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Objective: Experiments were undertaken to compare morbidity and mortality from brief inhalation exposures to high levels of hydrogen fluoride (HF) and carbonyl fluoride (COF2).Methods: Rats from both sexes were exposed for durations of 5 and 10 min to nominal concentrations of 10,000 to 57,000 ppm HF or 500 to 10,000 ppm COF2. Respiration was monitored before, during, and after exposure. Animals were observed up to 6 days post-exposure. Terminal blood samples were collected for routine clinical chemistry and hematology. Post-mortem lung fluoride concentrations and lung weights were measured, and gross pathology noted.Results: Both gases produced respiratory depression independent of concentration or exposure duration with minute ventilation decreasing to approximately 50% of baseline. Estimated mixed-gender HF and COF2 10-min LC50’s were 48,661 ppm and 1083 ppm, respectively. HF mortalities were generally delayed 3 to 4 days post-exposure, while COF2 mortalities occurred during or briefly after ex...

Purpose: To report on the presentation, radiography, histology, and treatment of 8 cases of extranodal Rosai–Dorfman disease involving the orbit. Methods: Multicenter retrospective case series. Results: Five males and 3 females had a median age of 10 years (range 2–78 years). Presenting signs and symptoms included proptosis, periorbital pain, palpable mass, blepharoptosis, decreased vision, diplopia, impaired extraocular motility, and afferent pupillary defect. Four patients had bilateral orbital disease, while 4 had unilateral disease. Six cases were extraconal, 1 was intraconal, and 1 was both intra- and extra-conal. Four cases had only extranodal disease without lymphadenopathy (3 of which had localized orbital disease). Diagnosis was confirmed by exam, orbital, and/or systemic radiography, and biopsy in all cases. Treatment strategies included excision or debulking, systemic corticosteroids, chemotherapy, radiotherapy, observation or a combination thereof. At last follow up, 4 patients were disease free, while 4 had residual improved disease. Conclusions: Rosai–Dorfman disease of the orbit is a rare clinical entity. Purely extranodal disease is rare, with isolated orbital disease being exceedingly rare. This study is unique in that 4 of 8 patients had strictly isolated extranodal disease of the orbit. A large majority of the cases had disease in the extraconal space, contrasting with previous reports. In addition, lacrimal gland disease, particularly bilateral involvement, was prominent in the current study. Although there is no consensus on treatment, surgical excision should be attempted if plausible in symptomatic patients especially if the orbit represents a localized site of disease.

Improving the provision of spiritual care to hospitalized patients requires understanding what patients look for from a hospital chaplain, and why. This qualitative study uses grounded theory methodology to analyze data from 25 interviews with adult patients and/or adult family members who received spiritual care in a large tertiary care hospital. Analysis reveals three key themes in chaplaincy care: the attributes valued in the chaplain's presence, the elements necessary to form relationship with the chaplain, and the role of the chaplain in helping patients to discover and express meaning in their experiences. The authors weave these three themes together into a grounded theory and propose an assessment model that incorporates psychological theory about human motivation, faith development, and the development of autonomy. An understanding of the proposed assessment model can guide chaplain interventions and benefit all members of the clinical care team.

Historically the US Army used Halon 1301 (bromotrifluoromethane), a chemical with high ozone depletion potential (ODP), to protect the crews of armored vehicles from the effects of peacetime and combat fires. Since the phase-out of Halon production the US Army has directed that zero ODP materials be used wherever possible. Subsequently, major new vehicle platforms have been deployed with automatic fire extinguishing systems (AFES) that rely on HFC-227BC, a zero-ODP-agent blend of HFC-227ea (heptafluoropropane) and sodium-bicarbonate-based dry chemical. Unfortunately, hydrofluorocarbons (HFCs) generally have high global warming potential (GWP)—thousands of times that of carbon dioxide on a weight basis. Hence, as part of a larger effort to reduce its carbon footprint, US Army Program Managers have asked that fire extinguishing agents that are more environmentally friendly be evaluated as part of ongoing vehicle modernization efforts. Several agents were investigated, including FK-5-1-12, water with additives, and dry chemicals. This report describes the findings of more than 150 live-fire tests using nine agents and four extinguisher technologies. The basic conclusion is that no alternate agent can yet be considered to be a practical replacement for Halon 1301 or HFC-227BC for this application. However, a blend of Halon 1301 and dry chemical has been found to be about twice as effective as Halon 1301 alone. Thus, pending confirmation tests on vehicles, it may be feasible to use less Halon in legacy systems without compromising fire protection performance.

