Failure to record delirium as a complication of intra-aortic balloon pump treatment: a retrospective study.

This study was conducted to determine whether or not diagnosis and treatment of delirium among patients treated with the intra-aortic balloon pump (IABP) correlates with the recording of this complication on discharge records. Since prior episodes of delirium are one of the few clear risk factors for future episodes of delirium, accurate recording of delirium on the discharge summary and list of discharge diagnoses is useful to clinicians. A retrospective review of the charts of all patients (N = 198) who underwent placement of an IABP during 1988; assessment of the type and frequency of medical and neuropsychiatric complications during IABP treatment; and comparison of chart review findings with the Massachusetts General Hospital's computer-generated lists of discharge diagnoses for the same IABP-treated patients was completed. Only 12% of patients diagnosed and treated for delirium had delirium recorded as a discharge diagnosis. In contrast, 44% and 52% of patients who had been diagnosed and treated for cerebrovascular accident and pneumonia, respectively, had these diagnoses recorded among the discharge diagnoses. Receiving a discharge diagnosis of organic brain syndrome increased the likelihood that delirium was recorded as a discharge diagnosis. Delirium is underdiagnosed as a complication associated with IABP-treatment and is under-reported on the list of discharge diagnoses, even when it is diagnosed. Further study is warranted to determine if making the diagnosis of delirium during a patient's hospital course and recording it is a complication at the time of discharge is translated into a higher level of preparedness by physicians during subsequent hospitalizations.

Ozone-Induced Ethylene Emission Accelerates the Loss of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase and Nuclear-Encoded mRNAs in Senescing Potato Leaves.

The relationships among O3-induced accelerated senescence, induction of ethylene, and changes in specific mRNA and protein levels were investigated in potato (Solanum tuberosum L. cv Norland) plants. When plants were exposed to 0.08 [mu]L L-1 O3 for 5 h d-1, steady-state levels of rbcS mRNA declined at least 5-fold in expanding leaves after 3 d of O3 exposure and ethylene levels increased 6- to 10-fold. The expression of OIP-1, a 1-aminocyclopropane-1-carboxylate synthase cDNA from potato, correlated with increased production of ethylene and decreased levels of rbcS mRNA in foliage of plants treated with O3. In plants exposed to 0.30 [mu]L L-1 O3 for 4 h, rbcS transcript levels were reduced 4-fold, whereas nuclear run-on experiments revealed that rbcS transcription declined an average of 50%. The loss of rbcS mRNA may be due, in part, to posttranscriptional regulation. The levels of transcripts for other chloroplast proteins, glyceraldehyde-3-phosphate dehydrogenase, and a photosystem II chlorophyll a/b-binding protein decreased in O3-treated plants, in parallel with the decrease in rbcS mRNA. The steady-state mRNA level of a cytosolic glyceral-dehyde-3-phosphate dehydrogenase increased in O3-treated plants. The induction of ethylene and changes in transcript levels preceded visible leaf damage and decreases in ribulose-1,5-bisphosphate carboxylase/oxygenase protein levels.

Molecular cloning of an ozone-induced 1-aminocyclopropane-1-carboxylate synthase cDNA and its relationship with a loss of rbcS in potato (Solanum tuberosum L.) plants.

Acute or chronic exposure of potato plants to ozone (O3) induces ethylene production. We isolated a 1586 bp cDNA (pOIP-1) encoding 1-aminocyclopropane-1-carboxylate (ACC) synthase from a cDNA library constructed with mRNA extracted from O3-treated leaves. The clone has a 1365 bp open reading frame and a 221 bp trailing sequence. The active site found in all ACC synthases and 11 of the 12 amino acid residues conserved in aminotransferases are found in pOIP-1. Northern analysis showed that the mRNA encoding ACC synthase was detectable 1 h after the onset of O3 exposure, and the message increased over time as did ethylene production. Concurrent with the increased ACC synthase mRNA was a decrease in the message for the Rubisco small subunit (rbcS) with no change in the large subunit (rbcL). When the plants were treated with aminooxyacetic acid (AOA), both ethylene production and level of ACC synthase transcript were inhibited. The decline in rbcS was also inhibited by AOA suggesting a correlation between ethylene production and loss of rbcS. Based on nuclear run-on studies it appears that the increase in ACC synthase mRNA may result from O3-induced transcriptional activity.

Genetically Programmed Chloroplast Dedifferentiation as a Consequence of Plastome-Genome Incompatibility in Oenothera.

