Investigation of the peak action wavelength of light-activated gene transduction.

Light-activated gene transduction (LAGT) is an approach to localize gene therapy via preactivation of cells with UV light, which facilitates transduction by recombinant adeno-associated virus vectors. Previous studies demonstrated that UVC induces LAGT secondary to pyrimidine dimer formation, whereas UVA induces LAGT secondary to reactive-oxygen species (ROS) generation. However, the empirical UVB boundary of these UV effects is unknown. Thus, we aimed to define the action spectra for UV-induced LAGT independent of DNA damage and determine an optimal wavelength to maximize safety and efficacy. UV at 288, 311 and 320 nm produced significant dose-dependent LAGT effects, of which the maximum (800-fold) was observed with 4 kJ m⁻² at 311 nm. Consistent with its robust cytotoxicity, 288 nm produced significantly high levels of DNA damage at all doses tested, whereas 311, 320 and 330 nm did not generate pyrimidine dimers and produced low levels of DNA damage detected by comet assay. Although 288 nm failed to induce ROS, the other wavelengths were effective, with the maximum (10-fold) effect observed with 30 kJ m⁻² at 311 nm. An in vivo pilot study assessing 311 nm-induced LAGT of rabbit articular chondrocytes demonstrated a significant 6.6-fold (P<0.05) increase in transduction with insignificant cytotoxicity. In conclusion, 311 nm was found to be the optimal wavelength for LAGT on the basis of its superior efficacy at the peak dose and its broad safety range that is remarkably wider than the other UV wavelengths tested.

Evaluation of the impact of highly active antiretroviral therapy on immune recovery in antiretroviral naive patients.

OBJECTIVES: To examine the extent of immune reconstitution in treatment-naive patients with CD4 T-cell counts <500 cells/microL following 48 weeks of highly active antiretroviral therapy (HAART). METHODS: Thirteen antiretroviral naive patients were evaluated longitudinally for 48 weeks on HAART utilizing immune functional and lymphocyte phenotyping assays, including lymphocyte proliferation assay, flow cytometric evaluation of cell surface markers, and delayed type hypersensitivity skin tests. Virologic responses were monitored using commercially available viral load assays and gag/pol mRNA quantification using simultaneous immunophenotyping/UltraSensitive fluorescence in situ hybridization (ViroTect In Cell HIV-1 Detection Kit; Invirion, Frankfort, MI). Thymic function was evaluated for a subset of four patients using real-time polymerase chain reaction (PCR) for T-cell receptor excision circle (TREC) quantification and thymic scans using computerized axial tomography (CT) of the thymus. RESULTS: HAART initiation resulted in a significant decline in plasma viremia and percentage of infected peripheral blood cells, and a rise in CD4 T cells from a baseline median of 207 cells/microL to a week-48 median of 617 cells/microL. The rise was predominately in CD4 memory cells. Naive T cells also increased in number, but at a slower rate. Activated (HLA-DR CD38) CD4 and CD8 T cells were elevated at baseline (24 and 62%, respectively) and declined by week 48 (17 and 36%, respectively) but did not reach normal levels. The number of Fas CD4 T cells increased from a baseline median of 169 to 381 cells/microL at week 48. Both soluble interleukin (IL)-2 and tumour necrosis factor (TNF) II receptors declined by week 48. HIV p24 lymphocyte proliferation assay responses were transiently detected in three patients. TREC values increased from a median 6400 copies/microg at baseline to a week-48 median value of 26 697 copies/microg. CONCLUSION: Immune functional reconstitution was not achieved in these HAART naive patients.

Trends in the use of selective serotonin reuptake inhibitors in nine Department of Veterans Affairs outpatient facilities.

The average daily dose and need for dose escalations for the drugs known as selective serotonin reuptake inhibitors (SSRIs) has frequently been a point of controversy. This study reports on the information gathered from nine Veterans Affairs hospitals over a two six-month periods. Average daily doses of fluoxetine, paroxetine, and sertraline started at 30.9 mg, 24.2 mg, and 87.8 mg and ended at 28.4 mg, 24.2 mg, and 89.8 mg, respectively. Cost, number of prescriptions, and dosage strength data is also presented.

Nefazodone: single versus twice daily dose.

This pilot study involved six men with major depression treated with nefazodone dosed either twice/day or once/day at bedtime. Depression was rated before nefazodone therapy and at 4 weeks by the self-report version of the Hamilton Depression Rating Scale. A simple 10-cm visual analog side effect scale for daytime drowsiness was completed at the latter time. Dosages of nefazodone were at least 400 mg/day. The results suggest that nefazodone given once/day at bedtime may be as effective as the currently accepted twice/day regimen, with less daytime drowsiness.

