A Prospective, Multicenter, Pilot Trial Of A Novel Homologous Skin Construct On Deep Partial-Thickness And Full-Thickness Burns.

Split-thickness skin grafting (STSG) is the standard of care for treating deep burns. They often contract, have unpredictable cosmetic outcomes, lack dermal appendages, and result in painful, conspicuous donor sites. An autologous homologous skin construct (AHSC) has been shown to produce full-thickness skin architecture. This study examined the safety profile, engraftment, and quality of healing of a pilot group of AHSC-treated burn wounds. Following IRB approval and informed consent, patients with deep-partial/full-thickness burns requiring grafting underwent side-by-side treatment with AHSC and STSG. A 2 cm2 fullthickness harvest was processed into AHSC at an FDA-registered facility, returned within 48 hours, and applied to a 4 cm2 area alongside a STSG. AHSC donor site was closed primarily. Wounds were evaluated for healing with digital photography and investigator assessments for 90 days. All adverse events (AEs) were recorded. Eight patients with average 13.3% TBSA [range 2-58%] burn wounds were treated: 5 Caucasian and 3 African American with an average body mass index (BMI) of 26.8. Injury was due to predominantly flame burn, with additional injury from grease, scald, contact, friction and flash. Mean time between injury and AHSC treatment was 11 days [range 5-35 days]. All patients had adequate engraftment and complete epithelialization by the end of the study. Patients required one application of AHSC and no other additional surgical procedures at the application sites. The most common AEs for STSG-treated wounds included hypertrophic scarring and pruritus. One non-infected AHSC harvest site experienced a dehiscence. There were no other AEs related to AHSC treatment. AHSC treatment is feasible in deep partial and fullthickness burn wounds warranting additional investigation.

La greffe dermo- épidermique (GDE) est le traitement de référence des brûlures profondes. La zone traitée est sujette aux brides, n'a pas d'appendices dermiques, a un aspect esthétique aléatoire et le site donneur est indéniablement douloureux. Un hybride cutané autologue- homologue (HCAH) a montré être architecturalement proche de la peau. Cette étude a pour but d'évaluer l'innocuité, la qualité de prise et la qualité cicatricielle obtenues sur un groupe pilote de brûlés profonds. Après autorisation des tutelles et consentement éclairé, les patients, nécessitant une greffe ont reçu, côte à côte, une GDE et un HCAH. Ce dernier est préparé à partir d'un prélèvement de 2 cm² de peau totale (auto- fermant), en 48 h, dans une structure approuvée par la FDA. On obtient une structure de 4 cm², installée à côté d'une GDE. Les brûlures ont été évaluées cliniquement et photographiées pendant 90 j. Tous les événements indésirables (EI) ont été répertoriés. Huit patients brûlés sur 13,3 % (2-58) de SCT ont été inclus. Il s'agissait de 5 blancs et 3 noirs (je dois traduire même ceci, qui me semble foncièrement non éthique- NDRLF) ayant un IMC de 26,8. Les brûlures étaient liées à un flamme mais aussi à de la graisse, par ébouillantement, contact, flash ou dermabrasion. Le délai moyen de mise en place de l'HCAH était de 11 jours (5-53). L'intégration de la greffe a été bonne et tous les patients étaient cicatrisés à la fin de l'étude, sans nécessité de nouvelle greffe. Les EI les plus fréquents observés sur les zones GDE étaient des cicatrices hypertrophiques et un prurit. Une zone HCAH s'est désunie (hors infection), seul EI observé dans ce groupe. L'HCAH semble utilisables sur les brûlures profondes et doit être étudié plus avant.

Shedding of the endothelial glycocalyx is quantitatively proportional to burn injury severity.