To study the cytotoxic effects and related signaling pathways of butein on human uveal melanoma cells in vitro.Three human uveal melanoma cell lines (M17, SP6.5, and C918), retinal pigment epithelial (RPE) cells and scleral fibroblasts were treated with butein at different dosages. The effects of butein on cell viability were assessed by using the MTT assay. Cell apoptosis was determined using annexin V-FITC/ethidium homodimer III flow cytometry. Mitochondrial transmembrane potential changes were assessed by using the JC-1 fluorescent reader, cytosol cytochrome c levels, and the activities of caspase-3, -8, and -9 were measured by using an enzyme-linked immunosorbent assay or colorimetric assay.Butein reduced the cell viability of cultured human uveal melanoma cells in a dose-dependent manner (10, 30, and 100 μM), with IC50 at 13.3 μM and 15.8 μM in SP6.5 and M17 cell lines, respectively. Similar effects were also found in a highly aggressive and metastatic C918 cell line (IC50 16.7 μM). Butein at lower concentrations (10-30 μM) selectively reduced the cell viability of uveal melanoma cells, without affecting cell viability of RPE cells and fibroblasts. Butein-induced apoptosis of melanoma cells, increased mitochondrial permeability and the level of cytosol cytochrome c, caspase-9 and -3 activities (but not caspase-8) in a dose-dependent manner.Butein has selectively potent pro-apoptotic effects on cultured human uveal melanoma cells via the intrinsic mitochondrial pathway.

Aims: To describe 2 cases of diffuse diabetic macular edema (DME) after diabetic vitrectomy caused by a taut internal limiting membrane (ILM), with clinicopathological correlation. Methods: Interventional case series with immunohistochemical analysis. Results: Two patients were referred for unresponsive diffuse DME after pars plana vitrectomy with removal of the posterior hyaloid. Clinically, a taut ILM was noted over the fovea, and its removal resulted in rapid and long-term resolution of the edema, confirmed by optical coherence tomography with visual acuity improvement. Histopathology with immunostaining revealed a segment of ILM with an inner monolayer of cytokeratin-positive (retinal pigment epithelial cells) and/or glial fibrillary acidic protein-positive cells with smooth muscle actin immunoreactivity. Conclusions: A taut ILM can cause diffuse DME after vitrectomy, and its removal can restore the normal foveal contour and improve visual acuity. Tangential tractional forces from contractile cells propagated across the fovea via the ILM appear to be the etiology.

Purpose To describe 4 patients with histopathologically confirmed tarsal conjunctival nevi and to review the literature regarding nevi in this location. Methods Clinicopathologic series of 4 cases diagnosed at the New York Eye and Ear Infirmary between January 1990 and April 2009. PubMed was searched for reported cases of tarsal conjunctival nevi. Results Four patients (mean age 44 years, median 50 years, range 5-70 years) with tarsal conjunctival nevi were identified (3.4% of all histopathologically diagnosed conjunctival nevi). These lesions presented in adulthood (3 cases) and were associated with elevation/nodularity (3 cases), gray-brown coloration (3 cases), physician-documented growth (1 case), madarosis (1 case), and intrinsic vessels (1 case). Histopathology revealed combined nevi centered in the posterior lamella of the eyelid, predominantly in peri-Meibomian gland distribution. Review of the literature identified 10 previously reported cases of tarsal nevi with detailed clinicopathologic correlation in 3 of these patients. Conclusions Tarsal conjunctival nevi are rare. Adult presentation; subjective and objective evidence of growth; and associated inflammation, vascularity, and eyelid margin changes may make clinical distinction of tarsal nevi from melanoma challenging. Biopsy, therefore, is prudent as histopathology alone allows documentation of the benign nature of these lesions.

Within inflammatory environments, B cells encountering foreign or self-Ag can develop tertiary lymphoid tissue expressing activation-induced cytosine deaminase (AID). Recently, this DNA-modifying enzyme was detected in nonlymphoid cells within several inflamed tissues and strongly implicated in malignant transformation. This study examines whether a cyclooxygenase 2 (COX-2) pathway, often linked to inflammation, influences AID expression in activated B lymphocytes. In this paper, we report that dividing human B cells responding to surrogate C3d-coated Ag, IL-4, and BAFF express AID, as well as COX-2. A progressive increase in AID with each division was paralleled by a division-related increase in a COX-2–linked enzyme, microsomal PGE2 synthase-1, and the PGE2R, EP2. Cells with the greatest expression of AID expressed the highest levels of EP2. Although COX-2 inhibitors diminished both AID expression and IgG class switching, exogenous PGE2 and butaprost, a selective EP2 agonist, augmented AID mRNA/protein and increased the numbers of IgG+ progeny. Despite the latter, the proportion of IgG+ cells within viable progeny generally declined with PGE2 supplementation. This was not due to PGE2-promoted differentiation to plasma cells or to greater downstream switching. Rather, because phosphorylated ataxia telangiectasia mutated levels were increased in progeny of PGE2-supplemented cultures, it appears more likely that PGE2 facilitates AID-dependent DNA double-strand breaks that block B cell cycle progression or promote activation-induced cell death, or both. Taken together, the results suggest that a PGE2 feed-forward mechanism for augmenting COX-2 pathway proteins promotes progressively increased levels of AID mRNA, protein, and function.

