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D Holland

Publications and source records attributed to D Holland.

At least 55 records · Page 3Linked to original sources
PubMed1994

A novel plant glutathione peroxidase-like protein provides tolerance to oxygen radicals generated by paraquat in Escherichia coli.

Citrus salt-stress associated protein (Cit-SAP) reveals significant sequence homology to mammalian glutathione peroxidase (GP). In an attempt to assign biological function to this protein, transformed E. coli cells expressing Cit-SAP were examined for their ability to tolerate free radicals formed by paraquat, an O2- radical forming agent. In the presence of paraquat, the survival rate of the transformed bacteria expressing Cit-SAP was much higher as compared to the wild-type bacteria. The results support the assumption that Cit-SAP is a plant GP-like protein which participate in the enzymatic system aimed at scavenging oxygen free-radicals in plants.

Drug Tolerance↗
PubMed1994

Two newly isolated genes from citrus exhibit a different pattern of diurnal expression and light response.

The diurnal expression of two newly isolated genes, ccr and clr, was studied in Duncan grapefruits (Citrus paradisi). The steady-state levels of mRNA corresponding to each of the genes reach their maximum at different time points during the day. The peak of ccr accumulation is at the end of the light period, while accumulation of mRNA corresponding to clr peaks already after 4-6 hours of light. The lowest steady-state levels of mRNA corresponding to both of the genes is at the dark period. ccr is highly homologous to lir1, a gene recently isolated from rice. Comparison of the deduced amino acid sequence of ccr to that deduced from lir1 indicates 49% of identity. The deduced proteins contain two imperfect repeats with two cystein residues that are positionally conserved in each of the repeats. The steady-state level of ccr transcripts continues to oscillate after the plants are transferred to darkness and displays a circadian rhythm similar to that of lir1. On the other hand, the level of clr transcripts is almost undetectable after more than 12 h in the dark. Artificial light supplement in the dark extends the length of time of clr expression during the day. clr does not show significant homology to any of the known circadian or light-regulated genes.

Amino Acid Sequence↗
PubMed1994

Diagnosis of enteroviral central nervous system infection by polymerase chain reaction during a large community outbreak.

Enteroviruses are common causes of localized and systemic infection in patients of all ages and are the most frequent cause of epidemic aseptic meningitis in the United States. We have developed a polymerase chain reaction (PCR) assay of cerebrospinal fluid (CSF) for rapid diagnosis of enteroviral meningitis. This assay was applied to 257 CSF specimens during a large community outbreak of enterovirus disease; 109 (97%) of 112 enterovirus culture-positive CSF samples contained enterovirus RNA. In addition 35 (66%) of 53 samples from patients with suspected central nervous system disease with negative or no CSF viral cultures were positive by enterovirus PCR. The enterovirus PCR detected 13 different enterovirus serotypes. PCR results are available within 24 hours compared with a mean of 6.8 days for enterovirus culture. The clinical characteristics of 141 patients with enterovirus central nervous system disease are presented. This study demonstrates the usefulness of enterovirus PCR for the rapid diagnosis of enterovirus central nervous system disease and the potential for PCR tests to shorten hospitalization.

Base Sequence↗
PubMed1994

Investigations of the 'active' edge of plaque psoriasis: vascular proliferation precedes changes in epidermal keratin.

We have investigated markers of epidermal proliferation and differentiation in terms of keratin expression, the morphology of the cutaneous vasculature, and numbers of cutaneous mast cells, in patients with chronic plaque psoriasis. Using the phenomenon of the 'active edge', we have studied these features in the psoriatic plaque itself, and in the clinically normal active and inactive edges of the same plaque. Our results confirm the anticipated changes in keratin profiles, mast cell numbers and psoriatic morphology of the vasculature within the plaque itself. They further indicate that the vascular changes precede the epidermal and mast cell features at the active edge, and that the inactive edge is inactive for all of these variables. Mediators responsible for the vascular proliferation and elongation must be present in increased amounts at the active edge when compared with the inactive, and include locally produced and circulating factors.

Cell Count↗
PubMed1993

The production of PCR products with 5' single-stranded tails using primers that incorporate novel phosphoramidite intermediates.

We have prepared several novel phosphoramidites and have synthesised oligonucleotides incorporating them internally. The presence of these residues in an oligonucleotide template presents an impossible barrier to primed synthesis by Taq DNA polymerase. When extended as polymerase chain reaction products, these oligonucleotides no longer serve as templates for the polymerase beyond the insertion sites of the modified intermediates, thereby producing single-stranded tails on amplification products. These tails can then be used for solid phase capture and colorimetric detection of PCR products.

