Detection and characterization of individual intermolecular bonds using optical tweezers.

The development of scanning probe techniques has made it possible to examine protein-protein interactions at the level of individual molecular pairs. A calibrated optical tweezers, along with immunoglobulin G (IgG)-coated polystyrene microspheres, has been used to detect individual surface-linked Staphylococcus protein A (SpA) molecules and to characterize the strength of the noncovalent IgG-SpA bond. Microspheres containing, on average, less than one IgG per contact area were held in the optical trap while an SpA-coated substrate was scanned beneath them at a distance of approximately 50 nm. This geometry allows the trapped bead to make contact with the surface, from bond formation to rupture, and results in an enhancement of the force applied to a bond due to leverage supplied by the bead itself. Experiments yielded median single-bond rupture forces from 25 to 44 pN for IgG from four mammalian species, in general agreement with predictions based on free energies of association obtained from solution equilibrium constants.

Psychosocial functioning in women with premenstrual dysphoric disorder before and after treatment with sertraline or placebo.

BACKGROUND: The objective of this study was to evaluate the pretreatment psychosocial functioning of women with premenstrual dysphoric disorder (PMDD) and the effect of sertraline treatment on psychosocial functioning in these patients. METHOD: Two hundred forty-three women recruited from 12 university-affiliated sites and meeting DSM-IV criteria for PMDD completed 1 cycle of single-blind placebo and were randomly assigned to flexible dose sertraline or placebo for 3 cycles. Psychosocial functioning was assessed by the Daily Record of Severity of Problems (DRSP), the Social Adjustment Scale (SAS), and the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q). RESULTS: SAS scores during the follicular phase were similar to SAS scores of community norms, whereas the pretreatment SAS and Q-LES-Q scores during the luteal phase were similar to scores of women with depressive disorders. Sertraline was significantly more effective than placebo in improving psychosocial functioning as measured by the SAS, the Q-LES-Q, and the 3 DRSP items of impaired productivity, interference with social activities, and interference with relationships with others. Improvement in psychosocial functioning assessed by SAS and Q-LES-Q correlated with improvement in symptomatology assessed by the Clinical Global Impressions-Improvement (CGI-I) scale and the Hamilton Rating Scale for Depression (HAM-D). Remitters (CGI-I score of 1) were more likely to function better at baseline and showed larger improvements in functioning and quality of life with treatment compared with nonremitters. CONCLUSION: Sertraline was superior to placebo in improving psychosocial functioning in women with PMDD as reflected by SAS, Q-LES-Q, and DRSP measures. Functional improvement correlated with improvement in premenstrual symptomatology and was apparent by the second cycle of treatment. Comparison of pretreatment SAS scores in women with PMDD with the scores of other populations of women documents the degree of luteal phase functional impairment in women with PMDD and a relative absence of follicular phase impairment.

Spontaneous recovery of fluorescence by photobleached surface-adsorbed proteins.

Fluorescence photobleaching of a carboxyfluorescein-labeled protein (erythrocyte cytoskeletal protein 4.1) immobilized on bare glass is found to be spontaneously reversible, provided that the sample is deoxygenated. After a short (hundredths of seconds) photobleaching laser flash, the subsequent fluorescence excited by a dim probe beam partly recovers on a long (tenths of second) time scale, even in the absence of chemical exchange or diffusion processes. Neither the fraction of the fluorescence that bleaches reversibly nor its recovery rate is a strong function of fluorophore surface concentration. At a fixed surface concentration, the reversibly photobleached fraction and its recovery rate decreases with increasing duration or intensity of the bleaching flash. On the other hand, nondeoxygenated air-equilibrated samples exhibit almost total irreversible bleaching on this time scale. Quantitative fluorescence microscopy experiments occasionally require deoxygenation to avoid photochemical crosslinking or photobleaching or to enhance the triplet state population. The observation presented here indicate that fluorescence recovery after photobleaching (FRAP) experiments performed under deoxygenated conditions for measuring diffusion or chemical kinetics should be interpreted with caution: fluorescence recoveries may be due to intrinsic photochemical processes rather than fluorophore mobility. The recovery effect appears too slow to be ascribed simply to a relaxation of a triplet state; other possible explanations are offered.

Reversible binding kinetics of a cytoskeletal protein at the erythrocyte submembrane.

