PubMed Health⌕ Search

Biomedical subjects

R Malkin

Publications and source records attributed to R Malkin.

At least 19 recordsLinked to original sources
PubMed1998

The relationship between symptoms and IgG and IgE antibodies in an office environment.

Airborne fungi have been postulated as a cause of symptoms among office workers. Using the MAST chemiluminescent system, this study evaluated 36 IgG and 36 IgE antibody levels in 47 office workers from an area with elevated airborne fungal concentrations and 44 office workers from an otherwise similar area with lower airborne fungal exposure. No difference was found in IgG antibody to fungi between the lower and higher exposure areas, but high IgG antibody to one or more of the fungi studied was detected in 67% of all the workers tested. IgE antibody to one or more antigens was detected in 40% of the participants. Workers who reported atopic symptoms (sneezing, runny nose, and itchy eyes) or "sick building" symptoms (any three of the following temporally related to work: headache, fatigue, stuffy nose, irritated eyes, or sore throat) were more likely to have one positive IgE antibody test. Type I hypersensitivity to aeroallergens besides fungi may play a role in some symptoms reported by some participants in this office building.

Adult↗
PubMed1998

Virtual electrode effects in transvenous defibrillation-modulation by structure and interface: evidence from bidomain simulations and optical mapping.

INTRODUCTION: Our goal in this combined modeling and experimental study was to gain insight into the transmembrane potential changes in defibrillation conditions, namely, when shocks are delivered by an implantable cardioverter defibrillator (ICD). Two hypotheses concerning the presence and characteristics of virtual electrode effects (VEE) during an ICD shock were tested numerically and experimentally: (H1) anisotropy-dependent VEE are induced over a considerable portion of the "bulk" myocardium; and (H2) surface (epicardial and endocardial) VEE are generated under special tissue bath conditions and are not fully anisotropy determined. METHODS AND RESULTS: Optical mapping was performed on Langendorff-perfused rabbit hearts (n = 4) stained with di-4-ANEPPS. Monophasic shocks were applied during the plateau phase of an action potential through a 9-mm long distal electrode in the right or left ventricle and a 6-cm proximal electrode positioned 3 cm posteriorly to the heart. We modeled the experiment using an ellipsoidal bidomain heart with transmural fiber rotation, placed in a perfusing bath, and subjected to defibrillation shocks delivered by an electrode configuration as described. Our numerical simulations demonstrated VEE occupying a significant portion of the myocardium in the conditions of unequal anisotropy ratios for the intra- and extracellular domains. Statistically significant differences in epicardial polarization patterns were predicted numerically and confirmed experimentally when the interface conditions varied. CONCLUSION: The present study concludes that VEE are present in transvenous defibrillation. They are shaped by the combined effect of cardiac tissue characteristics and interface conditions. Because of their size, VEE might contribute significantly to defibrillation outcome.

Animals↗
PubMed1996

Reconstitution of the 2Fe-2S center and g = 1.89 electron paramagnetic resonance signal into overproduced Nostoc sp. PCC 7906 Rieske protein.

The Rieske 2Fe-2S protein is a distinguishing subunit of the photosynthetic electron transport cytochrome b6f complex in chloroplast and cyanobacterial thylakoid membranes. We have constructed plasmids for overproduction in Escherichia coli of fusion, full-length, and truncated forms of the Rieske (PetC) protein from the cyanobacterium Nostoc sp. PCC 7906. A glutathione S-transferase/Rieske fusion protein was used to prepare specific chicken egg-yolk antibodies against the Rieske protein. Expression of the nonfusion petC gene in a T7 RNA polymerase promoter vector produced copious quantities of the full-length Rieske protein predominantly as inclusion bodies. The highly enriched, Rieske protein from inclusion bodies has been denatured in guanidine hydrochloride and refolded and the characteristic 2Fe-2S cluster reconstituted in vitro by incubation with iron and sulfide under reducing conditions. Purification by chromatography on Whatman DE52 cellulose and ultrafiltration through a 30000 molecular weight cutoff membrane yielded pure and predominantly monomeric Rieske protein. Reconstituted Rieske preparations showed intense and highly characteristic gx = 1.74, gy = 1.89, and gz = 2.03 "Rieske-type" electron paramagnetic resonance signals at 15 K. Two methods of reconstitution yielded Rieske preparations in which 20-60% of the protein contained 2Fe-2S clusters as determined by EPR spin quantitation. The reconstituted Rieske protein was soluble and stable at 4 degrees C in buffers containing nonionic detergents and showed a redox midpoint potential of +321 mV at pH 7.0 as determined by optical circular dichroism (CD) spectroscopy. These data demonstrate the in vitro restoration of a Cys and His liganded 2Fe-2S cluster and provide the basis for mutational and structural analysis of a PetC Rieske protein of oxygenic photosynthesis.