Limited information exists regarding endothelial dysfunction following burn injury. This project aims to evaluate whether thermal injury results in shedding of the endothelial glycocalyx in a manner quantitatively proportional to injury severity, and whether theloss of intact glycocalyx is measurable in end organs. C57BL/6 mice were grouped as uninjured controls, 10% or 25% Total Body Surface Area (TBSA) scald burns. Blood and tissue sampling was performed over a specific time course. Plasma levels of shed syndecan-1, a marker of glycocalyx damage, were quantified by ELISA. Lung and spleen sections were stained with immunofluorescent anti-syndecan-1 antibodies to evaluate intact glycocalyx. Plasma syndecan-1 levels were higher in injured versus uninjured animals. Normalized levels of syndecan-1 in burned mice were significantly increased compared to hour 0 (p<0.05) at hours 4 and 8 post-injury in the 10% TBSA, and at hour 4 in the 25% TBSA group. Levels in the 10% and 25% TBSA groups peaked at hour 4 with fold change of 2.3 and 2.4 respectively. There was less pulmonary syndecan-1 immunostaining in burned animals compared to controls, and the levels inversely correlated with systemic shed syndecan- 1, beginning at hour 4 in the 10% TBSA injury group and at all time points in the 25% TBSA injury group, (0.27±0.06 and 0.14±0.04 respectively for hour 4). Similarly, there was less spleen syndecan-1 immunostaining in burned animals compared to controls at all time points. Burn injury causes shedding of syndecan-1 in a murine model, with levels correlated to injury severity and loss of the glycocalyx in lung and spleen. This work provides further insight into quantification and temporality of glycocalyx damage and systemic response to burn.

Les données concernant la dysfonction endothéliale après brûlure sont parcellaires. Les buts de cette étude étaient d'établir une corrélation entre la perte de glycocalyx et la gravité de la brûlure et si cette perte était mesurable au niveau des organes. Des souris C57BL/6 ont été réparties en groupes contrôle, brûlure 10% et brûlure 25% de SCT. Des prélèvements de sang et de tissus ont été réalisés à intervalles prédéterminés. Les taux plasmatiques de syndecan 1 (S1), marqueur de lésion du glycocalyx, ont été mesuré par méthode ELISA. Des échantillons de poumon et de rate ont été mis en présence d'anticorps anti S1, afin d'évaluer le glycocalyx intact. Les taux plasmatiques de S1 étaient plus élevés que ceux du groupe contrôle. Chez les souris brûlées sur 10% de SCT, les taux de S1 à 4h et 8 h étaient supérieurs au taux avant brûlure, ceci n'étant observé qu'à h4 chez les souris brûlées sur 25% de SCT. Le pic de S1 se produisait à h4, avec un rapport de x2,3 (10%) et x2,4 (25%) par rapport à la valeur de base. A partir de h4, on observait une baisse de complexes S1-antiS1 dans les poumons des souris brûlées sur 10% (0,27 +/- 0,06), inversement corrélée aux taux plasmatiques de S1. Cette observation se répétait lors de tous les dosages chez les 25% (0,14 +/- 0,04 à h4). Les mêmes constatations étaient faites sur les échantillons de rate. La brûlure cause des lésions du glycocalyx, parallèles à sa gravité. Ces travaux ouvrent le champ à des recherches futures sur les lésions du glycocalyx et la réponse inflammatoire aux brûlures.

In situ biodistribution and residency of a topical anti-inflammatory using fluorescence lifetime imaging microscopy.

BACKGROUND: GSK2894512 is a topically delivered investigational drug being developed for treatment of atopic dermatitis and psoriasis. OBJECTIVES: To investigate, in a phase I clinical trial, the spatial biodistribution and residency of GSK2894512 within the epidermis and dermis of healthy human participants noninvasively using fluorescence lifetime imaging microscopy (FLIM). METHODS: Two topical drug formulations containing GSK2894512 1% were applied to the right and left forearms of six participants for seven consecutive days, followed by seven days of observation for residency. FLIM images were obtained daily throughout the study, approximately every 24 h. During the treatment phase of the study, images were collected from each participant pretreatment, reflecting the residual dose from the previous day. Three punch biopsies from each participant of one formulation was obtained from the treated region during the post-treatment follow-up period between days 8 and 14 for comparison with FLIM results. RESULTS: Cellular and subcellular features associated with different epidermal and dermal layers were visualized noninvasively, down to a depth of 200 µm. Results yielded three-dimensional maps of GSK2894512 spatial distribution and residency over time. This fluorescence data provided a marker that was used as a monitor for day-to-day variance of drug presence and residency postapplication. CONCLUSIONS: The results suggest FLIM could be a viable alternative to skin biopsies without the usual patient discomfort and limitations, thereby enabling the direct measurement of skin distribution through longitudinal monitoring. These results are the first step in establishing the unique capabilities that multiphoton imaging could provide to patients through noninvasive drug detection.

