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.     

Anaphylaxis to topical bacitracin zinc ointment

This report describes a case of systemic anaphylaxis to bacitracin zinc ointment in a 24-year-old man who was injured in a motorcycle accident. Extensive abrasions on the patient's extremities were cleaned with Shurclens before application of viscous Xylocaine and bacitracin zinc ointment. Five minutes later, the patient exhibited symptoms of severe anaphylaxis and required the administration of epinephrine, antihistamines, intravenous fluids, and corticosteroids. Two weeks later, he underwent prick/puncture skin testing to Shurclens and bacitracin zinc ointment as well as prick/puncture, intracutaneous, and subcutaneous challenge with Xylocaine. Only the result of the prick test to bacitracin zinc ointment was positive. Although bacitracin is considered to be a safe topical antibiotic, physicians should be aware of the potential not only for delayed hypersensitivity but also for acute IgE-mediated allergic reactions and life-threatening anaphylaxis.     

Addisonian crisis provoked by levothyroxine substitution therapy

A women aged 36 with a positive family anamnesis for autoimmune endocrine diseases and a history of thyroid diseases, developed major complaints of general malaise, orthostatic hypotension and loss of appetite after the start of a treatment with levothyroxin because of (sub)clinical hypothyroidism. She was found to suffer from primary adrenocortical insufficiency masked by excessive use of liquorice and a lowered metabolism, but which via the suppletion with thyroid hormone had led to an addisonian crisis.     

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.     

Ultrasonic measurements of the elastic stiffness constants of oriented polyethylene

High density polyethylene (Rigidex 9), a high density copolymer of polyethylene and poly(butene-1) (Rigidex 2000), and low density polyethylene (Alkathene WJG 11) were oriented by hot drawing. The crystalline texture, as determined by wide- and low-angle X-ray diffraction, was a highly oriented chain axis in the draw direction with random orientation transversely and with lamellae surfaces perpendicular to the draw direction. Elastic stiffness constants were measured by a contact-probe ultrasonic pulse technique at 2.5 MHz both before and after annealing in a temperature range which did not significantly alter the crystalline texture. Assuming orthorhombic symmetry the nine stiffness constants of Rigidex 2000 and the three longitudinal and three shear stiffness constants of Rigidex 9 were measured after drawing and after subsequent annealing. Only the longitudinal constants of Alkathene were measured, as shear waves could not be transmitted. The longitudinal stiffness in the draw direction was markedly affected by drawing and by annealing, while the crystalline texture remained substantially unchanged; by comparison the other stiffness constants showed little change. Drawn Rigidex 9 reached a tensile modulus in the draw direction of 69 GPa. The results are compared with ‘static’ and low frequency measurements reported in the literature.     

A study on the orientation effects in polyethylene in the light of crystalline texture

Special sample types established and characterised in Parts 1 and 2 [1,2] were examined systematically under different modes of external deformation at constant temperatures. The structural changes recorded by means of wide- and low-angle X-ray patterns fully substantiated the bodily rotation of the otherwise unaltered crystallites in accordance with the interlamellar slip mechanism postulated in Parts 1 and 2, both in tension and under two modes of compression. Dimensional changes, however, were significantly in excess of those expected from the rotation of crystallites, but were uniquely correlated with this relation in the different samples under different conditions of deformation. This points to a unique structural and mechanistic interrelation between the crystallites and more compliant amorphous regions. As the X-ray long periods were unaffected by the deformation, the more extensible material cannot be identical to that associated with the lamellar periodicity, a fact which points to a previously unsuspected superlamellar regularity in the texture. A simple structural suggestion is made.     

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.