Diagnostic accuracy of an exhaled breath test for TB in hospitalized patients with cough or risk.

Closing the TB diagnostic gap is an urgent priority, for which non-sputum-based tests are needed. We evaluated the diagnostic accuracy of Aeonose, an exhaled breath test (EBT), as a TB triage test.Patients with cough or TB risk factors admitted to a tertiary hospital in Lima, Peru, were prospectively enrolled and underwent EBT. We evaluated EBT sensitivity and specificity for diagnosing pulmonary TB using culture and Xpert as primary and secondary reference standards and conducted stratified analyses based on risk factors.EBT sensitivity was 85% (95% CI 72.9-93.4), and specificity was 51% (95% CI 46.0-56.6) in the training cohort (n = 417). EBT sensitivity was 70% (95% CI 47.1-86.8), and specificity was 54% (95% CI 44.8-63.6) in the validation cohort (n = 139) using the culture reference standard, with higher sensitivity (78%) when using the Xpert reference standard (n = 156). Sensitivity (60%) and specificity (48%) were lower when patients with prior TB were included. In a subset of participants randomly selected for interviews, 94% (15/16) preferred EBT to sputum-based testing.EBT had moderate sensitivity and low specificity as a TB triage test in this hospitalised cohort with cough or risk factors. Diagnostic accuracy was lower in people with prior TB..

FAST implementation in Bangladesh: high frequency of unsuspected tuberculosis justifies challenges of scale-up.

SETTING: National Institute of Diseases of the Chest and Hospital, Dhaka; Bangladesh Institute of Research and Rehabilitation in Diabetes, Endocrine and Metabolic Disorders, Dhaka; and Chittagong Chest Disease Hospital, Chittagong, Bangladesh. OBJECTIVE: To present operational data and discuss the challenges of implementing FAST (Find cases Actively, Separate safely and Treat effectively) as a tuberculosis (TB) transmission control strategy. DESIGN: FAST was implemented sequentially at three hospitals. RESULTS: Using Xpert® MTB/RIF, 733/6028 (12.2%, 95%CI 11.4-13.0) patients were diagnosed with unsuspected TB. Patients with a history of TB who were admitted with other lung diseases had more than twice the odds of being diagnosed with unsuspected TB as those with no history of TB (OR 2.6, 95%CI 2.2-3.0, P < 0.001). Unsuspected multidrug-resistant TB (MDR-TB) was diagnosed in 89/1415 patients (6.3%, 95%CI 5.1-7.7). Patients with unsuspected TB had nearly five times the odds of being diagnosed with MDR-TB than those admitted with a known TB diagnosis (OR 4.9, 95%CI 3.1-7.6, P < 0.001). Implementation challenges include staff shortages, diagnostic failure, supply-chain issues and reliance on external funding. CONCLUSION: FAST implementation revealed a high frequency of unsuspected TB in hospitalized patients in Bangladesh. Patients with a previous history of TB have an increased risk of being diagnosed with unsuspected TB. Ensuring financial resources, stakeholder engagement and laboratory capacity are important for sustainability and scalability.

Multi-screen electronic alerts to augment venous thromboembolism prophylaxis.

Venous thromboembolism (VTE) prophylaxis in high-risk patients is frequently underutilised. We previously devised a one-screen computer alert program that identified hospitalised patients at high risk for VTE who were not receiving prophylaxis and advised their physicians to prescribe prophylaxis. While this strategy reduced the 90-day incidence of symptomatic VTE by 41%, the majority of electronic alerts were ignored. We have now developed a serial three-screen alert computer program designed to educate physicians who initially declined to order prophylaxis after a single screen alert. Of a total cohort of 880, the responsible physicians for 425 patients received a single electronic alert, whereas 455 who declined prophylaxis after the first screen received the second and third screens of the novel three-screen alert. Our enhanced serial three-screen alert program generated VTE prophylaxis orders for 58.4% of the 455 patients whose physicians initially declined to order prophylaxis following the one-screen alert. There was no significant difference in symptomatic 90-day VTE rates between the two cohorts (2.8% for the one-screen vs. 2.2% for the three-screen, p=0.55). We conclude that our three-screen computer alert program can markedly increase prophylaxis among physicians who decline an initial single screen alert.

Vitamin D-deficiency and post-fracture changes in lower extremity function and falls in women with hip fractures.