Manometric evaluation of the motilin receptor agonist camicinal (GSK962040) in humans.

BACKGROUND: The gut hormone motilin stimulates gastrointestinal motility by inducing gastric phase III of the migrating motor complex (MMC) and enhancing the rate of gastric emptying. Camicinal (GSK962040), a small molecule motilin receptor agonist, has been shown to increase gastrointestinal motility. METHODS: In this proof of concept study the effects of camicinal on MMC activity, esophageal and gastric pH was evaluated in eight healthy volunteers as a secondary endpoint. Doses of 50 and 150 mg were compared to placebo for a period of 24 hours in a double-blinded randomized crossover trial. KEY RESULTS: The 50 mg dose (n=4) of camicinal had no significant impact on gastroduodenal manometry or pH parameters. A single dose of 150 mg (n=4) induced a gastric phase III after 0:34 h (0:25-0:58), which was significantly faster compared to placebo (18:15 h (4:32-22:16); P=.03). Moreover, the high dose significantly increased the occurrence of gastric phase III contractions compared to placebo (12% vs 39%; P=.0003). This increase in gastric phase III contractions during a period of 24 hour was due to an increased occurrence of gastric phases III during the daytime (5% vs 50%; P=.0001). The same dose however did not affect small bowel manometry parameters or esophageal and gastric pH. CONCLUSIONS AND INFERENCES: Considering its stimulating effect on the MMC and gastric emptying, camicinal is an attractive candidate for the treatment of gastroparesis and gastroesophageal reflux disease. This trial was registered at clinicaltrials.gov as NCT00562848.

Impact of microbiology cascade reporting on antibiotic de-escalation in cefazolin-susceptible Gram-negative bacteremia.

Cascade reporting (CR) involves reporting the susceptibilities of broad-spectrum agents only when the organism is resistant to more narrow-spectrum agents. The purpose of this study is to evaluate the impact of CR on antibiotic de-escalation practices and to characterize the impact of CR on clinical outcomes. CR rules were implemented in the microbiology laboratory at Atlantic Health System (AHS) in June 2013. A retrospective chart review was conducted at two community teaching hospitals in adult patients who had a blood culture positive for a Gram-negative organism susceptible to cefazolin and who were empirically treated with broad-spectrum beta-lactam (BSBL) antibiotics. De-escalation practices were compared in the pre-CR (July 2012-December 2012) and post-CR (July 2013-December 2013) periods. The primary endpoint was the percentage of patients whose BSBL agent was de-escalated to agents listed on the post-CR antibiotic susceptibility report within 48 h of the final report. Secondary endpoints include the difference in pre-CR and post-CR periods in terms of hospital length of stay, in-hospital mortality, 30-day readmission, Clostridium difficile infections, and re-initiation of a BSBL agent within 7 days. A total of 73 patients were included; 31 in the pre-CR and 42 in the post-CR period. Patients had similar baseline characteristics. Therapy was de-escalated in 48 % of pre-CR vs 71 % of post-CR patients (p = 0.043). No significant differences were observed in secondary endpoints between patients in the pre-CR and post-CR periods. CR resulted in significant improvements in de-escalation practices without affecting safety outcomes.

A Translational Animal Model for Scar Compression Therapy Using an Automated Pressure Delivery System.

BACKGROUND: Pressure therapy has been used to prevent and treat hypertrophic scars following cutaneous injury despite the limited understanding of its mechanism of action and lack of established animal model to optimize its usage. OBJECTIVES: The aim of this work was to test and characterize a novel automated pressure delivery system designed to deliver steady and controllable pressure in a red Duroc swine hypertrophic scar model. METHODS: Excisional wounds were created by dermatome on 6 red Duroc pigs and allowed to scar while assessed weekly via gross visual inspection, laser Doppler imaging, and biopsy. A portable novel automated pressure delivery system was mounted on developing scars (n = 6) for 2 weeks. RESULTS: The device maintained a pressure range of 30 ± 4 mm Hg for more than 90% of the 2-week treatment period. Pressure readings outside this designated range were attributed to normal animal behavior and responses to healing progression. Gross scar examination by the Vancouver Scar Scale showed significant and sustained (>4 weeks) improvement in pressure-treated scars (P < .05). Histological examination of pressure-treated scars showed a significant decrease in dermal thickness compared with other groups (P < .05). Pressure-treated scars also showed increased perfusion by laser Doppler imaging during the treatment period compared with sham-treated and untreated scars (P < .05). Cellular quantification showed differential changes among treatment groups. CONCLUSION: These results illustrate the applications of this technology in hypertrophic scar Duroc swine model and the evaluation and optimization of pressure therapy in wound-healing and hypertrophic scar management.