To study the cytotoxic effects and related signaling pathways of curcumin on human uveal melanoma cells in vitro.Two human uveal melanoma cell lines (M21 and SP6.5), scleral fibroblasts, and choroidal melanocytes were treated with curcumin. The effects of curcumin on cell viability were assessed by using the MTT (3-[4,5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium bromide) assay. Cytosol cytochrome c levels and the activities of caspase-9 and caspase-3 were measured by using an enzyme-linked immunosorbent assay.Curcumin induced cell death of cultured human uveal melanoma cells in a dose-dependent manner (10, 30, and 100 microM) and time-dependent manner (3-48 hr), with IC50 at 19.05 microM and 22.39 microM in M21 and SP6.5 cell lines, respectively. Curcumin at lower concentrations (10-30 microM) selectively reduced the cell viability of uveal melanoma cells, without affecting cell viability of fibroblasts and choroidal melanocytes. Curcumin significantly increased the level of cytosol cytochrome c (2-fold greater than the controls after 2 hr treatment), caspase-9 and caspase-3 activities (approximately 4.5- and 6-fold greater than the controls after 2-6 hr treatment, respectively) in a dose-dependent manner.Curcumin has selectively potent cytotoxic effects on cultured human uveal melanoma cells. This effect is associated with the release of cytochrome c from the mitochondria and the activation of caspase-9 and caspase-3 in uveal melanoma cells after treatment with curcumin.

Levels of eumelanin (EM) and pheomelanin (PM) of uveal melanoma cells have not been measured and compared with those of normal uveal melanocytes. EM and PM amounts in four immortal human uveal melanoma cell lines were measured by chemical degradation and microanalytical high-performance liquid chromatography and compared with those from 39 normal human uveal melanocyte cell lines reported earlier by us. Uveal melanoma cells had a very low EM/PM ratio (0.41), which was very significantly lower than that from normal melanocytes isolated both from eyes with light-colored irides (1.31) or dark-colored irides (7.32). The low EM/PM ratio was caused by a low level of EM in melanoma cells, which was only 1/8 and 1/31 of that in melanocytes from eyes with light-colored irides and dark-colored irides, respectively. The PM level in uveal melanoma cells was not statistically different from normal melanocytes from eyes with light-colored irides or dark-colored irides. The total quantity of EM and PM in uveal melanoma cells was significantly less than that in normal melanocytes. This difference was because of the low level of EM in uveal melanoma cells. The results of these studies indicate that the changes of melanin content in uveal melanoma cells mainly relate to the decrease of EM content. Low melanin and EM content may make melanoma cells more susceptible to mutagenic effects of ultraviolet radiation and oxidative stress, which may enhance the proliferation of melanoma cells and accelerate progression of melanoma.

Purpose To study the efficacy of oral montelukast, a cysteinyl leukotriene receptor antagonist, in combination with cetirizine, a histamine-1 receptor antagonist, in the treatment of thyroid eye disease. Methods Patients considering surgical correction of eyelid retraction for inflammatory symptoms of thyroid eye disease were offered a preoperative medical regimen of oral montelukast/cetirizine. Exclusion criteria included prior use of oral montelukast (i.e., for seasonal allergy or asthma), compressive optic neuropathy, severe ophthalmopathy requiring systemic corticosteroids, and orbital and/or muscle surgery. A 6-week course of oral cetirizine (10 mg every morning) and oral montelukast (10 mg every evening) was administered and patients subjectively rated their ocular surface dryness, tearing, itching, injection, eyelid swelling, eyelid retraction, double vision, proptosis, and visual clarity, at baseline, after 3 weeks and 6 weeks of medical therapy, and after 3 weeks off of the medications. Results Six of the 12 patients recruited for the study reported a subjective improvement in tearing, dryness, and itching. Less effect on diplopia and proptosis was noted after 6 weeks of medical therapy. Two of the patients who did not report response chose to proceed with eyelid retraction surgery and both had evidence of mast cell infiltration in their Muller muscle specimens. Conclusion The response observed in this open-label trial suggests that oral montelukast and cetirizine may be an effective medical regimen for patients with thyroid eye disease who experience mild to moderate orbital congestion and inflammation.