Alkaline Phosphatase↗
PubMed1993

Molecular characterization of salt-stress-associated protein in citrus: protein and cDNA sequence homology to mammalian glutathione peroxidases.

A gene encoding for a citrus salt-stress-associated protein (Cit-SAP) was cloned from Citrus sinensis salt-treated cell suspension. The gene, designated csa, was isolated from a cDNA expression library. The partial amino acid sequence of the protein, as well as that deduced from the nucleotide sequence of csa, revealed a considerable homology to mammalian glutathione peroxidase (GP), and to clone 6P229 from tobacco protoplasts. The increased expression of Cit-SAP in NaCl-treated cultured citrus cells and in citrus plants irrigated with saline water, and its similarity to GP, raise the possibility that one of the effects of salt stress in plants may be the increase of the level of free radicals.

Amino Acid Sequence↗
PubMed1993

Amplified expression of a transcriptional pattern formed during development of Anabaena.

The cyanobacterium Anabaena responds to nitrogen deprivation by producing heterocysts, cells specialized for nitrogen fixation, at well-spaced intervals along its filaments. The gene hepA, required for heterocyst maturation, is expressed in response to nitrogen deprivation, prior to visible differentiation. A spatial pattern of hepA expression indistinguishable from the eventual pattern of heterocysts was made visible by fusing the hepA promoter to luxAB, which encodes bacterial luciferase. Because the resulting signal did not greatly exceed instrumental background, T7 RNA polymerase was used to increase luminescence. The hepA promoter was fused to the gene for that polymerase, and a promoter recognized by that polymerase was fused to luxAB. Filaments containing these two fusions showed spaced luminescing cells many hours before differentiation became discernible morphologically.

ATP-Binding Cassette Transporters↗
PubMed1993

Role of femoral vessel catheterization and altered hemostasis in the development of extraperitoneal hematomas: CT study in 44 patients.

OBJECTIVE: To ascertain the role of femoral vessel catheterization and altered hemostasis in the development of extraperitoneal hematomas, we evaluated CT scans to study the locations of extraperitoneal hematomas in three distinct clinical settings: (1) after femoral vessel catheterization with concurrent altered hemostasis due to anticoagulant, thrombolytic, and/or antiplatelet therapy (catheterized-altered hemostasis group); (2) after femoral vessel catheterization without altered hemostasis (catheterized group); and (3) with no history of femoral vessel catheterization, with or without altered hemostasis caused by a bleeding diathesis or pharmacotherapy (uncatheterized group). MATERIALS AND METHODS: Forty-four patients were identified who had CT evidence of extraperitoneal hematomas. Twenty-four were in the catheterized-altered hemostasis group, one was in the catheterized group, and 19 were in the uncatheterized group. CT scans were evaluated for anatomic subspace involvement by hemorrhage and for the presence or absence of contiguity between the hematoma and the femoral vessels. RESULTS: Bilateral hematomas were present in six of 24 patients in the catheterized-altered hemostasis group and in four of 19 patients in the uncatheterized group. We found no significant difference between the catheterized-altered hemostasis and the uncatheterized groups regarding the proportion of cases involved at any individual retroperitoneal site (p > .05). The mean number of hematoma sites in the catheterized-altered hemostasis group (4.3) was not significantly different from the mean number in the uncatheterized group (3.7) (p > .05). Contiguity between the sites of hematoma and the punctured femoral vessels was present in 75% of the patients in the catheterized-altered hemostasis group; contiguity between the hematoma and a groin was seen in 58% in the uncatheterized group. Twenty-two of 24 patients in the catheterized-altered hemostasis group had CT findings of subcutaneous fat infiltration in the groin; however, six of 19 patients in the uncatheterized group also had this finding, indicating that its presence does not always represent changes caused by catheterization. CONCLUSION: Retroperitoneal hematomas in patients with altered hemostasis and femoral vessel catheterization do not show a unique distribution attributable solely to contiguous spread of blood from the vessel puncture site. Many of these hematomas probably arise at sites distant from the femoral vessel puncture. When the hematoma is not contiguous with the punctured femoral vessels, altered hemostasis is most likely the cause of the hemorrhage.