Epidemiology, clinical manifestations, and outcomes of Scedosporium infections among solid organ transplant recipients.

BACKGROUND: Few studies of Scedosporium infections following solid organ transplantation have been performed in the era of induction immunosuppression and widespread antifungal prophylaxis. METHODS: We performed a single-center, retrospective study of transplant recipients from 2000 through 2010 who had a positive Scedosporium culture. RESULTS: Among 27 patients, 67% (n = 18) and 33% (n = 9) were infected with Scedosporium apiospermum and Scedosporium prolificans, respectively. A total of 67% received induction immunosuppression and 74% received prior antifungal therapy. Isolates were broadly resistant to antifungals. Of these patients, 59% (n = 16) were colonized by Scedosporium, and 41% (n = 11) had disease (scedosporiosis). No significant clinical differences were seen between species. Colonization occurred exclusively in the lungs of lung transplant recipients (LTR). Scedosporiosis followed lung transplantation in 55%, and other organ transplants (multivisceral [18%]; and heart, liver, small intestine [9% each]) in 45%. Scedosporiosis was preceded by colonization in 36%. Diseases included pneumonia (64%), mediastinitis (18%), and fungemia/disseminated infections (18%). The 6-month outcomes were death in 55%, progression in 18%, stability in 9%, and resolution in 18%. Patients who died had earlier onset scedosporiosis post transplant (median: 80.5 vs. 1388 days; P = 0.04), and were more likely to have mediastinitis or disseminated infections than pneumonia (100% vs. 29%; P = 0.06). The 3 patients who developed scedosporiosis >1 year post transplant survived. All patients who survived were treated with a voriconazole-containing regimen. CONCLUSIONS: LTR were most susceptible to Scedosporium colonization and scedosporiosis, particularly within the lungs. Death was common with scedosporiosis in the first year after all types of organ transplants, consistent with profound immunosuppression and antifungal resistance, but not encountered thereafter.

Sustained reduction in tuberculosis incidence following a community-based participatory intervention.

BACKGROUND: Rates of latent tuberculosis infection (LTBI) and tuberculosis (TB) disease are elevated in the rural southeastern United States and among US- and foreign-born Black residents. To prevent TB and reduce TB transmission, community-based strategies are essential. OBJECTIVE: To describe a community-based participatory intervention for improving the detection and treatment of LTBI and TB and reducing TB incidence. DESIGN: In rural Florida, we carried out a community educational TB campaign from 1997 to 2000, including presentations at community events, a media campaign and working with local community groups to develop culturally appropriate prevention messages. The campaign was implemented concurrently with a population-based LTBI survey. RESULTS: The annual TB incidence rate in the intervention area decreased from 81 per 100 000 in 1994-1997, to 42/ 100 000 in 1998-2001, and to 25/100 000 in 2002-2005 (P = 0.001). This decrease was not observed in communities where the intervention was not implemented. There was no decrease in the TB incidence rate ratio between Blacks and non-Blacks in either region during the study period. CONCLUSIONS: We conclude that community participation in LTBI screening and TB education was associated with a substantial reduction in TB rates. Although the TB incidence rate ratio did not decrease between Blacks and non-Blacks, TB incidence fell in all racial groups.

Dual skin tests with Mycobacterium avium sensitin and PPD to detect misdiagnosis of latent tuberculosis infection.

