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Biomedical subjects

Z Du

Publications and source records attributed to Z Du.

At least 73 records · Page 4Linked to original sources

Structure of the autoregulatory pseudoknot within the gene 32 messenger RNA of bacteriophages T2 and T6: a model for a possible family of structurally related RNA pseudoknots.

A 36-nucleotide RNA with a sequence corresponding to the 5' end region of the gene 32 mRNA of bacteriophages T2 and T6 was analyzed by one- and two-dimensional NMR methods. NMR results provide clear evidence that the RNA is folded into a pseudoknot structure with two coaxial stems connected by two loops, in a classic pseudoknot topology. The pseudoknot is unusual in that one of the loops consists of only one nucleotide, which spans the major groove of a seven base pair helical stem. Imino proton resonances indicate the hydrogen bonding pattern within the pseudoknot, and two-dimensional NOE spectra provide information that describes many of the structural features. The temperature dependence of the UV absorption and imino proton exchange rates provides insight into the stability of the pseudoknot. A three-dimensional model of the pseudoknot that is consistent with our NMR data is presented, and features that may be important for stabilizing the pseudoknot structure are discussed. A substantial number of other putative RNA pseudoknots described in the literature have sequences and topologies that appear to be related to the T2 and T6 pseudoknots. We propose that these RNAs may be members of a family of pseudoknots related by a similar structural motif, which we refer to as "common pseudoknot motif 1" or CPK1. The bacteriophage T2/T6 pseudoknot can be considered a structural model for the CPK1 family. The common features of the CPK1 pseudoknots are a stem 2 with six or seven base pairs, a loop 1 consisting of a single adenosine, and a variable length stem 1 and loop 2. The first "dangling" nucleotide at the 3' end of the molecule probably stabilizes stem 2. The CPK1 family includes several of the retroviral pseudoknots associated with mRNA frameshifting and readthrough. The work presented here describes the first detailed NMR analysis of an RNA pseudoknot with an entirely natural nucleotide sequence.

Bacteriophages↗

Effect of isothermal radiofrequency radiation on cytolytic T lymphocytes.

Previous in vitro studies provide evidence that RF electromagnetic radiation modulates proliferation of human glioma, lymphocytes, and other cell types. The mechanism of RF radiation cell proliferation modulation, as well as mechanisms for effects on other cell physiologic endpoints, are not well understood. To obtain insight regarding interaction mechanisms, we investigated effects of RF radiation exposure on interleukin 2 (IL-2) -dependent proliferation of cytolytic T lymphocytes (CTLL-2). After exposure to RF radiation in the presence or absence of IL-2 cells were cultured at various physiological concentrations of IL-2. Treatment effects on CTLL-2 proliferation were determined by tritiated thymidine incorporation immediately or 24 h after exposure. Exposure to 2450 MHz RIF radiation at specific absorption rates (SARs) of greater than 25 W/kg (induced E-field strength 98.4 V/m) induced a consistent, statistically significant reduction in CTLL-2 proliferation, especially at low IL-2 concentrations. At lower SARs, 2450 MHz exposure increased CTLL-2 proliferation immediately after exposure but reduced 24 h postexposure proliferation. RF radiation effects depended on the mitotic state of the cells at the time of exposure. Comparison of the effects of temperature elevation and RF radiation indicated significant qualitative and quantitative differences.

Animals↗

Requirements for lymphocyte activation by unusual strains of simian immunodeficiency virus.

