Retrofitting of a satellite repeat DNA-based murine artificial chromosome (ACes) to contain loxP recombination sites.

A satellite DNA-based mammalian artificial chromosome (ACes) was generated and subsequently modified by targeting of a loxP-red fluorescent protein (RFP) expression cassette via homologous recombination into a ribosomal DNA (rDNA)-containing locus. Clones containing correctly targeted ACes were identified by PCR from populations of RFP-expressing cells enriched by FACS sorting and were further characterized by fluorescent in situ hybridization. The targeted ACes maintained its ability to be purified to near homogeneity. Studies are currently underway to further characterize the functionality, carrying capacity, stability and transfectability of this modified ACes.

The effect of changes in the health care environment on rehabilitation research: a survey of rehabilitation physicians.

OBJECTIVE: To assess what effect organizational, funding, and system changes in the health care environment may have on rehabilitation research. DESIGN: National survey. SETTING: Academic and clinical research programs. PARTICIPANTS: A total of 138 physicians participated in the survey. INTERVENTION: Mailed instrument requesting information on demographics, research activities, and indicators of change. MAIN OUTCOME MEASURES: Percentage of respondents reporting specific perceptions on (1) how academic and research programs are affected by organizational changes, (2) availability of research funds, and (3) the overall impact that health care changes have on research; between-group comparisons on survey responses. RESULTS: Usable responses were obtained from 138 physicians (response rate, 42.5%). Responding physicians reported workplace changes that included organizational restructuring (32.6%), affiliation with managed care plans (24%), and staff decreases (45.9%). Over half (54.8%) indicated that facility changes had detracted from their abilities to conduct research. A third (34.8%) reported declines in numbers of inpatient beds, and 89.6% reported decreased length of stay (LOS). Decreased LOS was cited as detracting from ongoing research by 36.6% and as discouraging new research by 33.3% of respondents. Although not reflected in measures of scholarly productivity, 53.6% reported having less time to devote to scholarship and 48.5% reported decreased professional activities. Over two thirds (67.4%) of responding physicians indicated that health care system changes had a negative impact on current research, and 54.5% indicated that such changes made it less likely that they would pursue new research. CONCLUSION: Changes in the health care system have had a dampening effect on rehabilitation physicians' research pursuits.

Disability, chronic illness, and risk selection.

As high-cost users of health care, people with disabilities or chronic conditions are particularly vulnerable to risk selection. Preferred risk selection, in which insurers avoid enrolling high-risk people, threatens their access to coverage. Adverse selection, in which high-risk people enroll in the most generous plans, compromises the financial viability of plans that are most responsive to their specific needs. The Americans with Disabilities Act prohibits some forms of risk selection, but does not prevent all disability-based distinctions in insurance practices. From a disability perspective, risk selection must be addressed in a manner that: (1) adequately reflects the health care costs of such individuals; (2) eliminates their need to engage in adverse selection; (3) does not stigmatize them; (4) preserves confidentiality of information; (5) uses substantial outcome measures to ensure quality; and (6) creates market conditions that discourage disability-based discrimination. A risk adjuster based on prior use/expenditures or on a diagnostic indicator sensitive to disability issues may be effective. Failure of reform to address risk selection may threaten the viability of a market-based health care system.

Production of intoxication states by actors--acoustic and temporal characteristics.