BACKGROUND: A positive tuberculin skin test (TST) may indicate cross-reacting immunity to non-tuberculous mycobacteria (NTM) and not latent tuberculosis infection (LTBI). OBJECTIVES: To assess misclassification of LTBI, as assessed by skin testing with Mycobacterium avium sensitin (MaS), and to determine how this misclassification affects the analysis of risk factors for LTBI. METHODS: In a population-based survey, participants underwent skin testing with M. tuberculosis purified protein derivative (PPD) and MaS. A PPD-dominant skin test was a reaction that was ≥ 3 mm larger than the MaS reaction; a MaS-dominant skin test was a reaction that was ≥ 3 mm larger than the PPD reaction. RESULTS: Of 447 randomly selected persons, 135 (30%) had a positive PPD test. Of these, 21 (16%) were MaS- dominant, and were therefore attributable to NTM and misclassified as LTBI. PPD reactions of 5-14 mm were more likely to be misclassified than those ≥ 15 mm (OR = 5.0, 95%CI 1.9-13.2). Adjusting for misclassification had only a small impact on the analysis of risk factors for LTBI. CONCLUSIONS: A substantial number of individuals who are diagnosed with LTBI are actually sensitized to NTM. Using dual skin testing would reduce misdiagnosis and prevent unnecessary treatment.

Racial disparities in primary and reactivation tuberculosis in a rural community in the southeastern United States.

SETTING: A rural section of a county in central Florida. BACKGROUND: Racial disparities in tuberculosis disease (TB) are substantial in the United States. OBJECTIVE: To determine if TB was attributable to primary infection, reactivation or both. DESIGN: A population-based survey of latent tuberculosis infection (LTBI), a case-control analysis of TB, and a cluster analysis of TB isolates were performed between 1997 and 2001. RESULTS: Of 447 survey participants, 135 (30%) had LTBI. Black race was strongly associated with LTBI among US-born (OR 2.6, 95%CI 1.3-5.5) and foreign-born subjects (OR 4.3, 95%CI 2.2-8.4). Risk factors for TB included human immunodeficiency virus (HIV; OR 27.4, 95%CI 10.1-74.1), drug use (OR 4.6, 95%CI 1.7-12.4) and Black race (OR 3.4, 95%CI 1.2-9.6). The population risk of TB attributable to Black race was 64%, while that attributable to HIV was 46%. Cluster analysis showed 67% of TB cases were clustered, but Blacks were not at a significantly increased risk of having a clustered isolate (OR 2.1, 95%CI 0.12-36.0). CONCLUSION: Both reactivation TB and recent TB transmission were increased among Blacks in this community. Therefore, LTBI screening and intensive contact tracing, both followed by LTBI treatment, will be needed to reduce TB in Blacks.

Retinoic acid induces sodium/iodide symporter gene expression and radioiodide uptake in the MCF-7 breast cancer cell line.

The sodium/iodide symporter (NIS) stimulates iodide uptake in normal lactating breast, but is not known to be active in nonlactating breast or breast cancer. We studied NIS gene regulation and iodide uptake in MCF-7 cells, an estrogen receptor (ER)-positive human breast cancer cell line. All-trans retinoic acid (tRA) treatment stimulated iodide uptake in a time- and dose-dependent fashion up to approximately 9.4-fold above baseline. Stimulation with selective retinoid compounds indicated that the induction of iodide uptake was mediated by retinoic acid receptor. Treatment with tRA markedly stimulated NIS mRNA and immunoreactive protein ( approximately 68 kDa). tRA stimulated NIS gene transcription approximately 4-fold, as shown by nuclear run-on assay. No induction of iodide uptake was observed with RA treatment of an ER-negative human breast cancer cell line, MDA-MB 231, or a normal human breast cell line, MCF-12A. The iodide efflux rate of tRA-treated MCF-7 cells was slow (t(1/2) = 24 min), compared with that in FRTL-5 thyroid cells (t(1/2) = 3.9 min), favoring iodide retention in MCF-7 cells. An in vitro clonogenic assay demonstrated selective cytotoxicity with (131)I after tRA stimulation of MCF-7 cells. tRA up-regulates NIS gene expression and iodide uptake in an ER-positive breast cancer cell line. Stimulation of radioiodide uptake after systemic retinoid treatment may be useful for diagnosis and treatment of some differentiated breast cancers.