UNLABELLED: We determined the prevalence of vitamin D deficiency and lower extremity function in women with hip fractures. Women with extremely low vitamin D levels had reduced lower extremity muscle function and increased falls 1 year later. Ensuring vitamin D sufficiency after a hip fracture may improve function and reduce falls. INTRODUCTION: Hip fractures are the most devastating of fractures, commonly leading to loss of independent ambulation and living. In this retrospective analysis we determined the prevalence of vitamin D deficiency in women with hip fractures and the association between 25-hydroxyvitamin D [25(OH)D] levels and functional impairment one year later. METHODS: One hundred ten community-dwelling women with hip fractures were recruited from Boston, MA (n = 30) and Baltimore, MD (n = 80) before 1998 and 25(OH)D levels were measured by radioimmunoassay. In a subset of women from Baltimore, a performance measure of the lower extremities using the lower extremity gain scale (LEGS) was measured at 2, 6, and 12 months. Falls, grip strength, chair rise time, walking speed, and balance were also determined. RESULTS: Vitamin D insufficiency defined as a 25(OH)D 9 ng/mL, those with 25(OH)D

Quantification of ankle articular cartilage topography and thickness using a high resolution stereophotography system.

OBJECTIVE: To describe the topography and to measure thicknesses, surface areas and volumes in the cartilage layers of the ankle. METHODS: Twelve cadaveric ankle joints were disarticulated and the cartilage surfaces of each bone were imaged with a highly accurate (+/-2 microm) stereophotography system (ATOS). The cartilage was then dissolved and the subchondral bone imaged. The geometric data were then used to measure the quantitative parameters in each cartilage layer. RESULTS: The mean cartilage volume across the 12 specimens ranged from 0.32+/-0.08 ml for the fibula to 2.44+/-0.48 ml for the talus. The mean thickness of both the talar (1.1+/-0.18 mm) and tibial (1.16+/-0.14 mm) cartilage was significantly thicker than the fibula (0.85+/-0.13 mm). The talus had the greatest mean maximum cartilage thickness (2.38+/-0.4 mm). CONCLUSIONS: The reported stereophotographic technique may be used as an independent gold standard for validation of the accuracy of quantitative cartilage measurements made using magnetic resonance imaging. The thickness distribution maps show that the thickest articular cartilage occurs over the talar shoulders where osteochondral lesions commonly occur and not in the centre of the talar dome as commonly believed.

Tibial plafond fractures treated by articulated external fixation: a randomized trial of postoperative motion versus nonmotion.

OBJECTIVES: Assess whether postoperative ankle motion after fixation of a fracture of the tibial plafond, treated with articulated external fixation, leads to a better outcome when compared with similar treatment without postoperative ankle motion. DESIGN: Multicenter randomized trial. SETTING: Three Level I trauma centers. PATIENTS/PARTICIPANTS: Fifty-five patients were enrolled and entered into a Web-based database and randomized into 1 of 2 groups. Forty-one patients were evaluated at a 1-year follow-up visit, and 31 were seen at 2 years or longer after injury. INTERVENTION: Patients were treated with a hinged external fixator and limited internal fixation of the articular surface. They were divided postoperatively into two groups, 1 of which had a locked hinge and the other had a mobile hinge and a motion protocol. MAIN OUTCOME MEASUREMENTS: A general health status questionnaire, the SF-36 (short-form 36); a joint-specific ankle questionnaire, the Ankle Osteoarthritis Score (AOS); and range of motion (ROM) of the ankle joint. RESULTS: There were no significant differences between the two groups at either follow-up interval in the ankle ROM measurement, the AOS pain and disability scale, or the SF-36 physical component summary (PCS) and mental component summary (MCS) scales. CONCLUSIONS: These results indicate that treatment protocols that use long periods of cross-joint external fixation that immobilizes the ankle as definitive treatment result in similar patient outcomes compared to otherwise identical treatment protocols that incorporate and use an articulated hinge for ankle motion. However, the results should be interpreted with caution because the patient numbers were too small to detect potentially meaningful differences in outcomes and the follow-up was too short to assess for differences in the development of arthrosis.

Prognosis of status epilepticus: role of aetiology, age, and consciousness impairment at presentation.

