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1.
Chemosphere ; 265: 129086, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33340834

ABSTRACT

The use of phytoextraction plant species to accumulate soil metals into harvestable plant parts is a method used for managing soils with high cadmium (Cd). We evaluated three Cd accumulating species recently recommended for such use in cacao farms where Cd removal is needed to maintain markets: Helianthus annuus (sunflower), Brassica napus (rapeseed), and Chyrsopogon zizanioides (vetiver). Plants were grown in two greenhouse pot experiments with different Cd-spiked growth media: (sand plus perlite) and a natural soil. Plant total Cd and Cd uptake in shoot biomass of all species, across both experiments, increased linearly with increasing amounts of added Cd. Rapeseed had the highest plant total Cd and sunflower had the highest Cd uptake in shoot biomass. The highest application of Cd corresponded to the highest plant total Cd and shoot biomass Cd uptake, regardless of species. The bioconcentration factor (BCF) for each species increased in a curvilinear manner with added Cd, with maximum BCF values for plants grown in the sand and perlite matrix at 2.5 mg kg-1 added Cd and in the natural soil at 5.0 mg kg-1 added Cd. We conclude that the Cd uptake (shoot biomass only) capability of the three species examined is greatest for sunflower given its increased uptake with Cd additions, its BCF value > 1, and lack of observed visual Cd toxicity symptoms, fungus and insect damage. Although these species had BCF >1, the potential annual removal of Cd would have been too small to support a meaningful phytoextraction practice.


Subject(s)
Brassica napus , Helianthus , Soil Pollutants , Biodegradation, Environmental , Cadmium/analysis , Soil , Soil Pollutants/analysis
2.
AJNR Am J Neuroradiol ; 40(10): 1759-1765, 2019 10.
Article in English | MEDLINE | ID: mdl-31558504

ABSTRACT

BACKGROUND AND PURPOSE: Selection of the correct flow-diverter size is critical for cerebral aneurysm treatment success, but it remains challenging due to the interplay of device size, anatomy, and deployment. Current convention does not address these challenges well. The goals of this pilot study were to determine whether computational modeling improves flow-diverter sizing over current convention and to validate simulated deployments. MATERIALS AND METHODS: Seven experienced neurosurgeons and interventional neuroradiologists used computational modeling to prospectively plan 19 clinical interventions. In each patient case, physicians simulated 2-4 flow-diverter sizes that were under consideration based on preprocedural imaging. In addition, physicians identified a preferred device size using the current convention. A questionnaire on the impact of computational modeling on the procedure was completed immediately after treatment. Rotational angiography image data were acquired after treatment and compared with flow-diverter simulations to validate the output of the software platform. RESULTS: According to questionnaire responses, physicians found the simulations useful for treatment planning, and they increased their confidence in device selection in 94.7% of cases. After viewing the simulations results, physicians selected a device size that was different from the original conventionally planned device size in 63.2% of cases. The average absolute difference between clinical and simulated flow-diverter lengths was 2.1 mm. In 57% of cases, average simulated flow-diverter diameters were within the measurement uncertainty of clinical flow-diverter diameters. CONCLUSIONS: Physicians found computational modeling to be an impactful and useful tool for flow-diverter treatment planning. Validation results showed good agreement between simulated and clinical flow-diverter diameters and lengths.


Subject(s)
Blood Vessel Prosthesis Implantation/methods , Blood Vessel Prosthesis , Computer Simulation , Intracranial Aneurysm/surgery , Female , Humans , Male , Pilot Projects , Software
3.
Sci Rep ; 7(1): 17102, 2017 12 06.
Article in English | MEDLINE | ID: mdl-29213103

ABSTRACT

The pervasive and unabated nature of global amphibian declines suggests common demographic responses to a given driver, and quantification of major drivers and responses could inform broad-scale conservation actions. We explored the influence of climate on demographic parameters (i.e., changes in the probabilities of survival and recruitment) using 31 datasets from temperate zone amphibian populations (North America and Europe) with more than a decade of observations each. There was evidence for an influence of climate on population demographic rates, but the direction and magnitude of responses to climate drivers was highly variable among taxa and among populations within taxa. These results reveal that climate drivers interact with variation in life-history traits and population-specific attributes resulting in a diversity of responses. This heterogeneity complicates the identification of conservation 'rules of thumb' for these taxa, and supports the notion of local focus as the most effective approach to overcome global-scale conservation challenges.


