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Biomedical subjects

W Lin

Publications and source records attributed to W Lin.

At least 271 records · Page 15Linked to original sources
PubMed1985

Linear sucrose transport in protoplasts from developing soybean cotyledons.

Previous studies with isolated soybean cotyledon protoplasts revealed the presence of a saturable, simple diffusion, and nonsaturating carrier-mediated uptake of sucrose into soybean cotyledon cells. A proton/sucrose cotransport may be involved in the saturable sucrose uptake (Lin et al. 1984 Plant Physiol 75: 936-940 and Schmitt et al. 1984 Plant Physiol 75: 941-946). In this study, we investigated the linear sucrose uptake mechanism by treating isolated protoplasts with 15 micromolar p-trifluoromethoxy-carbonylcyanide phenylhydrazone (FCCP) or 100 micromolar p-chloromecuribenzenesulfonic acid to eliminate the saturable uptake. We found: (a) increasing external pH decreases the linear sucrose uptake; (b) fusicoccin at 20 micromolar stimulates and FCCP at 15 micromolar inhibits this linear sucrose uptake; and (c) the ratio of the initial influx of proton to sucrose is close to one in both saturable and nondiffusive linear (difference between the total linear and diffusive components) uptakes. The results suggest that a proton/sucrose cotransport is also involved in the nondiffusive linear sucrose uptake into soybean cotyledon cells.

Journal Article↗
PubMed1984

Excretion of RNA catabolites by rats with hepatoma transplants.

Rats with transplants of Morris Hepatoma 5123 excreted in their urine greater than normal amounts of modified nucleosides and bases, catabolites of RNA. Despite rapid growth of the neoplasm, the elevated levels did not appear until 22 days after inoculation with the tumor. With tumor progression, there were increased levels and number of these catabolites. This study also suggests that the source of the elevated RNA catabolites is mainly from the host RNA rather than from tumor tissue and also that mRNA and rRNA as well as tRNA may contribute to the urinary levels.

Animals↗
PubMed1984

Resistance of chickens immunized against Mycoplasma gallisepticum is mediated by bursal dependent lymphoid cells.

Newly hatched chickens were significantly protected against challenge by the virulent S6 strain of Mycoplasma gallisepticum after vaccination with the TS 100 mutant. Removal of the thymus did not abolish the protective effect of the vaccine, but removal of the bursa of Fabricius did. The results suggest that the resistance induced by vaccine is mediated by the bursal-dependent lymphoid cells.

Animals↗
PubMed1984

Urinary catabolites of ribonucleic acid as cancer markers: a preliminary report of their use in patients with lung cancer.

Rats with aflatoxin-B1-induced hepatomas and dimethylnitrosamine-induced nephroblastomas excreted greater than normal amounts of urinary modified nucleosides and bases, catabolites of ribonucleic acid (RNA). Although both neoplasms caused increased excretions of the same catabolites, their quantitative profiles differed, suggesting that it may be possible to distinguish between tumors. Rats with transplanted tumors (e.g., hepatomas and osteogenic sarcomas) did not excrete elevated levels of urinary RNA catabolites until approximately 20 days after transplantation despite rapid growth of the tumor for the first 15 days. These data suggest that the source of the elevated levels of these excretory products may be the host's tissue RNA. Preliminary studies in human beings with lung cancer showed marked elevation of one or more urinary RNA catabolites. Resection of the diseased tissue in 2 patients caused a drop in levels. The measurement of urinary RNA catabolites may be useful in the diagnosis, prognosis, and evaluation of therapy in patients with lung cancer.

Aflatoxin B1↗
PubMed1984

Further Characterization on the Transport Property of Plasmalemma NADH Oxidation System in Isolated Corn Root Protoplasts.

Recent experiments show that exogenous NADH increases the O(2) consumption and uptake of inorganic ions into isolated corn (Zea mays L. Pioneer Hybrid 3320) root protoplasts (Lin 1982, Proc Natl Acad Sci USA 79: 3773-3776). A mild treatment of protoplasts with trypsin released most of the NADH oxidation system from the plasmalemma (Lin 1982 Plant Physiol 70: 326-328). Further studies on this system showed that exogenous NADH (1.5 millimolar) tripled the proton efflux from the protoplasts thus generating a greater electrochemical proton gradient across the plasmalemma. Trypsin also released ubiquinone (11.95 nanomoles per milligrams protein) but not flavin or cytochrome from the system. Kinetic analyses showed that 1.5 millimolar NADH quadrupled V(max) of the mechanism I (saturable) component of K(+) uptake, while K(m) was not affected. Diethylstibestrol and vanadate inhibited basal (ATPase-mediated) K(+) influx and H(+) efflux, while NADH-stimulated K(+) uptake was not or only slightly inhibited. p-Chloromercuribenzene-sulfonic acid, N,N'-dicyclohexylcarbodiimide, ethidium bromide, and oligomycin inhibited both ATPase- and NADH-mediated H(+) and K(+) fluxes. A combination of 10 millimolar fusicoccin and 1.5 millimolar NADH gave an 11-fold increase of K(+) influx and a more than 3-fold increase of H(+) efflux. It is concluded that a plasmalemma ATPase is not involved in the NADH-mediated ion transport mechanism. NADH oxidase is a -SH containing enzyme (protein) and the proton channel is an important element in this transport system. Fusicoccin synergistically stimulates the effect of NADH on K(+) uptake.