Reversible binding among components of the cellular submembrane cytoskeleton and reversible binding of some of these components with the plasma membrane likely play a role in nonelastic morphological changes and mechanoplastic properties of cells. However, relatively few studies have been devoted to investigating directly the kinetic aspects of the interactions of individual components of the membrane skeleton with the membrane. The experiments described here investigated whether one component of the erythrocyte membrane cytoskeleton, protein 4.1, binds to its sites on the membrane reversibly and if so, whether the different 4.1-binding sites display distinct kinetic behavior. Protein 4.1 is known to stabilize the membrane and to mediate the attachment of spectrin filaments to the membrane. Protein 4.1 previously has been shown to bind to integral membrane proteins band 3, glycophorin C, and to negatively charged phospholipids. To examine the kinetic rates of dissociation of carboxymethyl fluorescein-labeled 4.1 (CF-4.1) to the cytofacial surface of erythrocyte membrane, a special preparation of hemolyzed erythrocyte ghosts was used, in which the ghosts became flattened on a glass surface and exposed their cytofacial surfaces to the solution through a membrane rip in a distinctive characteristic pattern. This preparation was examined by the microscopy technique of total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP). Four different treatments were employed to help identify which membrane binding sites gave rise to the multiplicity of observed kinetic rates. The first treatment, the control, stripped off the native spectrin, actin, 4.1, and ankyrin. About 60% of the CF-4.1 bound to this control binded irreversibly (dissociation time > 20 min), but the remaining approximately 40% binded reversibly with a range of residency times averaging approximately 3 s. The second treatment subjected these stripped membranes to trypsin, which presumably removed most of the band 3. CF-4.1 binded significantly less to these trypsinized membranes and most of the decrease was a loss of the irreversibly binding sites. The third treatment simply preserved the native 4.1 and ankyrin. CF-4.1 binded less to this sample too, and the loss involved both the irreversible and reversible sites. The fourth treatment blocked the gycophorin C sites on the native 4.1-stripped membranes with an antibody. CF-4.1 again binded less to this sample than to a nonimmune serum control, and almost all of the decrease is a loss of irreversible sites. These rest suggest that 1) protein 4.1 binds to membrane or submembrane sites at least in part reversibly ; 2) the most reversible sites are probably not proteinaceous and not glycophorin C, but possibly are phospholipids (especially phosphatidylserine); and 3) TIWRFRAP can successfully examine the fast reversible dynamics of cytoskeletal components binding to biological membranes.

Impact of patient-controlled compression on the mammography experience.

The authors tested the hypothesis that giving women control over the compression portion of the mammography examination results in a less painful experience, greater overall patient satisfaction, and a radiographic image as good as that produced by means of technologist-controlled compression. One hundred nine women undergoing screening mammography at a hospital-based outpatient clinic were studied. Each underwent two-view, screen-film mammography performed in routine fashion except that, by random assignment, one breast was compressed by the technologist and the other breast, by the patient. Patient-controlled compression was significantly (P = .003) less painful than technologist-controlled compression. Overall patient satisfaction (96% [105 of 109]) and willingness to repeat the experience were extremely high. The majority of images (93.5% [202 of 216]) were rated as having good to excellent compression. With minimal patient education, self-compression resulted in an image at least as good as that produced with technologist-applied compression. Further study of this technique is warranted.

Reduced platelet tritium-labeled imipramine binding sites in women with premenstrual syndrome.

OBJECTIVE: We studied the possible role of serotonergic systems in the cause of premenstrual affective symptoms. STUDY DESIGN: The binding of tritium-labeled imipramine to platelets is thought to parallel central nervous system binding and to indicate serotonergic activity. We measured platelet tritium-labeled imipramine binding sites in the follicular and luteal phases in 12 controls and in 9 women with well-documented late luteal phase dysphoric disorder. In statistical analyses we used repeated measures analysis of variance, with Student-Newman-Keuls and Duncan's one-tailed t tests, and Pearson's r. RESULTS: The values of subjects with late luteal phase dysphoric disorder were lower than those of controls (F [1,39] = 5.13, p = 0.03). Both follicular and luteal phase level were lower in subjects with late luteal phase dysphoric disorder but reached statistical significance only in the follicular phase. CONCLUSION: Lower platelet tritium-labeled imipramine binding in women with late luteal phase dysphoric disorder supports the hypothesis that alteration of central serotonergic systems may contribute to premenstrual dysphoric symptoms.

Resolution of chronic pelvic pain after laparoscopic lysis of adhesions.

Thirty women undergoing laparoscopic lysis of adhesions for the treatment of chronic pelvic pain were prospectively evaluated for the presence of a chronic pain syndrome. At follow-up, of those with chronic pain syndrome (10), four (40%) reported continued improvement or resolution of pain during daily activities of dyspareunia, whereas of those without chronic pain syndrome (20), 15 (75%) were better (p = 0.06). When these two complaints are evaluated individually, both pain during daily activities (p less than 0.05) and dyspareunia (p less than 0.05) are more likely to improve after lysis of adhesions in women without chronic pain syndrome. Prognosis was not related to the number of previous operations, adhesion score, or other physical parameters. Laparoscopic lysis of adhesions is generally worthwhile in the treatment of chronic pelvic pain, although the presence of psychosocial compromise warrants preoperative evaluation and concomitant treatment.

