A phase I study of an adeno-associated virus-CFTR gene vector in adult CF patients with mild lung disease.
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Adolescent↗
Biomedical subjects
T Flotte
Publications and source records attributed to T Flotte.
Damage to hair follicles by normal-mode ruby laser pulses.
BACKGROUND: Although many temporary treatments exist for hirsutism and hypertrichosis, a practical and permanent hair removal treatment is needed. OBJECTIVE: Our purpose was to study the use of normal-mode ruby laser pulses (694 nm, 270 microseconds, 6 mm beam diameter) for hair follicle destruction by selective photothermolysis. METHODS: Histologically assessed damage in ex vivo black-haired dog skin after the use of different laser fluences was used to design a human study; 13 volunteers with brown or black hair were exposed to normal-mode ruby laser pulses at fluences of 30 to 60 J/cm2, delivered to both shaved and wax-epilated skin sites. An optical delivery device designed to maximize light delivery to the reticular dermis was used. Hair regrowth was assessed at 1, 3, and 6 months after exposure by counting terminal hairs. RESULTS: Fluence-dependent selective thermal injury to follicles was observed histologically. There was a significant delay in hair growth in all subjects at all laser-treated sites compared with the unexposed shaven and epilated control sites. At 6 months, there was significant hair loss only in the areas shaved before treatment at the highest fluence. At 6 months, four subjects had less than 50% regrowth, two of whom showed no change between 3 and 6 months. Transient pigmentary changes were observed; there was no scarring. CONCLUSION: Selective photothermolysis of hair follicles with the normal-mode ruby laser produces a growth delay consistent with induction of prolonged telogen with apparently permanent hair removal in some cases.
Animals↗
Flow cytometric DNA ploidy and cells phase fractions in recurrent human pituitary adenomas. A correlative study of flow cytometric analysis and the expression of proliferating cell nuclear antigen.
Flow cytometric analysis was applied to embedded tissue to measure the proliferative activity and the DNA ploidy of 16 recurrent and 17 nonrecurrent pituitary adenomas. The results were compared with data from a previous study which demonstrated that proliferating cell nuclear antigen (PCNA) labeling index was higher in recurrent adenomas than in nonrecurrent adenomas. Flow cytometric analysis as a tool for predicting aggressive behavior has been useful in a variety of human tumors; however, its prognostic value in pituitary adenomas is controversial. Therefore, we decided to explore the relationship of the results of flow cytometry and proliferating cell nuclear antigen labeling indices with the prognosis of pituitary adenomas. Three out of 16 recurrent adenomas and five out of 17 nonrecurrent adenomas demonstrated a DNA aneuploid pattern. All the nonfunctional recurrent adenomas had a diploid pattern, while only 40% of the functional recurrent adenomas had a diploid pattern. The GO/G1 phase fraction was higher in the recurrent adenomas, than in the nonrecurrent ones (p = 0.0005). In contrast, the S-phase fraction and the coefficient of variation were higher in the nonrecurrent adenomas (5.9 +/- 1.0%, 7.0 +/- 0.75, respectively) than in the recurrent ones (2.5 +/- 0.6%, 4.0 +/- 0.2%, respectively) (p = 0.003 and p = 0.001, respectively). The proliferating cell nuclear antigen labeling indices were higher in the recurrent adenomas (18.9 +/- 4.5%) than in the nonrecurrent adenomas (2.6 +/- 1.6%) (p = 0.003). The S-phase of flow cytometry correlated weakly with the proliferating cell nuclear antigen labeling indices when the recurrent and the nonrecurrent adenomas were considered as one group. (r = -0.356, p = 0.033). But no significant correlations were observed when the groups of recurrent (r = -0.311, p = 0.195) and nonrecurrent tumors (r = -0.019, p = 0.942) were compared separately. The results of flow cytometric analysis suggest that recurrent adenomas may have a higher proportion of cells in the presynthetic phase than the nonrecurrent adenomas. This study suggests that flow cytometric analysis is of limited value in predicting recurrence of pituitary adenomas.
Adenoma↗
Photodynamic therapy of nonmelanoma skin cancer with topical aminolevulinic acid: a clinical and histologic study.
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Administration, Cutaneous↗
In vivo uptake of liposomal benzoporphyrin derivative and photothrombosis in experimental corneal neovascularization.
BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) has been used successfully to occlude neovascularizations experimentally. We evaluated the vasoocclusive potential of benzoporphyrin derivative (BPD), a new photosensitizer currently in clinical trials. Since liposomally formulated BPD strongly binds to endogenous low density lipoproteins (LDL) after i.v. injection, LDL act as carrier to deliver BPD preferentially to proliferating endothelial cells. STUDY DESIGN/MATERIALS AND METHODS: Corneal neovascularizations in rabbits were used as model. Time-dependent uptake and retention of liposomal BPD were measured in vivo by monitoring the laser-induced fluorescence (LIF). Photothrombosis was induced using a dye laser emitting at 692 nm. RESULTS: A maximal BPD concentration was measured at 60-90 minutes postinjection determining the optimal time interval for treatment. Exposures as low as 10 J/cm2 allowed complete and irreversible neovascular occlusion as documented angiographically. Histology revealed selective endothelial damage, adjacent corneal stroma, or iris vessels, remained intact. Identical results were obtained using BPD directly complexed with LDL suggesting use of a LDL-mediated pathway. CONCLUSION: We suggest BPD-PDT for a selective treatment of neovascular diseases.
Animals↗
Acute and chronic response of articular cartilage to holmium:YAG laser irradiation.
A Ho:YAG (holmium-yttrium-aluminum-garnet) laser system operating at a wavelength of 2.1 microns has been introduced recently for use in arthroscopic surgery. The acceptability of this new tool will be determined not only by its ability to resect tissue, but also by its long-term effects on articular surfaces. Two studies were performed to evaluate the acute and chronic effects of the laser on cartilaginous tissue. The acute in vitro effects of 2.1-micron laser irradiation were evaluated, including measurement of ablation efficiency, ablation threshold, and thermal damage in meniscal and articular cartilage. To document the chronic effects on articular cartilage in vivo, a 10-week healing study was performed. Eight sheep weighing 30 to 40 kg underwent bilateral arthrotomy procedures. Multiple full-thickness and partial-thickness defects were created. Animals were euthanized at 0, 2, 4, and 10 weeks. The results of the healing study showed (1) no healing of full- or partial-thickness defects in hyaline cartilage at 10 weeks; (2) fibrocartilaginous granulation tissue filling full-thickness defects at 2 and 4 weeks that was no longer evident at 10 weeks; (3) chondrocyte necrosis extending to > 900 microns distal to ablation craters at 4 weeks, with no evidence of repair at later dates; (4) chondrocyte hyperplasia at the borders of the damage zone at 2 weeks but not at euthanization occurring at later periods.
Animals↗
Biodistribution and phototoxicity of 5-aminolevulinic acid-induced PpIX in an orthotopic rat bladder tumor model.
5-Aminolevulinic acid (ALA) is a precursor of heme biosynthesis. In the penultimate step of this biosynthesis, protoporphyrin IX (PpIX), an effective photosensitizer, is generated. In this study, the biodistribution and photodynamic effect of ALA-induced PpIX were investigated in an orthotopic rat bladder tumor model. A quantitative comparison of PpIX biodistribution by extraction and fluorescence spectroscopy following intravenous and intravesical administration of ALA was made. The tumor to normal bladder wall ratio was 2:1 at 4 hours for both delivery modes. Fluorescence microscopy demonstrated predominantly cellular rather than stromal PpIX fluorescence. Phototoxicity, evaluated 4 hours after ALA administration, was light dose-dependent with the most efficient tumor necrosis being observed upon 150 J/cm.2 of 630 nm. irradiation. These data suggest that optimized photodynamic therapy with ALA-induced PpIX may be an alternative treatment for superficial bladder carcinoma.
Aminolevulinic Acid↗
Both CFTR and outwardly rectifying chloride channels contribute to cAMP-stimulated whole cell chloride currents.