Comparision of chloroplast from plants with one of four plastome types (I, II, III, IV) in the nuclear background of Oenothera elata strain Johansen addressed the effects of plastome-genome incompatibility with respect to leaf pigmentation, plastid ultrastructure, chlorophyll a/chlorophyll b ratio, and photosynthetic electron transport. Previous observations of plastomes I, II, and IV in this nuclear background have revealed no indications of incompatibility, but the studies reported here demonstrate that chloroplasts of plastome IV have subtle alterations in their photosynthetic abilities, in particular, deficiencies in photosystem II. The well-characterized "hybrid bleaching" of plants with the AA genotype and plastome III involves leaves that become bleached in the center while remaining green at the tips, edges, and veins. Electron transport assays performed on fractionated bleached and green tissue from the same plants show photosynthetic defects in both the green and bleached regions, although defects in the latter are more severe. Ultrastructural studies show that chloroplasts in the bleached areas enlarge, thylakoid membranes become swollen and vesiculated, and production of new thylakoids is blocked, with chloroplasts appearing to undergo a programmed senescence. A time course revealed that the senescence is actually a reversible dedifferentiation. Alterations in the composition of medium to which AA/III seedlings were transferred showed that the presence of auxin can prevent the development of the typical incompatibility response, with leaf tissue remaining green rather than bleaching. It is proposed that differences in concentrations of plant growth regulators may be responsible for the persistence of normal chloroplasts near the vascular tissue and leaf blade edges and that seasonal fluctuations in auxin levels could explain the periodic bleaching that occurs in older plants.

Biochemical and molecular basis for impairment of photosynthetic potential.

Ozone induces reductions in net photosynthesis in a large number of plant species. A primary mechanism by which photosynthesis is reduced is through impact on carbon dioxide fixation. Ozone induces loss in Rubisco activity associated with loss in concentration of the protein. Evidence is presented that ozone may induce oxidative modification of Rubisco leading to subsequent proteolysis. In addition, plants exposed to ozone sustain reduction in rbcS, the mRNA for the small subunit of Rubisco. This loss in rbcS mRNA may lead to a reduced potential for synthesis of the protein. The regulation of O3-induced loss of Rubisco, and implications of the decline in this protein in relation to accelerated senescence are discussed.

Dynamics of Photosystem Stoichiometry Adjustment by Light Quality in Chloroplasts.

Long-term imbalance in light absorption and electron transport by photosystem I (PSI) and photosystem II (PSII) in chloroplasts brings about changes in the composition, structure, and function of thylakoid membranes. The response entails adjustment in the photosystem ratio, which is optimized to help the plant retain a high quantum efficiency of photosynthesis (W.S. Chow, A. Melis, J.M. Anderson [1990] Proc Nat Acad Sci USA 87: 7502-7506). The dynamics of photosystem ratio adjustment were investigated upon the transfer of pea {Pisum sativum} plants from a predominantly PSI-light to a predominantly PSII-light environment and vice versa. The concentration of functional components (primary electron accepting plastoquinone of PSII [QA], P700) and that of constituent proteins were monitored during acclimation by A difference spectrophotometry and immunoblot analysis, respectively. Fully reversible changes in photosystem ratio occurred with a half-time of about 20 h. They involved closely coordinated changes in the concentration of the QA, reaction center protein D1, D2, and the 9-kD apoprotein of the cytochrome b559 for PSII. Similarly, closely coordinated changes in the relative concentration of P700 and reaction center proteins of PSI were observed. The level of chlorophyll b and that of the light-harvesting complex II changed in accordance with the concentration of PSII in the acclimating thylakoids. Overall, adjustments in the photosystem ratio in response to PSI- or PSII-light conditions appeared to be a well-coordinated reaction in the chloroplast. The response was absent in the chlorophyll b-less chlorina f2 mutant of barley (Hordeum vulgare) and in a phycobilisomeless mutant of Agmenellum quadruplicatum, suggesting that photosystem accessory pigments act as the light-quality perception molecules and that PSI and PSII themselves play a role in the signal transduction pathway.

Large unidentified open reading frame in plastid DNA (ORF2280) is expressed in chloroplasts.