BACKGROUND: Identification of outcome-predictive factors could lower risk of under- or over-treatment in status epilepticus (SE). Older age and acute symptomatic aetiology have been shown to predict mortality, but other variables are controversial and level of consciousness has received relatively little attention. The objective of this study was to assess variables predictive of mortality, particularly those available at presentation. METHODS: The discharge database (1997-2004) of two university hospitals was screened for adult patients with EEG confirmed SE, excluding cerebral anoxia. Outcome at discharge (mortality, return to baseline clinical conditions) was analysed in relation to demographics, clinical features, and aetiology. Aetiologies were also classified based on whether or not they were potentially fatal independently of SE. RESULTS: Mortality was 15.6% among 96 patients with a first SE episode, 10 of whom also experienced recurrent SE during the study period. Eleven other patients had only recurrent SE. Mortality was 4.8% among these 21 patients with recurrent SE. Return to baseline condition was more frequent after recurrent than incident SE (p=0.02). For the first SE episode, death was associated with potentially fatal aetiology (p=0.01), age>or=65 (p=0.02), and stupor or coma at presentation (p=0.04), but not with gender, history of epilepsy, SE type, or time to treatment>or=1 h. CONCLUSIONS: At initial evaluation, older age and marked impairment of consciousness are predictive of death. Surviving a first SE episode could lower the mortality and morbidity of subsequent episodes, suggesting that underlying aetiology, rather than SE per se, is the major determinant of outcome.

Retrospective analysis of malleolar fractures in an impact environment.

To protect against malleolar fractures in frontal crashes it is important to understand the mechanisms of injury. We have investigated the accuracy of Orthopaedic Specialists in deducing the injury mechanisms of experimentally generated malleolar fractures from radiographs; and the applicability of classic descriptors of injury mechanisms, such as the Lauge-Hansen classification system, in analysing impact induced trauma. Orthopaedic Specialists did not consistently deduce the mechanism of ankle injuries suggesting there may not be a unique fracture pattern for every injury mechanism and that the Lauge-Hansen classification system does not reliably describe ankle fractures created in the impact environment.

Role of individual predisposition in orthostatic intolerance before and after simulated microgravity.

Orthostatic intolerance (OI) is a major problem after spaceflight. Its etiology remains uncertain, but reports have pointed toward an individual susceptibility to OI. We hypothesized that individual predisposition plays an important role in post-bed rest OI. Twenty-four healthy male subjects were equilibrated on a constant diet, after which they underwent tilt-stand test (pre-TST). They then completed 14-16 days of head-down-tilt bed rest, and 14 of the subjects underwent repeat tilt-stand test (post-TST). During various phases, the following were performed: 24-h urine collections and hormonal measurements, plethysmography, and cardiovascular system identification (a noninvasive method to assess autonomic function and separately quantify parasympathetic and sympathetic responsiveness). Development of presyncope or syncope defined OI. During pre-TST, 11 subjects were intolerant and 13 were tolerant. At baseline, intolerant subjects had lower serum aldosterone (P < 0.01), higher excretion of potassium (P = 0.01), lower leg venous compliance (P = 0.03), higher supine parasympathetic responsiveness (P = 0.02), and lower standing sympathetic responsiveness (P = 0.048). Of the 14 subjects who completed post-TST, 9 were intolerant and 5 were tolerant. Intolerant subjects had lower baseline serum cortisol (P = 0.03) and a higher sodium level (P = 0.02) compared with tolerant subjects. Thus several physiological characteristics were associated with increased susceptibility to OI. We propose a new model for OI, whereby individuals with greater leg venous compliance recruit compensatory mechanisms (activation of the renin-angiotensin-aldosterone system and sympathetic nervous system, and withdrawal of the parasympathetic nervous system) in the face of daily postural challenges, which places them at an advantage to face orthostatic stress. With head-down-tilt bed rest, the stimulus to recruit compensatory mechanisms disappears, and differences between the two subgroups attenuate.

A randomized study comparing Crinone 8% and intramuscular progesterone supplementation in in vitro fertilization-embryo transfer cycles.