From whole cell patch-clamp recordings at 35 degrees C utilizing either nystatin perforation or conventional methods with 5 mM MgATP in the pipette solution, it was demonstrated that both cystic fibrosis transmembrane conductance regulator (CFTR) chloride (Cl-) channels and outwardly rectifying Cl- channels (ORCC) contribute to adenosine 3',5'-cyclic monophosphate (cAMP)-activated whole cell Cl- currents in cultured human airway epithelial cells. These results were similar whether recordings were performed on two normal human cell lines or on two cystic fibrosis (CF) cell lines stably complemented with wild-type CF gene. These results were obtained by exploiting dissimilar biophysical properties of CFTR and ORCC currents such as the degree of rectification of the current-voltage relationship, the difference in sensitivity to Cl- channel-blocking drugs such as 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), calixarenes, and diphenylamine carboxylic acid (DPC), and the opposing Cl- relative to I- permeabilities of the two channels. In normal cells or complemented CF cells, 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate stimulated outwardly rectifying whole cell Cl- currents. Addition of DIDS in the presence of cAMP inhibited the outwardly rectifying portion of the cAMP-activated Cl- current. The remaining cAMP-activated, DIDS-insensitive, linear CFTR Cl- current was inhibited completely by DPC. Additional results showed that not only do ORCC and CFTR Cl- channels contribute to cAMP-activated Cl- currents in airway epithelial cells where wild-type CFTR is expressed but that both channels fail to respond to cAMP in delta F508-CFTR-containing CF airway cells. We conclude that CFTR not only functions as a cAMP-regulated Cl- channel in airway epithelial cells but also controls the regulation of ORCC.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
[Selective vascular occlusion by repetitive short laser pulse].
The occlusion of arterioles and venules (30 microns in diameter) by means of repeated dye laser pulses (577 nm, 160 microseconds, 0.5 Hz) was studied in the hamster cheek pouch model. Microscopically visible changes during and after irradiation were recorded and monitored by a video system. The energy necessary per pulse for vessel occlusion with a single pulse (arterioles 5 J/cm2, venules 3.5 J/cm2) can be lowered more than 50% with 100 pulses (1.8 J/cm2, 1.2 J/cm2). Rupture of vessels, which regularly occurs with a single pulse, can be totally avoided with 100 repetitive laser pulses. Investigations of the temperature dependence of the occlusion rate of arterioles showed different interaction mechanisms with one pulse and 100 pulses (mechanical and thermal respectively). Light and electron microscopic investigations supported this concept of selective occlusion using repeated short laser pulses. Possible clinical applications are portwine stains of the eyelid, neovascularisation on the cornea and subretinal neovascularisation.
Animals↗
[Photodynamic therapy of experimental, intraocular tumors with benzoporphyrin-lipoprotein].
Photodynamic therapy (PDT) with its potential for precise localization and absence of severe side effects such as radiation retinopathy may be particularly appropriate for the treatment of intraocular tumors. Benzoporphyrin (BPD), a potent photosensitizer currently in clinical trial, absorbs light at 692 nm, thus allowing sufficient tissue penetration due to minor light absorption in melanin and hemoglobin. The efficiency and selectivity of BPD are significantly pronounced through preassociation with low-density lipoprotein (LDL), since proliferating cells exhibit an increased metabolism of lipoproteins. As an experimental model Greene's melanomas were implanted either into the iris or choroid of albino rabbits. Irradiation at a radiation energy of 80 for iris and 100 J/cm2 for choroidal tumors 3 h after the i.v. injection of BPD-LDL (2 mg/kg) was administered via a laser arrangement with argon-pumped dye-laser, using the slit-lamp. Angiographies and LM/EM histologies were done immediately, and 1, 3, and 14-21 days post-exposure. All 16 treated tumors demonstrated complete regression with a remaining avascular, fibrotic scar. Immediate vascular occlusion within the tumor was seen angiographically, suggesting a direct vascular mechanism. Histologically, two primary mechanisms could be detected: destruction of neovascular endothelial cells and intracellular tumor cell damage. These results indicate that PDT using BPD-LDL complexes may provide an efficient and selective modality for the management of intraocular neoplasms. The availability of new and potent photosensitizers may also lead to broader clinical applications.
Animals↗
Microphotocoagulation: selective effects of repetitive short laser pulses.
Repetitive exposure to short laser pulses is shown to cause selective damage to absorbing structures (cells, organelles, or enzymes) with pulse energies below the threshold energy for single-pulse damage. Directly adjacent structures are spared in vivo. Additivity of (presumably nonphotochemical) subthreshold effects is demonstrated. Selective damage to the retinal pigment epithelium with sparing of the neural retina is shown (514 nm, 5 microseconds, 1-500 pulses at 500 Hz, 2- to 10-microJ pulse energy). A melanin granule model has been developed and applied to the experimental situation. Histological results as well as the basic mechanism for these effects are discussed.
Animals↗
Injury and adhesion formation following ovarian wedge resection with different thermal surgical modalities.
