Development of a human cell line by selection and drug-metabolizing gene transfection with increased capacity to activate promutagens

We have isolated a human lymphoblastoid cell line with higher levels of native cytochrome P450IA1 activity and by DNA transfection introduced human cDNAs for a putative cytochrome P450IIA2 and epoxide hydrolase (E.C. 3.3.2.3). The resultant cell line, designated MCL-1, was substantially more sensitive to the mutagenicity of dimethylnitrosamine and benzo[a]pyrene than the AHH-1 cell line and was found to have increased metabolism of benzo[a]pyrene to dihydrodiols. The increase in native cytochrome P450IA1 activity was achieved by mutation and selection based on resistance to the phototoxicity of benzo[ghi]perylene. One resistant clone, designated L3, was used for subsequent studies. Two complete cDNAs, one encoding a putative cytochrome P450IIA2 and the other a microsomal epoxide hydrolase, were isolated from a human liver cDNA library. After introduction of the cDNAs into an expression vector and transfection into AHH-1 cells, gene expression was detected at the level of enzyme activity (epoxide hydrolase) or by increased sensitivity to dimethylnitrosamine cytotoxicity/mutagenicity (putative P450IIA2). A vector containing both cDNAs was then constructed and transfected into L3 cells to produce MCL-1 cells. The potential usefulness of drug-metabolizing gene transfection and of the MCL-1 cell line, in particular, for genetic toxicity testing is discussed.     

Retinal pigment epithelial change and partial lipodystrophy

Cuticular drusen and retinal pigment epithelial changes were found incidentally in a 27 year old Lebanese woman during assessment of partial lipodystrophy. Her vision was normal despite involvement of both maculae. The patient had hypocomplementaemia, but serum C3 nephritic factor was absent and renal function was normal. She had impaired glucose tolerance and a continuous infusion of glucose with model assessment (CIGMA) test revealed low normal tissue insulin sensitivity and high normal pancreatic beta cell function. Mild fasting hypertriglyceridaemia (2.0 mmol/l) may have been secondary to impaired insulin sensitivity. Endocrine function was otherwise normal apart from a completely absent growth hormone response to adequate hypoglycaemia. The simultaneous occurrence of partial lipodystrophy and retinal pigmentary epithelial and basement membrane changes appears to be a newly recognized syndrome.     

Forensic psychiatrists' and psychologists' understanding of insanity: misguided expertise?

The ability of mental health experts to consult on insanity cases is highly dependent on their accurate understanding of the current Canadian standard. The study surveyed experienced forensic psychiatrists and psychologists and found the majority (88%) had erroneous beliefs regarding the insanity standard. Results of a discriminant analysis suggest that experience with insanity evaluations and courtroom testimony may improve experts' understanding. Implications of these results are discussed with respect to professional training and public policy.     

LONG-TERM OUABAIN ADMINISTRATION DOES NOT ALTER BLOOD PRESSURE IN CONSCIOUS SPRAGUE-DAWLEY RATS

1. We tested the ability of ouabain to cause chronic hyper tension by continuously infusing ouabain for 28 days (mini-osmotic pump implantation; i.p.). The blood pressure and metabolic effects of sham (150 mmol/L NaCI; n= 12) or ouabain infusion (10 μg/kg per day; n= 14; 100 μg/kg per day; n = 14) were examined in conscious Sprague-Dawley rats. 2. Plasma ouabain concentrations measured after 28 days of ouabain infusion were as follows: sham, not detectable (n= 11); ouabain 10 μg/kg per day, 0.60 ± 0.07 nmol/L (n= 14); and ouabain 100 μg/kg per day, 7.17 ± 0.57 nmol/L (n= 14; P < 0.001). 3. Sham or ouabain infusion did not alter food intake, bodyweight, water intake or urine output in conscious rats. 4. Blood pressure was not altered by sham treatment. Ouabain at 10 μg/kg per day or 100 μg/kg per day did not produce consistent rises in blood pressure. Ouabain at 10 μg/kg per day increased blood pressure on treatment day 12 only (+ 6mmHg; P < 0.05), while at 100μg/kg per day blood pres sure increased on treatment days 16 (+ 9 mmHg; P < 0.05) and day 18 (+ 8mmHg; P < 0.05) only. There was no significant difference in blood pressure between sham and ouabain groups. 5. Renal blood flow was decreased in rats infused with ouabain at 10 μg/kg per day (2.0 ± 0.3 mL/min per 100 g body-weight; n= 5; P < 0.01) and 100 μg/kg per day (2.2 ± 0.4 mL/ min per 100 g bodyweight; n= 7; P < 0.05) compared with sham treatment (3.5 ± 0.2 mL/min per 100 g bodyweight; n= 6). Renal vascular resistance was increased in rats treated with ouabain at 10 μg/kg per day (65.5 ± 12.6 mmHg/mL per min per 100 g bodyweight; n= 5; P < 0.01) and 100 μg/kg per day (66.0 ± 15.6 mmHg/mL per min per 100 g bodyweight; n= 7; P < 0.05) compared with sham treatment (32.6 ± 2.5 mmHg/mL per min per 100 g bodyweight; n= 6). 6. High plasma concentrations of ouabain do not cause consistent increases in blood pressure in conscious Sprague-Dawley rats.     