This paper is the second of a series; the first has been published (J Forensic Sci, 1998;43:1153-62). The goal in the initial pair of experiments was to determine if speakers (actors) could effectively mimic the speech of intoxicated individuals and also volitionally reduce the degradation to their speech that resulted from severe inebriation. To this end, two highly controlled experiments involving 12 actor-speakers were carried out. It was found that, even when sober, nearly all of them were judged drunker (when pretending) than when they actually were severely intoxicated. In the second experiment, they tried to sound sober when highly intoxicated; here most were judged less inebriated than they were. The goal of this second paper is to identify some of the speech characteristics that allowed the subjects to achieve the cited illusions. The focus here is on four paralinguistic factors: fundamental frequency (F0), speaking rate, vocal intensity, and nonfluency level. For the simulation of intoxication study, it was found that F0 was raised along with increased intoxication but raised even more when this state was feigned. A slowing of speaking rate was associated with increasing intoxication, but this shift also was greater when the speaker simulated intoxication. The most striking contrast was found for the nonfluencies; they were doubled for actual intoxication, but quadrupled when intoxication was simulated. On the other hand, the shifts exhibited by the subjects when they attempted to sound sober were not as clear cut. Indeed, no systematic relationships were found here for either F0 or vocal intensity. Both speaking rate and the number of nonfluencies shifted appropriately, but these changes were not statistically significant. In sum, discernable suprasegmental relationships occurred for both studies (but especially the first); further, it is predicted that useful cues also will be found embedded in the segmentals (the sounds of speech).

Effects of ethanol intoxication on speech suprasegmentals.

The effects of ingesting ethanol have been shown to be somewhat variable in humans. To date, there appear to be but few universals. Yet, the question often arises: is it possible to determine if a person is intoxicated by observing them in some manner? A closely related question is: can speech be used for this purpose and, if so, can the degree of intoxication be determined? One of the many issues associated with these questions involves the relationships between a person's paralinguistic characteristics and the presence and level of inebriation. To this end, young, healthy speakers of both sexes were carefully selected and sorted into roughly equal groups of light, moderate, and heavy drinkers. They were asked to produce four types of utterances during a learning phase, when sober and at four strictly controlled levels of intoxication (three ascending and one descending). The primary motor speech measures employed were speaking fundamental frequency, speech intensity, speaking rate and nonfluencies. Several statistically significant changes were found for increasing intoxication; the primary ones included rises in F0, in task duration and for nonfluencies. Minor gender differences were found but they lacked statistical significance. So did the small differences among the drinking category subgroups and the subject groupings related to levels of perceived intoxication. Finally, although it may be concluded that certain changes in speech suprasegmentals will occur as a function of increasing intoxication, these patterns cannot be viewed as universal since a few subjects (about 20%) exhibited no (or negative) changes.

Bax expressed from a herpes viral vector enhances the efficacy of N,N'-bis(2-hydroxyethyl)-N-nitrosourea treatment in a rat glioma model.

N,N'-bis(2-hydroxyethyl)-N-nitrosourea (BCNU) is a commonly used agent for treatment of malignant gliomas. The mechanisms of cell death and the role of Bcl-2 and Bax in a BCNU-treated rat glioma cell line were investigated. Our results indicate that apoptosis occurs only at a high concentration of BCNU with elevated levels of Bax and a reversed ratio of Bax/Bcl-2. Overexpression of Bax delivered by a herpes simplex viral vector in combination with BCNU chemotherapy enhanced the efficacy of BCNU in a rat glioma model. These findings suggest that conventional treatment with BCNU may be combined with gene therapy that delivers a bax gene into the glioma cells to achieve a high level of Bax, facilitating BCNU-induced cytotoxicity.

Novel generation of human satellite DNA-based artificial chromosomes in mammalian cells.

An in vivo approach has been developed for generation of artificial chromosomes, based on the induction of intrinsic, large-scale amplification mechanisms of mammalian cells. Here, we describe the successful generation of prototype human satellite DNA-based artificial chromosomes via amplification-dependent de novo chromosome formations induced by integration of exogenous DNA sequences into the centromeric/rDNA regions of human acrocentric chromosomes. Subclones with mitotically stable de novo chromosomes were established, which allowed the initial characterization and purification of these artificial chromosomes. Because of the low complexity of their DNA content, they may serve as a useful tool to study the structure and function of higher eukaryotic chromosomes. Human satellite DNA-based artificial chromosomes containing amplified satellite DNA, rDNA, and exogenous DNA sequences were heterochromatic, however, they provided a suitable chromosomal environment for the expression of the integrated exogenous genetic material. We demonstrate that induced de novo chromosome formation is a reproducible and effective methodology in generating artificial chromosomes from predictable sequences of different mammalian species. Satellite DNA-based artificial chromosomes formed by induced large-scale amplifications on the short arm of human acrocentric chromosomes may become safe or low risk vectors in gene therapy.