When residues 17 and 18 in nef of simian immunodeficiency virus strain SIVmac239 were changed from RQ to YE, the resultant virus was able to replicate in peripheral blood mononuclear cell cultures without prior lymphocyte activation and without the addition of exogenous interleukin-2, caused extensive lymphocyte activation in these cultures, and produced an acute disease in rhesus and pigtail macaques (Z. Du, S. M. Lang, V. G. Sasseville, A. A. Lackner, P. 0. Ilyinskii, M. D. Daniel, J. U. Jung, and R. C. Desrosiers, Cell 82:665-674, 1995). These properties are similar to those of the acutely lethal pathogen SIVpbj14 but dissimilar to those of the parental SIVmac239. We show here that the single change of R to Y at position 17 in nef of SIVmac239 is sufficient to confer the full, unusual phenotype. Conversely, the lymphocyte-activating properties of SIVpbj14 were lost by the single change of Y to R at position 17 of nef. The change of R17F or Q18E in SIVmac239 nef did not confer the unusual in vitro properties. Since SIVpbj14 has a duplication of the NF-kappaB binding sequence in the transcriptional control region, we also constructed and tested strains of SIVmac239/Rl7Y with zero, one, and two NF-kappaB binding elements. We found no difference in the properties of SIVmac239/R17Y, either in cell culture or in vivo, whether zero, one, or two NF-kappaB binding sites were present. Thus, tyrosine at position 17 of nef is absolutely necessary for the unusual phenotype of SIVpbj14 and is sufficient to convert SIVmac239 to a virus with a phenotype like that of SIVpbjl4. Multiple NF-kappaB binding sites are not required for the in vitro properties or for acute disease.

Amino Acid Sequence↗

Utilization of copper in copper proteinate, copper lysine, and cupric sulfate using the rat as an experimental model.

One hundred twenty male Sprague-Dawley rats, averaging 108.6 g initial body weight, were used in two feeding experiments to evaluate the utilization of Cu in Cu proteinate, Cu lysine, and cupric sulfate. In Exp. 1, 60 rats were randomly assigned to 12 treatments in a 2 x 2 x 3 factorial arrangement of treatments, with Zn supplementation at 0 or 1000 mg/kg diet, Cu supplementation at 5 or 15 mg/kg diet, and Cu form of CuSO4.5H2O, Cu proteinate, or Cu lysine. The purified basal diet contained .81 mg Cu, 20 mg Zn, and 60 mg Fe/kg diet. Experiment 2 was similar to Exp. 1 except Zn was replaced by Fe. In Exp. 1, feed intake of Cu proteinate (15.74 g/d) and Cu lysine (15.74 g/d) treatments was higher (P < .05) than that of CuSO4 (15.33 g/d). Body weight gain and feed intake were increased by high dietary Cu at either requirement or high levels of dietary Zn (P < .05). There were no differences in feed intake or body weight gain among the treatment groups in Exp. 2 (P > .05). The Cu utilization of Cu proteinate and Cu lysine were higher (P < .05) based on the liver Cu content. The rats fed Cu complexes had a higher liver Fe or Zn content (P < .05) than the rats fed CuSO4, suggesting that Cu complexes are absorbed via another mechanism that differs from that of inorganic Cu and does not interfere with Zn and Fe. Spleen Cu content may be a sensitive indicator of Cu status. High dietary Zn decreased Cu utilization, but this effect was overcome by high dietary Cu.

Animals↗

Copper metabolism of holstein and jersey cows and heifers fed diets high in cupric sulfate or copper proteinate.

Eight Holstein and 8 Jersey primiparous cows (3 d postcalving) and 8 Holstein and 8 Jersey growing heifers were randomly assigned to 1 of 8 treatments in a 2 x 2 x 2 factorial arrangement to compare Cu metabolism between Holsteins and Jerseys and the bioavailabilities of Cu in Cu proteinate and CuSO4. The variables were Holstein or Jersey, Cu supplementation at 5 or 80 mg/kg of DM, and supplements of CuSO4 or Cu proteinate. Jerseys had higher hepatic Cu concentrations than did Holsteins on d 60 (346 vs. 303 micrograms/g of DM). At the high Cu supplementation, hepatic Cu increased more rapidly, and content was higher in Jerseys than in Holsteins by d 60 (520 vs. 439 micrograms/g of DM). On d 0, plasma Cu concentrations were 0.99 and 0.80 microgram/ml, and, on d 60, concentrations were 0.96 and 0.88 microgram/ml for Jerseys and Holsteins, respectively. Overall, serum ceruloplasmin oxidase activity was greater for Jerseys than for Holsteins. Jersey cows and heifers also had greater hepatic Fe (208 vs. 173 micrograms/g of DM) and lower hepatic Zn (82 vs. 91 micrograms/g of DM) than did Holstein cows and heifers at d 60. The bioavailability of Cu in Cu proteinate and CuSO4 was the same. Plasma Cu concentration and ceruloplasmin have limited value as indicators of Cu status and availability to dairy cows and heifers.