Money as a tool to extinguish conditioned responses to cocaine in addicts.

Classical conditioning principles offer a nondrug way to treat cocaine dependence. Eleven male subjects with the primary diagnosis of cocaine dependence were placed into one of two groups. The experimental group was asked to handle $500 cash in a mock budgetary task. The control group was asked to just imagine handling and budgeting the money. The subjects rated their craving-related feelings before and after each task. The experimental group showed significantly more craving after the money-handling task as compared to the control group, and the scores improved with time and as more tasks were completed. These data show that craving induced by handling cash is powerful and can be attenuated, at least on a short-term basis, using classical extinction procedures.

A validational study of the Structured Interview of Symptoms Associated with Sexual Abuse (SASA) using three samples of sexually abused, allegedly abused, and nonabused boys.

OBJECTIVE: With the rapid rise of sexual abuse allegations, there is a growing need to develop instruments to help clinicians determine the likelihood that sexual abuse has occurred. METHOD: This study evaluated the discriminant validity of a structured parent interview regarding emotional, behavioral, and physical symptoms by comparing results among three subsamples of age matched boys: 22 sexually abused boys whose perpetrator confessed, 47 boys evaluated in a sexual abuse clinic but without a history of perpetrator confession and 52 nonabused boys selected after rigorous screening. RESULTS: In comparison with the nonabused (NA) boys, the sexually abused boys were significantly more likely to demonstrate sudden emotional and behavioral changes, frequent stomach aches, more knowledge about sex and sexual activities than expected for age, unusual aggressiveness toward playmate or toy's private parts, crying easily, difficulty getting to sleep, and a change to poor school performance. The internal reliability of the SASA was determined to be .83 and scoring of an abbreviated 12-item scale demonstrated a sensitivity of 90.9% and a specificity of 88.5%. CONCLUSIONS: The findings suggest that Structured Interview of Symptoms Associated with Sexual Abuse (SASA) is an effective tool which may help in the comprehensive assessment of boys who may have been sexually abused.

Emotional, behavioral, and physical symptoms reported by parents of sexually abused, nonabused, and allegedly abused prepubescent females.

There have been few studies that have examined emotional, behavioral, and physical symptoms that discriminate between well-defined groups of sexually abused and nonabused children. This study examined the results of a structured parent interview (SASA) completed on three matched samples of prepubescent females: 68 who were selected for nonabuse (NA Group), 68 from a sexual abuse clinic in which a perpetrator confessed (SA Group) and 68 seen at the same clinic who did not have a perpetrator confession (AA Group). Parents of girls in both the SA and AA groups reported increased sleep problems, fearfulness, emotional and behavioral changes, concentration problems, and sexual curiosity and knowledge. When contrasting the known (SA) with the allegedly abused sample (AA), self-consciousness, nightmares, and fearfulness of being left alone emerged significantly more frequently in the SA sample.

Daily cocaine craving in a 3-week inpatient treatment program.

Fifteen male veteran cocaine addicts took simple tests of cocaine craving, mood, energy, and perceived health status for 23 consecutive days during an inpatient treatment center. Results show they experienced very little cocaine craving or other symptomatology. Multiple regression analyses show that health rating alone was the best predictor of cocaine craving. These results question solely internal sources of craving and the oft-touted craving-depression connection. The study delineates further implications of the findings.

Perianal injuries resulting from sexual abuse: a longitudinal study.

Four children who incurred perianal injuries as a result of a sexual assault were followed on a longitudinal basis to document the anatomical changes that ensued. The subjects, whose ages ranged from 4 to 8 years, were followed from 1 week to 14 months. They were examined in both supine and prone knee-chest positions and a 35-mm camera mounted on a colposcope was used to record their injuries. At the time of the initial examination, there were a variety of findings including erythema of the tissues, edema of the skin folds, localized venous engorgement, dilation of the external anal sphincter, and lacerations of different depths. Superficial lacerations reepithelized within 1 to 11 days. The second-degree wounds in two of the children were healed by the 1- and 5-week return visits, leaving narrow bands of scar tissue. In the two subjects who were followed the longest, signs of both a second-degree laceration and a surgically repaired third-degree injury had virtually disappeared by 12 to 14 months after the assaults. The wounds in one subject, infected with a herpes simplex type 2 virus, remained erythematous for a longer period of time than did similar injuries in the other children. A skin tag created by the avulsion of the tissues in one subject persisted, although it became less obvious as it retracted into the redundant folds of the perianal tissues over time.

