Biomedical subjects

J Zheng

Publications and source records attributed to J Zheng.

At least 19 recordsLinked to original sources

PubMed1992

A vibrational analysis of the catalytically important C4-H bonds of NADH bound to lactate or malate dehydrogenase: ground-state effects.

We have measured the frequency of the carbon-hydrogen stretching mode of the pro-R and pro-S C4-H bonds of NADH in solution and when bound to pig heart lactate (LDH) or mitochondrial malate (mMDH) dehydrogenases. This is achieved by specifically deuterating the C4 pro-R or pro-S hydrogens of NADH and determining the frequencies of the resulting C4-D stretches by Raman difference spectroscopy. We find that the frequencies of the two C4-D stretching modes for the two bonds are essentially the same for the unliganded coenzyme. On the other hand, the position of the pro-S-[4-2H]NADH stretch shifts upward by about 23-30 cm-1 in its binary complex with either lactate or malate dehydrogenase relative to that observed in solution, while that for the bound pro-R-[4-2H]NADH is relatively unchanged. The fact that the frequency of the pro-R hydrogen is not significantly affected during complex formation suggests that the rate enhancements for reaction of substrate with NADH brought about by both pig heart LDH and mMDH apparently do not involve either stabilization or destabilization of the pro-R hydrogen of NADH in enzyme-coenzyme binary complexes, in agreement with previous chemical studies. That these proteins are able to regulate the frequencies of the two C4-D bonds differentially, and hence the electronic distributions in these bonds, has important implications for the stereochemical reactions catalyzed by the NAD dehydrogenases, and this is discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

The structure of hepadnaviral core antigens. Identification of free thiols and determination of the disulfide bonding pattern.

A set of wild-type and mutant human, woodchuck, and duck hepatitis viral core proteins have been prepared and used to study the free thiol groups and the disulfide bonding pattern present within the core particle. Human (HBcAg) and woodchuck (WHcAg) core proteins contain 4 cysteine residues, whereas duck (DHcAg) core protein contains a single cysteine residue. Each of the cysteines of HBcAg has been eliminated, either singly or in combinations, by a two-step mutagenesis procedure. All of the proteins were shown to have very similar physical and immunochemical properties. All assemble into essentially identical core particle structures. Therefore disulfide bonds are not essential for core particle formation. No intra-chain disulfide bonds occur. Cys107 is a free thiol buried within the particle structure, whereas Cys48 is present partly as a free sulfhydryl which is exposed at the surface of the particle. Cys61 is always and Cys48 is partly involved in interchain disulfide bonds with the identical residues of another monomer, whereas Cys183 is always involved in a disulfide bond with the Cys183 of a different monomer. WHcAg has the same pattern of bonding, whereas DHcAg lacks any disulfide bonds, and the single free sulfhydryl, Cys153 which is equivalent to Cys107 of HBcAg, is buried.

Amino Acid Sequence↗

PubMed1992

Spatial buffering of extracellular potassium by Müller (glial) cells in the toad retina.

We examined the role of Müller (glial) cells in buffering light-evoked changes in extracellular K+ concentration, [K+]o, in the isolated retina of the toad, Bufo marinus. We found evidence for two opposing Müller cell current loops that are generated by a light-evoked increase in [K+]o in the inner plexiform layer. These current loops, which are involved in the generation of the M-wave of the electroretinogram (ERG), prevent the accumulation of K+ in the inner plexiform layer by transporting K+ both to vitreous and to distal retina. In addition, under dark-adapted conditions, we found evidence for a Müller cell current loop that is generated by a light-evoked decrease in [K+]o in the receptor layer. This current loop, which is involved in the generation of the slow PIII component of the ERG, helps to buffer the light-evoked decrease in [K+]o throughout distal retina by transporting K+ from vitreous. The spatial buffering fluxes of K+ can be abolished by blocking Müller cell K+ conductance with 200 microM Ba2+. The separate contributions of the M-wave and slow PIII currents to Müller cell spatial buffering were isolated by various pharmacological treatments that were designed to enhance or suppress light-evoked activity in specific retinal neurons. Our results show that Müller cell K+ currents not only buffer light-evoked increases in [K+]o, but also buffer light-evoked decreases in [K+]o, and thereby diminish any deleterious effects upon neuronal function that could arise in response to large changes in [K+]o in the plexiform layers. Moreover, our results emphasize that spatial buffering currents generate many components of the electroretinogram.