Variance in iris color is related to the incidence of several important ocular diseases, including uveal melanoma and age-related macular degeneration. The purposes of this study were to determine the quantity and the types of melanin in cultured human uveal melanocytes in relation to the iris color. Sixty-one cell cultures of pure uveal melanocytes were isolated from donor eyes with various iris colors. The amount of eumelanin (EM) and pheomelanin (PM) of these cells was measured by chemical degradation and microanalytical high-performance liquid chromatography (HPLC) methods. The total amount of melanin was measured by both microanalytical methods and spectrophotometry. Total melanin content, measured by HPLC and spectrophotometry, correlated well with r = 0.872 (P 0.05). When cells became senescent, the levels of EM, PM and total melanin were significantly increased. In both growing and senescent melanocytes, the quantity and type of melanin were closely correlated to the iris color. In cells from eyes with dark-colored irides (dark brown and brown), the amount of EM, the ratio of EM/PM and total melanin were significantly greater than that from eyes with light-colored irides (hazel, green, yellow-brown and blue) (P 0.05). The present study shows that iris color is determined by both the quantity and the type of melanin in uveal melanocytes. These results suggest a possibility that uveal melanin in eyes with dark-colored irides is eumelanic at the surface and acts as an antioxidant while that in eyes with light-colored irides exposes pheomelanic core and behaves as a pro-oxidant.

This study was designed to review the clinical experience at our institution with fungal keratitis during a 16-year period.A review of the clinical and microbiology records of the New York Eye and Infirmary identified 61cases of fungal keratitis in 57 patients between January 1, 1987 and June 1, 2003. The medical records of all patients were retrospectively reviewed to better delineate patient demographics, risk factors, etiologic organisms, treatment, and outcomes.A total of 5083 positive corneal cultures were recorded'from January 1, 1987 to June 1, 2003. Sixty-one eyes in 57 patients (37 women) were positive for fungus (1.2%). Three'patients had bilateral simultaneous infections. Candida albicans accounted for 29 of 61 cases (48%). Human immunodeficiency virus (HIV) seropositivity (15 eyes), chronic ocular surface disease (14 eyes), and trauma (7 eyes) were the most commonly associated risk factors.Our experience with fungal keratitis in the northeastern United States appears to be different than those reported from other areas of the United States. Serologic positivity for HIV and chronic ocular surface disease were the most common associated risk factors followed by trauma, herpes simplex keratitis, and contact lens use. Candida species predominated, whereas filamentous fungi were uncommon.

Foci of autoantigen-specific B lymphocytes in nonlymphoid tissues have been associated with development of autoimmune disease. To better understand the genesis of such ectopic lymphoid tissue, this study investigated whether several B cell-tropic innate immune system molecules, known to be elevated in response to inflammatory stimuli, can cooperate in fostering the T cell-independent clonal expansion of mature human B2 cells under conditions of limiting BCR engagement. Notable synergy was observed between BCR coligation with the C3dg-binding CD21/CD19 costimulatory complex, B cell-activating factor belonging to the TNF family (BAFF), and IL-4 in generating B cell progeny with sustained CD86 and DR expression. The synergy was observed over a wide range of BCR:ligand affinities and involved: 1) cooperative effects at promoting early cell cycle progression and viability; 2) BCR:CD21 coligation-promoted increases in BAFF receptors that were highly regulated by IL-4; 3) reciprocal effects of IL-4 and BAFF at dampening daughter cell apoptosis typical of stimulation by BCR:CD21 and either cytokine alone; and 4) BAFF-sustained expression of antiapoptotic Mcl-1 within replicating lymphoblasts. The results suggest that significant clonal proliferation of recirculating B2 cells occurs upon limited binding to C3dg-coated Ag in an inflammatory in vivo milieu containing both BAFF and IL-4. When rare autoantigen-presenting B cells undergo such expansions, both B cell and T cell autoimmunity may be promoted.

