Comparative study of genetic activity of chlorambucil's active metabolite steroidal esters: the role of steroidal skeleton on aneugenic potential.

p-N,N-bis(2-chloroethyl)aminophenylacetic acid (PHE), a nitrogen mustard analogue and chlorambucil's active metabolite used as chemotherapeutic agent, has been shown that, in addition to its clastogenic activity, induces chromosome delay. In the present study an efford has been made (a) to investigate if the steroidal analogues of PHE (EA-92, EA-97, AK-333, AK-409 and AK-433) exert the same genetic activity as the parent compound, (b) to further analyze the aneugenic activity of nitrogen mustard analogues, (c) to investigate the mechanism by which they exert aneugenic potential and (d) to correlate the genetic activity with chemical structure. For this purpose the Cytokinesis Block Micronucleus (CBMN) assay was conducted in human lymphocytes in vitro and the micronucleus (MN) frequency was determined to investigate their genetic activity. The mechanism of micronucleation was determined in combination with Fluorescence In Situ Hybridization (FISH) using pancentromeric DNA probe. Since one of the mechanisms that chemicals cause aneuploidy is through alterations in the mitotic spindle, we also investigated the effect of the above compounds on the integrity and morphology of the mitotic spindle using double immunofluorescence of beta- and gamma-tubulin in C(2)C(12) mouse cell line. We found that PHE and its steroidal analogues, EA-92, EA-97, AK-333, AK-409 and AK-433, affect cell proliferation in human lymphocytes and C(2)C(12) mouse cells. All studied compounds are capable of inducing chromosome breakage events, as indicated by the enhanced C(-)MN frequencies. The less lipophilic compounds are the most genetically active molecules. PHE and only two of the studied analogues, AK-409 and AK-433, the most hydrophilic ones, showed aneugenic potential, by increasing the frequencies of MN containing a whole chromosome. The aneugenic potential of the above referred analogues is associated with amplification of centrosome number, since they caused high multipolar metaphase frequencies.

The Behavior Observation Instrument: a method of direct observation for program evaluation.

The background and development of a multicategory direct observation system, the Behavior Observation Instrument (BOI), is described. This time-sampling procedure for recording the behavior of persons is demonstrated in several treatment settings and the results applied to issues of program evaluation. Elements that have prevented direct observation from being widely adopted, such as costs, manpower, and training requirements, are systematically analyzed. A basic psychometric analysis of the instrument is used to determine optimum frequency and duration of observation intervals as well as observer agreement. The results imply that direct observation methods, once assumed by some to belong to the special province of the single-subject design, can be used to assess the effects of programs on groups of psychiatric clients in an efficient and economic manner.

Behavioral Effects of Reducing the Daily Frequency of Phenothiazine Administration.

After an 11-day base line of behavioral observations, 24 chronic female schizophrenics were assigned to two groups matched for alertness. In the first treatment phase, the administration of phenothiazine medication of one group was switched from a multiple-dose schedule (three to four times per day) to a single daily administration, while the total dosage per day was held constant. The second group continued on a multiple administration schedule for 11 days and then was switched to a single daily dosage. A multivariate analysis of variance showed that there was no overall effect (positive or negative) due to the schedule change; however, preplanned t tests showed transitory decreases in nonfunctional behavior. The results are discussed in terms of implications for the administration of phenothiazines and the experimental analysis of drug effects.