Relationship of laparoscopic findings to self-report of pelvic pain.

An assessment battery including standardized measures of behavioral and psychosocial factors associated with other chronic pain conditions was administered to 102 women scheduled for laparoscopic surgery. Surgeons who were blinded to the patient's self-reported pain data completed the American Fertility Society classification for endometriosis and adhesions on the basis of observed physical disease. Although American Fertility Society classification scores were significantly related to self-assignment into pain or no-pain groups, the extent of physical disease evaluated by this procedure was not significantly correlated with ratings of pain levels or a number of indexes of impairment. The group of patients with laparoscopically diagnosed pathologic conditions reported higher pain levels and greater interference than the group who reported pain and had negative laparoscopic results; however, some women with observable pathologic conditions reported no pain symptoms.

Evanescent field excitation of fluorescence by epi-illumination microscopy.

By simple modification of the pattern of fluorescence excitation light in an epi-illumination inverted microscope, one can achieve conditions that produce total internal reflection fluorescence (TIRF) by evanescent wave excitation. Though traditionally requiring a collimated beam traversing through a special prism, TIRF also can be achieved by epi-illumination through the periphery of a 1.4 numerical aperture objective. An opaque disk of appropriate size is placed in the illumination path external to the microscope so as to cast a sharp, real-image shadow at the objective's back focal plane. This shadow allows a hollow cone of epiillumination rays traveling at only super-critical angles to reach the glass/water interface at the sample plane. Three kinds of TIRF illumination patterns can be produced by variations of this scheme: (1) a small spot of illumination of 1.5 microm radius by use of a laser light source, (2) a large region of illumination by use of a laser-illuminated diffusing screen located upbeam from the opaque disk, and (3) a large region of illumination by use of a conventional mercury arc.

Cluster analysis of insomniacs' MMPI profiles: relation of subtypes to sleep history and treatment outcome.

The Minnesota Multiphasic Personality Inventory (MMPI) profiles of 100 insomniacs who presented for treatment at a university medical center were analyzed by a hierarchical clustering procedure. This analysis revealed two major personality types that accounted for 88% of the sample. The two types referred to as "Type 1" and "Type 2" insomniacs differed significantly in regard to sleep history questionnaire responses and responsivity to treatment. MMPI scales and sleep history questionnaire items suggested that Type 1 insomniacs were less defended, more aroused/activated, and had sleep histories characterized by more childhood sleep problems and greater difficulties with sleep-disruptive cognitions than did Type 2 patients. Further, Type 1 patients had a poorer response to behavioral treatment as measured by change in sleep-onset latency than did Type 2 patients. These results suggest that the MMPI may be useful in identifying distinctive personality subgroups among insomniacs. Treatment implications are discussed, and mechanisms underlying the group differences are considered.

Premenstrual symptoms in black and white community samples.

Premenstrual syndrome specialty clinics are reported to be almost exclusively attended by white women. This racial discrepancy has raised the question of whether there is a lower prevalence or severity of symptoms during the premenstruum among black women. The authors evaluated selected premenstrual symptoms in a representative community-based sample and found no difference in the prevalence or severity of premenstrual symptoms reported by black and white women, except for a higher prevalence of food cravings among blacks. Exploration of broader sociocultural factors may explain the observed racial difference in seeking help for premenstrual complaints.

Patient satisfaction in Scott and Small-Carrion penile implant recipients: a study of 52 patients.

Physical penile sensations and levels of sexual satisfaction and activity were assessed in a questionnaire survey of 52 penile implant recipients. Although technical surgical success rates were high and 90% of recipients would elect to have surgery again, overall sexual adjustment did not approach premorbid conditions. Reasons for incomplete satisfaction included altered sensations of erection, decreased penile dimensions when compared with recalled preoperative dimensions, and decreased sensations during ejaculation. Some of these changes may have been due to decreased arousal during intercourse. Potential implant recipients should be counseled regarding realistic expectations of the benefits of penile prosthesis implant surgery.

Bioavailability of nasally administered progesterone.