To evaluate the modulation of wound healing and soft tissue ingrowth in synthetic and allogeneic implants with platelet gel. Attempts to influence wound healing with exogenous growth factors are highly dependent on the timing and dosing of treatment. Platelet gel made from autologous platelet concentrate (PC) and activated with calcium thrombin is increasingly used to enhance healing of surgical and chronic wounds, based on the assumption that proteins found in the blood can promote healing.Adult New Zealand white rabbits underwent phlebotomy, and the blood was used to produce nonconcentrated autologous blood clot, platelet-poor plasma (PPP), and PC for each animal. Disks of porous high-density polyethylene (PHDPE) and acellular dermal graft (ADG) were implanted into each animal in a subcutaneous location. Implants of each type were treated with isotonic sodium chloride solution, PPP, PPP followed immediately with PC, or autologous blood clot (by means of manual impregnation). Animals were killed at 2, 7, 14, and 21 days after implantation. Implants were harvested with surrounding soft tissue and examined by means of light microscopy for evidence of acute and chronic inflammatory cells and vascular and fibroblast invasion.A platelet gel with platelet concentrations averaging 5.8 times greater than those of peripheral blood significantly improved wound healing and soft tissue ingrowth in surgically implanted grafts. Early inflammatory infiltrates were enhanced in PHDPE and ADG implants by PC and autologous blood clot compared with control implants, as evidenced by significantly increased neutrophil and macrophage counts at day 2. Compared with controls, statistically significant increases in fibroblast and endothelial cell counts were noted at day 7 in PC-treated implants (fibroblasts, 61% increase [P.001] in PHDPE implants and 52% increase [P.001] in ADG implants; capillaries, 95% increase [P.05] in PHDPE and 97% increase [P.001] in ADG implants). Lymphocyte counts were increased by PC in PHDPE and ADG implants (71% [P.001] and 100% [P.05], respectively). There were no statistically significant differences in any cell count variables beyond 7 days.Treatment with PC prepared at 5 times the baseline platelet count significantly accelerated maturation of experimental wounds. By 14 days, the degree and quality of wound cellularity were equivalent among all treatment groups. Rapid wound healing was expected with this surgical model, which was chosen to observe the biological effects on early wound healing of a platelet gel in a noncompromised wound. Treatment with PC may be useful in scenarios in which enhancement and acceleration of early wound healing is desired.

The aim of our study was to explore the wavelength dependence of welding efficacy. Ex vivo samples of human and porcine aorta and skin tissues were investigated using a tunable Cr(4+):yttrium aluminum garnet (YAG) laser.Tissue welding is possible using laser light in the NIR spectral range. Collagen bonding in the tissue induced by thermal, photothermal, and photochemical reactions-or a combination of all of these-is thought to be responsible for tissue welding. Laser tissue welding (LTW) has gained success in the laboratory using animal models. Transition from laboratory to clinical application requires the optimization of welding parameters.A near-infrared (NIR) Cr(4+):YAG laser was used to weld ex vivo samples of human and porcine aorta and skin at wavelengths from 1430 to 1470 nm. Welding efficacy was monitored by measuring the tensile strength of the welded tissue and the extent of collateral tissue damage. Tensile strengths were measured using a digital force gauge. Changes in tissue morphology were evaluated using optical and scanning electron microscope (SEM). Fluorescence imaging of the welded areas was also used to evaluate molecular changes following tissue welding.Full-thickness tissue bonding was observed with porcine aorta samples. No collateral damage of the aorta samples was observed. Tissue denaturation was observed with human aorta, human skin, and porcine skin samples. The optimum tensile strength for porcine and human aorta was 1.33 +/- 0.15 and 1.13 +/- 0.27 kg/cm2, respectively, at 1460 nm, while that for porcine and human skin was 0.94 +/- 0.15 and 1.05 +/- 0.19 kg/cm2, respectively, achieved at 1455 nm. The weld strength as a function of wavelength demonstrated a correlation with the absorption spectrum of water. Fluorescence imaging of welded aorta and skin demonstrated no significant changes in collagen and elastin emission at the weld site.The observation that welding strength as a function of wavelength follows the absorption bands of water suggests that absorption of light by water plays a significant role in laser tissue welding.

Light extinction and angular scattering measurements were performed on three species of bacteria with different sizes and shapes ( Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis). The Gaussian ray approximation of anomalous diffraction theory was used to determine the average bacteria size from transmission measurements. A rescaled spectra combining multiple angular data was analyzed in the framework of the Rayleigh-Gans theory of light scattering. Particle shape and size distribution is then obtained from the rescale spectra. Particle characteristics (size and/or shape) retrieved from both methods are in good agreement with size and shape measured under scanning electron microscopy. These results demonstrate that light scattering may be able to detect and identify microbial contamination in the environment.

