Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 3 de 3
Filter
Add more filters










Database
Language
Publication year range
1.
West J Emerg Med ; 15(2): 142-4, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24672600

ABSTRACT

We present an interesting case of a patient with a previously known diaphragmatic hernia in which the colon became incarcerated, ischemic and finally perforated. She had no prior history of abdominal pain or vomiting, yet she presented with cardiovascular collapse. She was quickly diagnosed with a tension pneumothorax and treated accordingly. To our knowledge, this is the only case report of a tension pneumothorax associated with perforated bowel that was not in the setting of trauma or colonoscopy.


Subject(s)
Hernia, Diaphragmatic/diagnosis , Intestinal Perforation/diagnosis , Pneumothorax/diagnosis , Diagnosis, Differential , Emergency Service, Hospital , Female , Hernia, Diaphragmatic/complications , Hernia, Diaphragmatic/diagnostic imaging , Humans , Intestinal Perforation/complications , Intestinal Perforation/diagnostic imaging , Middle Aged , Pneumothorax/diagnostic imaging , Sickle Cell Trait/complications , Tomography, X-Ray Computed
2.
J Am Chem Soc ; 130(38): 12568-9, 2008 Sep 24.
Article in English | MEDLINE | ID: mdl-18759389

ABSTRACT

Inclusion bodies are insoluble aggregates that are formed by bacteria to store excess recombinant protein produced during expression. The structure of the protein in inclusion bodies is poorly understood but it has been hypothesized that the protein may form misfolded beta sheet aggregates. This paper presents an isotopic labeling and solid-state nuclear magnetic resonance approach to determine the secondary structure of individual residues within a recombinant influenza virus "FHA2" protein in inclusion bodies. The inclusion bodies were studied either in the context of the unlysed hydrated E. coli cells or in the hydrated pellet formed from centrifugation of the material insoluble in the cell lysate. The native structure of FHA2 is predominantly helical and native helical structure was also observed for several specific residues in the inclusion body FHA2. This approach will be applicable to structural analysis of many inclusion body proteins and should provide useful information for optimizing solubilization and purification protocols of these proteins.


Subject(s)
Hemagglutinins, Viral/chemistry , Inclusion Bodies/chemistry , Nuclear Magnetic Resonance, Biomolecular/methods , Carbon Isotopes , Escherichia coli/genetics , Escherichia coli/metabolism , Hemagglutinins, Viral/analysis , Hemagglutinins, Viral/biosynthesis , Hemagglutinins, Viral/genetics , Inclusion Bodies/metabolism , Protein Structure, Secondary , Recombinant Proteins/analysis , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/genetics
3.
Protein Expr Purif ; 61(2): 212-9, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18640277

ABSTRACT

This paper describes methods to produce an isotopically labeled 23 kDa viral membrane protein with purified yield of 20 mg/L of Escherichia coli shake flask culture. This yield is sufficient for NMR structural studies and the protein production methods are simple, straightforward, and rapid and likely applicable to other recombinant membrane proteins expressed in E. coli. The target FHA2 protein is the full ectodomain construct of the influenza virus hemagglutinin protein which catalyzes fusion between the viral and the cellular endosomal membranes during infection. The high yield of FHA2 was achieved by: (1) initial growth in rich medium to A(600) approximately 8 followed by a switch to minimal medium and induction of protein expression; and (2) obtaining protein both from purification of the detergent-soluble lysate and from solubilization, purification, and refolding of inclusion bodies. The high cell density was achieved after optimization of pH, oxygenation, and carbon source and concentration, and the refolding protocol was optimized using circular dichroism spectroscopy. For a single residue of membrane-associated FHA2 that was obtained from purification and refolding of inclusion bodies, native conformation was verified by the (13)CO chemical shifts measured using solid-state nuclear magnetic resonance spectroscopy.


Subject(s)
Escherichia coli/metabolism , Membrane Fusion , Protein Folding , Viral Fusion Proteins , Amino Acid Sequence , Carbon Isotopes , Cell Proliferation/drug effects , Cells, Cultured , Circular Dichroism/methods , Cloning, Molecular , Dose-Response Relationship, Drug , Escherichia coli/chemistry , Escherichia coli/growth & development , Glucose/pharmacology , Glycerol/pharmacology , Hydrogen-Ion Concentration , Isotope Labeling/methods , Magnetic Resonance Spectroscopy/methods , Magnetic Resonance Spectroscopy/standards , Molecular Sequence Data , Protein Structure, Tertiary , Reference Standards , Solubility , Time Factors , Viral Fusion Proteins/biosynthesis , Viral Fusion Proteins/chemistry , Viral Fusion Proteins/isolation & purification
SELECTION OF CITATIONS
SEARCH DETAIL
...