Genital injuries resulting from sexual abuse: a longitudinal study.

Three children who incurred genital injuries as a result of sexual assaults were followed up on a longitudinal basis to document the anatomical changes which ensued. The subjects, who were 4 months, 4 years 5 months, and 9 years of age, were followed up for periods ranging from 14 months to 3 years. A multi-method examination approach and a 35-mm camera mounted on a colposcope were used to examine and record their injuries. Signs of the acute damage disappeared rapidly, and the wounds healed without complications. Following the resolution of the acute injuries, the changes created by the trauma remained relatively stable throughout the prepubertal years. The most persistent findings were irregular hymenal edges and narrow rims at the point of the injury. Over time the jagged, angular margins smoothed off. Disruption of the hymen exposed underlying longitudinal intravaginal ridges whose hymenal attachments created mounds or projections. There was little apparent scar formation. Even the injuries to the posterior fourchettes healed with minimal scar tissue and left only the slightest evidence of the trauma. With the onset of puberty, the hymenal changes in the oldest subject were obscured by the hypertrophy of this membrane. An examination technique which used a Q-tip to separate the redundant tissues demonstrated that the signs of trauma had survived.

Neuroleptic malignant syndrome associated with carbamazepine.

We have described the case of a patient who had symptoms of the neuroleptic malignant syndrome after treatment with carbamazepine. The symptoms resolved after carbamazepine therapy was stopped, which suggests carbamazepine as their cause.

A postdischarge adverse drug reaction capture mechanism using medical record technicians.

This article describes a system of adverse drug reaction (ADR) reporting implemented at WJB Dorn Veterans Affairs (VA) Hospital, Columbia, South Carolina, which involves reviewing all inpatient charts by medical record technicians for possible ADRs and notifying the chief pharmacist for investigation. Cases are assigned to a clinical or staff pharmacist who completes an ADR data collection form. All ADR forms are presented to and reviewed by the pharmacy and therapeutic committee where a decision is made on whether to forward the ADR to the U.S. Food and Drug Administration via the VA Central Office for compilation. In the program's second quarter of 1990, ADR reporting increased approximately fivefold.

A simple test of cocaine craving and related responses.

This study evaluated a self-rated instrument for assessing perceived craving, mood, energy, and health levels in patients who are withdrawing from cocaine addiction. The instrument consisted of four 20-centimeter line scales. Twenty-five patients in a 28-day inpatient addictions treatment unit were chosen at random points in their treatment and given a chance to rate themselves on each scale by making a mark on a line that corresponded to their self-perception. Each patient performed two self-assessment, one day apart. The scores for each scale were correlated and found to be highly reliable. Surprisingly, none of the other three scale scores had a strong relationship with craving, and possible reasons for this result are discussed.

Genital findings in prepubertal girls selected for nonabuse: a descriptive study.

This project was designed to collect normative data on the genital anatomy from a sample of nonabused prepubertal girls. A total of 114 girls between the ages of 10 months and 10 years were examined and photographed with a colposcope. After screening for the onset of puberty and the possibility of undetected abuse, 93 subjects remained. Examination techniques included a supine labial separation approach, a supine labial traction method, and a prone knee-chest position. Common genital finding included erythema of the vestibule (56%), periurethral bands (50.6%), labial adhesions (38.9%), lymphoid follicles on the fossa navicularis (33.7%), posterior fourchette midline avascular areas (25.6%), and urethral dilation with labial traction (14.9%). The hymenal orifice opened more frequently during the knee-chest (95.2%) and the labial traction (90.5%) methods than with the supine separation (79.3%) approach. Hymenal findings included mounds (33.8%), projections (33.3%), and "septal remnants"/midline hymenal tags (18.5%). Intravaginal findings of vaginal ridges (90.2%) and rugae (88.7%) were found primarily through the labial traction approach. The cervix was visualized without the use of speculum in 69% of the children during the knee-chest examination. Unusual findings included posterior fourchette friability (4.7%), anterior hymenal clefts (1.2%), and notches of the hymen (6%). One child had an imperforate hymen (1.2%) and 2 (2.5%) had hymenal septa. A vaginal discharge was detected in 2 (2.6%) and a foreign body in 1 (1.3%). Tables that include the vertical and horizontal hymenal orifice diameters by age group and by method are presented.