The chloroplast DNA encodes genes for components of photosynthesis and the transcription-translation machinery; a number of unidentified open reading frames (ORFs) are also present. To determine whether a large ORF in the inverted repeat of chloroplast DNA of tobacco (ORF2280) encodes a chloroplast protein, a conserved region of the ORF was expressed in Escherichia coli. An antibody against the ORF protein was prepared using the purified fusion protein as an antigen. When incubated with proteins from the soluble fraction of tobacco, spinach and Oenothera chloroplasts, the antiserum detects relatively labile polypeptides, which have apparent molecular weights of 170 to 180 kDa. The ORF in tobacco and spinach is large enough to encode a protein of 240-250 kDa, thus it is possible that post-transcriptional or post-translational processing reduces the size of the expression product. Analysis of Oenothera chloroplasts representing four different plastome types revealed endonuclease restriction fragment length polymorphisms in chloroplast DNA indicative of insertion/deletion events in a region of the chloroplast DNA that shared significant sequence similarity with ORF2280. The ORF2280 antiserum was used to demonstrate that there are qualitative differences in the ORF proteins from different Oenothera plastome types.

Changes in Photosynthetic Capacity and Photosynthetic Protein Pattern during Tomato Fruit Ripening.

Levels of polypeptides participating in the photosynthetic light and dark reactions have been measured during fruit ripening in tomato. Photosynthetic proteins were identified by Western blot analysis with heterologous antibodies. The concentrations of proteins of photosystem (PS) I (14 kilodaltons), of PSII (47-kilodalton reaction center protein, 32-kilodalton ;Q(B) binding' protein and light harvesting complex proteins), of the photosynthetic electron transport chain (ferredoxin-NADP-oxidoreductase and plastocyanin), and of the stroma (ribulose-1,5-bisphosphate carboxylase) decrease during the ripening process. The 32-kilodalton protein and plastocyanin were detectable in pericarp protein preparations of ripe tomato fruits. Absorbance difference spectrophotometry provided information on the relative concentrations of PSII and PSI reaction centers in leaf and green fruit tissue of tomato. These results indicate that green fruit pericarp of tomato is photosynthetically active. Photosynthetic activity decreases during chloroplast/chromoplast differentiation. This is consistent with changes that occur at the transcript level of photosynthesis-specific proteins during the differentiation process.

Light quality regulates expression of chloroplast genes and assembly of photosynthetic membrane complexes.

The concentrations of photosystem I (PSI) and photosystem II (PSII) reaction centers and the level of chloroplast reaction center gene transcripts were determined in pea plants grown under different light-quality regimes. In plants grown in light primarily absorbed by PSI ("red" light), the PSII/PSI reaction center ratio was 2-fold greater than that in plants grown in PSII-sensitizing ("yellow") light. In addition, the ratio of a PSII gene (psbB) transcript to a PSI gene (psaA) transcript was 2.6 times greater in red-grown plants relative to yellow-grown plants. Thus, a differential reaction-center concentration in the thylakoid membrane was accompanied by a differential expression of reaction center genes, suggesting that the synthesis of chloroplast membrane complexes and the assembly of photosystems are regulated by light quality at the transcriptional and/or post-transcriptional level.

Freeze-fracture analysis of thylakoid membranes and photosystem I and II enriched fractions from Phormidium laminosum.

Thylakoid membranes of the thermophilic cyanobacterium Phormidium laminosum have been fractionated into photosystem II and photosystem I particles. These fractions have been characterized by their partial electron transport activities, and biochemical and spectral properties. Exoplasmic fracture face and protoplasmic fracture face particles in the unfractionated thylakoid membranes were shown to correspond in size to particles in freeze-fractured photosystem II and photosystem I fractions, respectively. Differences between the histograms of the thylakoid membrane protoplasmic fracture face particles and the isolated photosystem I particles suggest that in addition to photosystem I complexes some of the particles on the thylakoid protoplasmic fracture face may be related to cytochrome b/f complexes, the hydrophobic component of the coupling factor, or respiratory complexes.

Photochemical Apparatus Organization in the Chloroplasts of Two Beta vulgaris Genotypes.

The composition and structural organization of thylakoid membranes of a low chlorophyll mutant of Beta vulgaris was investigated using spectroscopic, kinetic and electrophoretic techniques. The data obtained were compared with those of a standard F1 hybrid of the same species. The mutant was depleted in chlorophyll b relative to the hybrid and it had a higher photosystem II/photosystem I reaction center (Q/P(700)) ratio and a smaller functional chlorophyll antenna size. Analysis of thylakoid membranes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the mutant lacked a portion of the chlorophyll a/b light-harvesting complex but was enriched in the photosystem II reaction center chlorophyll protein complex. Comparison of functional antenna sizes and of photosystem stoichiometries determined electrophoretically were in good agreement with those determined spectroscopically. Both approaches indicated that about 30% of the total chlorophyll was associated with photosystem I and about 70% with photosystem II. A greater proportion of photosystem II(beta) was detected in the mutant. The results suggest that a higher photosystem II to photosystem I ratio in the sugar beet mutant has apparently compensated for the smaller photosystem II chlorophyll light-harvesting antenna in its chloroplasts. Moreover, a lack of chlorophyll a/b light-harvesting complex correlates with the abundance of photosystem II(beta). It is proposed that a developmental relationship exists between the two types of photosystem II where photosystem II(beta) is a precursor form of photosystem II(alpha) occurring prior to the addition of the chlorophyll a/b light-harvesting complex and grana formation.