Marital satisfaction in patients with cancer: does support from intimate partners benefit those who need it the most?

This cross-sectional study assessed 3 ways of providing spousal support. Active engagement means involving the patient in discussions and using constructive problem-solving methods; protective buffering means hiding one's concerns; and overprotection refers to underestimation of the patient's capabilities, resulting in unnecessary help and excessive praise for accomplishments. Ratings of received spousal support by 68 patients with cancer revealed findings similar to those of partners' ratings of provided support. The positive association between active engagement and the patient's marital satisfaction was stronger for patients with a rather poor psychological and physical condition than for those with a rather good condition. Furthermore, protective buffering and overprotection were negatively associated with marital satisfaction only when patients experienced relatively high levels of psychological distress or physical limitations.

The rehabilitation marketplace: economics, values, and proposals for reform.

This article examines whether the ideals, goals, and values of medical rehabilitation can be realized in a market-based health care system. The article observes that rehabilitation is greatly disadvantaged in today's health care marketplace, which violates virtually all the assumptions of a perfectly competitive one. Nevertheless, the authors argue that rehabilitation goals and market economics are not inherently incompatible and call for several market reforms that are congruent with both rehabilitation goals and market theory. These reforms will clarify and facilitate providers' fiduciary responsibilities to patients as well as their accountability to payers. The authors conclude that while the marketplace is an inevitable medium for realizing rehabilitation goals, the vision and value of rehabilitation will not derive from the internal workings of the marketplace but ultimately from committed individuals and socially responsible institutions outside the marketplace.

Mammalian artificial chromosome pilot production facility: large-scale isolation of functional satellite DNA-based artificial chromosomes.

BACKGROUND: A pilot production facility has been established to isolate mammillian artificial chromosomes at high purity by using flow cytometric techniques. Dicentric chromosomes have been generated by the targeted amplification of pericentric heterochromatic and centromeric DNA by activating the "megareplicator." Breakage of these dicentric chromosomes generates satellite DNA-based artificial chromosomes (SATAC) from 60 to 400 megabases. METHODS: For large-scale production, we have developed cell lines capable of carrying one or two SATACs. A SATAC, because of a high adenine-thymine (AT) composition, is easily identified and sorted by using chromomycin A3 and Hoechst 33258 stains and a dual laser high-speed flow cytometer. A prototype SATAC (60 megabases) has been characterized. The prototype SATAC has been isolated from an original rodent/human hybrid cell line and transferred by using modified microcell fusion into a CHO production cell line. RESULTS: Metaphase chromosomes from this production cell line were isolated in a modified polyamine buffer, stained, and sorted by using a modified sheath buffer that maintains condensed chromosomes. SATACs are routinely sorted at rates greater than 1 million per hour. Sorted SATACs have been transferred to a variety of cells by using microcell fusion technology and were found to be functional. CONCLUSIONS: By developing new SATAC containing cell lines with fewer numbers of chromosomes in conjunction with operating a high speed flow sorter we have effectively generated an efficient production facility geared purely for the isolation of SATACs.

Cell surface expression of MHC molecules in glioma cells infected with herpes simplex virus type-1.

9L glioma cells consistently expressed major histocompatibility complex (MHC) class I but not class II molecules. Herpes simplex type-1 virus (HSV-1) infection significantly reduced the expression of MHC I on the cell surface. Recombinant interferons could enhance the cell-surface expression of MHC I but had no effect on MHC II. This enhancement was partially inhibited by HSV-1 infection. HSV-1 mutants with deletions in ICP4, ICP6, ICP27, ICP47 and UL41 genes do not affect the infection induced inhibition, suggest that a different mechanism may be employed in the inhibition of cell-surface expression of MHC molecules.