Action research sets the stage to improve discharge preparation.

An action research project was conducted to explore parents' perceptions about discharge preparation for a child with a respiratory problem. The action research approach was selected to provide bedside nurses with an opportunity to participate in research activities and actively change practice. Seven bedside nurses were recruited and prepared to conduct home interviews of 14 urban families and telephone interviews of six rural families. Families not participating in interviews were asked to complete questionnaires. Data analysis revealed four major themes: (a) the parent as learner, (b) the content of teaching activities, (c) timing of discharge, and (d) the impact of hospitalization after discharge. All nurses were positive about their experience. Action research provided a nonthreatening opportunity to involve bedside nurses in a research project.

Initial clinical experience with a video-based patient positioning system.

PURPOSE: To report initial clinical experience with an interactive, video-based patient positioning system that is inexpensive, quick, accurate, and easy to use. METHODS AND MATERIALS: System hardware includes two black-and-white CCD cameras, zoom lenses, and a PC equipped with a frame grabber. Custom software is used to acquire and archive video images, as well as to display real-time subtraction images revealing patient misalignment in multiple views. Two studies are described. In the first study, video is used to document the daily setup histories of 5 head and neck patients. Time-lapse cine loops are generated for each patient and used to diagnose and correct common setup errors. In the second study, 6 twice-daily (BID) head and neck patients are positioned according to the following protocol: at AM setups conventional treatment room lasers are used; at PM setups lasers are used initially and then video is used for 1-2 minutes to fine-tune the patient position. Lateral video images and lateral verification films are registered off-line to compare the distribution of setup errors per patient, with and without video assistance. RESULTS: In the first study, video images were used to determine the accuracy of our conventional head and neck setup technique, i.e., alignment of lightcast marks and surface anatomy to treatment room lasers and the light field. For this initial cohort of patients, errors ranged from sigma = 5 to 7 mm and were patient-specific. Time-lapse cine loops of the images revealed sources of the error, and as a result, our localization techniques and immobilization device were modified to improve setup accuracy. After the improvements, conventional setup errors were reduced to sigma = 3 to 5 mm. In the second study, when a stereo pair of live subtraction images were introduced to perform daily "on-line" setup correction, errors were reduced to sigma = 1 to 3 mm. Results depended on patient health and cooperation and the length of time spent fine-tuning the position. CONCLUSION: An interactive, video-based patient positioning system was shown to reduce setup errors to within 1 to 3 mm in head and neck patients, without a significant increase in overall treatment time or labor-intensive procedures. Unlike retrospective portal image analysis, use of two live-video images provides the therapists with immediate feedback and allows for true 3-D positioning and correction of out-of-plane rotation before radiation is delivered. With significant improvement in head and neck alignment and the elimination of setup errors greater than 3 to 5 mm, margins associated with treatment volumes potentially can be reduced, thereby decreasing normal tissue irradiation.

Adolescent sexuality and sexually transmitted diseases: attitudes, beliefs, knowledge, and values.

This study described rural adolescents' attitudes, beliefs, knowledge, and values with regard to sexuality and sexually transmitted diseases (STDs). Rotter's Social Learning Theory (1954) provided the theoretical framework for this descriptive, correlational design. The convenience sample consisted of 170 students from one rural high school. Consistent with past studies, results included the following: participants had more correct than incorrect knowledge related to sexual intercourse and STDs; the majority had positive attitudes toward condom use and believed it was OK for peers to have sex with a "steady;" the value of an exciting life correlated positively with attitudes toward sex; knowledge of sexual intercourse correlated positively with attitudes toward condom use; and the value health correlated positively with knowledge of sex and attitudes toward condom use, and negatively with attitudes toward sex. The findings in this study suggest the need for ongoing research with adolescents in the area of sexuality and STDs. Additionally, the findings support past studies, which revealed that knowledge of sexual intercourse and STDs has little impact on attitudes toward sexual intercourse. With the serious nature of some of the undesired consequences of adolescent sexual behavior, current and accurate information on this population is needed to assist health educators in developing interventions in this area.