Subject(s)
Amphibians/physiology , Conservation of Natural Resources , Animals , Climate Change , Europe , North America , Population Dynamics , Seasons , Urodela/physiology
4.
Opt Lett ; 41(22): 5318-5321, 2016 Nov 15.
Article in English | MEDLINE | ID: mdl-27842122

ABSTRACT

Imperfections in integrated photonics manufacturing have a detrimental effect on the maximal achievable visibility in interferometric architectures. These limits have profound implications for further technological developments in photonics and in particular for quantum photonic technologies. Active optimization approaches, together with reconfigurable photonics, have been proposed as a solution to overcome this. In this Letter, we demonstrate an ultrahigh (>60 dB) extinction ratio in a silicon photonic device consisting of cascaded Mach-Zehnder interferometers, in which additional interferometers function as variable beamsplitters. The imperfections of fabricated beamsplitters are compensated using an automated progressive optimization algorithm with no requirement for pre-calibration. This work shows the possibility of integrating and accurately controlling linear-optical components for large-scale quantum information processing and other applications.

5.
BMC Pediatr ; 16(1): 168, 2016 10 21.
Article in English | MEDLINE | ID: mdl-27765020

ABSTRACT

BACKGROUND: Global childhood mortality rates remain high. Millennium Development Goal 4 focused efforts on reducing rates by two-thirds between 1990 and 2015. In Ethiopia, child mortality rates dropped 71 % from 1990 to 2015, however it is estimated that 184,000 Ethiopian children die each year. There is limited information about pediatric hospital admissions in Ethiopia. Our aims were to examine the temporal relationship of mortality to admission, describe the demographics, and identify cause mortality of children admitted to the Zewditu Memorial Hospital (ZMH). METHODS: A four-year retrospective review of pediatric admissions was conducted at the pediatric emergency room and pediatric hospital ward at ZMH in Addis Ababa, Ethiopia. Admission entries from 2011-2014 of children age 29 days-14 years were reviewed. Age, gender, admission date, disease classification, discharge status and date were obtained. Patient gender was compared using Chi-square analysis. A descriptive analysis was used for age and cause mortality. RESULTS: A total of 6866 patient entries were reviewed. The proportion of admissions younger than age 5 was 0.747 (95 % CI 0.736-0.757). Overall mortality was 0.042 (95 % CI, 0.037-0.047). The proportion of recorded deaths occurring within 2 days of admission was 0.437 (95 % CI 0.380-0.494). The proportion of male admissions was significantly higher than female admissions in all age groups (male 0.575, p < 0.0001, 95 % CI 0.562-0.586). The main causes of mortality were pneumonia (0.253, 95 % CI, 0.203-0.303), severe acute malnutrition (0.222, 95 % CI 0.174-0.27), HIV/AIDS-related complications (0.056, 95 % CI 0.029-0.083), spina bifida (0.049, 95 % CI 0.024-0.074), and hydrocephalus (0.045, 95 % CI 0.021-0.069). CONCLUSIONS: Our study revealed a lower mortality rate than previously reported in Ethiopia. Despite this, 44 % of pediatric hospital mortality occurred early during hospitalization, higher than reported at other Ethiopian hospitals. This adds further evidence that systematic efforts should be dedicated to improve pediatric emergency care. Admissions included 58 % male patients, similar to other reports in Ethiopia implying that this may be a nation-wide phenomenon. The observed disparity may be due to societal factors regarding care-seeking behaviors or male predilection for respiratory illness warranting further investigation. Cause mortality patterns were similar to reports in analogous settings.