Animals↗

PubMed1992

cAMP-dependent protein kinase and the protein kinase family.

The structure of the catalytic subunit of cAMP-dependent protein kinase, the first protein kinase structure to be solved, is reviewed. The general architecture of the enzyme is described as well as the active site regions associated with substrate binding and catalysis. In particular, the unique features of the protein kinase nucleotide fold are outlined. While the catalytic subunit is one of the simplest of the protein kinases, it nevertheless serves as a structural framework for the catalytic core of the entire protein kinase family which now includes over 200 important regulatory enzymes. The essential and conserved features of this core are summarized, and a preliminary model of myosin light-chain kinase, based on the structure of the catalytic subunit, is also discussed.

Amino Acid Sequence↗

PubMed1992

A cytomechanical investigation of neurite growth on different culture surfaces.

We have examined the relationship between tension, an intrinsic stimulator of axonal elongation, and the culture substrate, an extrinsic regulator of axonal elongation. Chick sensory neurons were cultured on three substrata: (a) plain tissue culture plastic; (b) plastic treated with collagen type IV; and (c) plastic treated with laminin. Calibrated glass needles were used to increase the tension loads on growing neurites. We found that growth cones on all substrata failed to detach when subjected to two to threefold and in some cases 5-10-fold greater tensions than their self-imposed rest tension. We conclude that adhesion to the substrate does not limit the tension exerted by growth cones. These data argue against a "tug-of-war" model for substrate-mediated guidance of growth cones. Neurite elongation was experimentally induced by towing neurites with a force-calibrated glass needle. On all substrata, towed elongation rate was proportional to applied tension above a threshold tension. The proportionality between elongation rate and tension can be regarded as the growth sensitivity of the neurite to tension, i.e., its growth rate per unit tension. On this basis, towed growth on all substrata can be described by the simple linear equation: elongation rate = sensitivity x (applied tension - tension threshold) The numerical values of tension thresholds and neurite sensitivities varied widely among different neurites. On all substrata, thresholds varied from near zero to greater than 200 mudynes, with some tendency for thresholds to cluster between 100 and 150 mudynes. Similarly, the tension sensitivity of neurites varied between 0.5 and 5.0 microns/h/mudyne. The lack of significant differences among sensitivity or threshold values on the various substrata suggest to use that the substratum does not affect the internal "set points" of the neurite for its response to tension. The growth cone of chick sensory neurons is known to pull on its neurite. The simplest cytomechanical model would assume that both growth cone-mediated elongation and towed growth are identical as far as tension input and elongation rate are concerned. We used the equation above and mean values for thresholds and sensitivity from towing experiments to predict the mean growth cone-mediated elongation rate based on mean rest tensions. These predictions are consistent with the observed mean values.

Animals↗

PubMed1992

Structural framework for the protein kinase family.

In this review, we have summarized the general structural features of the catalytic subunit of cAMP-dependent protein kinase, emphasizing those features that will very likely be conserved in all members of the protein kinase family. The overall secondary structure of the catalytic core will probably be conserved throughout the catalytic core, as will the active site regions associated with MgATP binding and catalysis. The mechanisms for activation and the role of protein phosphorylation are unique for each kinase. The structure of the catalytic subunit now provides a general framework for modeling other protein kinases. Although this is no substitute for a crystal structure for each protein kinase, this one structure, nevertheless, does provide major insights to the molecular organization of each of these enzymes.

Adenosine Triphosphate↗

PubMed1992

PII component of the toad electroretinogram.

