The previously described hybrid plasmid pC7 which carries lacI+ O+Δ(Z)Y+ A+ on a 12.3 × 106‐Mr DNA fragment [Teather et al. (1978) Mol. Gen. Genet. 159, 239–248] was partially digested with the restriction endonuclease EcoRI under conditions reducing the recognition sequence to ↓ d(A‐A‐T‐T) and ligated to the vector pBR322. lacY‐carrying inserts of various sizes (Mr, 1.5–4.7 × 106) were obtained.
Hybrid plasmid pTE18 (2300‐base‐pair insert) carries part of the I (repressor) gene, the promotor‐operator region, part of the Z (β‐galactosidase) gene, the Y (lactose carrier) gene and part of the A (transacetylase) gene. Upon induction of pTE18‐harbouring strains the Y‐gene product is expressed at a nearly constant rate for several generations and accumulates to a level of 12–16% of the total cytoplasmic membrane protein. Integration into the membrane leads to active carrier as judged by binding and transport measurements.
In the Intraoperative Hypothermia for Aneurysm Surgery Trial, neither systemic hypothermia nor supplemental protective drug affected short- or long-term neurologic outcomes of patients undergoing temporary clipping.
There has been much speculation about the mechanism by which cholera toxin exerts its effect on the cytoplasmic side of the membranes with which it interacts. After the pentamer of B subunits (5B) binds to membrane receptors, particularly the monosialylganglioside GM1, the disulphide-linked dimer A1SSA2 (which together with 5B constitutes the complete toxin) is thought to penetrate the membrane, perhaps through a channel formed by 5B and become reduced so that A1SH units reach the cytoplasm and stimulate adenylate cyclase. Evidence for this mechanism is circumstantial. If it is correct, a compound which will specifically label intramembranous sections of the toxin should label the channel-forming B subunits but not the channel-contained A1 subunit. We have tested this prediction with a photoreactive glycolipid compound and have obtained the opposite result. Therefore, we propose that only the A1 subunit enters the membrane and we provide here data on the kinetics of that process.
Purpose: Lumbar subarachnoid catheters for cerebrospinal fluid (CSF) drainage (lumbar drains) are indicated for several medical and surgical conditions. A number of complications can occur from the placement of this type of catheter, including catheter breakage from excessive traction or shearing over the Tuohy needle.
Clinical features:Five cases of lumbar subarachnoid catheter breakage/shearing and catheter fragment retention, as well as one near miss, were identified over a one-year period at a single institution. All (n = 6) patients were undergoing neurosurgical procedures. Four patients required surgical retrieval of the catheter fragments. No patient experienced log-term neurological sequelae.Discussion: From these experiences, the following risks factors for catheter rupture are identified: 1) intentional or accidental retraction of the catheter through the needle during placement; 2) faulty use of the guidewire; or 3) use of excessive force during removal of the catheter. Methods to prevent such complications are suggested, including minimal use, or complete avoidance of a guidewire.
There are distinct differences in the molecular packing of phospholipid molecules in the inner and outer membrane monolayers of small lipid vesicles; a small radius of curvature imparts an asymmetry to the interface between these two monolayers. I have used an amphiphilic fluorescent probe, N-[5-(dimethylamino)naphthalenyl-1-sulfonyl]glycine (dansylglycine), to determine if this asymmetry in molecular packing leads to the existence of different environments for fluorescent probes resident in the membrane. Dansylglycine is highly sensitive to the dielectric constant of its environment, and the fluorescence signal from membrane-bound dye is distinct from that in the aqueous medium. When dansylglycine is first mixed with vesicles, it rapidly partitions into the outer monolayer; the subsequent movement of dye into the inner monolayer is much slower. Because of the time lag between the initial partitioning and the subsequent translocation, it is possible to measure the emission spectrum from membrane-bound dye before and after translocation, thus distinguishing the two potential environments for dansylglycine molecules. In the outer membrane monolayer of small dipalmitoylphosphatidylcholine vesicles, dye fluorescence emission is maximal at 530 nm, corresponding to a dielectric constant of 7 for the medium surrounding the fluorophore. For dye in the inner monolayer, emission is maximal at 519 nm, corresponding to a dielectric constant of 4.7. The results suggest that water molecules are excluded more efficiently from the dye binding sites of the inner membrane monolayer than they are from those of the outer monolayer.(ABSTRACT TRUNCATED AT 250 WORDS)
Background
Perioperative hypothermia has been reported to increase the occurrence of cardiovascular complications. By increasing sympathetic nervous system activity, perioperative hypothermia also has the potential to increase cardiac injury and dysfunction associated with subarachnoid hemorrhage.
Methods
The Intraoperative Hypothermia for Aneurysm Surgery Trial randomized patients undergoing cerebral aneurysm surgery to intraoperative hypothermia (n = 499, 33.3 ± 0.8°C) or normothermia (n = 501, 36.7 ± 0.5°C). Cardiovascular events (hypotension, arrhythmias, vasopressor use, myocardial infarction, etc.) were prospectively followed until 3 month follow-up and were compared between hypothermic and normothermic patients. A subset of 62 patients (hypothermia, n = 33; normothermia, n = 29) also had preoperative and postoperative (within 24 h) measurement of cardiac troponin-I and echocardiography to explore the association between perioperative hypothermia and subarachnoid hemorrhage-associated myocardial injury and left ventricular function.
Results
There was no difference between hypothermic and normothermic patients in the occurrence of any single cardiovascular event or in composite cardiovascular events. There was no difference in mortality (6%) between groups and there was only a single primary cardiovascular death (normothermia). There was no difference between hypothermic and normothermic patients in post- vs. preoperative left ventricular regional wall motion or ejection fraction. Compared with preoperative values, hypothermic patients had no postoperative increase in cardiac troponin-I (median change 0.00 μg/L) whereas normothermic patients had a small postoperative increase (median change + 0.01 μg/L, P = 0.038).
Conclusion
In patients undergoing cerebral aneurysm surgery, perioperative hypothermia was not associated with an increased occurrence of cardiovascular events.
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