Animals↗

Induction of lymphocyte proliferation and severe gastrointestinal disease in macaques by a nef gene variant SIVmac239.

The molecularly cloned virus known as SIVmac239/YEnef causes extensive lymphocyte activation in unstimulated peripheral mononuclear cell cultures and induces an acute disease syndrome in macaque monkeys. Here we describe the histopathological and immunophenotypic changes and viral localization in peripheral lymph nodes, spleen, and gastrointestinal tract (including the gut-associated lymphoid tissue (GALT) in rhesus monkeys inoculated with SIVmac239/YEnet beginning at day 3 postinoculation (pi). The findings are compared with those of rhesus monkeys inoculated with the same dose of parental SIVmac239. Histopathological examination of peripheral lymphoid tissue and GALT demonstrated marked hyperplasia of T-cell-dependent regions and involution of germinal centers as early as day 7 pi. The most striking lesions were multifocal areas of lymphohistiocytic gastroenteritis and colitis. Cellular infiltrates peaked between day 7 and 14 pi and were composed primarily of CD3+ T lymphocytes and HAM-56+ monocyte/macrophages. Many of these inflammatory cells were also strongly immunoreactive for teh nuclear proliferation antigen Ki-67. Despite the presence of severe gastrointestinal pathology by day 7 pi, no significant difference in the numbers of virus-positive cells in the gastrointestinal tract was observed between these animals and SIVmac239-infected animals examined at the same time point. However, the distribution of virus in the gastrointestinal tract was markedly different, with virus localized to lymphoid nodules of GALT in SIVmac239-infected animals and restricted to areas of lymphohistiocytic gastroenteritis and colitis in animals infected with SIVmac239/YEnef. Our data indicate that the acute disease syndrome induced by SIVmac239/YEnef is not simply related to increased viral replication in the gastrointestinal tract but is likely due to inappropriate virus-induced T lymphocyte activation and proliferation in GALT and subsequent mucosal destruction.

Animals↗

Target imaging diagnosis of human brain glioma. Clinical analysis of 40 cases.

OBJECTIVE: To establish a method for localization and qualitative diagnosis of glioma. METHODS: The monoclonal antibody SZ-39 against human glioma was labelled with 131I and injected intravenously into 40 patients with intracranial occupying lesions proved by X-CT. 72 hours after 131I-McAb-SZ39 administration, cranial bone nuclide-imaging agent 99mTc-MDP was injected intravenously, and the patients were examined by SPECT scan with dual nuclide double channel tomography and special software. On radio-immuno-image, the immunocomplex formed by specific conjugation of 131I-McAb-SZ39 with target tissue was red, while the cranial bone incorporated with 99mTc-MDP was green. The location of the immunocomplex area could be identified by the superimposition of the images. RESULTS: 21 patients with positive targeting diagnosis of glioma showed the accuracy rate checked by SPECT was 81%(17/21) and by X-CT 48% (10/21). Seven patients with negative targeting diagnosis showed the accuracy rate checked by SPECT was 86%(6/7) and by X-CT 14% (1/7). Pathologically, the sensitivity of targeting SPECT to glioma was 94.5%(17/18) and that of X-CT 55%(10/18). CONCLUSIONS: Targeting SPECT examination could be used for localization and qualitative diagnosis of glioma and make up the inadequacy of X-CT in the qualitative diagnosis of atypical incipient and recurrent glioma, meningioma, metastatic carcinoma, and inflammatory lesions of brain.