Subject(s)
Cause of Death/trends , Child Mortality/trends , Hospital Mortality/trends , Adolescent , Child , Child, Preschool , Developing Countries/statistics & numerical data , Emergency Service, Hospital/statistics & numerical data , Emergency Service, Hospital/trends , Ethiopia/epidemiology , Female , Hospitals, Pediatric/statistics & numerical data , Hospitals, Pediatric/trends , Humans , Infant , Infant, Newborn , Male , Retrospective Studies
7.
Biol Cybern ; 104(3): 185-95, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21394539

ABSTRACT

Studying neurons from an energy efficiency perspective has produced results in the research literature. This paper presents a method that enables computation of low energy input current stimuli that are able to drive a reduced Hodgkin-Huxley neuron model to approximate a prescribed time-varying reference membrane voltage. An optimal control technique is used to discover an input current that optimally minimizes a user selected balance between the square of the input stimulus current (input current 'energy') and the difference between the reference voltage and the membrane voltage (tracking error) over a stimulation period. Selecting reference signals to be membrane voltages produced by the neuron model in response to common types of input currents i(t) enables a comparison between i(t) and the determined optimal current stimulus i*(t). The intent is not to modify neuron dynamics, but through comparison of i(t) and i*(t) provide insight into neuron dynamics. Simulation results for four different bifurcation types demonstrate that this method consistently finds lower energy stimulus currents i*(t) that are able to approximate membrane voltages as produced by higher energy input currents i(t) in this neuron model.


Subject(s)
Action Potentials/physiology , Electric Stimulation/methods , Membrane Potentials/physiology , Models, Neurological , Neurons/physiology , Mathematics
8.
J Perinatol ; 31(5): 368-72, 2011 May.
Article in English | MEDLINE | ID: mdl-21151008

ABSTRACT

OBJECTIVE: The objective of this study was to compare alterations in the middle cerebral artery (MCA) pulsatility index (PI) and mean velocity (V (mean)) after laser surgery for twin-twin transfusion syndrome (TTTS). STUDY DESIGN: MCA Doppler studies were conducted 1 day before and after laser surgery for TTTS. The pre- and postoperative mean (standard deviation) of the MCA PI and V (mean) z-scores of the recipient and donor fetuses were calculated and compared. Data were analyzed using paired testing and multivariable linear regression models. RESULTS: A total of 103 patients met the study criteria. Recipients' MCA PI increased from -1.29 (1.20) preoperatively to 0.14 (1.52) postoperatively (P < 0.0001), whereas the donors' PI did not change significantly (-0.31 (1.67) to -0.67 (1.29); P = 0.12). There was no significant difference between preoperative and postoperative MCA V (mean) in donors (0.39 (0.83) and 0.38 (0.93), respectively; P = 0.5048) or recipients (0.60 (0.74) and 0.63 (0.90), respectively; P = 0.5324). CONCLUSIONS: Despite the changes in the MCA PI after laser surgery for TTTS, the MCA V (mean) remained constant. These findings may suggest some autoregulatory capacity in the cerebral vessels of the mid-trimester fetus.


Subject(s)
Blood Flow Velocity , Middle Cerebral Artery/physiopathology , Ultrasonography, Doppler, Color , Ultrasonography, Prenatal/methods , Female , Fetofetal Transfusion/physiopathology , Fetofetal Transfusion/surgery , Fetus , Homeostasis , Humans , Laser Therapy , Perioperative Care , Placental Circulation , Pregnancy , Pregnancy Complications, Hematologic/physiopathology , Pregnancy Complications, Hematologic/surgery , Pregnancy Trimester, Second
9.
AJNR Am J Neuroradiol ; 30(9): 1787-91, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19541779