PIP: This study examines adolescent attitudes, knowledge, and values on sexuality and sexually transmitted diseases (STDs). Using the Rotter's Social Learning Theory, 170 students were interviewed from one rural high school. Three hypotheses were analyzed in this study: 1) a higher value of an exciting life has a higher attitude toward sexual intercourse scores; 2) a higher knowledge score on sexual intercourse will have a lower attitude toward sexual intercourse scores; and 3) a higher knowledge score about AIDS/STDs will have a higher attitude toward condom use scores. Results revealed that 1) participants have higher accurate knowledge on sexual intercourse and STDs; 2) that a positive attitudes toward condom use and sexual intercourse with the girl/boyfriend is normal; and 3) that health values have a positive correlation between an exciting life and the knowledge and attitude about sex and condom use, and a negative correlation toward sex. Findings of minimal impact in knowledge of sexual intercourse and STDs on sexual behavior attitudes among adolescents are consistent with past studies.

Performance of a video-image-subtraction-based patient positioning system.

PURPOSE: We have developed and tested an interactive video system that utilizes image subtraction techniques to enable high precision patient repositioning using surface features. We report quantitative measurements of system performance characteristics. METHODS AND MATERIALS: Video images can provide a high precision, low cost measure of patient position. Image subtraction techniques enable one to incorporate detailed information contained in the image of a carefully verified reference position into real-time images. We have developed a system using video cameras providing orthogonal images of the treatment setup. The images are acquired, processed and viewed using an inexpensive frame grabber and a PC. The subtraction images provide the interactive guidance needed to quickly and accurately place a patient in the same position for each treatment session. We describe the design and implementation of our system, and its quantitative performance, using images both to measure changes in position, and to achieve accurate setup reproducibility. RESULTS: Under clinical conditions (60 cm field of view, 3.6 m object distance), the position of static, high contrast objects could be measured with a resolution of 0.04 mm (rms) in each of two dimensions. The two-dimensional position could be reproduced using the real-time image display with a resolution of 0.15 mm (rms). Two-dimensional measurement resolution of the head of a patient undergoing treatment for head and neck cancer was 0.1 mm (rms), using a lateral view, measuring the variation in position of the nose and the ear over the course of a single radiation treatment. Three-dimensional repositioning accuracy of the head of a healthy volunteer using orthogonal camera views was less than 0.7 mm (systematic error) with an rms variation of 1.2 mm. Setup adjustments based on the video images were typically performed within a few minutes. The higher precision achieved using the system to measure objects than to reposition them suggests that the variability in repositioning is dominated by the ability of the therapist to make small, controlled changes in the position of the patient. CONCLUSION: Using affordable, off-the-shelf technology, we have developed a patient positioning system that achieves repositioning accuracy normally associated with fractionated stereotactic systems. The technique provides real-time guidance and can be used to easily and quickly correct patient setup before every treatment, thus significantly reducing overall random positioning error. This improved positioning capability provides the precision required to realize the potential gains of conformal radiotherapy.

A diffusible cytotoxin of Haemophilus ducreyi.

Little is known about the virulence mechanisms employed by Haemophilus ducreyi in the production of genital ulcers. This Gram-negative bacterium previously has been shown to produce a soluble cytotoxic activity that kills HeLa and HEp-2 cells. We have now identified a cluster of three H. ducreyi genes that encode this cytotoxic activity. The predicted proteins encoded by these genes are most similar to the products of the Escherichia coli cdtABC genes that comprise the cytolethal distending toxin (CDT) of this enteric pathogen. Eleven of 12 H. ducreyi strains were shown to possess this gene cluster and culture supernatants from these strains readily killed HeLa cells. The culture supernatant from a single strain of H. ducreyi that lacked these genes was unable to kill HeLa cells. When the H. ducreyi cdtABC gene cluster was cloned into E. coli, culture supernatant from the recombinant E. coli clone killed HeLa cells. A monoclonal antibody that neutralized this soluble cytotoxic activity of H. ducreyi was shown to bind to the H. ducreyi cdtC gene product. This soluble H. ducreyi cytotoxin may play a role in the development or persistence of the ulcerative lesions characteristic of chancroid.

Beta-particle dosimetry in radiation synovectomy.

