ABSTRACT
Small-angle neutron scattering (SANS) is a powerful technique for the characterisation of macromolecular structures and interactions. Its main advantage over other solution state approaches is the ability to use D2O/H2O solvent contrast variation to selectively match out specific parts of a multi-component system. While proteins, nucleic acids, and lipids are readily distinguished in this way, it is not possible to locate different parts of a protein-protein system without the introduction of additional contrast by selective deuteration. Here, we describe new methods by which 'matchout labelled' proteins can be produced using Escherichia coli and Pichia pastoris expression systems in high cell-density cultures. The method is designed to produce protein that has a scattering length density that is very close to that of 100% D2O, providing clear contrast when used with hydrogenated partner proteins in a complex. This allows the production of a single sample system for which SANS measurements at different solvent contrasts can be used to distinguish and model the hydrogenated component, the deuterated component, and the whole complex. The approach, which has significant cost advantages, has been extensively tested for both types of expression system.
Subject(s)
Deuterium/chemistry , Escherichia coli/genetics , Genetic Engineering/methods , Neutron Diffraction , Pichia/genetics , Recombinant Proteins/genetics , Scattering, Small Angle , Cell Count , Cells, Cultured , Complement Factor H/biosynthesis , Complement Factor H/chemistry , Complement Factor H/genetics , Complement Factor H/isolation & purification , Gene Expression , Isotope Labeling , Maltose-Binding Proteins/biosynthesis , Maltose-Binding Proteins/chemistry , Maltose-Binding Proteins/genetics , Maltose-Binding Proteins/isolation & purification , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purificationABSTRACT
We present the case of a 58-year old female with de novo dialysis-dependent acute kidney injury (AKI) secondary to myeloma cast nephropathy. The patient underwent extended high cut-off haemodialysis (HCO-HD), in conjunction with bortezomib-based chemotherapy, and soon became dialysis independent with normal renal function. To our knowledge, this is the first time this treatment strategy has been employed successfully in an Irish centre.
Subject(s)
Acute Kidney Injury/therapy , Antineoplastic Agents/therapeutic use , Boronic Acids/therapeutic use , Multiple Myeloma/complications , Pyrazines/therapeutic use , Renal Dialysis/methods , Acute Kidney Injury/blood , Acute Kidney Injury/etiology , Anti-Bacterial Agents/therapeutic use , Biopsy , Bortezomib , Combined Modality Therapy , Female , Humans , Ireland , Kidney Function Tests , Middle Aged , Multiple Myeloma/drug therapyABSTRACT
We report a fatal case of meningitis caused by Salmonella Enteritidis phage type 14b in a middle-aged man who had no history or findings to suggest he was immunocompromised. To our knowledge, this is the first reported case of Salmonella meningitis in an adult in Ireland, and the first case of meningitis in an adult caused by phage type 14b. This case was associated with a nationwide cluster of salmonellosis which is still under investigation at the time of writing.
Subject(s)
Immunocompromised Host , Meningitis, Bacterial/drug therapy , Salmonella Infections/drug therapy , Salmonella enteritidis/isolation & purification , Sepsis , Fatal Outcome , Food Microbiology , Humans , Ireland , Male , Middle Aged , Salmonella enteritidis/drug effects , Sepsis/physiopathology , Severity of Illness IndexABSTRACT
OBJECTIVES: To determine patient preferences for information and for participation in decision-making, and the determinants of these preferences in patients recently admitted to an acute hospital. DESIGN: Prospective questionnaire-based study. SETTING: Medical wards of an acute teaching hospital. PARTICIPANTS: One hundred and fifty-two consecutive acute medical inpatients, median age 74 years. MEASUREMENTS: Standardised assessment included abbreviated mental test and subjective measure of severity of illness. Patients' desire for information was assessed using a 5-point Likert scale, and their desire for a role in medical decision-making using the Degner Control of Preferences Scale. RESULTS: Of the 152 patients, 93 (61%) favoured a passive approach to decision-making (either "leave all decisions to the doctor" or "doctor makes final decision but seriously considers my opinion." In contrast, 101 (66%) patients sought "very extensive" or "a lot" of information about their condition. No significant effects of age, sex, socio-economic group or severity of acute illness on desire for information or the Degner scale result were found. There was no agreement between patients' preferences on the Degner scale and their doctors' predictions of those preferences. CONCLUSIONS: Acute medical inpatients want to receive a lot of information about their illness, but most prefer a relatively passive role in decision-making. The only way to determine individual patient preferences is to ask them; preferences cannot be predicted from clinical or sociodemographic data.