Journal Article↗
PubMed1984

Sugar transport into protoplasts isolated from developing soybean cotyledons : I. Protoplast isolation and general characteristics of sugar transport.

A procedure is described to isolated functional protoplasts from developing soybean (Glycine max L. Merr. cv Wye) cotyledons. Studies of sucrose and hexose uptake into these protoplasts show that the plasmalemma of cotyledons during the stage of rapid seed growth contains a sucrose-specific carrier which is energetically and kinetically distinct from the system(s) involved in hexose transport. For example, sucrose, but not hexose uptake: (a) is inhibited by alkaline pH and the nonpermeant SH modifier, p-chloromercuribenzene sulfonic acid; (b) is stimulated by fusicoccin; (c) shows both a saturable and a linear component of uptake in response to substrate concentration; and (d) displays a sharp temperature response (high Q(10) value and high activation energies).

Journal Article↗
PubMed1984

Sugar Transport into Protoplasts Isolated from Developing Soybean Cotyledons : II. Sucrose Transport Kinetics, Selectivity, and Modeling Studies.

The effects of metabolic inhibitors, pH, and temperature on the kinetics of sucrose uptake protoplasts isolated from developing soybean Glycine max L. cv Wye cotyledons were studied. Structural requirements for substrate recognition by the sucrose carrier were examined by observing the effects of potential alternate substrates for the saturable component on sucrose uptake.Uptake by the three components (saturable, sulfhydryl reagent-sensitive nonsaturable, and diffusive) was calculated over a range of sucrose concentrations. The saturable component dominated uptake at external sucrose concentrations below 12 millimolar and was approximately equal to the nonsaturable and diffusive components at 44 and 22 millimolar external sucrose, respectively. The three uptake components showed different temperature sensitivities.Increasing external pH decreased both the linear component and the V(max) calculated for the saturable component. Conversely, increasing pH increased the calculated K(m) (sucrose) for the saturable component.Sucrose uptake by the saturable component was insensitive to several mono- and divalent cations. Competition for uptake of 0.5 millimolar sucrose by several sugars suggested that the beta-d-fructofuranoside bond and molecular size of sucrose were particularly important in sugar recognition by the saturable component carrier.

Journal Article↗
PubMed1984

Sugar transport in isolated corn root protoplasts.

Isolated corn (Zea mays L.) root protoplasts were used to study sucrose and hexose uptake. It is found that glucose was preferentially taken up by the protoplasts over sucrose and other hexoses. Glucose uptake showed a biphasic dependence on external glucose concentration with saturable (K(m) of 7 millimolar) and linear components. In contrast, sucrose uptake only showed a linear kinetic curve. Sucrose and glucose uptake were linear over a minimum of 1 hour at pH 6.0 and 1 millimolar exogenous sugar concentration. Glucose uptake showed a sharp 42 degrees C temperature optimum, while sucrose uptake showed a lower temperature sensitivity which did not reach a maximum below 50 degrees C. Uptake of both sugars was sensitive to several metabolic inhibitors and external pH. Differences between sucrose and glucose uptake in two different sink tissue (i.e. protoplasts from corn roots and soybean cotyledons) are discussed.

Journal Article↗
PubMed1984

Subcellular distribution of viral structural proteins during simian virus 40 infection.