ABSTRACT

BACKGROUND AND PURPOSE: To date, very limited attention has been given to ocular abnormalities or growth parameters detected by fetal MR imaging. Our objective was to retrospectively determine the relationship between different parameters of eye development and estimated gestational age in the human fetus by use of fetal MR imaging. MATERIALS AND METHODS: A retrospective study was performed to measure the transverse diameter, interocular distance, and lens diameter of the globes of 127 fetuses who had a morphologically normal central nervous system. Multiple single-shot T2 fast spin-echo images were obtained with a 1.5T magnet by use of contiguous 3-mm intervals in at least 2 orthogonal planes. Loess curves were fitted to explore the relationship between gestational age and each of the 3 measurements of interest. Different models were compared statistically to determine the model of best fit. RESULTS: For each variable of interest, the "best" model of eye growth was a quadratic function. Specifically, lens growth seems to plateau after 36 weeks of gestation, interocular distance plateaus after 36 weeks of gestation, and globe growth plateaus after 42 weeks of gestation. CONCLUSIONS: The lens, orbit, and interocular distance growth of the fetus can be demonstrated on fetal MR imaging. All 3 measurements suggest a quadratic model of growth, which indicates slowing of growth toward the end of gestation.


Subject(s)
Eye/anatomy & histology , Eye/embryology , Gestational Age , Magnetic Resonance Imaging/methods , Prenatal Diagnosis/methods , Eye/growth & development , Female , Humans , Male
10.
Biomed Khim ; 54(2): 218-22, 2008.
Article in Russian | MEDLINE | ID: mdl-18522224

ABSTRACT

Blood plasma phospholipids from 45 patients with chronic gastritis and 20 healthy volunteers have been investigated. Chronic gastritis exacerbation is accompanied by essential change in the spectrum and concentration of the phospholipids. The absolute concentrations of the phospholipids and such fractions as phosphatidylcholine, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine and sphingomyelin significally increased while concentrations of phosphatidilinositol and lysophosphatidylserine decreased. The period of the remission of the disease was accompanied by a certain normalization of the level and spectrum phospholipids and literature data suggest that phospholipids and their particular fractions are involved into pathogenesis of the inflammatory process in gastric mucosa and in the process of reparative regeneration aswell.


Subject(s)
Gastritis/metabolism , Phospholipids/blood , Adolescent , Adult , Chronic Disease , Female , Humans , Male , Plasma
11.
Eksp Klin Gastroenterol ; (3): 130-4, 2008.
Article in Russian | MEDLINE | ID: mdl-19145893
12.
Interv Neuroradiol ; 9(1): 7-19, 2003 Mar 30.
Article in English | MEDLINE | ID: mdl-20591298

ABSTRACT

SUMMARY: Thrombus organization in canine carotid arteries occluded with platinum microcoils was studied to determine if endothelial injury created with a Xenon Chloride Excimer Laser (XEL) could acclerate endovascular fibrosis. Ten common carotid artery stumps were created in ten dogs. Each of four stumps were schematically divided into four longitudinally contiguous injury zones (thermal ablation injury, non-ablative injury, proximal and distal non-injury zones) to test the effects of ablative and non-ablative injury and to establish a set of internal controls that would account for proximity to circulating blood at the ostium of the occluded artery. Following XEL irradiation of the endothelium through an arteriotomy, each stump was embolized with microcoils. Four control stumps were subjected to sham laser procedures, and embolized in an identical fashion. Two additional stumps were embolized in the absence of sham surgery. Angiographic, gross and histologic analysis was performed after four weeks. Specimens of freshly clotted whole blood mixed with microcoils were used as an additional control. In irradiated stumps and non-irradiated stumps (sham and embolization only), angiography revealed no evidence of coil compaction or recanalization. In all irradiated stumps the thermal ablation zone contained fibrous tissue and neovascularity without unorganized thrombus. The other zones in the irradiated stumps were indistingnishable from each other and from all zones in the non-irradiated sham stumps, containing primarily unorganized thrombus. Stumps embolized in the absence of sham surgery were filled with material that was grossly and microscopically identical to specimens of freshly clotted whole blood containing microcoils. The results indicate that thermal ablation injury of the endothelium accelerates thrombus organization in canine carotid arteries occluded with platinum microcoils.