Adolescent↗

[Stability of jingu tongxial pills].

The stability of Jingu Tongxiao Pills was studied on the basis of accelerated destruction and sample-reserving observation at room temperature. According to the changes of tanshinone IIA in the sample, the time of efficacy determined by the two methods is 1.89 and 2.30 years respectively. Based on the analysis of other indices, the effective period is tentatively determined as two years.

Abietanes↗

Identification of a nef allele that causes lymphocyte activation and acute disease in macaque monkeys.

Residues 17 and 18 in nef of SIVmac239 were changed from RQ to YE to create a translated sequence of SRPSGDLYERLLRARGETYGRLLGEVEDGYSQSP from residues 10-43. The YXXL motifs in this context match very well with consensus sequences for SH2 binding domains and are similar to ones present in nef of the acutely lethal pathogen SIVpbj14. The YE variant of SIVmac239, unlike SIVmac239 but like SIVpbj14, replicated well in resting peripheral blood mononuclear cell cultures, caused extensive T lymphocyte activation, and produced an acute disease in rhesus and pigtailed monkeys characterized by severe diarrhea, rash, and extensive lymphoid proliferation in the gastrointestinal tract. The YEnef gene transformed NIH 3T3 cells in culture. Both 239nef and YEnef were found to associate with src in cotransfected COS cells, and both 60 kDa src and 34 kDa nef were phosphorylated at tyrosine in these cells. The extent of tyrosine phosphorylation of 239nef was considerably less than that of YEnef in these assays. These findings identify an important determinant of the SIVpbj14 phenotype, and they provide evidence of a role for nef in signal transduction and cellular activation.

3T3 Cells↗

[Investigation and experimental study on the standards and quality of leech pieces].

It has been confirmed by investigation that Whitmania pigra is the main medicinal breed of leech, and filiform pieces are the best for extraction. The quality of leech processed by 3 different methods has been studied qualitatively and quantitatively in terms of amounts of extraction, TLC, electrophoresis of protein and anti-thrombin action.

Animals↗

Complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII.

The complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII reveals that it contains 269 predicted or known genes (300 base pairs or larger). Fifty-nine of these genes (22 percent) were previously identified. Of the 210 novel genes, 65 are predicted to encode proteins that are similar to other proteins of known or predicted function. Sixteen genes appear to be relatively recently duplicated. On average, there is one gene approximately every 2 kilobases. Although the coding density and base composition across the chromosome are not uniform, no regular pattern of variation is apparent.

Base Composition↗

Structure of 1,1,5,5-tetranitro-[4]peristylane. Structure solution from molecular packing analysis.

The structure of the title compound, decahydro-2,2,5,5-tetranitro-1,6:3,4-dimethanocyclobuta[1,2: 3, 4]dicyclopentene, C12H12N4O8, was solved with the molecular packing program MOLPAK, starting with an AM1-geometry-optimized model of an isolated molecule. The 20 best predicted crystal structures from the MOLPAK procedure were subjected to lattice energy refinement with the WMIN program. A structure-factor calculation with the top MOLPAK/WMIN derived structure and the 197 Fo data to sin theta/lambda = 0.324 A-1 (theta = 30 degrees) gave an R value of 0.29. Final R = 0.057, wR = 0.067 for 730 reflections with I > 3 sigma (I). There are no unusual intermolecular distances and crystal packing is normal.

Bridged-Ring Compounds↗

Kinetic characterization of phosphoenolpyruvate carboxylase extracted from whole-leaf and from guard-cell protoplasts of Vicia faba L. (C3 plant) with respect to tissue pre-illumination.