Production of intoxication states by actors: perception by lay listeners.

The effects of ingesting ethanol have been shown to be somewhat variable in humans; there appear to be but few universals. Yet, questions about intoxication often are asked by law enforcement personnel (especially relative to DUI), clinicians and various individuals in social settings. A key question: Is it possible to determine if a person is intoxicated by observing them in some manner? A closely associated one: Can speech be used for that purpose? Two of the many issues related to the second of these questions involve the possibility that (1) speakers, especially actors, can effectively mimic the speech of intoxicated individuals, and (2) they may be able to volitionally reduce any speech degradation which results from intoxication. The approach used to test these two questions tasked auditors to determine if these simulations were possible. To this end, young, healthy actors chosen on the basis of a large number of selection criteria were asked to produce several types of controlled utterances (1) during a learning phase, (2) when sober, (3) at three simulated levels of intoxication (mildly, legally and severely drunk), (4) during actual, and parallel, levels of intoxication, and (5) at the highest intoxication level attained but when attempting to sound completely sober. Two aural-perceptual studies were conducted; both involved counterbalanced ABX procedures where each subject was paired with him/herself. Listeners were normally hearing university students drawn from undergraduate phonetics and linguistics courses. In the first study, they rated the actors as being more intoxicated--when they actually were sober but simulating drunkenness--88% more often than when they actually were intoxicated. In the second study, they were judged as sounding less inebriated when attempting to sound sober (than they actually were) 61% of the time. These relationships would appear to impact a number of situations; one of special importance would be the detection of intoxication in motorists.

Managed care and people with disabilities: framing the issues.

A physical, cognitive, or mental disability presents significant challenges to an individual in gaining access to a coordinated program of preventive, primary, and secondary health care services. This article describes the health care needs of people with disabilities and discusses how the financial incentives in managed care may threaten access to the health care services they need to maintain their health and functional independence. We argue that despite the shortcomings of present models, managed care has the potential to improve the health care of people with disabilities. Moreover, as health plans become increasingly accountable to consumers (and begin to compete on the basis of quality), they will not be able to ignore the distinct health care needs of people with disabilities.

Personal assistance for people with physical disabilities: consumer-direction and satisfaction with services.

OBJECTIVE: To determine whether people who receive consumer-directed personal assistance services (PAS) are more satisfied with the services they receive than those receiving PAS that are not consumer-directed. DESIGN: A quasi-experimental survey comparison of long-term outcomes among people receiving consumer-directed PAS in Virginia and persons on the waiting list to receive those services. Surveys were conducted by mail and telephone. SETTING: The general community in Virginia. PARTICIPANTS: Ninety-two Virginia residents with physical disabilities living in the community. Sixty individuals were receiving consumer-directed PAS, and 32 were on the waiting list for consumer-directed PAS and were receiving paid personal assistance that was not consumer-directed. MAIN OUTCOMES MEASURES: The Personal Assistance Satisfaction Index (PASI); chi2 and t tests were conducted. RESULTS: People receiving consumer-directed PAS scored significantly higher on the PASI than the waiting-list control group and were consistently more likely to report high levels of satisfaction on the majority of individual PASI items. CONCLUSION: Consumer-directed PAS are associated with high levels of satisfaction relative to PAS that are not consumer-directed. Individual PASI items related to control over PAS and flexibility of services showed the greatest differences in satisfaction.

Family interaction and migration decision making.

Using data from the Residential Preference and Migration Survey, a panel study of Pennsylvania households on migration intention and behavior, the authors examine some aspects of the decision to migrate. "We first examine the impact of the presence or absence of household interaction on the desire to move, migration expectations, and actual migration behavior. We hypothesize that the presence of household interaction is associated with migration behavior, while the absence of household interaction on migration-related issues is a predictor of staying in the present residence. Second, we test an expanded residential satisfaction migration decision model.... The expanded model permits us to test the thesis that household interaction frequency and consensus/conflict are moderating factors in explaining and predicting the outcome of migration decision making."