Animals↗
PubMed1996

Active photosynthesis in cyanobacterial mutants with directed modifications in the ligands for two iron-sulfur clusters on the PsaC protein of photosystem I.

The PsaC protein of the Photosystem I (PSI) complex in thylakoid membranes coordinates two [4Fe-4S] clusters, FA and FB. Although it is known that PsaC participates in electron transfer to ferredoxin, the pathway of electrons through this protein is unknown. To elucidate the roles of FA and FB, we created two site-directed mutant strains of the cyanobacterium Anabaena variabilis ATCC 29413. In one mutant, cysteine 13, a ligand for FB was replaced by an aspartic acid (C13D); in the other mutant, cysteine 50, a ligand for FA was modified similarly (C50D). Low-temperature electron paramagnetic resonance studies demonstrated that the C50D mutant has a normal FB center and a modified FA center. In contrast, the C13D strain has normal FA, but failed to reveal any signal from FB. Room-temperature optical studies showed that C13D has only one functional electron acceptor in PsaC, whereas two such acceptors are functional in the C50D and wild-type strains. Although both mutants grow under photoautotrophic conditions, the rate of PSI-mediated electron transfer in C13D under low light levels is about half that of C50D or wild type. These data show that (i) FB is not essential for the assembly of the PsaC protein in PSI and (ii) FB is not absolutely required for electron transfer from the PSI reaction center to ferredoxin.

Amino Acid Sequence↗
PubMed1996

Nucleotide sequence of the PetM gene encoding a 4 kDa subunit of the cytochrome b6f complex from Chlamydomonas reinhardtii.

We have determined the nucleotide sequence of the PetM gene from the single celled alga Chlamydomonas reinhardtii. The gene encodes a recently characterized, small protein of the cytochrome b6f complex, and based on this sequence, it is proposed that this protein spans the membrane by a single alpha-helix. Comparison of the nucleotide sequence with the deduced amino acid sequence reveals a 60-amino-acid presequence similar to a stroma-targeting peptide.

Amino Acid Sequence↗
PubMed1996

N-terminal mutants of chloroplast cytochrome f. Effect on redox reactions and growth in Chlamydomonas reinhardtII.

The N-terminal tyrosine of cytochrome f, which provides the sixth ligand to the heme group, has been changed by site-directed mutagenesis in Chlamydomonas reinhardtii to evaluate the role of this amino acid in assembly and function. The second and third residues, proline and valine, respectively, have also been mutated. Y1P is the only strain that did not grow photoautotrophically. The other strains show cytochrome b6f complex/photosystem I reaction center chlorophyll, photosystem I unit size and chlorophyll a+b/cell ratios comparable with wild-type cells. Rates of cytochrome f photooxidation in all strains were similar (t1/2 approximately = 300 microsec), whereas the rate of re-reduction sensitive to stigmatellin (at Eh = 0 mV, (where Eh is the ambient redox potential) for wild-type, Y1W, Y1F, Y1S, P2V, and V3P had a tl/2 of 3, 4, 5, 9, 40, and 2 ms, respectively. Rates of oxygen evolution by whole cells of P2V, Y1F, and Y1S were 67, 80, and 80% of wild-type rates, respectively. At low light intensity, all competent strains had the same growth rate whereas at saturating intensities, only P2V showed a significant inhibition. These results are considered in relation to structure-function relationships in the cytochrome f molecule.

Animals↗
PubMed1996

1-Hydroxypyrene levels in coal-handling workers at a coke oven.

An environmental and medical survey was conducted at the coal-handling area of a coke oven, where workers came in contact with coal-tar sludge. The purpose of the study was to determine if skin contact with coal-tar sludge was an important route of exposure to pyrene because workers were observed to have substantial contact with the sludge. Environmental monitoring revealed minimal airborne exposure to pyrene, a byproduct of the coke distillation process; only one personal breathing zone sample detected pyrene, and at least of 0.001 mg/m3. However, the mean preshift urinary 1-hydroxypyrene concentration was 1.00 mumol/mol creatinine (range, 0.16 to 2.96 mumol/mol creatinine) and the mean postshift level was 1.7 mumol/mol creatinine (range, 0.24 to 4.85 mumol/mol creatinine) (P < 0.01). These levels probably reflect absorption as a result of skin exposure.