Beta-particle dosimetry of various radionuclides used in the treatment of rheumatoid arthritis was estimated using Monte Carlo radiation transport simulation coupled with experiments using reactor-produced radionuclides and radiachromic film dosimeters inserted into joint phantoms and the knees of cadavers. Results are presented as absorbed dose factors (cGy-cm2/MBq-s) versus depth in a mathematical model of the rheumatoid joint which includes regions of bone, articular cartilage, joint capsule, and tissue (synovium) found in all synovial joints. The factors can be used to estimate absorbed dose and dose rate distributions in treated joints. In particular, guidance is provided for those interested in (a) a given radionuclide's therapeutic range, (b) the amount of radioactivity to administer on a case-by-case basis, (c) the expected therapeutic dose to synovium, and (d) the radiation dose imparted to other, nontarget components in the joint, including bone and articular cartilage.

Mitogenic effects of propoxur on male rat bladder urothelium.

Propoxur produces bladder tumors in rats, but not other species. The hyperplastic and tumorigenic effects do not occur if urinary pH is lowered by administering propoxur in a semi-synthetic diet or co-administering it with ammonium chloride (NH4Cl). We fed propoxur at 8000 p.p.m. in Altromin 1321 diet to male Wistar rats for 4 weeks, with or without NH4Cl as 10,000 p.p.m. of the diet. The urine of rats fed control diet with or without propoxur had a relatively high urinary pH (approximately 8); the addition of NH4Cl lowered the urinary pH by approximately 0.5-1.0 units. There was no evidence of urinary calculi or amorphous precipitate nor was there an increase in microcrystals or formation of different crystals than occur in normal rat urine. Propoxur produced hyperplasia of the urothelium, as observed by light and scanning electron microscopy, and increased the labeling index for proliferating cell nuclear antigen. These effects were significantly inhibited by co-administration with NH4Cl. There was no evidence of urothelial necrosis. Thus, the hyperplasia appears to result from a direct mitogenic effect of propoxur or a metabolite on the urothelium, rather than from toxicity and consequent regeneration. Based on the present study and previous investigations, the urothelial effects of propoxur in the rat are dependent on high urinary pH and high administered doses, factors which need to be incorporated into any mechanistic model for the chemical and into any extrapolation to potential effects in humans.

A comparison of the effects of sodium saccharin in NBR rats and in intact and castrated male F344 rats.

High doses of sodium saccharin (NaSac) increase proliferation in the bladder of the rat, with a male preponderance. The possibility that alpha 2u-globulin is involved in its mechanism of action was evaluated by feeding it at 7.5% of the diet to NCI-Black-Reiter (NBR) male rats, which do not synthesize liver-derived alpha 2u-globulin. NaSac affected urinary parameters similarly in F344 and NBR male rats, but NBR rats consumed more water leading to greater urinary volume. NaSac produced less proliferation in NBR than in intact F344 rats, with intermediate changes in castrated F344 males, which had intermediate urinary alpha 2u-globulin levels.

Slowed receptor trafficking in mutant CHO lines of the End1 and End2 complementation groups.

A mutant Chinese hamster ovary cell line with nonconditional kinetic defects in receptor internalization and recycling was isolated, based on selection for resistance to a transferrin-diphtheria toxin conjugate and screening for aberrant receptor trafficking. The 12-4 cell line internalizes transferrin at approximately 75% of the parental rate and recycles transferrin back to the cell surface at approximately 55% of the parental rate. Internalization of low density lipoprotein is also reduced to approximately 70% of the parental cell rate, demonstrating that the mutant phenotype affects the trafficking of multiple receptors. Characterization of somatic cell hybrids indicated that the 12-4 phenotype is recessive, and complementation analysis determined that the 12-4 cell line is a member of the End2 complementation group. End2 mutants have previously been described as defective in endosomal acidification but have not been known to be defective in receptor trafficking. We have found similar defects in another End2 mutant cell line, suggesting that slowed receptor trafficking is characteristic of End2 mutants. Interestingly, transferrin receptor recycling and internalization are also slowed in another complementation group of mutants, End1, that is also defective in endosomal acidification. This study demonstrates altered receptor trafficking in End1 and End2 cell lines, a novel aspect of the mutant phenotypes. These findings provide evidence, based on a cellular genetic approach, that proper endosome acidification is necessary for maintenance of normal receptor recycling.