Cloning of a mu-class glutathione S-transferase complementary DNA and characterization of its glucocorticoid inducibility in a smooth muscle tumor cell line.

A cDNA (designated hGSTYBX) encompassing the complete coding sequence of a hamster mu-class glutathione S-transferase (GST) subunit was cloned from a lambda ZAP library constructed with mRNA isolated from triamcinolone acetonide-treated smooth muscle tumor cells (DDT1 MF-2). Analysis of its nucleotide and deduced amino acid sequences demonstrated highest homology to the rat mu-class GST YB2 subunit. In proliferating subconfluent cells, in which constitutive expression of hGSTYBX mRNA was undetectable, glucocorticoid treatment induced hGSTYBX expression after a time lag of 3 h, and maximal induction occurred at 10 h. Nuclear run-on analysis showed that glucocorticoid induction resulted at least in part from an increased rate of transcription. Simultaneous treatment with glucocorticoid and cycloheximide prevented glucocorticoid induction, but had little effect on basal expression in confluent cells. In contrast, cycloheximide treatment 3 h after glucocorticoid treatment resulted in nearly full induction. These results taken together suggest that hGSTYBX induction may be a secondary glucocorticoid response.

The Python pit organ: imaging and immunocytochemical analysis of an extremely sensitive natural infrared detector.

The Python infrared-sensitive pit organ is a natural infrared imager that combines high sensitivity, ambient temperature function, microscopic dimensions, and self-repair. We are investigating the spectral sensitivity and signal transduction process in snake infrared-sensitive neurons, neither of which is understood. For example, it is unknown whether infrared receptor neurons function on a thermal or a photic mechanism. We imaged pit organs in living Python molurus and Python regius using infrared-sensitive digital video cameras. Pit organs were significantly more absorptive and/or emissive than surrounding tissues in both 3-5 microns and 8-12 microns wavelength ranges. Pit organs exhibited greater absorption/emissivity in the 8-12 microns range than in the 3-5 microns range. To directly test the relationship between photoreceptors and pit organ infrared-sensitive neurons, we performed immunocytochemistry using antisera directed against retinal photoreceptor opsins. Retinal photoreceptors were labeled with antisera specific for retinal opsins, but these antisera failed to label terminals of infrared-sensitive neurons in the pit organ. Infrared-receptive neurons were also distinguished from retinal photoreceptors on the basis of their calcium-binding protein content. These results indicate that the pit organ absorbs infrared radiation in two major atmospheric transmission windows, one of which (8-12 microns) matches emission of targeted prey, and that infrared receptors are biochemically distinct from retinal photoreceptors. These results also provide the first identification of prospective biochemical components of infrared signal transduction in pit organ receptor neurons.

Behaviorally based measures for evaluating the nonclinical performance of dentists in team settings.

Retranslations of expectations technique was used to develop behaviorally anchored scales for evaluating dentists' utilization of expanded duty dental auxiliaries. To enhance the reliability and validity of such instruments the project focused on specific acts and decisions of the dentist which are either effective or ineffective in accomplishing the health team's tasks.

Development of scales to measure provider attitudes toward expanded duty dental auxiliaries.

Much of the research on attitudes toward utilizing EDDAs has been rather haphazard in its measurement aspects. Examining responses to single items or summing across a series of items without any empirical rationale for such summing cannot be justified given current knowledge about measuring attitudes. The scales developed here are appreciably better from a psychometric point of view than scales reported in the dental literature to date. The use of standardized measuring instruments would facilitate cross-study comparisons. Such a process is presently not feasible because each researcher is using his own idiosyncratic scales of unknown psychometric characteristics. The three scales developed here to measure (1) the value of expanding auxiliary functions, (2) patient reaction to the use of EDDAs, and (3) satisfaction with changed dentist role may permit more standardized measurement of at least some aspects of provider and future provider attitudes toward the utilization of expanded duty dental auxiliaries.

Symptomatology, pulmonary function and response, and T lymphocyte beta 2-receptors during smoking cessation in patients with chronic obstructive pulmonary disease.

STUDY OBJECTIVES: To characterize the effect of smoking cessation and nicotine replacement on pulmonary symptomatology, baseline pulmonary function and response to terbutaline, and purified T lymphocyte beta 2-receptor regulation; and the relationship between T lymphocyte beta 2-receptor density and pulmonary function. DESIGN: Open-label, longitudinal, 28-week study. SETTING: A university clinical research center. PATIENTS: Eighteen long-term smokers with mild to moderate chronic obstructive pulmonary disease (COPD) were enrolled and seven completed the study. INTERVENTIONS: Subjects stopped smoking with the aid of nicotine substitution and behavioral counseling. Pulmonary response (FEV1) to subcutaneous terbutaline and T lymphocyte beta 2-receptor density (Bmax) and function (cAMP) were measured prior to smoking cessation (week 0), during nicotine replacement (week 8), and after nicotine cessation (week 28). MEASUREMENTS AND MAIN RESULTS: Serum cotinine concentrations, plasma epinephrine concentrations, and day and night cough decreased significantly after smoking cessation, whereas basal cAMP concentrations increased (p < 0.05). No significant change was seen in baseline FEV1, pulmonary response to terbutaline, or Bmax over the 28 weeks; however, intrasubject changes in Bmax between visits correlated significantly (p < 0.05) with intrasubject changes in pulmonary response between visits. CONCLUSIONS: Our data indicate that smoking cessation is associated with a significant decrease in the symptomatology of COPD, and that change in T lymphocyte beta 2-receptor density is a good marker of change in pulmonary response to beta 2-agonists.