Adult↗
PubMed1995

The deletion of petG in Chlamydomonas reinhardtii disrupts the cytochrome bf complex.

The 4-kDa protein encoded by chloroplast petG copurifies with the cytochrome bf complex of spinach and is found in a number of other photosynthetic organisms, including the eukaryotic alga Chlamydomonas reinhardtii. To determine whether petG is involved in the function or assembly of the cytochrome bf complex, the gene was cloned from C. reinhardtii, excised from the DNA fragment, and replaced with a spectinomycin resistance cassette. A petG deletion strain of C. reinhardtii was then obtained by biolistic transformation. The resulting homoplasmic petG deletion strains are unable to grow photosynthetically, and immunoblot analysis shows markedly decreased levels of cytochrome b6, cytochrome f, the Rieske iron-sulfur protein, and subunit IV. To verify that this phenotype was due to the removal of petG, we also constructed a strain with a deletion in the open reading frame (ORF56), which is found 25 base pairs downstream of petG. The ORF56 deletion strain grew photosynthetically and had wild-type levels of the four major cytochrome bf subunits. We conclude that the absence of the PetG protein affects either the assembly or stability of the cytochrome bf complex in C. reinhardtii.

Amino Acid Sequence↗
PubMed1995

Occupational and environmental lead and PCB exposure at a scrap metal dealer.

Blood lead levels (BPb) and serum polychlorinated biphenyl levels (PCB) were obtained from 17 employees at two adjacent scrap metal dealers. One facility was located outdoors, directly on top of soil known to be contaminated with lead and PCBs, and the other was located indoors with a concrete floor. BPbs ranged from 4.0 to 39.8 microgram/dl (mean 19.9 microgram/dl, geometric mean 17.5 microgram/dl) and PCB levels ranged from <1 to 65.3 ppb (mean 7.5 ppb). There was no significant difference in either BPb or serum PCB between the two sites. BPb was significantly correlated with the number of cigarettes smoked at work, and both BPb and serum PCB were significantly related to eating lunch outside the lunchroom, suggesting hand-to-mouth contact as a source of exposure. The lack of difference in BPb between employees of the two scrap metal dealers suggests an ongoing source of lead exposure at the sites, other than the soil.

Analysis of Variance↗
PubMed1992

Specific release of a 9-kDa extrinsic polypeptide of photosystem I from spinach chloroplasts by salt washing.

The newly reported 9-kDa polypeptide in photosystem I [(1991) FEBS Lett. 280, 332-334] is an extrinsic component located on the lumenal side of the thylakoid membrane. This subunit can be solubilized with high salt buffer and does not bind any cofactors. The photosystem I electron transfer chain remains intact and functional in the absence of this component as characterized by the photoreduction of NADP+.

Amino Acid Sequence↗
PubMed1992

Blood lead levels in incinerator workers.

Questions have been raised concerning the safety of mass burn incineration and its role in solid waste management. In 1989, the New York City Office of Occupational Safety and Health examined air levels of metals in New York City incinerators and found that workers were exposed to air lead levels as high as 2500 micrograms/m3 while cleaning the electrostatic precipitators in the plant. In order to determine the biologic significance of these exposures to the workers, blood samples were taken from 56 incinerator workers and 25 controls and analyzed for lead and erythrocyte protoporphyrin levels. Incinerator workers were found to have a mean blood lead of 11.0 micrograms/dl as compared to the control group level of 7.4 micrograms/dl. Risk factors for increased blood lead levels were analyzed using multiple regression analyses. Wearing a personal protective device "always" or not and the interaction of smoking and cleaning the precipitator more than seven times in the past year were found to be significant predictors for blood lead. These results indicate that lead in municipal incinerator ash from electrostatic precipitators is bioavailable and that the effects of such exposure can be minimized by wearing personal protective devices, not smoking, and rotating the work force to minimize precipitator ash contact.

Air Pollutants, Occupational↗
PubMed1992

Organization and topology of photosystem I subunits.