13.
Eksp Klin Gastroenterol ; (6): 76-9, 155, 2003.
Article in Russian | MEDLINE | ID: mdl-15065533

ABSTRACT

The study of systemic hemostasis in 254 patients with chronic gastritis has revealed that the exacerbation of the disease proceeds with marked disorders of blood coagulation typical for the first and second stages of the chronic relapsing thrombohemorrhagic syndrome and thrombophilia. They manifested in intensification of coagulability, lowering of anticoagulative and fibrinolytic potentials of blood, pathological intravascular coagulation. The disorders of hemostasis were strongly expressed during the period of marked exacerbation of the disease and in patients with erosive gastritis.


Subject(s)
Gastritis/blood , Gastrointestinal Hemorrhage/blood , Helicobacter Infections/blood , Hemostasis , Thrombosis/blood , Adult , Chronic Disease , Female , Gastric Mucosa/blood supply , Gastric Mucosa/pathology , Gastritis/complications , Gastritis/microbiology , Gastritis/pathology , Gastritis, Atrophic/blood , Gastritis, Atrophic/complications , Gastritis, Atrophic/microbiology , Gastritis, Atrophic/pathology , Gastrointestinal Hemorrhage/etiology , Gastrointestinal Hemorrhage/pathology , Helicobacter Infections/complications , Helicobacter Infections/microbiology , Helicobacter Infections/pathology , Helicobacter pylori/isolation & purification , Humans , Male , Thrombosis/etiology , Thrombosis/pathology
14.
Eksp Klin Gastroenterol ; (4): 14-7, 130, 2002.
Article in Russian | MEDLINE | ID: mdl-12503266

ABSTRACT

The study of microvasculature in bioptic specimens of gastric mucosa and of conjunctival microcirculation from 254 patients with chronic gastritis revealed that exacerbation of the disease proceeds with severe disorders of terminal blood flow which are typical for the chronic relapse trombohemorrhagic syndrome and have generalized character. The microvascular disorders is manifested by plasma- and hemorrhages, by microvascular spasm and dilatation, intravascular hemostasis and microthrombosis. These manifestations were more expressed over the period of bright marked exacerbation of the disease and in patients with erosive affection of gastric mucosa.


Subject(s)
Gastritis/pathology , Gastritis/physiopathology , Stomach/blood supply , Adult , Chronic Disease , Female , Gastric Mucosa/blood supply , Gastric Mucosa/pathology , Humans , Male , Microcirculation , Middle Aged , Stomach/pathology
15.
Eksp Klin Gastroenterol ; (3): 22-5, 118, 2002.
Article in Russian | MEDLINE | ID: mdl-12353381

ABSTRACT

The study of the microvessels in bioptates of gastric mucosa and micro haemocirculation in the conjunctiva of 254 patients with chronic gastritis revealed that exacerbation of the gastric process is going on the background of hard terminal bloodstream disorders. They have the generalized character and picture of the typical chronic relapsing trombohaemorrhagic syndrome. The use of Prodectin (250 mg), Teonicolum (150 mg), Redergin (1 tab.) and Aescuzan (25 dr.) 4 times per day during 3 weeks helps to eliminate the microcirculatory disorders and exacerbation of the chronic gastritis.