1. The PII component of the electroretinogram (ERG) is comprised of the b-wave and the DC component and is thought to reflect bipolar cell activity. Although the b-wave is generated in large part by a K+/Müller cell mechanism, the origin of the DC component is unclear. In this paper we detail our investigation of the origin of the DC component. We hypothesize that the DC component is generated by a K+/Müller cell mechanism identical to that involved in b-wave generation. 2. We studied the ERG in the dark-adapted, isolated retina preparation of the toad, Bu fo marinus. We used K+ ion-sensitive microelectrodes (K+ISM), as well as conventional intra- and extracellular microelectrodes, to record [K+]o changes, the vitreal ERG, and Müller cell responses. 3. We used the excitatory amino acid receptor agonist N-methyl-DL-aspartate (NMDLA) to inhibit light responses of third-order neurons and thereby eliminate most of the ERG M-wave. In the absence of the M-wave, the ERG consisted of PII and PIII. We then superfused the retina with a solution containing both kynurenic acid (KYN) and 2-amino-4-phosphonobutyric acid (APB), which together inhibit all retinal responses proximal to the photoreceptors. In the presence of KYN and APB, the ERG consisted only of PIII. Using digital subtraction, we reconstructed PII. To our knowledge, this is the first report of the isolation of a PII component in the ERG of a nonmammalian species. 4. Using K+ISMs, we recorded the distal K+ changes in the outer plexiform layer (OPL).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

[Studies on negative conversion of schistosome-infected snails in mountainous region of Yunnan].

The monthly cercaria shedding of 90 artificially infected Oncomelania snails was observed under a field condition in mountainous region, Shitoudi Village, Weishan County, Yunnan Province. The results showed that 20% of teh snails shed cercaria monthly, 42.2% could shed irregularly and 37.8% stopped releasing cercariae after several times of shedding. Final direction of the snails showed that none cercaria or sporocyst could be found in part of the snails. A total of 304 naturally infected snails were observed individually every month. Samples were taken each month from the snails which did not shed cercaria, then dissected and examined. The negative conversion rate was calculated in the second, third, fourth, fifth and sixth month after the first shedding, which were 36.1, 50.0, 41.0, 39.8 and 2.6% respectively.

Animals↗

PubMed1991

The determination of the pKa of histidine residues in proteins by Raman difference spectroscopy.

Sensitive Raman difference spectroscopy was used to monitor the protonation and deprotonation of histidine residues in apo-transferrin. We have shown previously that the behavior of small molecules and/or small molecular groups bound to proteins or other large macromolecules can be studied by Raman difference spectroscopy (Yue, K.T. et al. (1989) J. Raman Spectrosc. 20, 541-545). Using this method, we have measured the Raman difference spectra of human transferrin at different pH values with respect to pH 8.9, titrating its various histidine residues. About 12 +/- 2 of the 19 residues were titrated. The pH difference spectrum of transferrin obtained is very similar to that of histidine in solution, but with clear differences in the 1200-1400 cm-1 region. A titration curve with pKa of 6.08 +/- 0.01 fit the data of histidine in solution and a value of 6.56 +/- 0.02 was found for the average value of the 12 histidine residues inside transferrin. The technique has enough sensitivity at present to monitor a single histidine residue in a 130 kDa molecule and to determine the titration curve of one residue in a 40 kDa protein.

Apoproteins↗

PubMed1991

Case of atypical cyclopia.

A case of atypical cyclopia, involving the right eye with double lens, cornea, iris, and ciliary body, is reported. The possible mechanism of the histogenesis is discussed, together with a review of the relevant literature.

Abnormalities, Drug-Induced↗

PubMed1991

A generic algorithm for finding restriction sites within DNA sequences.

This paper describes a generic algorithm for finding restriction sites within DNA sequences. The 'genericity' of the algorithm is made possible through the use of set theory. Basic elements of DNA sequences, i.e. nucleotides (bases), are represented in sets, and DNA sequences, whether specific, ambiguous or even protein-coding, are represented as sequences of those sets. The set intersection operation demonstrates its ability to perform pattern-matching correctly on various DNA sequences. The performance analysis showed that the degree of complexity of the pattern matching is reduced from exponential to linear. An example is given to show the actual and potential restriction sites, derived by the generic algorithm, in the DNA sequence template coding for a synthetic calmodulin.

Algorithms↗

PubMed1991

Premercapturic acid metabolites of bromobenzene derived via its 2,3- and 3,4-oxide metabolites.

