRPV), protected against challenge with the virus. Neutralizing antibodies were elicited by both the L1 and L2 trpE fusion proteins. Neutralization with anti-L1 serum, however, was more efficient than with anti-L2 serum. In contrast, when tested on Western blots the immune response to L2 was stronger than to L1. Rabbits were also protected against CRPV infection by immunization with L1 expressing recombinant vaccinia virus. Sera from two of three rabbits immunized with recombinant vaccinia virus were negative on Western blots but all three were positive in ELISA's with nondenatured fusion protein or in immunoprecipitations. The results suggest that both the viral structural proteins, L1 and L2, merit consideration in the development of a vaccine against papillomavirus.

Animals↗

Identification of three transforming proteins encoded by cottontail rabbit papillomavirus.

Cottontail rabbit papillomavirus (CRPV) provides an animal model for human papillomaviruses associated with a high risk of cancer development. So far, nothing is known about the transforming functions of CRPV genes because of the lack of an assay system. We have recently developed two systems to assay for CRPV transforming functions. One is based on the finding that transformation of NIH 3T3 cells by CRPV is considerably increased by deleting sequences in open reading frame L2. The second one is based on the use of a cottontail rabbit skin epithelial cell line, sf1Ep (C. Meyers and F. O. Wettstein, Virology 181:637-646, 1991). Mutations were introduced which abolished expression of the full-length E6 protein (LE6), the short E6 protein (SE6) initiated at the second ATG of E6, the E7 protein, or the E5 protein. Mutations affecting LE6 or E7, but not SE6, reduced transformation of NIH 3T3 and sf1Ep cells. Transformed NIH 3T3 cell lines with mutations in LE6 and E7 did not grow in soft agar, while those with mutations in SE6 and E5 grew with a reduced efficiency. The cell lines with mutations in LE6, SE6, or E7 still did induce tumors in nude mice. These mutations, however, abolished the ability to induce papillomas in rabbits. When expressed individually with a retroviral vector, LE6, SE6, or E7, but not E5, conferred anchorage-independent growth. The level of viral protein expression in these cell lines was generally low, and a comparison of the abundance of virus-specific mRNA showed that cell lines contained 20 to 50 times less mRNA than a cottontail rabbit papilloma. These data demonstrate that CRPV encodes at least three transforming proteins.

3T3 Cells↗

Antitumor effects of new-type recombinant interleukin-2.

Two new human recombinant interleukin-2 (rIL-2), 125-Ser-rIL-2, and 125-Ala-rIL-2, were generated by protein engineering technique. Both of them maintained the proliferation of natural killer (NK) cells, CTLL-2 cells and their long-term propagations. The mutated new rIL-2 also enhanced the bioactivity of NK cells and the cytotoxicity of tumor-infiltrating lymphocytes (TIL) against the target tumor cells. The above results were all compared with that of the native rIL-2 and a similarity between them was found, which indicates that new type rIL-2 could be used for adoptive immunotherapy of malignant diseases.

Animals↗

Proopiomelanocortin (POMC)-derived peptides and sleep in the rat. Part 2--Aminergic regulatory processes.

Apomorphine (Apo), a D1/D2 Dopamine (DA) agonist, at high doses (500 micrograms/kg) induces a short-lasting insomnia, antagonized by a secondary injection of corticotropin-like intermediate lobe peptide (CLIP, 10 ng); these effects are also observed with hypophysectomized (hypoX) rats. The administration of the serotonin (5-HT) agonist 8-hydroxy-2-di-n-propylamino-tetralin (8-OHDPAT, 0.3 mg/kg) induces also an insomnia which, unlike Apo, is followed by a significant PS rebound. CLIP, again, antagonizes the 8-OHDPAT-induced insomnia. Finally, Bromocriptine (5 mg/kg), an agonist for both DA and 5-HT, induces first an insomnia (antagonized by CLIP), followed by a PS rebound; these effects persist in hypoX rats.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

[Immunological screening of clonorchiasis].

Crude antigens of Clonorchis sinensis adult worms were purified by Sephadex G-200 column chromatography. A purified antigen was then applied to detect clonorchiasis patients and parasite-free subjects by an enzyme-linked immunosorbent assay (ELISA) and an intradermal test (IDT). The results of the ELISA showed that specific antibody levels of IgG, IgM, IgA, IgE and total IgE in 371 infected patients were significantly higher than those of 181 parasite-free subjects (Student's t-test, p less than 10(-6) or p less than 0.005). The sensitivity was 92.8% and the specificity was 89.0%. Significant correlations between the intensity of infection, expressed as egg per gram feces, and specific IgG, IgA and total IgE concentration were observed (linear regression, p less than 0.0005). Among the 309 subjects tested intradermally with Clonorchis antigen, 128 showed a positive reaction while all of the 60 parasite-free subjects showed negative results. By this test, the sensitivity was 49% and the specificity of IDT was 100%. The specific IgG, IgA and total IgE antibody levels of the IDT positive patients were significantly higher than those of the negative subjects (Student's t-test, p less than 0.0005). There was no correlation between the induced swelling diameter and the intensity of infection in IDT positive clonorchiasis patients. Cross reactions in ELISA were observed in patients infected with Capillaria philippinensis, Sparganum proliferum and Toxocara canis, when the antigens prepared in the present study were used.