OBJECTIVE: To compare the efficacy of Crinone 8% intravaginal progesterone gel vs. IM progesterone for luteal phase and early pregnancy support after IVF-ET. DESIGN: Randomized, open-label study. SETTING: Academic medical center. PATIENT(S): Two hundred and one women undergoing IVF-ET. INTERVENTION(S): Women were randomized to supplementation with Crinone 8% (90 mg once daily) or IM progesterone (50 mg once daily) beginning the day after oocyte retrieval. MAIN OUTCOME MEASURE(S): Pregnancy, embryo implantation, and live birth rates. RESULT(S): The women randomized to luteal phase supplementation with IM progesterone had significantly higher clinical pregnancy (48.5% vs. 30.4%; odds ratio [OR], 2.16; 95% confidence interval [CI], 1.21, 3.87), embryo implantation (24.1% vs. 17.5%; OR, 1.89; 95% CI, 1.08, 3.30), and live birth rates (39.4% vs. 24.5%; OR, 2.00; 95% CI, 1.10, 3.70) than women randomized to Crinone 8%. CONCLUSION(S): In women undergoing IVF-ET, once-a-day progesterone supplementation with Crinone 8%, beginning the day after oocyte retrieval, resulted in significantly lower embryo implantation, clinical pregnancy, and live birth rates compared with women supplemented with IM progesterone.

Retrospective review of eighteen patients who underwent transtibial amputation for intractable pain.

BACKGROUND: Amputations are rarely performed solely for pain relief because of concerns regarding the persistence of pain and disability after the procedure. The purpose of this study was to assess the outcome of below-the-knee amputations performed to relieve intractable foot and ankle pain. METHODS: A chart review was conducted to identify all below-the-knee amputations that had been performed for the treatment of chronic foot and ankle pain by three orthopaedic foot and ankle specialists at three institutions. The inclusion criteria included (1) intractable foot or ankle pain as the surgical indication, (2) failure of maximal medical therapy, (3) failure of prior surgical reconstruction, and (4) a minimum follow-up period of twenty-four months after below-the-knee amputation. Patients with diabetes mellitus, peripheral vascular occlusive disease, or peripheral neuropathy were excluded. Each participant completed a two-part questionnaire with regard to the levels of disability, function, pain, and recreational activity both before and after the amputation. RESULTS: Twenty patients met the inclusion criteria, and eighteen completed the study. The study group included four women and fourteen men who had an average age of forty-two years (range, twenty-six to sixty-one years) and were followed for an average of forty-one months (range, twenty-five to eighty-five months) after the amputation. When asked whether they would have the below-the-knee amputation done again under similar circumstances, sixteen patients said yes, one was unsure, and one said no. The same distribution was observed when the patients were asked whether they were satisfied with the outcome: sixteen said yes, one was unsure, and one said no. Disability, pain, and recreational status were assessed with a 10-cm visual analog scale. After the amputation, the patients reported a decrease in both pain frequency (with the average score improving from 9.8 to 1.7; p < 0.0001) and pain intensity (with the average score improving from 8.4 to 2.6; p < 0.0001). Ten patients discontinued the use of narcotics, and seven decreased the level and/or dosage. Three patients worked before the amputation, and eight worked after the amputation. The average walking distance increased from 0.3 to 0.8 mile (p = 0.0034). CONCLUSION: In selected patients, a below-the-knee amputation may be a good salvage procedure for intractable foot and ankle pain that is unresponsive to all medical and local surgical reconstructive techniques.

Adrenal response to angiotensin II in black hypertension: lack of sexual dimorphism.

Adrenal responsiveness to angiotensin (Ang) II is markedly blunted in black hypertensive patients compared with white hypertensive patients. One characteristic of this blunted adrenal response in whites is a powerful sexual dimorphism: premenopausal white women rarely show blunted responses. This abnormality, most evident when the system is activated by a low-salt diet, is a cardinal feature of the syndrome of nonmodulation, affecting a large percentage of white hypertensive patients. Nonmodulation is also marked by an increase in cardiovascular risk beyond that from hypertension itself. This study investigated whether young black women are likewise spared its expression or whether the adrenal unresponsiveness common among black hypertensive patients is unaccompanied by a gender bias. We compared the adrenal response to Ang II in 382 hypertensive patients (313 white, 69 black; 238 male, 144 female). Ang II was infused when subjects were in balance on a 10-mmol Na(+) intake. As anticipated, white hypertensive patients showed a very strong sexual dimorphism, with women having twice the aldosterone response of men (P=0.0001). Blacks, on the other hand, showed no gender difference (P=0.9). Increasing age had the dramatic effect of reducing responsiveness in white women but not in blacks. Young black women demonstrated the same blunting of adrenal responsiveness as older black women and black men of all ages. Mechanisms protecting against a blunted adrenal response to Ang II in young white women are absent in blacks. These differences may contribute to the markedly increased prevalence of hypertension in young black women.