PIP: 8 healthy women volunteers between the ages of 22-38 participated in a study designed to explore the relationship between endogenous estradiol (E2) levels and progesterone (P) absorption. Physical and pelvic examinations and laboratory screening tests revealed no abnormalities. All women had regular menstrual cycles, at 24-32-day intervals, and all were free of a significant menstrual cycle symptoms. 2 studies were performed at least 72 hours apart in the follicular phase of the menstrual cycle. The following medications were administered in random order: nasal P (Pronasone), 20 mg and nasal P (Pronasone), 30 mg. Serum P levels were drawn at the following times: 0, 3, 6, 10, 20, 30, 60, 120, 180, 240, 360, and 480 minutes. Serum for E2 assay was taken from the 0 time sample. The women were examined with a nasal speculum after each nasal absorption study. Serum was separated and frozen for subsequent assay. The data were analyzed using the CLINFO system from the National Institutes of Health. All of the women complained of a mildly unpleasant taste within several minutes following Pronasone administration. No evidence of nasal irritation was observed in any woman. The similar absorption curves obtained with Pronasone 20 mg and 30 mg doses and the aberrantly high values and delayed peaks ob tained in 2 subjects with the 30 mg dose imply that further work on dosage range, ointment formulation, and the method of application may be necessary before dependable clinical utility can be demonstrated. The peak levels of P that were reached compare favorably with results using similar doses (25 mg) in cocoa butter rectal or vaginal suppositories but are somewhat lower than those seen with polyethylene glycol base suppositories. The apparent inverse relationship between serum E2 levels and P levels obtained with Pronasone in the 20-mg dose was not expected. Alterations of nasal vascularity, interstitial hydration, and mucous blanket production all might influence absorption. The study demonstrates that the intranasal route is a potentially useful approach for the administration of unmodified sex steroid medications and is likely to be clinically safe and acceptable to patients.^ieng

Comparison of lifetime psychiatric diagnoses in Premenstrual Syndrome Clinic and community samples.

The purpose of this investigation was to obtain information about lifetime psychiatric diagnoses of women seeking treatment for premenstrual syndrome. The National Institute for Mental Health Diagnostic Interview Schedule (DIS) was administered to 223 women attending a premenstrual syndrome clinic. Rates of symptoms and psychiatric disorders were then compared with DIS data collected from an Epidemiologic Catchment Area (ECA) program community sample of 923 women in the same age group from the same geographic location. Women in the Premenstrual Syndrome Clinic sample met DIS/DSM-III criteria for dysthymia, phobia, obsessive-compulsive disorder, alcohol abuse/dependence, and drug abuse/dependence with a greater frequency than did women from the community sample. There appears to be much overlap between the symptoms for which women seek help from a specialty premenstrual syndrome clinic and symptoms related to several specific affective, anxiety, and substance abuse disorders. Further investigation is needed to determine whether premenstrual syndrome is strongly associated with DSM-III psychiatric diagnoses or whether current psychiatric classification systems are inadequate for differentiation.

Relationships among premenstrual symptoms and menstrual cycle characteristics.

The severity of premenstrual syndrome symptoms has been described by different investigators as varying directly or inversely with the severity of dysmenorrhea. In a large population of women undergoing comprehensive psychological and medical evaluation for premenstrual syndrome complaints, the authors examined the relationship between menstrual cycle parameters and premenstrual syndrome. The severity of premenstrual symptoms varied directly with the severity of dysmenorrhea and premenstrual spotting but was not associated with any other menstrual cycle parameter. Multivariate analysis revealed that social and psychological variables are more strongly associated with variations in premenstrual symptoms than are any menstrual cycle parameters.

Psychological assessment of women seeking treatment for premenstrual syndrome.

One hundred (100) women who sought evaluation for premenstrual syndrome were administered a battery of standardized psychological tests, including a Minnesota Multiphasic Personality Inventory (MMPI), Beck Depression Inventory (BDI) and Short Marital Adjustment Scale (MAS), during the follicular phase of the menstrual cycle. A BDI was repeated in the luteal phase of the menstrual cycle. MMPI profiles were classified by code type using Lachar system as follows: 'Normal'--36%; 'Neurotic'--31%; 'Characterological'--11%, 'Psychotic'--5%, 'Unclassified'--17%. A common feature of MMPI profiles was a low Scale 5 (Mf). BDI scores had several patterns; however, a significant group (25%) appeared to have clinically significant continuous depression regardless of superimposed premenstrual changes. About half of the women (42%) reported marital distress (MAS scores less than 100) at the time of evaluation.

Responses by graduates to memory of their internship in clinical psychology.

This study is a two-part investigation of clinical psychology internship training at the William S. Hall Psychiatric Institute. The first part surveyed 25 intern graduates to identify the relationship between experiences as an intern and current professional activities. Results are related to findings from other research in training clinical psychologists. The second part pertains to suggestions of intern graduates for specific additions and revisions of intership training at the Institute.