Animals↗

Morphological changes in the hypothalamic neurons of female rats exposed to continuous illumination.

The morphological changes of neurons in the paraventricular nucleus (PVN), suprachiasmatic nucleus (SCN), ventromedial nucleus (VMN), preoptic area (POA), supraoptic nucleus (SON) and arcuate nucleus (ARN) were studied in 16-18 month-old Long-Evans female rats which had been exposed to continuous illumination (LL) for 11 months. Most neurons in those hypothalamic areas showed compact cytoplasm with abundant Nissl substances and evenly distributed chromatin. These morphological features were not found in the same hypothalamic regions of rats placed under a regular alternative light-dark regimen (14h light and 10h dark). These findings suggest that decreased melatonin and elevated estrogen levels induced by long-term exposure to LL might be involved in the morphological changes in hypothalamic neurons of female rats.

Animals↗

Influence of long-term continuous illumination on the nuclear volume of hypothalamic neurons in female rats.

A light microscopic quantitative analysis was performed for studying the effects of long-term exposure to continuous illumination on hypothalamic morphology in female rats. Nuclear volumes of neurons in the suprachiasmatic nucleus, paraventricular nucleus, ventromedial nucleus and preoptic area were measured in 16-18 month-old Long-Evans female rats which had been exposed to continuous light for 11 months. Significant nuclear volume shrinkage could be observed in all investigated regions except for the preoptic area in these animals when compared to that of control rats kept under a photoperiod of 14h light and 10h dark. The relationship between morphological changes of hypothalamic neurons and the marked alteration of melatonin and estrogen levels induced by long-term exposure to continuous light is discussed.

Animals↗

An acute pleuropneumonia in a pig caused by Chromobacterium violaceum.

A 2.5-month-old, 30 kg Duroc pig died 10 days after showing clinical signs of dyspnoea and diarrhoea. Acute necrotizing and fibrinous pleuropneumonia with locally extensive lesions was found. Chromobacterium violaceum was isolated from pneumonic lung tissues and intratracheal inoculation of a pure culture into two SPF pigs reproduced lesions similar to those found in the natural infection.

Acute Disease↗

Effect of transmural versus nontransmural myocardial infarction on inducibility of ventricular arrhythmias during sympathetic stimulation in dogs.

Transmural myocardial infarction interrupts sympathetic nerves and denervates viable muscle distal to myocardial infarction. The effect of sympathetic stimulation on responses to programmed ventricular stimulation was studied in dogs without myocardial infarction (Group I: n = 5), with transmural anterior wall myocardial infarction (Group II: n = 6) and with nontransmural anterior wall myocardial infarction (Group III: n = 9). Ventricular effective refractory period during sympathetic stimulation decreased by 16 +/- 18, 1 +/- 2 and 12 +/- 8 ms (mean +/- SD) in viable muscle of the inferoapical left ventricle in Groups I, II and III, respectively, suggesting efferent sympathetic denervation by transmural myocardial infarction only. Sustained ventricular tachycardia or fibrillation was induced more easily during sympathetic stimulation in six of the six dogs with transmural infarction, but in only two of the nine dogs with nontransmural infarction (p less than 0.01). It is concluded that the partial sympathetic denervation produced by transmural myocardial infarction enhances the ease of induction of ventricular tachycardia and fibrillation during sympathetic stimulation. A similar mechanism may lead to increased risk for lethal arrhythmias during periods of high sympathetic tone in patients with transmural myocardial infarction.

Animals↗

Transformation of Heat-Treated Clostridium acetobutylicum Protoplasts with pUB110 Plasmid DNA.

Heat treatment of Clostridium acetobutylicum SA-1 protoplasts at 55 degrees C for 15 min before transformation resulted in expression in this microorganism of the kanamycin resistance determinant associated with plasmid pUB110. No heat treatment, or heat treatment at 65 or 44 degrees C for various time intervals, resulted in no kanamycin resistance transformants being recovered on selective kanamycin-containing regeneration medium. DNase plate assay indicated that treatment at 55 degrees C for 15 min completely inactivated the DNase activity associated with SA-1 protoplasts. Treatment of protoplasts at 65 or 55 degrees C for various periods under simulated transformation conditions had an inhibitory effect, although prolonged treatment at 55 or 44 degrees C appeared to stimulate DNase activity. Inactivation of protoplast-associated DNase activity by heat treatment at 55 degrees C for 15 min correlated with successful expression of kanamycin resistance and suggests that an extremely active, heatsensitive, protoplast-associated DNase may be a factor in the polyethylene glycol-induced transformation of C. acetobutylicum SA-1 protoplasts. Plasmid pUB110 DNA was isolated from C. acetobutylicum SA-1 kanamycin-resistant (Km) transformant cultures by a modification of the procedure used for C. perfringens plasmids. Detection of pUB110 DNA was possible only when diethyl pyrocarbonate was incorporated into isolation protocols to inactivate DNase activity. Restriction studies further verified the presence of pUB110 DNA in C. acetobutylicum SA-1 Km transformants.

Journal Article↗