Effect of light quality on chloroplast-membrane organization and function in pea.

The effect of light quality during plant growth of chloroplast membrane organization and function in peas (Pisum sativum L. cv. Alaska) was investigated. In plants grown under photosystem (PS) I-enriched (far-red enriched) illumination both the PSII/PSI stoichiometry and the electrontransport capacity ratios were high, about 1.9. In plants grown under PSII-enriched (far-red depleted) illumination both the PSII/PSI stoichiometry and the electron-transport capacity ratios were significantly lower, about 1.3. In agreement, steady-state electron-transport measurements under synchronous illumination of PSII and PSI demonstrated an excess of PSII in plants grown under far-red-enriched light. Sodium dodecylsulfate polyacrylamide gel electrophoretic analysis of chlorophyll-containing complexes showed greater relative amounts of the PSII reaction center chlorophyll-protein complex in plants grown under farred-enriched light. Additional changes were observed in the ratio of light-harvesting chlorophyll a/b protein to PSII reaction center chlorophyll-protein under the two different light-quality regimes. The results demonstrate the dynamic nature of chloroplast structure and support the notion that light quality is an important factor in the regulation of chloroplast membrane organization and-function.

Role of the colorless polypeptides in phycobilisome reconstitution from separated phycobiliproteins.

A phycoerythrin (PE) and phycocyanin (PC) mixture was separated from allophycocyanin on calcium phosphate chromatography from completely dissociated phycobilisomes of the blue-green alga, Nostoc sp. After dialysis of the PE-PC mixture in 0.75 m potassium phosphate, pH 7, which allows reassociation of the dissociated pigment-proteins, complexes of PE and PC in a 2:1 m ratio (PE/PC complex) as well as complexes predominantly of PC (PC/PE complex) were then separated by sedimentation on linear sucrose gradients. These complexes resemble the rods of intact phycobilisomes and transfer energy efficiently from PE to PC. They contain the Group II colorless polypeptides described by Tandeau de Marsac and Cohen-Bazire (1977 Proc Natl Acad Sci USA 74: 1635 61639). Phycobilisomes can be reconstituted by combining the allophycocyanin pool with (a) the PE-PC mixture, (b) the PE/PC complex, or (c) the PC/PE complex. Successful reconstitution is measured by absorption, fluorescence, circular dichroism, and electron microscopy. The major requirement for reconstitution is the 29-kilodalton colorless polypeptide. In its absence, no phycobilisomes are formed. It is the only colorless polypeptide common to both the PE/PC complex and the PC/PE complex, and appears to be the polypeptide responsible for rod attachment to the allophycocyanin. In addition, high phosphate concentrations and 20 degrees C temperatures are needed for reconstitution.

Noncovalent Intermolecular Forces in Phycobilisomes of Porphyridium cruentum.

Using sensitized fluorescence as a measure of intactness of phycobilisomes isolated from Porphyridium cruentum, the effects of various environmental perturbations on phycobilisome integrity were investigated. The rate of phycobilisome dissociation in 0.75 ionic strength sodium salts proceeds in the order: SCN(-) > NO(3) (-) > Cl(-) > C(6)H(5)O(7) (3-) > SO(4) (2-) > PO(4) (3-), as predicted from the lyotropic series of anions and their effects on hydrophobic interactions in proteins. Similarly, increasing temperature (to 30 C) and pH values approaching the isoelectric points of the biliproteins stabilize phycobilisomes. Deuterium substitution at exchangeable sites on the phycobiliproteins decreases the rate of phycobilisome dissociation, while substitution at nonexchangeable sites increases rates of dissociation. It is concluded that hydrophobic intermolecular interactions are the most important forces in maintaining the phycobilisome structure. Dispersion forces also seem to contribute to phycobilisome stabilization. The adverse effects of electrostatic repulsion must not be ignored; however, it seems that the requirement of phycobilisomes of high salt concentrations is not simply countershielding of charges on the proteins.