Evaluation of a flow cytometry-based assay for natural killer cell activity in clinical settings.

Natural killer (NK) cells are not only important in first line defence against viral and bacterial infections, but also in immune surveillance of malignant cells and thus NK cell cytotoxicity is primary indicator of immune function. Although chromium release assay is recognized as 'gold standard' for measuring NK cell activity, it has disadvantages like use of radioactive compounds, poor loading and high spontaneous release. It is difficult to perform this assay in clinical laboratory because of difficulties with disposal of radioactive waste and standardization problems. We describe a flow cytometry-based assay for the measurement of NK cell activity by incorporating fluorescent dye, DiO, into membranes of target cells. NK cell activity was measured at baseline, 1 and 4 weeks follow-up in 20 normal healthy individuals on a dietary supplement immunomodulator to enhance NK cell function. Mean baseline NK cell activity percentage (21.5; SD = 9.3) increased significantly to a maximum level at 1 week (31.3%; SD = 7.9; P = 0.007) and then returned to baseline level at 4 weeks (21.5; SD = 8.3). An important feature of flow cytometry-based assays is its ability to discriminate effector cells from target cells, and potential for explaining molecular interactions underlying target cell lysis. Under clinical settings, this assay will be of interest for frequently monitoring immunological status of patients on treatment for various diseases that affect their immune status. The assay is easy to perform without using radioactive material and thus could become a tool for monitoring pathogenesis and immune reconstitution.

ABC relaxation theory and the factor structure of relaxation states, recalled relaxation activities, dispositions, and motivations.

ABC Relaxation Theory proposes 15 psychological relaxation-related states (R-States): Sleepiness, Disengagement, Physical Relaxation, Mental Quiet, Rested/Refreshed, At Ease/At Peace, Energized, Aware, Joy, Thankfulness and Love, Prayerfulness, Childlike Innocence, Awe and Wonder, Mystery, and Timeless/Boundless/Infinite. The present study summarizes the results of 13 separate factor analyses of immediate relaxation-related states, states associated with recalled relaxation activities, relaxation dispositions, and relaxation motivations on a combined sample of 1,904 individuals (group average ages ranged from 28-40 yr.). Four exploratory factor analyses of Smith Relaxation Inventories yielded 15 items that most consistently and exclusively load (generally at least .70) on six replicated factors. These items included happy, joyful, energized, rested, at peace, warm, limp, silent, quiet, dozing, drowsy, prayerful, mystery, distant, and indifferent. Subsequent factor analyses restricted to these items and specifying six factors were performed on 13 different data sets. Each yielded the same six-factor solution: Factor 1: Centered Positive Affect, Factor 2: Sleepiness, Factor 3: Disengagement, Factor 4: Physical Relaxation, Factor 5: Mental Quiet, and Factor 6: Spiritual. Implications for ABC Relaxation Theory are discussed.

Quantitative analysis of a directed behavior in the medicinal leech: implications for organizing motor output.

The local bend is a directed behavior produced by the leech, Hirudo medicinalis, in response to a light touch. Contraction of longitudinal muscles near the touched location results in a bend directed away from the stimulus. We quantify the relationship between the location of touch around the body perimeter and the behavioral output by using video analysis, muscle tension measurements, and electromyography. On average, the direction of the behavioral output differed from the touch location by <8% of the total body perimeter. We discuss our results in the context of two contrasting behavioral strategies: a Continuous strategy, in which the local bend is directed exactly opposite to stimulus location, and a Categorical strategy, in which there are four distinct bend directions, each elicited by stimuli given in a single quadrant of the body perimeter. To distinguish between these strategies, we delivered two competing stimuli simultaneously. The resulting behavioral output is best described by an average of the effects of each stimulus given alone and thus provides support for the Continuous strategy. We also use a simple model, based on anatomical and physiological data, to predict the responses of the known motor neurons to different stimulus locations. The model shows that the activation of two of the motor neurons (D and V) is inconsistent with a Categorical strategy. However, these neurons are known to be active during the local bend behavior. This result, along with our experimental observations, suggests that the local bend network uses a Continuous strategy to encode stimulus location and produce directed behavioral output.

Using reflexive behaviors of the medicinal leech to study information processing.

The interneuronal network that produces local bending in the leech is distributed, in the sense that most of the interneurons involved are activated in all forms of local bending, even those in which their outputs would produce inappropriate movements. Such networks have been found to control a number of different behaviors in a variety of animals. This article reviews three issues: the physiological and modeling observations that led to the conclusion that local bending in leeches is controlled by a distributed system; what distributed processing means for this and other behaviors; and why the leech interneuronal network may have evolved to be distributed in the first place.

How big should an integrated health care delivery system be at an academic medical center?

The author defines integrated health care delivery systems and comments that there are few such systems now but many in various stages of development. The size of such a system can be described in terms of the number of patients it serves, including their health status and utilization of care, the geographic configuration of the served area, the number of physicians, and the scope and extent of the facilities network. There are a variety of factors that influence a system's size; the author concentrates on the factors that an academic medical center must consider when formulating system-size goals. He discusses (1) the influence of how the institution structures itself to survive; (2) the effects of technology, innovation, and health care costs on the size and organization of these systems; and (3) the effects of the specific characteristics of the institutions' missions of education, research, and patient care. Real numbers describing integrated systems are presented as they relate to three possible institutional goals: economic viability, academic viability, and academic leadership. The author explains why academic medical centers should not try to be only tertiary or quaternary care providers for other integrated health care delivery systems, but at the same time emphasizes that there is no easy or inexpensive way for centers to develop their own systems. Alternative structures for integrated systems are discussed, such as "disease management systems" being developed by the pharmaceutical industry, an "end-to-end linkage" approach, and various ideas to include "captive markets," such as prisoners, university students, and members of communities for the elderly.(ABSTRACT TRUNCATED AT 250 WORDS)

An amphipathic alpha-helical decapeptide in phosphatidylcholine is an effective synthetic lung surfactant.

An idealized model amphipathic alpha-helical decapeptide was synthesized and tested for efficacy as a totally synthetic lung surfactant in simple mixtures with dipalmitoylphosphatidylcholine (DPPC). Quasi-static lung compliance was restored to 92 +/- 3% of the unlavaged value at a pressure of 5 cm H2O in an in vitro lavaged rat lung model. A sustained improvement in gas exchange was also observed when guinea pigs were treated with the synthetic lung surfactant in an in vivo lavaged lung model. DPPC/peptide mixtures rapidly formed low surface tension films in the pulsating bubble surfactometer consistent with a mechanism in which the lipid and peptide mixture spreads rapidly in the lavaged lung to minimize the surface tension at the air/tissue interface. This decapeptide sequence is active in mixtures with DPPC whether the residues are in the all L or all D conformation. However, a peptide with identical sequence, but with alternating D and L amino acid residues, is relatively inactive. Positive charge interactions are not important since a peptide with formylated lysine residues is active. The activity of these decapeptides, with sequences unrelated to any of those in natural lung surfactants, shows that the classic amphipathic alpha-helical hypothesis may be useful in designing peptides that will be effective synthetic lung surfactants in binary mixtures with DPPC. The data demonstrate that a small water-soluble synthetic peptide containing an amphipathic alpha-helical structure combined solely with the major lipid of natural lung surfactant is effective in restoring lung compliance and gas exchange in surfactant-deficient lungs and may be useful in treatment of the respiratory distress syndromes.