Mechanism-based inactivation of human liver cytochrome P450 2A6 by 8-methoxypsoralen

The P450 2A6 catalyzed 7-hydroxylation of coumarin proceeded with a mean Km of 0.40 (+/-0.13) microM and Vmax of 6.34 nmol/nmol P450/min (36-fold variation) in microsomal preparations from a panel of 12 human livers. Substrate depletion was avoided during the kinetic determinations. 8-Methoxypsoralen (8-MOP) is a potent mechanism-based inactivator of human liver P450 2A6 and reconstituted purified recombinant P450 2A6 based on the following evidence: 1) 8-MOP causes time, concentration, and NADPH-dependent loss of P450 2A6 activity that is not reversed by potassium ferricyanide or extensive dialysis, 2) loss of P450 2A6 activity is associated with a loss of spectrally observable P450, 3) addition of nucleophiles or reactive oxygen scavengers to the incubations does not prevent inactivation of P450 2A6, and 4) 8-MOP-dependent P450 2A6 inactivation is inhibited (concentration dependent) by the addition of a competitive inhibitor (pilocarpine). Inactivation is selective for P450 2A6 at low concentrations of 8-MOP (2.5 microM) after short incubation time periods (3 min) and was characterized by a KI of 0.8 and 1.9 microM in a reconstituted and microsomal system, respectively, and a kinact of 1 min-1 and 2 min-1 in a reconstituted and microsomal system, respectively. A substrate depletion partition ratio of 21 was calculated for the inactivation of recombinant P450 2A6. Potency and selectivity suggest that 8-MOP could be a useful tool in vitro for evaluating P450 2A6 activity in various enzyme preparations.     

The min K channel underlies the cardiac potassium current IKs and mediates species-specific responses to protein kinase C

A clone encoding the guinea pig (gp) min K potassium channel was isolated and expressed in Xenopus oocytes. The currents, gpIsK, exhibit many of the electrophysiological and pharmacological properties characteristic of gpIKs, the slow component of the delayed rectifier potassium conductance in guinea pig cardiac myocytes. Depolarizing commands evoke outward potassium currents that activate slowly, with time constants on the order of seconds. The currents are blocked by the class III antiarrhythmic compound clofilium but not by the sotalol derivative E4031 or low concentrations of lanthanum. Like IKs in guinea pig myocytes, gpIsK is modulated by stimulation of protein kinase A and protein kinase C (PKC). In contrast to rat and mouse IsK, which are decreased upon stimulation of PKC, myocyte IK and gpIsK in oocytes are increased after PKC stimulation. Substitution of an asparagine residue at position 102 by serine (N102S), the residue found in the analogous position of the mouse and rat min K proteins, results in decreased gpIsK in response to PKC stimulation. These results support the hypothesis that the min K protein underlies the slow component of the delayed rectifier potassium current in ventricular myocytes and account for the species-specific responses to stimulation of PKC.     

Segregation distortion in myotonic dystrophy

Myotonic dystrophy (DM) is an autosomal dominant disease which, in the typical pedigree, shows a three generation anticipation cascade. This results in infertility and congenital myotonic dystrophy (CDM) with the disappearance of DM in that pedigree. The concept of segregation distortion, where there is preferential transmission of the larger allele at the DM locus, has been put forward to explain partially the maintenance of DM in the population. In a survey of DM in Northern Ireland, 59 pedigrees were ascertained. Sibships where the status of all the members had been identified were examined to determine the transmission of the DM expansion from affected parents to their offspring. Where the transmitting parent was male, 58.3% of the offspring were affected, and in the case of a female transmitting parent, 68.7% were affected. Studies on meiotic drive in DM have shown increased transmission of the larger allele at the DM locus in non-DM heterozygotes for CTGn. This study provides further evidence that the DM expansion tends to be transmitted preferentially.     

Persistent acute renal failure with renal artery stenosis: cure following reconstructive arterial operation

A case of acute renal failure is reported in which renal function returned after surgical correction of renal artery stenosis to the sole functioning kidney. Acute renal failure associated with renal vascular lesions is reviewed. Although often clinically apparent vascular obstruction may be asymptomatic and should be considered in cases of prolonged acute renal failure.     

Silicon diode detectors used in radiological physics measurements. Part II: Measurement of dosimetry data for high-energy photons

Initial calibration of a linear accelerator requires physics instruments to measure accurately central axis depth-dose and off-axis data, both in and out of the beam. These data for an 8- MeV unit were first measured using film, a Farmer 0.6-cm3 ion chamber, a 0.3-cm3 ion chamber, and a 0.1-cm3 silicon diode. Both small probes and film gave a high response compared to the Farmer probe, which has a uniform energy response. Measurements with the diode interfaced to an X-Y recorder required only a fraction of the time required with the chambers, minimizing error due to change in machine output, and permitted resolution of isodose lines in the penumbra. However, corrections required at points in depth due to nonuniform energy response of the unshielded diode were laborious. Construction of a partially shielded diode which duplicates the response of the Farmer probe eliminated the necessity for corrections, permitting rapid accumulation of a wide range of depth-dose and off-axis data.     

Respiration and the generation of rhythmic outputs in insects

In insects gas exchange may be: 1) entirely passive, when metabolic rate is low; 2) enhanced automatically by muscle contractions that produce movements, e.g., wing movements in flight; or 3) produced by ventilatory movements, particularly of the abdomen. In terrestrial insects such as locusts and cockroaches ventilatory movements are governed by a dominant oscillator in the metathoracic or anterior abdominal ganglion. The dominant oscillator overrides local oscillators in the abdominal ganglia and thus sets the rhythm for the entire abdomen, and it also controls spiracle opening and closing in several thoracic and abdominal segments. This ventilatory control mechanism appears to be different from that generating metachronal rhythms such as occur in the ventilatory and locomotory movements of aquatic arthropods. There are now several examples of rhythms, both ventilatory and locomotory, that can be generated by the central nervous system in the absence of phasic sensory feedback, but the mechanism of rhythm production is not known. Studies of ganglionic output suggest that neuronal oscillators can produce a range of frequencies and that some oscillators may be employed in more than one function or behavior. The mechanisms by which central oscillators are coupled to the output motorneurons are also not known; large phase changes suggest that in some cases different coupling interneurons are active. Intracellular recordings from identified neurons have begun to clarify the important roles of interneurons in the production of motor patterns.