Comparison of genital examination techniques in prepubertal girls.

The results of three separate methods used to examine prepubertal girls are compared and a technique for measuring hymenal orifice diameters from colposcopic photographs is presented. A total of 172 girls who were examined by three techniques during their evaluation in a clinic for suspected child sexual abuse victims were studied. Their ages ranged from 10 months to 11 years with a mean of 5 years, 8 months. The examination techniques used were the supine position with labial separation, the supine position with labial traction, and the knee-chest position. The knee-chest position (98%) and the supine traction method (96%) proved to be superior to the supine separation technique (86%) in opening the vaginal introitus. The largest vertical transhymenal diameters were produced in the knee-chest position, whereas the greatest transverse horizontal spans were generated by the supine traction procedure. Other soft tissue changes were noted but not quantified. A multimethod approach to the examination of the sexually abused child is recommended to take advantage of the strengths of each technique.

Perianal findings in prepubertal children selected for nonabuse: a descriptive study.

The results of the perianal portion of a project designed to collect normative data of the anogenital anatomy from a representative sample of prepubertal children is presented. A total of 318 children were examined by three physicians from a child sexual abuse evaluation program. After screening for the onset of puberty and the possibility of undetected abuse, 267 subjects remained. The sample included 161 girls and 106 boys ranging in age from 2 months to 11 years. The perianal findings that were encountered with the greatest frequency included erythema (41%), increased pigmentation (30%), and venous engorgement (52%) after two minutes in the knee-chest position. Wedge-shaped smooth areas in the midline, with or without depressions, were found both anterior and posterior to the anus in 26% of the children. Anal skin tags/folds were discovered anterior to the anus in 11%. In 49% of the children there was some dilatation of the anus which opened and closed intermittently in 62%. Flattening of the anal verge and rugae occurred during dilatation by the midpoint of the examination in 44% and 34%, respectively. Perianal findings that were found infrequently in all subgroups included skin tags/folds (0%) and scars (1%) outside the midline, anal dilatation greater than 20 mm without the presence of stool in the rectal ampulla (1.2%), irregularity of the anal orifice after complete dilatation (3%), and prominence of the anal verge (3%). No abrasions, hematomas, fissures, or hemorrhoids were encountered. Less commonly detected findings within specific subgroups included perianal erythema in girls (32%) as compared to boys (57%), pigmentation in the lighter skinned white children (22%) when compared to black (53%) and Hispanic (58%) children, and venous congestion at the beginning of the examination (7%) when compared to the same findings after four minutes in the knee-chest position (73%). There were no perianal skin tags/folds found in the boys. The relatively high incidence of perianal soft tissue changes that were found in this study, when compared to the frequency of similar observations in children suspected of having been sexually abused, reemphasizes the caution medical examiners must exercise in rendering an opinion as to the significance of medical findings.

Development of Pseudomonas aeruginosa resistance to norfloxacin during therapy.

Two elderly patients diagnosed with Pseudomonas aeruginosa urinary tract infections were treated with oral norfloxacin in the recommended dose of 400 mg q12h. Initially, antimicrobial susceptibility data indicated the organisms were sensitive to norfloxacin. Six to eight days into therapy urine cultures became positive for P. aeruginosa once again; this time, however, susceptibility reports indicated the organisms were now resistant to norfloxacin. Since cross-resistance among norfloxacin, other quinolones, and cephalosporins can occur, we recommend repeated urine cultures during and after norfloxacin therapy in elderly patients with complicated P. aeruginosa urinary tract infections.

Labial adhesions and posterior fourchette injuries in childhood sexual abuse.

Six cases involving six sisters, all of whom were sexually molested, are presented to illustrate the association between labial adhesions and posterior fourchette injuries in sexually abused children. Although labial adhesions are a relatively common lesion, they only recently have been associated with childhood sexual abuse. Labial adhesions are ordinarily found in girls between 2 months and 7 years of age and are usually very superficial and disappear spontaneously by the time the child approaches puberty. We encountered six sisters, all of whom had been sexually abused, who had labial adhesions. Four of the six had changes in the area of the posterior fourchette that were consistent with previous trauma. In addition, four of the girls' hymens had thickened, irregular edges or other changes that were considered abnormal. The father, the grandfather, and an uncle confessed to lewd and lascivious misconduct with the children. Although labial adhesions alone are not reason enough to make a report of sexual abuse, the physician is obligated to inquire as to the possible cause if the adhesions do not fit the usual pattern or if there are other suspicious findings.