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Biomedical subjects

P Horton

Publications and source records attributed to P Horton.

At least 55 records · Page 3Linked to original sources

A probabilistic classification system for predicting the cellular localization sites of proteins.

We have defined a simple model of classification which combines human provided expert knowledge with probabilistic reasoning. We have developed software to implement this model and have applied it to the problem of classifying proteins into their various cellular localization sites based on their amino acid sequences. Since our system requires no hand tuning to learn training data, we can now evaluate the prediction accuracy of protein localization sites by a more objective cross-validation method than earlier studies using production rule type expert systems. 336 E. coli proteins were classified into 8 classes with an accuracy of 81% while 1484 yeast proteins were classified into 10 classes with an accuracy of 55%. Additionally we report empirical results using three different strategies for handling continuously valued variables in our probabilistic reasoning system.

Cells↗

A branch and bound algorithm for local multiple alignment.

A Branch and Bound Algorithm has been developed to find a set of window positions in a compilation of sequences with globally maximal information content. We have also developed an algorithm for brute force evaluation of solutions which is faster by a factor of the length of the windows than the naïve brute force algorithm. The combination of these two algorithms allows us to solve problems to optimality that were previously amenable only to heuristic algorithms.

Algorithms↗

Resonance Raman spectroscopy of the photosystem II light-harvesting complex of green plants: a comparison of trimeric and aggregated states.

Resonance Raman spectroscopy was performed on photosystem II light-harvesting complex (LHCII) in trimeric and oligomeric states with various excitation conditions. From these studies, it can be concluded that the structure of LHCII is altered during the trimer/oligomer transition. These structural changes affect the conformation of a population of carotenoid within the complex, which becomes twisted in the oligomeric form. Moreover, the interactions assumed by a chlorophyll a and a chlorophyll b are modified during the oligomerization process. This induces the formation of an H-bond to a formyl group of a chlorophyll b molecule and to a keto group of a chlorophyll a molecule. The extent to which these H-bonds to chlorophyll relate to the formation of the quencher cannot yet be precisely established. However, the structural changes they evidence may play a role in the control of the energy flux by LHCII complexes.

Carotenoids↗

Acclimation of Arabidopsis thaliana to the light environment: regulation of chloroplast composition.

The regulation by light of the composition of the photosynthetic apparatus was investigated in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta. When grown in high- and low-irradiance white light, wild-type plants and photomorphogenic mutants showed large differences in their maximum photosynthetic rate and chlorophyll a/b ratios; such changes were abolished by growth in red light. Photosystem I (PSI) and PSII levels were measured in wild-type plants grown under a range of light environments; the results indicate that regulation of photosystem stoichiometry involves the specific detection of blue light. Supplementing red growth lights with low levels of blue light led to large increases in PSII content, while further increases in blue irradiance had the opposite effect; this latter response was abolished by the hy4 mutation, which affects certain events controlled by a blue-light receptor. Mutants defective in the phytochrome photoreceptors retained regulation of photosystem stoichiometry. We discuss the results in terms of two separate responses controlled by blue-light receptors: a blue-high-fluence response which controls photosystem stoichiometry; and a blue-low-fluence response necessary for activation of such control. Variation in the irradiance of the red growth light revealed that the blue-high-fluence response is attenuated by red light; this may be evidence that photosystem stoichiometry is controlled not only by photoreceptors, but also by photosynthetic metabolism.

Acclimatization↗

Acclimation of Arabidopsis thaliana to the light environment: changes in photosynthetic function.

Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta was grown under light regimes of differing spectral qualities, which results in differences in the stoichiometries of the two photosynthetic reaction centres. The acclimative value of these changes was investigated by assessing photosynthetic function in these plants when exposed to two spectrally distinct actinic lights. Plants grown in an environment enriched in far-red light were better able to make efficient use of non-saturating levels of actinic light enriched in long-wavelength red light. Simultaneous measurements of chlorophyll fluorescence and absorption changes at 820 nm indicated that differences between plants grown under alternative light regimes can be ascribed to imbalances in excitation of photosystems I and II (PSI, PSII). Measurements of chlorophyll fluorescence emission and excitation spectra at 77 K provided strong evidence that there was little or no difference in the composition or function of PSI or PSII between the two sets of plants, implying that changes in photosynthetic stoichiometry are primarily responsible for the observed differences in photosynthetic function.