Catheterization, Peripheral↗
PubMed1991

1.9-A structures of ternary complexes of citrate synthase with D- and L-malate: mechanistic implications.

The structures of four isomorphous crystals of ternary complexes of chicken heart citrate synthase with D- or L-malate and acetyl coenzyme A or carboxymethyl coenzyme A have been determined by X-ray crystallography and fully refined at 1.9-A resolution. The structures show that both L-malate and D-malate bind in a very similar way in the presence of acetylCoA and that the enzyme conformation is "closed". Hydrogen bond geometry is suggested to account for the difference in binding constants of the two stereoisomers. The structures suggest that steric hindrance can account for the observation that proton exchange of acetyl coenzyme A with solvent is catalyzed by citrate synthase in the presence of L-malate but not D-malate. The ternary complexes with malate reveal the mode of binding of the substrate acetylCoA in the ground state. The carbonyl oxygen of the acetyl group is hydrogen bonded to a water molecule and to histidine 274, allowing unambiguous identification of the orientation of this group. The structures support the hypothesis that carboxymethyl coenzyme A is a transition-state analogue for the enolization step of the reaction (Bayer et al., 1981) and additionally support proposed mechanisms for the condensation reaction (Karpusas et al., 1990; Alter et al., 1990).

Animals↗
PubMed1991

Blood level monitoring of antipsychotics and antidepressants.

This chapter begins with an introduction to the usefulness of plasma levels in general, addressing their utility in monitoring compliance and toxicity. Particular classes of psychotropic medications and the specific medications within them are addressed, particularly antipsychotics and antidepressants.

Antidepressive Agents↗
PubMed1991

Atypical antipsychotics.

This chapter discusses the clinical application of the currently studied atypical antipsychotics in the United States and abroad. A description of the proposed mode of action of these medications and the most relevant studies is included.

Animals↗
PubMed1990

Effects of oligo sequence and chemistry on the efficiency of oligodeoxyribonucleotide-mediated mRNA cleavage.

Using the endogenous histone H4 mRNA of Xenopus oocytes as a target, we have previously shown that 20mer oligos complementary to different parts of this sequence vary in their effectiveness at causing mRNA cleavage in vivo, and that some of the RNA can never be cleaved. In this paper we show that the resistant RNA is not localised within one part of the oocyte, and that the relative resistance in vivo of endogenous or synthetic H4 mRNA to the different oligos is preserved in an in vitro assay system using deproteinised RNA. If an prior annealing step is included in vitro, all resistance is abolished. Chemical modification of one oligo by end substitution with methylphosphonate or phosphorothioate residues did not improve cleavage efficiency. Oligos with complete phosphorothioate substitution cause slower cleavage in vivo but persist for longer. Consequently phosphorothioate oligos are effective at lower doses than phosphodiester ones, provided that the incubation time is long enough (24 hours). Increasing oligo length from 20nt to 30nt increases phosphorothioate oligo efficiency over long reaction times in vivo, but decreases efficiency during short in vitro assays. Similar increases in length did not affect phosphodiester oligo performance in vivo, but caused a decrease in efficiency in vitro which was overcome by an annealing step.

Animals↗
PubMed1990

Identification and characterization of hetA, a gene that acts early in the process of morphological differentiation of heterocysts.

Envelope polysaccharide is a major early diagnostic of differentiating heterocysts. The mutation in mutant EF116 of Anabaena sp. strain PCC 7120 reduces the cohesiveness of this polysaccharide. A 3.5-kilobase fragment of DNA cloned from the wild type of this Anabaena sp. was previously shown to complement this mutation. We present the nucleotide sequence of a 2,555-base-pair portion of this fragment containing an open reading frame (ORF) of 601 amino acids. Complementation analysis using deletion derivatives of the 3.5-kilobase fragment showed that the gene mutated in EF116, which we designate hetA, lies within this ORF. Transcription of hetA was induced as a result of deprivation for nitrate and yielded a monocistronic mRNA that was present at greatest abundance 7 h after nitrogen stepdown. At that time, proheterocysts could not be distinguished by light microscopy; transcription of nifHD, structural genes of nitrogenase, peaked much later. Situated 3' to hetA are 4 identical repeats of the sequence 5'-TTCAAAA-3' and 12 repeats (10 identical) of the sequence 5'-CCCCAAT-3'. The 12 repeats, present within and near the 5' end of a second ORF, are almost identical to repeats that have been reported to be present in the region between the petC and petA genes of a related cyanobacterium.