The amounts of simian virus 40 structural polypeptides Vp1, Vp2, and Vp3 in different subcellular fractions at various times after lytic infection were determined by a quantitative immunoblotting procedure. Simian virus 40-infected cells were lysed with a buffer containing Nonidet P-40 to yield a soluble fraction. The Nonidet P-40-insoluble fraction was further fractionated in the presence of deoxycholate and Tween 40 to yield a soluble fraction (cytoskeletal) and an insoluble fraction (Nuc), which is primarily cell nuclei. At 33 h postinfection, the majority of viral structural proteins was found in the cell nucleus, whereas, at 48 to 65 h postinfection, Vp1 was distributed evenly among all cell fractions and Vp2 and Vp3 were found predominantly in the cytoskeletal and Nuc fractions. Thus, not all of the viral polypeptides synthesized in the cytoplasm migrated into the cell nucleus. Throughout infection, the molar ratio (Vp3/Vp2) was rather constant in all subcellular fractions, indicating that the synthesis or processing or both of Vp2 and Vp3 are coordinately regulated. The molar ratio of Vp1/(Vp2 + Vp3) varied among the fractions. The Vp1/(Vp2 + Vp3) molar ratio in the soluble fraction varied during the course of infection; however, constant ratios were maintained in the cytoskeletal and Nuc fractions. Thus, the mechanism which controls the movement of Vp1 to different compartments of the cell appears to be different from that of Vp2 and Vp3. The Vp1/(Vp2 + Vp3) value in the Nuc fraction was similar to the ratio found in virus particles. The constant molar distribution of Vp1, Vp2, and Vp3 in the Nuc fraction throughout infection suggests that there is a specific mechanism which regulates the transport of viral structural proteins. These results support the hypothesis that the structural proteins of simian virus 40 are transported into the cell nucleus in precise proportions.

Animals↗
PubMed1984

Utilization of temperature-sensitive mutants of Mycoplasma gallisepticum to prevent air sac infections.

Three stable temperature-sensitive (ts) mutants were obtained by treating the S6 strain of Mycoplasma gallisepticum with 50 micrograms/ml of nitrosoguanidine. These mutants were morphologically and serologically indistinguishable from the parent S6 strain. Mutants ts 37 and ts 102 were apathogenic, and ts 100 was moderately pathogenic to chickens when inoculated directly into the air sac. To turkeys, ts 37 remained apathogenic, ts 102 was slightly pathogenic, and ts 100 was highly pathogenic, Intranasal immunization of newly hatched chickens with any of the mutants resulted in antibody production, but only ts 100 and ts 102 protected chickens against experimental M. gallisepticum infections 3 weeks later. Vaccination of 2-day-old turkeys with ts resulted in erratic antibody response; however, 5 weeks after immunization, turkeys were able to resist challenge with the virulent S6 strain at a highly significant level.

Agglutination Tests↗
PubMed1983

On the electrotransfer of polypeptides from gels to nitrocellulose membranes.

The conditions which affect the elution of polypeptides from polyacrylamide gels by electrophoresis and polypeptide-nitrocellulose interactions have been studied. The rate of elution of polypeptides from a 15% sodium dodecyl sulfate-polyacrylamide gel is dependent on the molecular weight of the individual polypeptides, which is in agreement with the results of W. N. Burnette (Anal. Biochem. 112, 195 (1981)). We also observed that current density affects the rate of elution. Polypeptides smaller than 20,000 daltons pass through pores of 0.45 microns, but not through the pores of 0.1-microns nitrocellulose membranes during electrophoresis. The nonionic detergent NP-40 inhibits the binding of polypeptides to nitrocellulose and removes prebound polypeptides from the membranes. Amido black and Coomassie blue staining and destaining processes do not remove the bound polypeptides from the membranes, but may affect the antigenicity of polypeptides. Polypeptides immobilized on nitrocellulose can be stored at -70 degrees C for future use.

Antigens, Viral↗
PubMed1983

Comparison of urinary modified nucleosides and bases in rats with hepatomas and nephroblastomas.

Hepatomas were induced in rats with aflatoxin B1, and nephroblastomas with dimethylnitrosamine. Microscopic examination of livers of aflatoxin-treated rats revealed multinodular hepatocyte hyperplasia at 8 months, and by 13 months all rats had hepatomas. Nephroblastomas were observed by 4 months and by 8 months all rats had developed them. The urinary excretion of several modified nucleosides and bases by normal rats is dependent on body weight and reflects, to a certain extent, their concentrations in tissue tRNA. Increased levels of several modified nucleosides and bases were found in all rats that had cancer. Rats with hepatomas excreted essentially the same modified nucleosides and bases as did those with nephroblastomas; the quantitative patterns of excretion were different, however, suggesting that the urinary modified nucleosides and bases may be used to differentiate between neoplasms. Although the increase in urinary modified nucleosides and bases by tumor-bearing animals results primarily from more rapid turnover of neoplastic tRNAs, the data indicate that increased turnover of mRNA and possibly rRNA may occur in neoplastic tissue. Preliminary data suggest that increases in urinary modified nucleosides and bases may occur during a precancerous stage. The urinary pattern of modified nucleosides and bases by rats with hepatomas is altered if another primary tumor is present. The results obtained from these studies support the use of modified nucleosides and bases in urine as biochemical markers of cancer.

