Dynamics of chromosome spreading

Consistency of optimum chromosome spreading during harvest of cytogenetic specimens remains a major concern. We have tested the idea that a precise control of the drying rate (the time with which metaphase cells dry), as fixed cell suspension is placed on a slide or an in situ culture in last fixation, may be the answer. Amniocyte and lymphocyte cultures were allowed to dry at defined combinations of relative humidity (RH) and temperature (T) in a modified Thermotron environmental control unit. We were able to demonstrate, based on 2,250 amniocytes and 1,650 lymphocytes, that the metaphase area after drying was a function of RH and T for both in situ and non-in situ culture systems. As the RH and T increase, the metaphase area increases until a threshold is reached. Also, as RH increases, the slide drying time increases. Data obtained using a response surface regression, proportional hazards regression analysis and slide drying time studies are consistent with our model of chromosome spreading. Optimum metaphase areas can be achieved at various combinations of RH and T. We propose that the use of an environmental control unit is a practical way of achieving optimum chromosome spreading routinely and in a highly consistent manner.     

Effects of time compression and time compression plus reverberation on the intelligibility of Northwestern University Auditory Test No. 6

The Tonal and Speech Materials for Auditory Perceptual Assessment, Disc 1.0 audio compact disc developed in 1992 includes several sets of degraded speech materials, two of which, time compression and reverberation, are described in this paper. The digital techniques used to compress the NU No. 6 materials (female speaker) on an 80386-based computer are described, along with a series of experiments on subjects with normal hearing that document the effects of the time compression on recognition performance. Experiment I examined at 70 dB SPL the effect on word recognition of 45, 55, 65, 70, and 75 percent compressions. Experiment II developed psychometric functions for the 45, 65, and 75 percent time-compression conditions. Experiment III defined the effects that time-compression degradation (45% and 65%) plus reverberation degradation (0.3 sec) had on the recognition performance on the NU No. 6 materials. Based on the experiments, four conditions (45% compression, 65% compression, 45% compression plus 0.3-sec reverberation, and 65% compression plus 0.3-sec reverberation) were selected and recorded on the compact disc. In the compact disc trials, normative data on the four conditions were developed from 120 listeners with normal hearing.     

Social contagion, adolescent sexual behavior, and pregnancy: a nonlinear dynamic EMOSA model

Nonlinear dynamic modeling has useful developmental applications. The authors introduce this class of models and contrast them with traditional linear models. Epidemic models of the onset of social activities (EMOSA models) are a special case, motivated by J. L. Rodgers and D. C. Rowe's (1993) social contagion theory, which predict the spread of adolescent behaviors like smoking, drinking, delinquency, and sexuality. In this article, a biological outcome, pregnancy, is added to an earlier EMOSA sexuality model. Parameters quantify likelihood of pregnancy for girls of different sexuality statuses. Five different sexuality/pregnancy models compete to explain variance in national prevalence curves. One finding was that, in the context of the authors' simplified model, adolescent girls have an approximately constant probability of pregnancy across age and time since virginity.     

Detection of soluble interleukin-2 receptor in the serum of patients with non-Hodgkin''s lymphoma

Falls are the most common type of injury among the elderly, and the source of both functional and psychological morbidity. The aim of this study was to validate the Elderly Fall Screening Test (EFST). In a community primary-care clinic, the members 60 years or older who were functionally independent were screened. Of the 568 elderly persons who met these criteria, 361 were interviewed once and 283 persons were re-interviewed a year later. The EFST, a five-item test, was used to divide participants into low- and high-risk groups. Concurrent criterion validity was assessed by physical examinations conducted by physicians who were blind as to the risk designation. Using data from the follow-up interview, predictive validity was assessed on both fall-related and general health measures. Based on the results of the EFST, 28% of the respondents were designated as being at high risk for falls (i.e. having a score of two or more risk items). The results of physicians' examinations corroborated the screening test results in 75% of the cases, with 83% sensitivity and 69% specificity. In the follow-up interview, the high-risk group, as compared to the low-risk group, was more likely to have high scores on EFST, a fall in the past month or year, frequent near falls, and an injurious fall. Those with high EFST scores were more likely to report four or more sick days in the past six months, a hospitalization in the past year, poor self-rated health, a decline in health in the past 6 months, and symptoms of depression. The EFST has both criterion and predictive validity. It can be useful in community-based prevention programmes with functionally independent elderly people.     