1. A new premercapturic acid metabolite of bromobenzene was isolated from the urine of beta-naphthoflavone-induced rats; using 1H-n.m.r., FAB mass spectrometry and chemical degradation it was identified as S-(2-hydroxy-3-bromocyclohexa-3,5-dienyl)-N-acetylcysteine. 2. Two regioisomeric premercapturic acids apparently derived from bromobenzene-3,4-oxide were isolated as an inseparable 1:1 mixture from the urine of phenobarbital-induced rats and characterized by similar means. 3. Acid dehydration of bromobenzene 3,4- and 4,3-premercapturic acids (mixture) afforded only p-bromophenylmercapturic acid, whereas acid dehydration of 3,2-premercapturic acid gave both o- and m-bromophenylmercapturic acids. This implies a shift of sulphur in acid dehydration of the 3,4- and 3,2- but not the 4,3-premercapturic acids. 4. Base dehydration of the 3,4- and 4,3-premercapturic acid mixture gave a mixture of p- and m-bromophenylmercapturic acids, whereas base dehydration of the 3,2-premercapturic acid gave only m-bromophenylmercapturic acid. This indicates these premercapturic acids dehydrate by direct elimination without rearrangment. 5. The 3,2-premercapturic acid was detected only in the urine of BNF-induced animals, whereas the 3,4- and 4,3-premercapturic acids were detected in the urines of untreated as well as PB- and BNF-induced animals. 6. Together with earlier reports of the isolation of the 2,3-dihydrodiol, the isolation of the 3,2-premercapturic acid as a urinary metabolite of bromobenzene implies that bromobenzene-2,3-oxide is a discrete metabolite of bromobenzene and not merely a hypothetical intermediate.

Acetylcysteine↗

PubMed1991

[Screening of active anti-inflammatory, immunosuppressive and antifertility components of Tripterygium wilfordii. III. A comparison of the antiinflammatory and immunosuppressive activities of 7 diterpene lactone epoxide compounds in vivo].

The half-effective dose(ED50), therapeutic index(TI) and certain safety factor(CSF) of 7 diterpene lactone epoxide compounds with antiinflammatory and immunosuppressive action extracted from Tripterygium wilfordii (TW) were assayed using croton oil-induced ear swelling and hemolysin-antibody formation mouse models. The results indicated that triptolide(T10), tripchlorolide(T4), triptonide(T7), tripdiolide(T8), triptolidenol(T9) and 16-hydroxytriptolide(L2) possessed both anti-inflammatory and immunosuppressive activities, while triptriolide(T11) had antiinflammatory activity only. The TI of the antiinflammatory action of 7 compounds were arranged in following order: T11(greater than 19), T10(17), T9(9.6), T4(9.0), T8(7.3), L2(6.6), T7(5.9), while the TI of the immunosuppressive action of 6 compounds were arranged in following order: T9(30.7), T4(16.7), L2(15.8), T10(13.7), T8(8.8), T7(7.5). The CSF parameters of both the activities of 7 compounds were all higher than 1. In which, the CSF of the immunosuppressive action of T9, T4 and L2 were 7.1, 5.1 and 3.6 respectively. These facts clearly demonstrate that the antiinflammatory and immunosuppressive compounds in TW are pluralistic. These differences among these compounds can be used as one of the bases for the evaluation and selection of these compounds. The above experiments conclude that the practical value of these compounds, however, will also depend on their content and yield rate in the herb, on the degree of difficulty in synthesizing these compounds, and on difficulty involved in the preparation of their derivatives. The future of these compounds will depend on the results of genetic toxicology studies as well.

Animals↗

PubMed1991

[Screening of active anti-inflammatory, immunosuppressive and antifertility components from Tripterygium wilfordii. IV. A comparison of the male antifertility activities of 7 diterpene lactone epoxide compounds].