The purpose of this study is to determine the role of bleeding, acute thermal damage, and charring in adhesion formation. Postoperative adhesions were compared following ovarian wedge resection in 48 rabbits using different lasers, electrosurgery, and scalpel. Twelve ovaries were sectioned per modality, in randomized pairs. Acute thermal injury as assessed by histology, bleeding, and charring differed among the modalities used. Adhesions were assessed 4 weeks later, by an investigator completely blinded of the treatment protocol. The adhesion scores were 11.6 +/- 8.0 with pulsed Er:YAG laser; 11.9 +/- 7.5 with scalpel; 8.3 +/- 9.3 with electrocautery; 6.7 +/- 8.8 with a continuous (c.w.) Nd:YAG laser; 5.3 +/- 4.8 with c.w. CO2 laser; 3.1 +/- 2.7 with pulsed CO2 laser; 1.7 +/- 1.8 with pulsed Ho:YAG laser; and 0.8 +/- 1.5 in the control (no resection) group. Ho:YAG, Nd:YAG, and electrocautery were completely hemostatic. Bleeding was minimal with the CO2 lasers. Er:YAG and scalpel caused maximum bleeding, requiring hemostatic measures to prevent exsanguination. Charring occurred with electrocautery, CO2 laser, and Nd:YAG laser. Bleeding and charring correlated with adhesion formation, but the histological depth of thermal damage did not. The Ho:YAG laser is a hemostatic, fiber-optic compatible laser causing significantly fewer adhesions (P < 0.04) than scalpel, electrocautery, Nd:YAG, Er:YAG, and c.w. CO2 lasers. Clinical use of the Ho:YAG laser, and the role of carbonization in promoting adhesions, deserve further study.
Adnexal Diseases↗
Effects of systemic indomethacin, meclizine, and BW755C on chronic ultraviolet B-induced effects in hairless mouse skin.
Chronic exposure of hairless mice to ultraviolet B (UVB) radiation is associated with inflammation as well as an altered macromolecular composition of the dermis. This study was designed to determine whether or not various systemic anti-inflammatory agents inhibit chronic UVB-induced changes in the macromolecular content of the dermis and, if so, whether each agent had the same or different effects. The agents and doses were chosen for their ability to inhibit the changes induced by a single exposure to UVB radiation (increased vasopermeability, neutrophil accumulation, and skin-fold thickness). Indomethacin, a cyclooxygenase inhibitor, and meclizine, an H1 histamine receptor antagonist, were administered from slow-release pellets. BW755C, a combined cyclooxygenase and lipoxygenase inhibitor, was administered intraperitoneally 30 min prior to UVB exposure. Animals were exposed to UVB three times per week for 20-26 weeks or were unirradiated. The elastin, glycosaminoglycan and collagen content of the skin were determined by measuring the desmosine, uronic acid, and hydroxyproline levels, respectively. The amount of each macromolecule per area of skin increased after chronic UVB exposure. The increase in desmosine was inhibited by indomethacin; the increase in hydroxyproline was inhibited by meclizine and BW755C. None of the agents inhibited the uronic acid increase. These results suggest that chronic inflammation contributes to the dermal changes seen in chronically UVB-exposed skin and that different inflammatory mediators are involved in the increases observed in elastin, glycosaminoglycans, and collagen.
4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗
[Histology of retinal lesions after continuous irradiation and selective micro-coagulation of the retinal pigment epithelium].
Mild continuous wave (CW) irradiation (100 ms, 20 mW, 514 nm) and irradiation with 100 repetitive 5 microseconds laser pulses (3 or 6 microJ, 514 nm) at a repetition rate of 500 Hz was performed to the regio macularis of chinchilla rabbits. The angiographically visible lesions were histologically followed up to 4 weeks. With both irradiation modalities the original retinal pigment epithelium (RPE) was replaced by a monolayer of new RPE cells. Only minimal immediate and no subsequent damage to the photoreceptors was found after selective RPE photocoagulation. Only minimal inflammatory response was found after selective RPE photocoagulation in contrast to CW photocoagulation where macrophages, RPE cells and lymphocytes regularly appear in the damaged photoreceptor layer.
Animals↗
Defective regulation of outwardly rectifying Cl- channels by protein kinase A corrected by insertion of CFTR.