Myocardial dysfunction in ankylosing spondylitis.

Echocardiographic evidence has suggested abnormalities of the myocardial function in patients with ankylosing spondylitis. In this work the cardiac function in patients with ankylosing spondylitis and in normal volunteers was evaluated. Twenty four normal volunteers and 21 patients with ankylosing spondylitis aged 18-45 were studied. None had overt cardiac disease. Cardiac function was assessed at rest with echocardiography, at rest and during supine bicycle exercise using radionuclide angiography in the left anterior oblique position following equilibration with 740 MBq of technetium-99. The subjects undertook supine bicycle exercise with 30 W increments every three minutes to the point of fatigue. Comparison of data from normal volunteers and patients with ankylosing spondylitis were made using Student's t test for independent samples or the Mann-Whitney non-parametric technique, as appropriate. Subjects were matched for age, sex, height, and weight. There were no echocardiographic differences; however, global nuclide left ventricular function showed several differences between normal volunteers and patients with ankylosing spondylitis. The peak filling rate during exercise was significantly lower in patients with ankylosing spondylitis: normal volunteers 6.5 (SD 1.2); patients with ankylosing spondylitis 5.7 (1.2). The time to reach peak filling during exercise was significantly lower in patients with ankylosing spondylitis: normal volunteers 102 (22); patients with ankylosing spondylitis 120 (23). Regional analysis also showed differences between patients with ankylosing spondylitis and normal volunteers both at rest and during exercise. In the anteroseptal region the filling fraction and peak filling rate were significantly lower in patients with ankylosing spondylitis. Most of the differences (although not all) were in the variables of diastolic function. This study shows that there are subtle abnormalities in cardiac function in patients with ankylosing spondylitis. The major abnormalities are in the diastolic function, suggesting a decrease in left ventricular compliance.     

Who is providing ambulatory care to Medicaid beneficiaries with acquired immunodeficiency syndrome in New York State?

This study examined the number of ambulatory care providers treating individuals with the acquired immunodeficiency syndrome who were Medicaid beneficiaries in New York State in 1988 and examined the distribution of this care across various practice settings. The study population was identified retrospectively in the New York State Medical HIV/AIDS Research Data Base and included a cohort of 5535 individuals with the acquired immunodeficiency syndrome who were enrolled in Medicaid in 1988 for at least 6 months after being diagnosed as having the disease and who had at least one ambulatory care encounter during the year. Ambulatory care for the study group was provided by more than 700 hospital or freestanding clinics and more than 3000 private physicians in 1988. Many sites had low caseloads; 47% of the clinics and 68% of the physicians treating this population saw only one or two patients with the acquired immunodeficiency syndrome who were enrolled in Medicaid. More than half the patients in the study group were seen most frequently in clinics for their ambulatory care during 1988. These data provide reassurance that a wide network of providers is involved in the care of patients with the acquired immunodeficiency syndrome who are Medicaid beneficiaries in New York.     

Object recognition versus object discrimination: comparison between human infants and infant monkeys.

Human infants (12-32 months old) and adults learned a delayed nonmatching-to-sample (DNMS) task and single- and multiple-pair discrimination tasks using nonverbal procedures previously used with monkeys. Infants learned discriminations rapidly and at a young age (12 months), but they required prolonged training and maturation before learning the DNMS task. Adults learned all tasks rapidly. After learning the DNMS task to criterion, memory performance declined systematically in an inverse relation to age. The dissociation in ability of infants on the DNMS versus discrimination tasks closely resembles the dissociation previously reported with infant monkeys (Bachevalier & Mishkin, 1984). Results from both infant humans and monkeys support a neurocognitive maturational model.     

Use of long catheters for multipatient anesthetic monitoring at high respiratory frequencies

Anesthetic gases from several patients can be monitored simultaneously with a centrally located mass spectrometer. Such monitoring requires catheters from patient to spectrometer that are several meters long. Scamman (J Clin Monit 1988;4:227–229) found that when the respiratory frequency is high, as with infants, the CO₂ signal from the patient is unacceptably distorted during passage down the catheter. This is due to Taylor dispersion of the input signal. An outline of the theory of Taylor dispersion is given. The equations describe the interaction between the velocity distribution (which, in laminar flow, is parabolic) and the radial diffusion of CO₂. This interaction keeps a tracer signal together in a pulse, as it moves down the tube with themean velocity, spreading somewhat as it proceeds. How much does an initially sharp signal become blurred? The spread of such a signal when it reaches the detector, measured in time, can be expressed in various ways. Measurement is complicated, however, by the fact that the gas pressure may fall by as much as a factor of 10 along the line. The resultant expansion and acceleration of the gas cannot be ignored. A full treatment of this complication is given elsewhere, but the following simple equation is described: {ie237-1} Typically, the spread time is up to a quarter of a second for catheters of 50 m, such as used by Scamman. This is comparable with the period of CO₂ rise and fall for infants and explains the serious distortion in wave form that Scamman found. Some distortion can be eliminated by reducing R to 0.1 or less, but the extent of this improvement is small. Ideally, for fast-breathing patients, the catheter length should be reduced to 20 m or less, if possible.