Plasmacytomas are plasma cell neoplasms that rarely involve ocular adnexal tissues as a primary lesion or secondary manifestation of plasma cell myeloma. Only one case of plasmacytoma involving the lacrimal drainage system, to our knowledge, is described in the literature. The clinical presentation of ocular adnexal primary plasmacytoma typically relates to its mass effect. In this clinicopathologic report, we describe an unusual presentation of primary plasmacytoma of the lacrimal canaliculus as infectious canaliculitis.

Melanoma cells produce tissue plasminogen activator (t-PA) that plays an important role in tumor invasion and metastasis. The production of t-PA by normal human uveal melanocytes has not been reported previously. In order to explore this possibility, we studied the production of t-PA by cultured human uveal melanocytes and compared that with the production by cultured human uveal melanoma cells and epidermal melanocytes. Human adult uveal melanocytes were isolated and cultured from donor eyes. The cells were cultured in serum-free medium for 48 h and the conditioned medium then collected for the plasminogen activator (PA) activity assay. Free PA activity was tested in an amidolytic assay using a t-PA standard curve. PA type was identified by fibrinography and antihuman t-PA and urokinase plasminogen activator (u-PA) blocking antibodies. Free PA activity was found in the conditioned medium of normal melanocytes and melanoma cells. The predominant PA activity was t-PA. Normal uveal melanocytes produced more t-PA (3.23 +/- 0.73 IU/105 cells/24 h) than that of epidermal melanocytes (1.25 IU/105 cells/24 h) but much less than uveal melanoma cells (11.0 +/- 3.39 IU/105 cells/24 h). Western blot analysis revealed that most t-PA in conditioned media were one-chain t-PA with molecular weight of 69 kDa. Our study indicates that uveal melanocytes may contribute to the free t-PA activity previously found in aqueous humor and choroidal eye cup superfusions. Therefore, this function of uveal melanocytes may play a role in intraocular matrix remodeling, fibrinolysis and aqueous humor outflow.

For a better understanding of genetic alterations in head and neck squamous cell carcinoma (HNSCC), we applied comparative genomic hybridization (CGH) in the analysis of 75 HNSCCs, comprised of 18 pharyngeal squamous cell carcinomas (PSCCs), 23 laryngeal squamous cell carcinomas (LSCCs), and 34 oral squamous cell carcinomas (OSCCs). The three subgroups of HNSCC showed significant differences in genetic alteration patterns. Overall, PSCC and LSCC had more copy number aberrations (CNAs) per tumor than did OSCC. Apparent differing patterns of high-level amplification were also observed. The smallest recurrent chromosomal regions of high-level amplification (≥15% of cases) were 7q22, 8q24.1, and 11q12–13 in PSCC and 3q26.1–29 in OSCC. According to single frequency and combined frequencies of CNAs, we concluded that the most important chromosomal events for progression of head and neck cancer were +3q, +5p, +8q, and −3p for all subgroups of HNSCC; additionally, +7q, +17q, −9p, and −13q for PSCC; +7p, +9q, +11q12–13, +14q, and +17q for LSCC; and +1p and +11q12–13 for OSCC. To identify further important genetic alterations and the relationships among the alterations, we constructed oncogenetic tree models for tumor progression of HNSCC from CGH data using branching and distance-based tree models. The tree models predicted that: (1) +3q21–29 was the most important early chromosomal event, and −3p, which occurred after +3q21–29, was also an important chromosomal event for all subsites of HNSCC; (2) +8q is the second most important early chromosomal event; (3) there may be at least three subgroups of HNSCC: one characterized by −3p, −9p, +7p, and −13q; another by +5p, +9qter, and +17p; and the other by +8q and +18p. These results suggest that different chromosomal aberrations may play a role in the initiation and/or progression of different subgroups of HNSCC. © 2002 Wiley-Liss, Inc.

Fluorescence images were acquired from ex vivo head and neck tumor specimens. The excitation and emission wavelength combinations were selected to image emission from native tissue fluorophores (collagen, tryptophan, elastin and nicotinamide adenine dinucleotide). Two dimensional intensity ratio maps were generated from the fluorescence images and compared to histology images. The ratio maps accurately distinguished the regions of tumor from normal tissue in the specimens. Additional features, such as collagen capsules, blood vessels, mucus glands and muscles were evident in the fluorescence images.