Base Sequence↗
PubMed1989

General pharmacology of nizatidine in animals.

Nizatidine (N-[2-[[[2-[(dimethylamino)methyl-4-thiazolyl]- methyl]thio]ethyl]-N'-methyl-2-nitro-1,1-ethenediamine, LY139037, Axid) is a novel, potent, and selective H2-antagonist. The potential secondary pharmacologic effects of this agent on the cardiovascular, respiratory, gastrointestinal, renal, hepatic, autonomic, and central nervous systems as well as effects on circulating blood glucose and the acute inflammatory response were examined. Nizatidine was generally inactive in the tests conducted in mice, rats, guinea pigs, rabbits, and dogs. Nizatidine and the reference H2-antagonist, cimetidine, both produced effects upon the cardiovascular and respiratory systems by intravenous administration in anesthetized dogs at doses in excess of the intended clinical exposure. In summary, these studies confirm the selective pharmacologic activity of nizatidine and indicate a low potential for secondary pharmacologic side effects to be encountered clinically.

Animals↗
PubMed1987

A quantitative approach to sequence comparisons of nitrogenase MoFe protein alpha- and beta-subunits including the newly sequenced nifK gene from Klebsiella pneumoniae.

The nucleotide sequence was determined for part of the Klebsiella pneumoniae nif gene cluster containing the 3' end of the nifD gene and the entire length of the nifK gene (encoding the alpha- and beta-subunits of the nitrogenase MoFe protein respectively), as well as the putative start of the nifY gene, a gene of as yet unknown function. A broad-based comparison of a number of MoFe protein alpha-subunits, beta-subunits and alpha-versus beta-subunits was carried out by the use of a computer program that simultaneously aligns three protein sequences according to the mutation data matrix of Dayhoff. A new kind of quantitative statistical measure of the similarity between the aligned sequences was obtained by calculating and plotting standardized similarity scores for overlapping segments along the aligned proteins. This calculation determines if a test sequence is similar to the consensus sequence of two other proteins that are known to be related to each other. The different beta-subunits compared were found to be significantly similar along most of their sequence, with the exception of two relatively short regions centred around residues 225 and 300, which contain insertions/deletions. The overall pattern of similarity between different alpha-subunits exhibits resemblance to the overall pattern of similarity between different beta-subunits, including regions of low similarity centred around residues 225 and 340. Comparison of alpha-subunits with beta-subunits showed that a region of significant similarity between the two types of subunits was located approximately between residues 120 and 180 in both subunits, but other parts of the proteins were only marginally similar. These results provide insights into likely tertiary structural features of the MoFe protein subunits.

Amino Acid Sequence↗
PubMed1987

Nitrogenase MoFe protein subunits from Klebsiella pneumoniae expressed in foreign hosts. Characteristics and interactions.

The expression of selected nitrogen fixation (nif) genes from Klebsiella pneumoniae in foreign hosts provides an approach to determine the pathway, minimal genetic requirements, and host dependence of nitrogenase assembly. In this study, we investigated the assembly of the alpha 2 beta 2 MoFe protein, responsible for substrate binding and reduction, by introducing nifD and nifK (encoding respectively, the alpha and beta subunits) into Escherichia coli and the yeast Saccharomyces cerevisiae. In E. coli, both genes were expressed from the nifHDKY operon; in yeast, the genes, separately fused to the yeast ADH1 promoter, were introduced on two different plasmids. Denaturing immunoblot analyses demonstrated the presence of significant amounts of NifD and NifK in both hosts. In E. coli, the level or perhaps modification of NifD depended on the growth medium of the bacteria. Nondenaturing, anaerobic immunoblot assays revealed in E. coli, nif-specific antigens of lower electrophoretic mobility than Kp1, which may represent assembly intermediates. In yeast, no putative assembled products were evident, and the predominant antigens corresponded to the monomeric forms of the polypeptides. These results indicate that, unlike NifH, the Fe protein subunit (Berman, J., Gershoni, J. M., and Zamir, A. (1985) J. Biol. Chem. 260, 5240-5243), NifD and NifK are insufficient for the assembly of an electrophoretically Kp1-like structure. Homodimerization of nifK and probably of nifD primary gene products does not appear to occur spontaneously and hence is unlikely to represent the initial step in the assembly. The difference between the two hosts suggests that the cellular environment or mode of expression could affect the interaction between the two subunits.

Escherichia coli↗