Aflatoxin B1↗
PubMed1983

Visualization of antigens attached to cytoskeletal framework in animal cells: colocalization of simian virus 40 Vp1 polypeptide and actin in TC7 cells.

Actin and the simian virus 40 viral structural polypeptide Vp1 are observed to be present on cytoskeletal fibers of virus-infected TC7 cells, when these antigens in detergent-extracted whole cell mounts were labeled by specific antibodies and colloidal gold particles coated with a second antibody. In both cases, actin and Vp1 were found associated with fibers and fiber-associated electron-dense materials. Patches or clusters of colloidal gold particles denoting the presence of either Vp1 or actin were found on fibers uniformly distributed throughout the cytoplasm. By using simultaneous decoration of the two antigens with colloidal gold particles of different diameters, it was shown that the majority of Vp1 appears attached to cytoskeletal fibers in association with cellular actin. When Vp1 and actin were decorated with Imposil and ferritin simultaneously in infected cells that were fixed first and then permeabilized with saponin, both labels were found in the same spatial domain of the cell cytoplasm. Thus, the colocalization of Vp1 and actin on the cytoskeletal framework seems to reflect their actual state in the living cells. The electron-dense material to which colloidal gold particles localize in our cytoskeletal preparations may be the remnants of subcellular structures with which actin and Vp1 are both associated in intact cells.

Actins↗
PubMed1983

Pathway of Phloem unloading of sucrose in corn roots.

The pathway of phloem unloading and the metabolism of translocated sucrose were determined in corn (Zea mays) seedling roots. Several lines of evidence show that exogenous sucrose, unlike translocated sucrose, is hydrolyzed in the apoplast prior to uptake into the root cortical cells. These include (a) presence of cell wall invertase activity which represents 20% of the total tissue activity; (b) similarity in uptake and metabolism of [(14)C]sucrose and [(14)C]hexoses; and (c) randomization of (14)C within the hexose moieties of intracellular sucrose following accumulation of [(14)C] (fructosyl)sucrose. Conversely, translocated sucrose does not undergo apoplastic hydrolysis during unloading. Asymmetrically labeled sucrose ([(14)C](fructose)sucrose), translocated from the germinating kernels to the root, remained intact indicating a symplastic pathway for unloading. In addition, isolated root protoplasts and vacuoles were used to demonstrate that soluble invertase activity (V(max) = 29 micromoles per milligram protein per hour, K(m) = 4 millimolar) was located mainly in the vacuole, suggesting that translocated sucrose entered via the symplasm and was hydrolyzed at the vacuole prior to metabolism.

Journal Article↗
PubMed1983

Isolation of mesophyll protoplasts from mature leaves of soybeans.

A procedure based on a combined cellulase-Pectolyase Y-23 enzyme digestion and metrizamide-sorbitol gradient purification protocol was developed for isolating mesophyll protoplasts from mature leaves of soybean (Glycine max L. Merr.). Based on chlorophyll content, this procedure results in a 10 to 15% protoplast yield from fully expanded mature leaves and a 20 to 30% yield from young (expanding) leaves within 3 hours. Isolated protoplasts displayed high rates of HCO(3) (-)-dependent photosynthesis; greater than 75 micromoles O(2) evolved per milligram chlorophyll per hour at 25 degrees C. This photosynthetic rate is comparable to that of mesophyll cells isolated mechanically from the same leaves.

Journal Article↗
PubMed1983

Enhanced Mineral Uptake by Zea mays and Sorghum bicolor Roots Inoculated with Azospirillum brasilense.

Inoculation of corn (Zea mays) seeds with Azospirillum brasilense strain Cd or Sp 7 significantly enhanced (30 to 50% over controls) the uptake of NO(3), K, and H(2)PO(4) into 3- to 4-day- and 2-week-old root segments. No gross changes in root morphology were observed; altered cell arrangement in the outer four or five layers of the cortex was seen in photomicrographs of cross sections of inoculated corn roots. The surface activity involved in ion uptake probably increased, as shown by the darker staining by methylene blue of the affected area. Shoot dry weight increased 20 to 30% in inoculated plants after 3 weeks, presumably by enhancement of mineral uptake. Corn and sorghum plants grown to maturity on limiting nutrients in the greenhouse showed improved growth from inoculation approaching that of plants grown on normal nutrient concentrations. Enhanced ion uptake may be a significant factor in the crop yield enhancement reported for Azospirillum inoculation.

Journal Article↗