Investigation of light-induced conformation changes in spiropyran-modified succinylated poly(L-lysine).

To determine the maximum range of coupling between side-chain photochromism and polypeptide conformation change, we modified the carboxylate side chains of succinylated poly(L-lysine) with a spiropyran to form polypeptide I. The extent of modification was determined to be 35.5%. The spacer group length between the polypeptide alpha-carbon and the dye was 12 atoms, providing minimum polypeptide-dye interaction. Conformation changes were monitored by circular dichroism as a function of light adaptation and solvent composition (hexafluoroisopropanol [HFIP] vs trifluoroethanol [TFE]). Under all solvent compositions, the dark-adapted dye was in the merocyanine form. Light adaptation by visible light converted the dye to the spiropyran form. When dissolved in TFE, I adopted a helical conformation insensitive to light adaptation. With increasing percentage HFIP, a solvent-induced helix-to-coil transition was observed around 80% (vol/vol) HFIP. At 100% HFIP, both light- and dark-adapted forms of I were in the coil state. Near the midpoint of the solvent-induced helix-to-coil transition, light adaptation caused conformation changes. Applying helix-to-coil transition theory, we measured a statistically significant difference in coil segment-HFIP binding constant for light- vs dark-adapted solutions (6.38 +/- 0.03 M-1 vs 6.56 +/- 0.03 M-1), but not for the nucleation parameter sigma (1.2 +/- 0.4 10(-3) vs 1.3 +/- 0.3 x 10(-3). The small binding constant difference translated to a light-induced binding energy difference of 17 cal/mol/monomer. Near the midpoint of the helix-to-coil transition, collective interactions between monomer units made possible the translation of a small energy difference (less than RT) into large macromolecular conformation changes.(ABSTRACT TRUNCATED AT 250 WORDS)

Kinetic study of the helix to coil dark reaction of poly(spiropyran-L-glutamate).

An investigation of kinetics of the helix to coil dark reaction of light adapted poly(spiropyran-L-glutamic acid) (PSLG) dissolved in hexafluoroisopropanol was performed. The reaction was associated with the spiropyran (SP) to merocyanine (MC) ring opening. The ring opening reaction monitored with UV/VIS spectroscopy showed first order kinetics. Chromophore and polypeptide backbone circular dichroism data fit to an expression consistent with a single intermediate series mechanism. By FTIR, we monitored the polypeptide alpha-helix amide I, the MC chromophore--C = C--stretch and the protonated unmodified carboxylate C = O stretch bands. During the first step of the series mechanism, changes in the hydrogen bonding of the unmodified carboxylate groups occurred, suggesting breakup of polypeptide aggregates. The second step of the proposed series mechanism was dominated by the helix to coil transition and the ring opening of SP to MC. The CD spectrum of MC in the dark adapted PSLG was red shifted and had a narrower bandwidth than the UV/VIS spectrum. The kinetic and spectroscopic data suggested that a fraction (population I) of the MC chromophores experienced optical activity induced by the chiral polypeptide environment, while the remainder of the MC chromophores (population II) were solvated and enantiomeric.

Free groin flap revisited.

Reported herein are 130 consecutive cases of free groin flap transfer performed by one surgeon over a 19-year period. Transplantation was performed for soft-tissue cover or augmentation of contour defects involving the head and neck (68 cases), trunk (4 cases), upper limb (14 cases), and lower limb (44 cases). Indications for flap coverage/augmentation were classified broadly into tumor, trauma, radiation induced, and miscellaneous. Specific reconstructive problems included augmentation for Romberg's hemifacial atrophy, external ear canal reconstruction after tumor ablation, and coverage of lower limb defects. There were nine failures (total flap loss), seven cases of partial flap loss, and two cases were abandoned intraoperatively. Of 15 cases that were urgently re-explored, 9 flaps were salvaged. The failure rate for the groin flap series (130 cases) was 8.5 percent compared with the failure rate of 4.2 percent for the other 517 cases of microvascular transfer performed over the same period by the same surgeon. Donor-site complications occurred in 24 cases and included hematoma or seroma formation, hypertrophic scars, nerve paresthesiae, infection, and dehiscence. Secondary debulking procedures were performed in 26 cases. The free groin flap, contrary to some reports, is a reliable flap that provides relatively thin pliable soft-tissue cover or augmentation, with minimal donor-site morbidity. The specific indications for its use have undergone an evolution since first described in 1973.