Expanding the orthopaedist's role in the treatment of foot and ankle disorders.

Many opportunities exist and more will become available for expanding the role of orthopaedic surgeons in the treatment of patients with foot and ankle conditions in the United States. The current authors present the main areas of opportunity: giving comprehensive foot care to patients who already are treated by orthopaedic surgeons and the application of advanced scientific biotechnology that will improve outcomes for patients with foot and ankle conditions who currently do not have satisfactory treatment. Orthopaedic surgeons must maintain a high level of commitment in their education and professional availability to patients and primary care physicians to expand their participation in foot and ankle care. Networking with physician and nonphysician primary care providers and volunteering educational services for fellow professionals in the evaluation and treatment of the entire spectrum of foot and ankle conditions is necessary. The general orthopaedist and the orthopaedic foot and ankle specialist can share in the future of treatment of patients with foot and ankle conditions in the United States.

Influence of sodium intake on the reliability of active renin as a measure of the renin-angiotensin system in essential hypertension.

Plasma renin activity (PRA), active renin (AR), prorenin, and angiotensinogen were assessed in 486 hypertensive and 175 normotensive subjects with a sodium intake of 10 or 200 mEq/d during supine and upright posture and after infusion of angiotensin II. PRA and AR levels were compared in hypertensive subjects in each condition. With low sodium intake, particularly while upright, there was a significant correlation between PRA and AR. In upright subjects with low sodium intake who had a PRA of 2.4 ng/mL per hour or less (1.85 nmol.L-1.h-1 or less), the correlation was also strong. With high sodium intake, the correlation was weaker. With intermediate sodium excretion, the correlation was intermediate. Prorenin was less predictive of PRA than was AR, and angiotensinogen had a marginal role. Using PRA during sodium restriction while upright as the standard for determining renin status, the precision of AR for predicting renin status was excellent. AR may be used for surrogate assessment of the renin-angiotensin system activity when the system is activated.

Adjunctive salvage therapy with inhaled aminoglycosides for patients with persistent smear-positive pulmonary tuberculosis.

A proportion of patients with drug-resistant and drug-susceptible tuberculosis (TB) have sputum that is smear and culture positive for Mycobacterium tuberculosis for a prolonged period of time, despite conventional therapy. Among such patients with refractory TB, an unblinded, observational study was undertaken that used conventional TB therapy and adjunctive aerosol aminoglycosides. Patients with persistent smear- and culture-positive sputum for M. tuberculosis (despite > or =2 months of optimal systemic therapy) were selected for adjunctive treatment via inhalation with aminoglycosides, and microbiological responses were monitored. Thirteen of 19 patients converted to smear negativity during the study: 6 of 7 with drug-susceptible TB and 7 of 12 with drug-resistant TB. Among patients with drug-susceptible TB, the median time to sputum conversion was 23 days, a shorter time than for a population of historical control patients. Recurrent infection was not observed. Adjunctive aerosol aminoglycosides may expedite sterilization of sputum among certain patients with refractory TB and diminish the risk of transmission.

Optimal antidepressant dosing. Practical framework for selection, titration, and duration of therapy.

Appropriate antidepressant dosing and trial duration are crucial for successful treatment of depression. Before prescribing an antidepressant, primary care physicians should take into account each patient's history, responses to previous antidepressants, depressive symptoms, coexisting illnesses, and current prescriptions. Physicians must be able to help patients manage side effects and know when to discontinue treatment, switch antidepressants, or refer patients to a psychiatrist.

Association of posterior tibial tendon abnormalities with abnormal signal intensity in the sinus tarsi on MR imaging.

OBJECTIVE: To evaluate the association of abnormal signal intensity within the sinus tarsi with abnormalities of the posterior tibial tendon (PTT) on MR imaging. DESIGN AND PATIENTS: Sinus tarsi abnormalities were identified on 30 ankle MR examinations in 29 patients. The PTT and anterior talofibular ligament were retrospectively analyzed for abnormalities in these same patients. RESULTS AND CONCLUSIONS: Tears of the anterior talofibular ligament were found in 13 of 30 (43%) ankles. PTT abnormalities (complete tear, partial tear or dislocation) were seen in 14 of 30 (47%) studies, and were distributed relatively equally between those patients with and without lateral ligament tears. Our results provide evidence of an association between abnormalities of the PTT and the sinus tarsi. The finding of abnormal signal intensity within the sinus tarsi on MR imaging should alert the radiologist to potential abnormalities of the PTT.