Purpose To describe four patients with an unusual presentation of metastatic carcinoma to the eye, diagnosed by histopathologic and immunohistochemical evaluation of vitrectomy material. Methods Retrospective case series of four patients with metastatic carcinoma to the retina/vitreous. The New York Eye and Ear Infirmary database was searched to identify patients with metastatic carcinoma to the retina or vitreous, diagnosed from 1995 to 2006. Diagnoses were established through histopathologic and immunohistochemical evaluation of vitreous fluid obtained via pars plana vitrectomy. Results The New York Eye and Ear Infirmary Pathology database search resulted in 19,149 vitrectomy specimens, 4 of which were positive for metastatic carcinoma. The diagnosis was clinically unsuspected in three patients. The oncologic work-up revealed the origin of metastases to be from the breast in two patients and from the lung in two patients. Conclusion Metastatic carcinoma to the eye can present with clinically innocuous metastases, masquerading as epiretinal membrane and macular hole, central serous chorioretinopathy, rhegmatogenous retinal detachment, retinal granuloma, and endophthalmitis. These findings suggest that submission of vitrectomy specimens for pathologic evaluation should be considered in patients with an oncologic history or in those with atypical clinical presentation.

Purpose Melanocytes are one of the major cellular components in the uvea. Interleukin-8/CXCL8 and monocyte chemoattractant protein-1 (MCP-1/CCL2) are the two most important proinflammatory chemokines. We studied the constitutive and lipopolysaccharide (LPS)-induced expression of IL-8 and MCP-1 in cultured human uveal melanocytes (UM) and explored the relevant signal pathways. Methods Conditioned media and cells were collected from UM cultured in medium with and without stimulation of LPS. Interleukin-8 and MCP-1 proteins and mRNAs were measured using an ELISA kit and RT-PCR, respectively. Nuclear factor (NF)-κB in nuclear extracts and phosphorylated p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases1/2 (ERK1/2), and c-Jun N-terminal kinase1/2 (JNK1/2) in cells cultured with and without LPS were measured by ELISA kits. Inhibitors of p38 (SB203580), ERK1/2 (UO1026), JNK1/2 (SP600125), and NF-κB (BAY11-7082) were added to the cultures to evaluate their effects. Results Low levels of IL-8 and MCP-1 proteins were detected in the conditioned media in UM cultured without serum. Lipopolysaccharide (0.01-1 μg/mL) increased IL-8 and MCP-1 mRNAs and proteins levels in a dose- and time-dependent manner, accompanied by a significant increase of phosphorylated JNK1/2 in cell lysates and NF-κB in nuclear extracts. Nuclear factor-κB and JNK1/2 inhibitors significantly blocked LPS-induced expression of IL-8 and MCP-1. Conclusions This is the first report on the expression and secretion of chemokines by UM. The data suggest that UM may play a role in the pathogenesis of ocular inflammatory diseases.

The effects of various prostanoids on the growth, melanogenesis and dendrification of cultured iridal melanocytes were studied. Iridal melanocytes were isolated and cultured with medium supplemented with cAMP elevating agents and basic fibroblast growth factor (bFGF) (complete medium). The iridal melanocytes were plated into multiple well plates and cultured with complete medium or various deleted media with or without various prostanoids at different concentrations. After 6 days, the numbers of cells and dendrites were counted and melanin content was measured and compared with controls. Prostaglandin E(2), an EP(2)receptor agonist (AH 13205) and AGN 192093 (thromboxane mimetic) stimulated growth, melanogenesis and dendrification of cultured iridal melanocytes in cAMP-deleted medium. A mixed EP(1)and EP(3)receptor agonist (sulprostone), a EP(4)receptor agonist (ONO-AE1-329), IP receptor agonists (cicaprost or iloprost) and a TP receptor agonist (U-46619) showed no effect. Prostaglandin D(2)showed stimulating effects. However, these stimulating effects could not be blocked by the addition of a DP receptor antagonist (BW A868C). Furthermore, a DP receptor agonist (BW 245C) showed no effects, indicating that the effect of prostaglandin D(2)may involve receptors other than the DP receptor subtype. The present study indicates that: (1) among various EP receptor agonists, only an EP(2)receptor agonist has stimulating effects on iridal melanocytes; (2) DP, IP and TP receptor agonists do not have stimulating effects; and (3) the mechanisms of action of prostaglandin D(2)and AGN 192093 need further study.