The effect of an implantable Doppler probe on the salvage of microvascular tissue transplants.

One hundred forty-seven flaps in 135 consecutive patients undergoing microvascular transplantation were monitored using a miniature Doppler ultrasonic probe. Using a modification of a technique described previously by Swartz, the probes were secured to the outflow vein of the flap with Vicryl mesh. Twenty instances of thrombosis or spasm were detected in 16 patients, and all flaps were salvaged (100 percent). There were four false positive and no false negative results. This probe allows for safe, continuous monitoring of flap blood flow, which permits the rapid detection and hence rapid treatment of postoperative complications. Our experience suggests that a significant improvement in the salvage rate of microvascular transplants may be attainable with the use of this device.

Maintaining vitality in the mature dental practice.

The dentists interviewed for this third article in the series report this is a satisfying time; they are private practitioners aged 40 to 54. These dentists are enthusiastic about their profession and enjoy using the new technological advances for better treatment of their patients. For these active practitioners, the practice does not need to be revitalized: they're enjoying fulfillment in what they do.

Stop-smoking program using nicotine reduction therapy and behavior modification for heavy smokers.

One hundred eighteen volunteers were studied in a hospital-based smoking cessation program in which nicotine polacrilex (Nicorette) was used as an adjunct to behavioral modification (group support). The "success" rate of 40% to 47% at 1 year was chemically verified. The regimen for using the nicotine polacrilex was "within labeling" but decidedly different from the regimen used in most smoking cessation programs involving this product.

Gloved and ungloved: performance time for two dental procedures.

This study examines the time required to perform two routine dental procedures requiring fine hand manipulative skills--a pin placement procedure and an endodontic procedure. No significant time difference was noted in performing the procedures while wearing gloves or not wearing gloves. Twelve dentists who had routinely worn gloves for the past 6 months participated, each performing four procedures.

Dentist-staff interpersonal relationships.

The growing demand for dental services has resulted in an expanded utilization of auxiliary personnel in an effort to increase productivity. Enlarged dental teams which have evolved from this effort have also magnified the potential for interpersonal conflicts in the office. Although such conflicts are sometimes unavoidable, they can be reduced through effective preventive management. Careful selection of personnel, well designed training programs, and attention to those factors which assist retention of qualified staff members are important in this process. Finally, the entire team must have common goals and work in a coordinated fashion to achieve ultimate success. The dentist of today must possess the skills of an effective manager as well as those of a competent clinician.

Soft tissue sarcoma of the extremity.

A retrospective review of 33 cases of soft tissue sarcoma of the extremity presenting over a 10 year period was undertaken. The history, patterns of referral, diagnostic investigations, procedures undertaken and outcomes were studied. We found there was a frequent delay in diagnosis and sometimes misinterpretation of biopsy specimens. Patients were seen by a variety of specialists from disciplines such as general surgery, plastic surgery, orthopaedic surgery and rheumatology. Considerable progress has been made in the treatment of soft tissue sarcomas, often allowing local control of the tumour without amputation. We believe there should be early referral of patients having these tumours to a centre where a combined multidisciplinary approach can be undertaken.

Faculty evaluation at the University of Kentucky.

Faculty at the University of Kentucky College of Dentistry are now evaluated through the use of a negotiated Distribution of Effort Agreement. This document, negotiated at the beginning of each academic year, must satisfy both department objectives and the professional and personal requirements of the individual faculty member. It may be modified during the year if both the faculty member and his department chairman agree to the change. This document states work objectives for the year similar to learning objectives in a course. At the end of the year, each faculty member is evaluated on how well he has met his individual objectives. Information relevant to teaching, university service, and student service is gathered by college administrators and forwarded to department chairmen. Together with information on research accomplishment, these data are used by the chairman in the evaluation of each faculty member. The college has clearly charged department chairmen with the direct responsibility for faculty evaluation. The Appointments, Promotions, and Tenure Committee also plays a role in faculty development and evaluation by recommending to the dean ways of improving individual faculty performance.

Reengineering managed behavioral healthcare.

Reengineering is an industrial method for rethinking and redesigning basic business processes in order to improve performance and lower cost. Just as many of the corporate clients of managed behavioral healthcare companies go through reengineering initiatives, behavioral healthcare needs to streamline its production processes, in order to deliver services that are high quality but affordable. Traditional, cumbersome, bureaucratic and labor-intensive work processes must be reexamined. This article reviews basic principles of reengineering in the context of the evolving managed behavioral healthcare industry, defines ten core processes for managed behavioral healthcare companies and then illustrates how reengineering methods can be applied to two intermediate processes: "manage access" and "manage network."