Whole leaves and guard-cell protoplasts of the C3 plant Vicia faba L. (broad bean) were separately extracted following a period of illumination or following a period of darkness. Kinetic parameters of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), Vmax and Km(PEP.Mg), were determined as a function of assay pH (7.0 or 8.1), the presence of 5 mM glucose-6-Pfree (Glc-6-P, an activator), and the presence of 5 mM malatefree (an inhibitor). On the basis of these parameters, guard-cell PEPC was distinguished from that of whole leaf, indicating either that guard cells contain a unique isoenzyme of PEPC or a different complement of isoenzymes or--and less likely--that the obligatorily different methodologies for the leaf (intact organ) and the guard-cell (protoplast) enzymes altered them specifically. The values of Vmax were relatively unchanged, regardless of assay conditions or tissue pretreatment. The values obtained for whole-leaf PEPC Vmax were restricted to a small range (52.4 +/- 5.9 (SD) to 64.4 +/- 4.8 (SD) mumol.g fresh mass-1.h-1; the high value coincided with the presence of Glc-6-P, and the low value was obtained in the presence of malate. Guard-cell PEPC Vmax was also restricted to a small range: 7.48 +/- 0.89 (SD) pmol.guard-cell pair-1.h-1 (pH 8.1, light, +Glc-6-P) to 5.79 +/- 0.60 (SD) pmol.guard-cell pair-1.h-1 (pH 7.0, dark, +malate). Depending on effectors, and particularly pH, large changes in Km(PEP.Mg) were calculated (whole-leaf PEPC: 0.03 to 3.84 mM; guard-cell PEPC: 0.06 to 3.43 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

Fabaceae↗

Influence of penning type and feeding level on sexual behavior and feet and leg soundness in boars.

Ninety-six Yorkshire males at 28 d of age were placed on a pre-test feeding regimen (22% CP) of either ad libitum (A) access to feed or a restricted (R, 85% of A) diet. All pigs were housed in groups of four until approximately 30 kg body weight. The boars were then placed under test and assigned to a group pen (GP) containing eight boards or to individual pens (IP) and fed a 16% CP diet at A or R levels. Feeding types during pre-test and test phases were, therefore, ad libitum followed by ad libitum (AA), ad libitum: restricted (AR), restricted: ad libitum (RA), and restricted: restricted (RR) for each of the two (IP and GP) housing types. All boars were weighed every 2 wk to determine the feeding level and ADG. The pen floors were partly slatted with water nipples in the slatted areas. Areas available for each board were 1.63 and 2.15 m2 in IP and GP, respectively. Sexual behavior, semen characteristics, and feet and leg scores were recorded between 150 and 240 d of age. The IP boards required longer (P < .05) contact with receptive gilts before mating, made more (P < .01) ano-genital sniffings, and attempted more (P < .10) incorrect mounts than GP boars. The GP boards had a higher (P < .01) mating score, younger (P < .001) age at completion of the mating test, and lower (P < .01) total sperm count and sperm concentration (P < .001) than IP boars. The AA boards exhibited less (P < .10) chomping and salivation than boars on other feeding regimens.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Automated fluorescent DNA sequencing of polymerase chain reaction products.

The methods described in this chapter provide some useful approaches for DNA sequencing of templates produced by PCR. These procedures have been employed successfully for large-scale DNA sequencing of cosmid fragments subcloned in plasmid or M13 vectors, and for sequence analysis of cDNAs cloned in bacteriophage lambda vectors. In addition, the method describing direct sequencing from PEG-precipitated PCR product has been used successfully for analysis of Caenorhabditis elegans genomic and cDNA sequences. It is important to reiterate that for every combination of amplification primer pair and target DNA, there is an optimal method for PCR amplification; the ability to sequence the products of any PCR experiment directly will also vary. A coupled PCR/DNA sequencing method that works well for one experimental system may work quite poorly with others. Hence, a few days or hours spent optimizing PCR amplification conditions and selecting the best DNA sequencing method for the target DNA of interest will be time well spent.

Animals↗