Cystic fibrosis (CF) is a lethal genetic disease resulting in a reduced Cl- permeability, increased mucous sulphation, increased Na+ absorption and defective acidification of lysosomal vesicles. The CF gene encodes a protein (the cystic fibrosis transmembrane conductance regulator, CFTR) that can function as a low-conductance Cl- channel with a linear current-voltage relationship whose regulation is defective in CF patients. Larger conductance, outwardly rectifying Cl- channels are also defective in CF and fail to activate when exposed either to cyclic AMP-dependent protein kinase A or to protein kinase C. The role of the outwardly rectifying Cl- channel in CF has been questioned. We report here that expression of recombinant CF genes using adeno-associated virus vectors in CF bronchial epithelial cells corrects defective Cl- secretion, that it induces the appearance of small, linear conductance Cl- channels, and restores protein kinase A activation of outwardly rectifying Cl- channels. These results re-establish an involvement of outwardly rectifying Cl- channels in CF and suggest that CFTR regulates more than one conductance pathway in airway tissues.
Adenosine Triphosphate↗
Dye-enhanced laser welding for skin closure.
The use of a laser to weld tissue in combination with a topical photosensitizing dye permits selective delivery of energy to the target tissue. A combination of indocyanine green (IG), absorption peak 780 nm, and the near-infrared (IR) alexandrite laser was studied with albino guinea pig skin. IG was shown to bind to the outer 25 microns of guinea pig dermis and appeared to be bound to collagen. The optical transmittance of full-thickness guinea pig skin in the near IR was 40% indicating that the alexandrite laser should provide adequate tissue penetration. Laser "welding" of skin in vivo was achieved at various concentrations of IG from 0.03 to 3 mg/cc using the alexandrite at 780 nm, 250-microseconds pulse duration, 8 Hz, and a 4-mm spot size. A spectrum of welds was obtained from 1- to 20-W/cm2 average irradiance. Weak welds occurred with no thermal damage obtained at lower irradiances: stronger welds with thermal damage confined to the weld site occurred at higher irradiances. At still higher irradiances, local vaporization occurred with failure to "weld." Thus, there was an optimal range of irradiances for "welding," which varied inversely with dye concentration. Histology confirmed the thermal damage results that were evident clinically. IG dye-enhanced laser welding is possible in skin and with further optimization may have practical application.
Animals↗
Endometrial ablation by means of photodynamic therapy with photofrin II.
OBJECTIVE: Photodynamic therapy is a technique in which tissue is irradiated with light after the use of a photosensitizing drug that produces singlet oxygen, which has a cytotoxic effect. The feasibility of using photodynamic therapy with photofrin II for endometrial ablation was studied. STUDY DESIGN: Fifty-eight rabbits were studied. Preferential uptake of photofrin II by endometrial tissue, compared with the myometrium, was established by drug extraction and fluorescence microscopy after administration of photofrin II intravenously. Dosimetry for endometrial ablation was established by administering photofrin II in 1, 2, 5, and 10 mg/kg doses and laser light (630 nm) at radiant exposures of 100 and 200 J/cm2. Histologic examination was performed at 24 hours, 5 days, and 10 days after treatment. There were two control groups. One group received laser light but no photofrin II, and the other received photofrin II without laser light. RESULTS: The concentration of photofrin II was three times higher in the endometrium than in the myometrium at doses of 1 and 2 mg/kg. Fluorescence microscopy of frozen sections of endometrium and myometrium showed a predominantly perivascular fluorescence from photofrin II. A dose of 1 and 2 mg/kg and a flow of 100 J/cm2 was adequate for endometrial ablation in rabbits. At 24 hours after treatment there was extensive hemorrhage and evidence of cell death in the entire endometrium and mild hemorrhage in 10% to 50% of the inner circular layer of the myometrium. At 5 days after treatment necrosis of the entire endometrium and the inner half of the myometrium was seen, but the outer half of the myometrium and the serosa were normal. There were no cases of uterine perforation. Similar results were seen at 10 days after treatment, except for the additional presence of inflammatory cells. Neither control group (drug without light, light without drug) showed any injury to the endometrium at 24 hours. CONCLUSION: We conclude that endometrial ablation can be effectively achieved in rabbits by means of photodynamic therapy with photofrin II without significant complications.
Animals↗
Optical coherence tomography.
A technique called optical coherence tomography (OCT) has been developed for noninvasive cross-sectional imaging in biological systems. OCT uses low-coherence interferometry to produce a two-dimensional image of optical scattering from internal tissue microstructures in a way that is analogous to ultrasonic pulse-echo imaging. OCT has longitudinal and lateral spatial resolutions of a few micrometers and can detect reflected signals as small as approximately 10(-10) of the incident optical power. Tomographic imaging is demonstrated in vitro in the peripapillary area of the retina and in the coronary artery, two clinically relevant examples that are representative of transparent and turbid media, respectively.
Coronary Disease↗