Intact spinach (Spinacia oleracea) thylakoid membranes were treated with various proteases and photosystem I (PSI) complexes were isolated from these membranes to define the membrane topology of specific PSI subunits. Trypsin treatment caused cleavage of the PSI-D and E subunits. Thermolysin treatment cleaved the PSI-D, E, H, and K subunits, and also caused limited degradation of the reaction center core PSI-A and B subunits. Pronase treatment produced the most dramatic results as the PSI-A and B subunits were cleaved to 47-, 45-, 26-, and 24-kilodalton products. In addition, pronase degraded the PSI-D, E, H, K, and L subunits. Proteolytic cleavage sites for several of the products were identified by amino acid sequencing. The results indicate that PSI-A, B, D, E, H, K, and L subunits all have stroma-exposed regions, and these findings are summarized in a model describing the subunit organization of PSI.

Journal Article↗
PubMed1991

Structural characterization of heme sites in spinach cytochrome b6f complexes: a resonance Raman study.

Resonance Raman spectra of cytochrome b6f complexes isolated from spinach chloroplasts have been obtained. Selective resonance enhancements and partial reductions of the complex by redox mediators were used to isolate and identify the contributions of heme b6 and heme f sites to the observed spectra. Corresponding spectra for turnip cytochrome f have also been obtained. Power-dependent photoreduction was observed in cytochrome f of the complex as well as in the isolated cytochrome f during the course of the Raman experiments.

Cytochromes↗
PubMed1991

Electron spin echo envelope modulation spectroscopy supports the suggested coordination of two histidine ligands to the Rieske Fe-S centers of the cytochrome b6f complex of spinach and the cytochrome bc1 complexes of Rhodospirillum rubrum, Rhodobacter sphaeroides R-26, and bovine heart mitochondria.

Electron spin echo envelope modulation (ESEEM) experiments performed on the Rieske Fe-S clusters of the cytochrome b6f complex of spinach chloroplasts and of the cytochrome bc1 complexes of Rhodospirillum rubrum, Rhodobacter sphaeroides R-26, and bovine heart mitochondria show modulation components resulting from two distinct classes of 14N ligands. At the g = 1.92 region of the Rieske EPR spectrum of the cytochrome b6f complex, the measured hyperfine couplings for the two classes of coupled nitrogens are A1 = 4.6 MHz and A2 = 3.8 MHz. Similar couplings are observed for the Rieske centers in the three cytochrome bc1 complexes. These ESEEM results indicate a nitrogen coordination environment for these Rieske Fe-S centers that is similar to that of the Fe-S cluster of a bacterial dioxygenase enzyme with two coordinated histidine ligands [Gurbiel, R. J., Batie, C. J., Sivaraja, M., True, A. E., Fee, J. A., Hoffman, B. M., & Ballou, D. P. (1989) Biochemistry 28, 4861-4871]. The Rieske Fe-S cluster lacks modulation components from a weakly coupled peptide nitrogen observed in water-soluble spinach ferredoxin. Treatment with the quinone analogue inhibitor DBMIB causes a shift in the Rieske EPR spectrum to g = 1.95 with no alteration in the magnetic coupling to the two nitrogen atoms. However, the ESEEM pattern of the DBMIB-altered Rieske EPR signal shows evidence of an additional weakly coupled nitrogen similar to that observed in the spinach ferredoxin ESEEM patterns.

Animals↗
PubMed1991

Biosynthesis of the chloroplast cytochrome b6f complex: studies in a photosynthetic mutant of Lemna.

The biosynthesis of the cytochrome b6f complex has been studied in a mutant, no. 1073, of Lemna perpusilla that contained less than 1% of the four protein subunits when compared with a wild-type strain. RNA gel blot analyses of the mutant indicated that the chloroplast genes for cytochrome f, cytochrome b6, and subunit IV (petA, petB, and petD, respectively) are transcribed and that the petB and petD transcripts undergo their normal processing. Analysis of polysomal polyA+ RNA indicated that the level of translationally active mRNA for the nuclear-encoded Rieske Fe-S protein (petC) was reduced by greater than 100-fold in the mutant. Immunoprecipitation of in vivo labeled proteins indicated that both cytochrome f and subunit IV are synthesized and that subunit IV has a 10-fold higher rate of protein turnover in the mutant. These results are discussed in terms of the assembly of the cytochrome complex and the key role of the Rieske Fe-S protein in this process.

Chloroplasts↗