Managed care and the transformation of the medical rehabilitation industry.

This article reports the findings of a study analyzing the impact of managed care on medical rehabilitation providers in three leading markets: San Diego, California; Minneapolis/St. Paul, Minnesota; and Worcester, Massachusetts. Changes in utilization patterns and the increasing pressure to reduce costs have compelled providers to make numerous strategic adjustments.

Primary care for persons with disabilities. An overview of the problem.

This article outlines the ongoing health care needs of people with disabilities and how organized health care, particularly primary care, often fails to address these needs in a timely fashion. The article's central argument is that managed care and the ferment present in health care today present enormous opportunities for rehabilitation providers and others to develop creative solutions to address the shortcomings of the present health care system.

Effects of exercise during head-down bed rest on postural control.

BACKGROUND: The effects of exercise on postural control during walking were evaluated following exposure to head-down bed rest (BR). METHODS: Two groups of male subjects (N = 18, mean age = 21.4 yr +/- SE 1.0) were exposed to 5 d of 6 degrees head-down bed rest. The experimental group (E) exercised 90 min.d-1 (n = 12 subjects) during the BR while the control group received no intervention (n = 6). The exercise treatment consisted of a combination of isotonic and isokinetic lower extremity exercise training. Electromyographic (EMG) activity in the tibialis anterior, gastrocnemius, vastus lateralis, and biceps femoris was measured during walking before and after BR. RESULTS: Following BR, EMG activity increased in both test subject groups for all muscles (p < 0.05). However, there were no significant differences in EMG activity between the C and E groups either before or after BR. CONCLUSIONS: These data suggest that the ability of the postural control system to adjust to the gravitational environment was compromised as a result of BR. In addition, we conclude that the exercise protocol used was not an effective countermeasure to the alterations in the postural control system.

Impact of a function-based payment model on the financial performance of acute inpatient medical rehabilitation providers: a simulation analysis.

OBJECTIVES: To operationalize research findings about a medical rehabilitation classification and payment model by building a prototype of a prospective payment system, and to determine whether this prototype model promotes payment equity. This latter objective is accomplished by identifying whether any facility or payment model characteristics are systematically associated with financial performance. DESIGN: This study was conducted in two phases. In Phase 1 the components of a diagnosis-related group (DRG)-like payment system, including a base rate, function-related group (FRG) weights, and adjusters, were identified and estimated using hospital cost functions. Phase 2 consisted of a simulation analysis in which each facility's financial performance was modeled, based on its 1990-1991 case mix. A multivariate regression equation was conducted to assess the extent to which characteristics of 42 rehabilitation facilities contribute toward determining financial performance under the present Medicare payment system as well as under the hypothetical model developed. PARTICIPANTS: Phase 1 (model development) included 61 rehabilitation hospitals. Approximately 59% were rehabilitation units within a general hospital and 48% were teaching facilities. The number of rehabilitation beds averaged 52. Phase 2 of the stimulation analysis included 42 rehabilitation facilities, subscribers to UDS in 1990-1991. Of these, 69% were rehabilitation units and 52% were teaching facilities. The number of rehabilitation beds averaged 48. MAIN OUTCOME MEASURE: Financial performance, as measured by the ratio of reimbursement to average costs. RESULTS: Case-mix index is the primary determinant of financial performance under the present Medicare payment system. None of the facility characteristics included in this analysis were associated with financial performance under the hypothetical FRG payment model. CONCLUSIONS: The most notable impact of an FRG-based payment model would be to create a stronger link between resource intensity and level of reimbursement, resulting in greater equity in the reimbursement of inpatient medical rehabilitation hospitals.