Radiographic fracture assessments: which ones can we reliably make?

OBJECTIVE: To identify the fracture characteristics that can be reliably assessed by analysis of plain radiographs of tibial plateau fractures. DESIGN: Radiographic review study. PARTICIPANTS: Five orthopaedic traumatologists served as observers. INTERVENTION: Observers made assessments based on the radiographs of fifty-six tibial plateau fractures. Precise definitions of the assessments to be made were agreed on by all observers. The tested assessments included raters' abilities to identify and locate fracture lines, identify the presence of fracture displacement and comminution, make quantitative measurements of displacement, and characterize qualitative features of fractures. For thirty-eight of the fractures that had a computed tomography (CT) scan available, assessments were repeated using both radiographs and CT scans. MAIN OUTCOME MEASURES: To characterize interobserver reliability, percentage agreement and kappa statistics were calculated for categorical variables, and intraclass correlation coefficients (ICC) were calculated for noncategorical variables. RESULTS: Reliability of the assessments varied widely. Determining the location of fracture lines had the greatest reliability, whereas the subjective assessments of fracture stability and energy showed the poorest reliability. Although the ICCs for quantitative measurements approached acceptable levels, the tolerance limits were extremely wide. The addition of a CT scan improved the reliability of most assessments, but not to a statistically significant degree. CONCLUSIONS: Many basic radiographic interpretations relied on in making treatment decisions are made variably by observers. Using experienced raters and precise definitions of fracture assessments does not guarantee a high level of agreement. Discrete assessments have higher interrater agreements than do more qualitative assessments. Quantitative measures have wide tolerance limits and, therefore, probably cannot be used reproducibly to classify fractures or make treatment decisions. We conclude the reliability of fracture classification is limited by raters' abilities to agree on basic radiographic assessments.

Alterations of cartilage and collagen expression during fracture healing in experimental diabetes.

We investigated alterations in the expression of mRNA for type II and type X collagen in fracture callus of experimentally induced diabetic animals compared with controls and performed radiographic, histological, immunocytochemical and biomechanical studies. Experimentally induced diabetic rats exhibited an alteration in the temporal expression of type II and type X collagen mRNA and a decrease in type X mRNA expression as compared to controls. Radiographs showed a more intense periosteal reaction and a more rapid reconstitution of cortices in control versus diabetic animals. Histologically there was a delay in chondrocyte maturation and hypertrophy seen in diabetics. Immunolocalization of type X collagen demonstrated a delay in type X collagen expression around the hypertrophic chondrocytes. Biomechanical analysis showed a decrease in the strength of healing fractures in diabetic animals. Fracture healing in diabetic patients is compromised and may lead to delays in bone union. Though the exact mechanisms are unknown, we present evidence of decreased mechanical strength of the fracture and suggest that associated changes in collagen expression and chondrocyte maturation are mechanisms leading to delayed healing in untreated and poorly controlled diabetes.

Use of enhanced green fluorescent protein to monitor retroviral-mediated gene therapy in human keratinocytes.

Keratinocytes have great promise as targets for gene therapy involving both skin as well as for systemic disorders due to their availability and potential long life span. Improvement of gene transfer into keratinocytes will be greatly facilitated by markers that will allow both rapid detection and efficient selection of transduced cells. For these purposes, a recombinant version of the Aequorea victoria green fluorescent protein that is enhanced for high-level expression in mammalian cells (EGFP) was placed into a replication-deficient retroviral vector. High-titer retrovirus was used to transduce both primary cultures of neonatal foreskin-derived human keratinocytes (HK) as well as the immortalized keratinocyte-derived cell line HaCaT. Both cell types stably expressed the EGFP, and this marker allowed rapid purification of transduced cells by fluorescence-activated cell sorting. EGFP expression was seen in HaCaT keratinocytes for at least 40 passages, and the presence of this construct did not effect cell growth, or apoptosis in response to UVB or etoposide. Transduced populations of HK were grafted into SCID mice, resulting in a functional epidermis. EGFP expression was readily seen in vivo by exposing the xenografts to an ultraviolet light source. These studies demonstrate the feasibility of using EGFP as a convenient and rapid marker to monitor keratinocyte gene transfer both in vitro and in vivo.