Adaptation, Physiological↗

An efficient route to N6 deoxyadenosine adducts of diol epoxides of carcinogenic polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons are metabolized to a wide variety of oxidized derivatives, including highly reactive diol epoxides which alkylate DNA. The reaction lacks regio- or stereospecificity but occurs primarily at the exocyclic amino groups of deoxyguanosine and deoxyadenosine. An efficient route to N6 adducts of deoxyadenosine is described using as examples those arising from trans opening of the anti-tetrahydrodiol epoxides of naphthalene, benzo[a]pyrene, and benzo[c]phenanthrene. The adducts were synthesized in 50-92% yields by reaction of 6-fluoropurine 2'-deoxyriboside with aminotriols formed by trans opening of racemic dihydrodiol epoxides using liquid NH3. The diastereomeric adducts were separated by HPLC and their absolute configurations were assigned by circular dichroism. 1H NMR studies revealed significant differences in conformation of the tetrahydroaromatic ring between the sterically unrestricted naphthalene derivative and the sterically congested derivatives of benzo[a]pyrene and benzo[c]phenanthrene. These differences may have a bearing on the higher carcinogenicity shown by the latter hydrocarbons. Undecadeoxyoligonucleotides bearing regio- and stereochemically defined adenine N6-anti-trans-benzo[a]pyrene adducts have been prepared.

Base Sequence↗

Higher plant light-harvesting complexes LHCIIa and LHCIIc are bound by dicyclohexylcarbodiimide during inhibition of energy dissipation.

We have investigated the binding to proteins of the photosynthetic apparatus of the carboxy-modifying agent dicyclohexylcarbodiimide, (cHxN)2C; this inhibits the protective dissipation of excess absorbed light energy (qE) by the light-harvesting apparatus of photosystem II (LHCII), suggesting that carboxyl amino-acid side chains within hydrophobic protein domains may be involved in qE. (cHxN)2(14)C was used to label thylakoids and photosystem II particles, so as to identify proteins which may be involved in the detection of lumen pH during qE induction. Of six thylakoid proteins labelled with (cHxN)2C under conditions where qE is efficiently induced, three are associated with photosystem I, and none with the bulk LHCII. PSII-associated label is bound to three minor components of LHCII, identified as LHCIIa (two species) and LHCIIc, as shown by protein sequencing of tryptic fragments of purified complexes. pH titration of qE formation and protein labelling in coupled thylakoids showed that both qE and labelling of LHCIIa increased at pH 7-8.

Amino Acid Sequence↗

The molecular mechanism of the control of excitation energy dissipation in chloroplast membranes. Inhibition of delta pH-dependent quenching of chlorophyll fluorescence by dicyclohexylcarbodiimide.

Non-radiative dissipation of absorbed excitation energy in chloroplast membranes is induced in the presence of the trans-thylakoid proton motive force; this dissipation is measured as high energy state quenching of chlorophyll fluorescence, qE. It has been suggested that this results from a low pH-induced structural alteration in the light harvesting complex of photosystem II, LHCII [(1991) FEBS Letters 292, 1-4]. The effect of the carboxyl-modifying agent, dicyclohexylcarbodiimide (DCCD), on energy dissipation in chloroplast membranes has been investigated. At concentrations below that required to inhibit electron transport, DCCD caused a decrease in the steady state delta pH, completely inhibited qE and also inhibited the low pH-dependent induction of qE. DCCD binding to polypeptides in the 22-28 kDa range correlated with inhibition of qE. It is suggested that DCCD reacts with amino acid residues in LHCII whose protonation is the primary event in the induction of energy dissipation. This LHCII domain may be identical to one forming a proton channel linking the site of PSII-dependent water oxidation to the thylakoid lumen [(1990) Eur. J. Biochem. 193, 731-736].

Chlorophyll↗

Control of the light-harvesting function of chloroplast membranes by aggregation of the LHCII chlorophyll-protein complex.

A new hypothesis is presented to explain the major molecular process that regulates the efficiency of light harvesting by chloroplast membranes. It is proposed that in excess light the decrease in the thylakoid lumen pH causes an increase in aggregation of the light harvesting complexes of photosystem II resulting in formation of an efficient pathway for non-radiative dissipation of excitation energy. The aggregation is potentiated by the conversion of violaxanthin to zeaxanthin. This hypothesis is based upon (i) similarity between the spectroscopic changes associated with energy dissipation and those observed upon aggregation of isolated light harvesting complex; and (ii) the link between changes in light scattering and increased energy dissipation.

Chlorophyll↗

Reflex sympathetic dystrophy syndrome and barbiturates. A study of 25 cases treated with barbiturates compared with 124 cases treated without barbiturates.