The male antifertility activities under equal toxic dosages 1/10-1/60 LD50 and hemolysin antibody formation parameters under effective threshold dosages respectively of 7 diterpene lactone epoxide compounds extracted from Tripterygium wilfordii (TW) were assayed in mice. The results indicated that among the 7 compounds, 6 (T10, T4, T7, T8, T9 and L2) showed antifertility activities, the effective thresholds of T7, T8, T10 and L2 were below 1/40 LD50, those of T4 and T9 were below 1/60 LD50, while the other one (T11) did not. The above results reconfirm that the antifertility and immunosuppressive activities of all 6 active compounds are inseparable, but the formation of hemolysin antibodies was not inhibited under the total dosages which resulted in antifertility. The lowest dosages which were positive in terms of antifertility activity were 5-28 times lower than those for antiinflammatory activity and 5-12 times lower than those for immunosuppressive activity. And the total dosages necessary for effective immunosuppression were 0.9-1.8 times than the total dosages needed for effective antifertility. Finally, the value for the use of antifertility activity of these compounds, the influence of antifertility on antiinflammatory and immunosuppressive therapy, and the use of mouse models for screening of male antifertility activity are discussed.

Animals↗

PubMed1991

[Retinoblastoma-culture in vitro and establishment of cell line].

60 cases of retinoblastoma cells have been cultured in vitro in recent 6 years. 10 of them grew more than 3 months, and one continuous retinoblastoma cell line (SO-Rb50) has been established for more than 3 years. In addition to the degree of malignancy of the tumor cells, selection of tumor tissues, components of culture media, optimal selection of seeding, passage and densities of cells play important roles in the establishment of immortal retinoblastoma cell line.

Culture Media↗

PubMed1991

Tensile regulation of axonal elongation and initiation.

Neurites of chick sensory neurons in culture were attached by their growth cones to glass needles of known compliance and were subjected to increasing tensions as steps of constant force; each step lasted 30-60 min and was 25-50 mu dyn greater than the previous step. After correcting for elastic stretching, neurite elongation rate increased in proportion to tension magnitude greater than a tension threshold. The value of the tension threshold required for growth varied between 25 and 560 mu dyn, with most between 50 and 150 mu dyn. The growth sensitivity of neurites to tension was surprisingly high: an increase in tension of 1 mu dyn increased the elongation rate an average of about 1.5 microns/hr. The linear relationship between growth rate and tension provides a simple control mechanism for axons to accommodate tissue expansion in growing animals that consistently maintains a moderate rest tension on axons. Styrene microspheres treated with polyethyleneimine were used to label the surface of neurites in order to determine the site and pattern of surface addition during the experimental "towed growth" regime. New membrane is added interstitially throughout the neurite, but different regions of neurite vary widely in the amount of new membrane added. This contrasts with membrane addition specifically at the distal end in growth-cone-mediated growth. The different sites for membrane addition in growth mediated by towing and by the growth cone indicate that the membrane addition process is sensitive to the mode of growth. We confirmed the finding of Bray (1984) that neurites can be initiated de novo by application of tension to the cell margin of chick sensory neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

A knowledge-based experimental design system for nucleic acid engineering.

Presented in this paper is a knowledge-based experimental design system that incorporates the domain expertise used in nucleic acid engineering, thus automating the processing of error-prone, laborious low-level work, and many decision-making steps, and guiding the biologist toward a workable plan. This allows the biologist to work at a higher abstraction level, concentrating on more fundamental, difficult and challenging problems directly related to protein structure - function relationships. Cassette-based site-directed mutagenesis and synthetic gene designs are used as examples to illustrate the utility of the knowledge-based system approach to experimental design.

Algorithms↗

PubMed1990

[Life span and cercaria shedding of schistosome-infected snails in mountain region of Yunnan].

The life span and cercaria shedding of infected Oncomelania snails in a mountain region of Shitoudi village, Weishan County, Yunnan Province were observed in simulated local ecological environments. 135 infected snails were isolated for observation 3 months after exposure to miracidia in August, 1987. The snail survival rate from the day of initial cercaria shedding to next June, July, August and September was 27.4, 16.3, 13.3 and 11.9% respectively, and the average number of cercariae shed was 139.9, 29.6, 39.2 and 75 per month respectively. The average life span of infected snails was 171.6 days. The average number of cercariae shed per snail in its whole life was 673.0. It was estimated that the average patent period of infected snails was over half a year. As this is the first report in our country in respect to the life span and cercariae shedding of infected snails in a mountain region, the result might be useful for quantitative analysis of epidemiological factors of schistosomiasis in this kind of endemic areas as well as for formulation of control strategy.

Animals↗