Subject(s)
Ergoloid Mesylates/therapeutic use , Escin/therapeutic use , Gastritis/drug therapy , Gastrointestinal Agents/therapeutic use , Pyridinolcarbamate/therapeutic use , Xanthinol Niacinate/therapeutic use , Drug Therapy, Combination , Ergoloid Mesylates/administration & dosage , Escin/administration & dosage , Female , Gastrointestinal Agents/administration & dosage , Humans , Male , Middle Aged , Pyridinolcarbamate/administration & dosage , Xanthinol Niacinate/administration & dosage
16.
Curr Opin Chem Biol ; 5(5): 525-34, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11578925

ABSTRACT

Nonribosomal peptide synthetases are large enzyme complexes that synthesize a variety of peptide natural products through a thiotemplated mechanism. Assembly of the peptides proceeds through amino acid loading, amide-bond formation and chain translocation, and finally thioester lysis to release the product. The final products are often heavily modified, however, through methylation, epimerization, hydroxylation, heterocyclization, oxidative cross-linking and attachment of sugars. These activities are the province of specialized enzymes (either embedded in the multidomain nonribosomal peptide synthetase structure or standalone).


Subject(s)
Bacteria/metabolism , Multienzyme Complexes/metabolism , Peptide Biosynthesis , Peptide Synthases/chemistry , Peptides/metabolism , Cytochrome P-450 Enzyme System/metabolism , Esters/chemical synthesis , Esters/chemistry , Esters/metabolism , Glycosyltransferases , Methyltransferases/metabolism , Models, Chemical , Peptide Chain Elongation, Translational , Peptide Synthases/metabolism , Peptides/chemistry , Peptides, Cyclic/biosynthesis , Protein Conformation , Racemases and Epimerases/metabolism , Stereoisomerism
17.
J Clin Psychiatry ; 62 Suppl 22: 38-42, 2001.
Article in English | MEDLINE | ID: mdl-11599647

ABSTRACT

Antidepressant medications are typically taken on a daily basis owing to both tradition and the pharmacokinetics of these agents. Because fluoxetine and its primary metabolite norfluoxetine have long half-lives and flat dose-response curves, we examined the tolerability of a weekly dose and its equivalence to daily dosing during the continuation phase of treatment for major depressive disorder (MDD). Open-label treatment with 20 mg of fluoxetine daily for 7 weeks began with 114 subjects. Subsequently, 70 subjects who met criteria for response were randomly assigned in a double-blind design to 1 of 3 treatment groups (20 mg of fluoxetine daily [N = 21], 60 mg of fluoxetine weekly [N = 28], or placebo [N = 21]) and followed for 7 weeks. No statistically significant differences were observed in several clinical measures. Tolerability in the 3 groups was similar; there was no difference in dropout rates or adverse events. Hence, weekly dosing of fluoxetine appears to be well tolerated and possibly as effective as daily dosing in the treatment of MDD. It is proposed that less frequent dosing could potentially benefit patients by enhancing adherence and minimizing the risk of side effects and drug-drug interactions.


Subject(s)
Depressive Disorder/drug therapy , Fluoxetine/administration & dosage , Selective Serotonin Reuptake Inhibitors/administration & dosage , Adult , Chronic Disease , Depressive Disorder/prevention & control , Double-Blind Method , Drug Administration Schedule , Drug Interactions , Female , Fluoxetine/adverse effects , Fluoxetine/pharmacokinetics , Follow-Up Studies , Humans , Male , Patient Dropouts/statistics & numerical data , Placebos , Selective Serotonin Reuptake Inhibitors/adverse effects , Selective Serotonin Reuptake Inhibitors/pharmacokinetics , Treatment Outcome
19.
Proc Natl Acad Sci U S A ; 98(14): 7712-7, 2001 Jul 03.
Article in English | MEDLINE | ID: mdl-11427730