Amongst 149 cases of reflex sympathetic dystrophy syndrome (RSDS), 25 (16.8%) were being treated with barbiturates at the time the RSDS symptoms began. This group is unusual by the frequent absence of provocative events (32%), a high number of diseased joints, bilaterality (76%), involvement of upper limbs (76%) and Dupuytren's disease (52%). Swift and complete recovery depends mostly on barbiturate withdrawal. These data support the idea that barbiturates may be the principal initiating event in some RSDS. Since unfavorable progress is seen with persistent use of barbiturates, this medication should be recognised early in the course of the disease in order to prevent severe sequels.

Adult↗

Relationships between protein phosphorylation and electron transport in the reconstituted chloroplast system.

Phosphorylation of the light-harvesting chlorophyll protein (LHCP) by the thylakoid protein kinase has been examined in the reconstituted chloroplast system. The level of phosphorylation by [32P]Pi was maximal at high light intensity and in the absence of 3-phosphoglycerate; dephosphorylation resulted from a subsequent decrease in light intensity or from the addition of 3-phosphoglycerate. Addition of ribose 5-phosphate, which acts as an ATP 'sink', also caused dephosphorylation. It is concluded that the degree of phosphorylation is dependent on the redox state and energy state of the system, thereby providing a mechanism for adapting light harvesting to the demands of carbon assimilation.

Chlorophyll↗

Characterization of photosystem ii electron acceptors in Phormidium laminosum.

Chlorophyll a fluorescence has been used to monitor the redox state of the primary electron acceptor of photosystem II (PS II) in the blue-green alga Phormidium laminosum during equilibrium titrations. The shape of induction curves measured in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU) have been analyzed. The induction curves were very similar in unfractionated thylakoid membranes and PS II particles. In both, the fast (alpha) phase was sigmoidal, and was followed by a slow (beta) exponential tail. Thus, the structural organization and complexity of the particles (J. M. Bowes and P. Horton, Biochim, Biophys. Acta 680, 127-133 (1982), as indicated by the occurrence of energy transfer between alpha centers and presence of beta centers, must preexist in the membranes. Redox titration of the initial level of fluorescence indicated the presence of a single quencher QH in the unfractionated thylakoids, midpoint potential: Em7.0 approximately -35 mV (n = 1). Thus, the occurrence of a single acceptor is characteristic for P. laminosum and the absence of a low potential acceptor in PS II particles (J.M. Bowes, P. Horton, and D.S. Bendall, FEBS Lett. 135, 261-264 (1981] was not the result of its removal during their preparation. The midpoint potential of Q varied by -60 mV/pH unit in PS II particles and membrane fragments, with a pK at pH greater than 8.5 (particles) and at pH 7.5 (fragments). In PS II particles, DCMU raised the pK by approximately 0.5 pH units. It is argued that the pH dependence of Q is conferred by protonation of a protein which accompanies its reduction rather than protonation of the semiquinone Q X itself.

Chlorophyll↗

Spontaneous drainage of cardiac tamponade.

The case of a 23-year-old man with life-threatening pericardial tamponade resulting from a cardiac stab wound is presented. After initial attempts at stabilization failed to reverse the progressively worsening status of the patient, he was prepared for surgery. Just prior to induction of anesthesia, blood began to flow from the patient's chest tube, resulting in a return to a near normal hemodynamic status. This was believed to be the result of spontaneous drainage of cardiac tamponade prior to cardiorrhaphy. The patient was discharged ten days post admission in excellent health and was doing well at a one-month follow-up visit. The discussion includes what comprises definitive care for cardiac tamponade, and the controversy in the timing and appropriateness of pericardiocentesis.

Adult↗

A rapid quantitative method for the preparation of 123I-iodo-hippuric acid.

The labelling of o-iodo-hippuric acid (hippuran) with 123I by several methods was investigated with a view to producing a simple one-step kit preparation. The requirements for the final product are high labelling efficiency and high radiochemical purity. A method which incorporates CuSO4 . 5H2O as a catalyst was found which gave promising preliminary results. Such variables as pH, mass of CuSO4 . 5H2O, volume of 0.02 N NaOH containing the 123I, reaction temperature and time were investigated in detail and optimum values obtained. The results have led to the production of a kit which can be stored for up to 3 months in liquid form and up to 9 months if freeze-dried. The kit has been used in several thousand patients and has yielded consistently good clinical results.

Humans↗