Purification from Bothrops lanceolatus (fer de lance) venom of a fibrino(geno)lytic enzyme with esterolytic activity

Bothrops lanceolatus venom has high caseinolytic, phospholipasic, esterolytic and hemorrhagic activities. In spite of having no coagulant effect on plasma, this venom contains a thrombin-like enzyme. Using gel filtration and ion-exchange chromatographies, we have purified an esterolytic fraction (F-II-1a) from this venom with a protein yield of 4% and a 58% recovery in enzyme activity. SDS-PAGE in the presence of beta-mercaptoethanol showed that the enzyme is a single chain polypeptide with a MW=38,100. Immunodiffusion and immunoelectrophoresis of fraction F-II-1a against serum from horses immunized with B. lanceolatus venom and against rabbit antiserum prepared using fraction F-II-1a both showed a single immunoprecipitin line. The Km and Vmax values for TAME hydrolysis were 0.85 mM and 38.6 micromol/min/mg, respectively. The esterolytic activity was completely inhibited by PMSF (10 mM) but not by EDTA (20 mM). Fraction F-II-1a hydrolyzed the alpha and beta chains of fibrinogen. Degradation of the alpha chain occurred within 10 min while that of the beta-chain was slower. The enzyme had no effect on the gamma-chain even after 4 h of hydrolysis.     

Penetration of intravesical doxorubicin in human bladders

The bladder wall penetration kinetics of intravesical doxorubicin were examined in radical cystectomy patients, to provide insight into drug concentrations at target tumor sites. The dosing solution (40 mg/20 ml) was instilled just prior to the start of surgery and maintained for 60-115 min until just prior to bladder excision. The data showed considerable inter-patient variability in the peak plasma concentration (24-fold), urine concentration (7- fold), and tissue concentration (28-fold). The urine concentration at the time of tissue harvest was about 17% of the concentration in the dosing solution. This was due to the dilution by post-catheterization residual urine and urine produced during treatment. The doxorubicin concentration dropped by 32-fold across the urothelium, and declined semi-logarithmically with respect to depth in the capillary-perfused tissues beneath the urothelium with a 50% decrease over about 500 micromole. In three of six patients from whom tumor tissue was obtained, the doxorubicin concentration was higher than the adjacent non-tumor-bearing tissues of comparable tissue depth, whereas the reverse was seen in the remaining three tumors. The plasma concentrations were 0.02, 0.03, 0.05, 0.27, and 0.69% of the concentrations in the tumors, urothelium, lamina propria, superficial and deep muscle layers, respectively. These data indicate: (a) a considerable intra- and inter-patient variability in bladder tissue concentrations, in part due to the variability in the urine concentration; (b) the urothelium is an effective barrier to doxorubicin penetration; and (c) a targeting advantage of intravesical therapy for the treatment of superficial bladder cancer yielding superficial bladder tissue concentrations at least 2000-fold higher than in the systemic circulation. A comparison of the data of doxorubicin with our previously published data on mitomycin C shows similar bladder tissue pharmacokinetics for the two drugs, suggesting that there is no pharmacokinetic preference for either drug.     

Hospital utilization of Saskatchewan people with fetal alcohol syndrome

Endometriosis is defined as the presence of both functioning endometrial glands and stroma outside their usual location lining the uterine cavity. It has been identified in virtually all tissues and organs of the female body with the exception of the spleen. There have been many theories proposed regarding the etiology and pathogenesis of endometriosis. One is the transport of cells through lymphatics and blood vessels. A 44-year-old female suffering from an episode of endometriosis of the umbilicus, left inguinal, uterus, and bilateral ovaries in association with a Múllerian anomaly is presented. Histopathological findings of a skin biopsy from her umbilicus showed aggregated cells within the lymphatic vessel of the upper dermis. This case provides evidence suggesting that cutaneous endometriosis could occur by transport of endometrial cells through lymphatics or blood vessels.     

Optimized release of dexamethasone and gentamicin from a soluble ocular insert for the treatment of external ophthalmic infections

The aziridinium ion of ethylcholine (AF64A), a cholinergic neurotoxin, was injected into the right striatum of a rat. The unilateral injection of 10 nmol AF64A reduced the activity of choline acetyltransferase (CAT) and the tissue content of acetylcholine (ACh) in the striatum. The striatal contents of dopamine (DA), norepinephrine (NE), 5-hydroxyindoleacetic acid (5-HIAA) and gamma-aminobutyric acid (GABA) were unchanged. These results suggest that the cholinospecificity in the striatal lesion was induced by the 10 nmol dose of AF64A. The number of N-methyl-D-aspartic acid (NMDA) receptors in the striatum treated with 10 nmol AF64A was determined by a specific binding assay using [3H](+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid ([3H]CPP), a selective ligand for NMDA receptors. The number of the NMDA receptors decreased significantly in the injected area. On the other hand, in a microdialysis using normal rats, the perfusion of 50 microM NMDA into the striatum increased ACh release. The perfusion of 100 microM MK801 which is the specific and non-competitive NMDA receptor antagonist, decreased the basal levels of ACh release and blocked NMDA-elicited ACh release. Taken together, the present results strongly suggest that a population of NMDA receptors exists on cholinergic interneurons within the striatum, and it directly regulates ACh release.