ABSTRACT

Microcin B17 (MccB17) is a 3.1-kDa Escherichia coli antibiotic that contains thiazole and oxazole heterocycles in a peptide backbone. MccB17 inhibits its cellular target, DNA gyrase, by trapping the enzyme in a complex that is covalently bound to double-strand cleaved DNA, in a manner similar to the well-known quinolone drugs. The identification of gyrase as the target of MccB17 provides an opportunity to analyze the relationship between the structure of this unusual antibiotic and its activity. In this report, steady-state parameters are used to describe the induction of the cleavable complex by MccB17 analogs containing modified bisheterocyclic sites. The relative potency of these analogs corresponds to the capacity of the compounds to prevent growth of sensitive cells. In contrast to previously reported experiments, inhibition of DNA gyrase supercoiling activity by wild-type MccB17 also was observed. These results suggest that DNA gyrase is the main intracellular target of MccB17. This study probes the structure-function relationship of a new class of gyrase inhibitors and demonstrates that these techniques could be used to analyze compounds in the search for clinically useful antibiotics that block DNA gyrase.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteriocins/pharmacology , Enzyme Inhibitors/pharmacology , Topoisomerase II Inhibitors , Anti-Bacterial Agents/metabolism , Bacteriocins/chemistry , Bacteriocins/metabolism , DNA Topoisomerases, Type II/metabolism , Enzyme Inhibitors/metabolism
20.
Biochemistry ; 40(17): 5313-21, 2001 May 01.
Article in English | MEDLINE | ID: mdl-11318656

ABSTRACT

The HMWP2 subunit of yersiniabactin (Ybt) synthetase, a 230 kDa nonribosomal peptide synthetase (NRPS) making the N-terminus of the Ybt siderophore of Yersinia pestis, has one cysteine-specific adenylation (A) domain, three carrier protein domains (ArCP, PCP1, PCP2), and two heterocyclization domains (Cy1, Cy2). The A domain loads the two PCP domains with cysteines that get heterocyclized by the Cy domains to yield a tricyclic hydroxyphenylthiazolinylthiazolinyl (HPTT) chain lodged in thioester linkage to the PCP2 domain. The interdomain recognition by the Cy1 and Cy2 domains for the three carrier proteins was tested using inactivating mutations at the conserved serine that is phosphopantetheinylated in each carrier domain (S52A, S1439A, and S1977A). These mutant forms of HMWP2 were tested for in trans complementation by carrier protein fragments: holo-ArCPs (S52A), holo-PCP1 and analogues (S1439A), and holo-PCP2 and analogues (S1977A). The S52A mutant tests the recognition of the Cy1 domain for donor acyl-ArCP substrates, while the S1439A mutant tests the specificity of the same Cy1 domain for downstream substrates presented by distinct PCPs. The S1439A likewise tests the recognition of Cy2 for its upstream PCP-tethered acyl donor. The S1977A mutant analogously tests the Cy2 domain for downstream Cys-PCP recognition. In all cases in trans complementation was successful with the carrier protein fragments, allowing kinetic probes of catalytic efficiency for PCP scaffolds and for uncoupling of the condensation and heterocyclization functions of Cy1 and Cy2. Overall, the Cy domains tested showed a definite selectivity for the upstream protein scaffold but were more relaxed toward the downstream acceptor protein. This work points to the importance of protein-protein interactions in mediating directional chain growth in NRPS and presents the first systematic exploration of how the protein scaffolds affect catalytic efficiency.


Subject(s)
Acyl Carrier Protein/chemistry , Bacterial Proteins/chemistry , Catalytic Domain , Coenzyme A Ligases , Peptide Chain Initiation, Translational , Peptide Synthases/chemistry , Phenols , Siderophores/biosynthesis , Thiazoles , Yersinia pestis/enzymology , Acyl Carrier Protein/genetics , Acyl Carrier Protein/metabolism , Alanine/genetics , Amino Acid Isomerases/metabolism , Amino Acid Substitution/genetics , Bacterial Outer Membrane Proteins , Bacterial Proteins/metabolism , Catalytic Domain/genetics , Enzyme Activation/genetics , Iron-Binding Proteins , Molecular Weight , Peptide Chain Initiation, Translational/genetics , Peptide Synthases/genetics , Peptide Synthases/metabolism , Periplasmic Binding Proteins , Protein Structure, Tertiary/genetics , Serine/genetics , Substrate Specificity/genetics , Yersinia pestis/genetics
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