PURPOSE: To evaluate the utility of ultrasound biomicroscopy in imaging cyclitic membranes. METHODS: Patients with hypotony and suspected or known cyclitic membrane underwent ultrasound biomicroscopic examination. Histopathology of cyclitic membrane was correlated with ultrasound biomicroscopy in three cases. RESULTS: Six eyes of six patients were enrolled. Mean patient age was 62.2 ± 18.4 (SD) years. The mean intraocular pressure in the affected eye was 4.3 ± 3.4 mmHg. Three eyes were pseudophakic and three eyes were aphakic. All eyes had undergone two or more previous intraocular surgeries. Ultrasound biomicroscopy imaged the cyclitic membrane in all six eyes. Histopathology revealed fibroproliferative cyclitic membranes with associated inflammatory cells. CONCLUSION: Ultrasound biomicroscopy is useful in detecting the presence of those cyclitic membranes that may not be identified on clinical examination.

Objective: To study the factor structure, internal consistency, concurrent validity, and sensitivity to detect change in patient report of problems of a structured interview in relationship with accepted outcome measures. Design: Outcome status of patients with severe traumatic brain injury participating in a randomized, phase III, multicenter clinical trial was assessed at 6 mo postinjury using the Glasgow Outcome Scale, the Disability Rating Scale, and the Severe Traumatic Brain Injury Outcome Interview. Results: Exploratory factor analysis of the Severe Traumatic Brain Injury Outcome Interview produced a meaningful five-factor model: (1) activities of daily living; (2) cognitive; (3) affective; (4) behavioral; and (5) instrumental activities of daily living. The internal consistency of the factors ranged from moderate (0.61 instrumental activities of daily living) to high (0.94 activities of daily living); the interfactor correlations were moderate. The summed factor scores were significantly correlated with measures of global outcome: the Glasgow Outcome Scale (r = 0.66; P < 0.0001) and the Disability Rating Scale (r = 0.61; P < 0.0001). Patient report of cognitive problems correlated moderately with the neuropsychological tests. The summed factor scores were sensitive to change over time. Conclusions: Overall, the interview assessed the major important features of outcome pertinent to traumatic brain injury and demonstrated greater sensitivity to subtle changes over time than the unidimensional approaches, such as the Glasgow Outcome Scale and Disability Rating Scale.

Endoscopic brow lift techniques using temporary fixation rely on rapid readherence of the periosteum to calvarial bone. Little is known about the histologic events that occur during the early postoperative period after these procedures. An animal study was designed to compare and contrast periosteal fixation to bone and unelevated periosteum, with endoscopic and bicoronal brow lift techniques. One method of temporary fixation is the use of absorbable (polylactic/polyglycolic acid copolymer) LactoSorb screws; a histologic analysis of implanted LactoSorb screws was also performed. Sixteen rabbits underwent brow lifts; eight underwent endoscopic brow lift and fixation with LactoSorb screws without skin excision, and another eight underwent traditional bicoronal brow lift with skin excision and closure under tension. Animals were killed 1, 2, 6, and 12 weeks after the procedures were performed to evaluate the interaction of periosteum and bone and the normal, unelevated periosteum/calvarium interface at a site distant from the operative area. Histologic specimens were examined for the degree of apposition of periosteum to bone and for any fibrous or bony reaction at this interface. Histologic analysis showed various degrees of periosteal fibrosis and fixation to calvarial bone. After an initial phase of minimal periosteal adherence and moderate inflammation, the periosteum became progressively more adherent to bone in both groups, with no significant differences between treatment groups in rates of fixation. Fixation required at least 6 weeks. LactoSorb screws were surrounded by an area of mild inflammation and were progressively hydrolyzed and digested. Periosteal fixation increases over time for bicoronal and endoscopic brow lifts with minimal differences between the two techniques. With this animal model, periosteal adherence to calvarium requires at least 6 weeks with complete adherence by 12 weeks. In addition, the use of absorbable fixation screws seems to be both effective and well tolerated. The histologic changes associated with periosteal healing observed in this study suggest that permanent or semipermanent fixation may improve the accuracy and early postoperative maintenance of forehead advancement.

To investigate the effect of direct application of biologic materials normally present in wounds (basic fibroblast growth factor [bFGF] and autologous blood clot [ABC]) to accelerate the bony and soft tissue ingrowth into porous high-density polyethylene implants.We conducted a prospective, blinded animal histological study. Disks made of porous high-density polyethylene impregnated with bFGF or ABC were implanted into adult Sprague-Dawley rats in both subcutaneous and subperiosteal locations. Animals were killed and implants were harvested at 2, 4, and 10 weeks postimplantation and examined histologically for fibroblast invasion, collagen deposition, and inflammatory reaction.The results were compared with control (untreated) implants.As a group, the histological results showed significantly more fibroblasts within the ABC-treated implants than control implants or bFGF-treated implants. This difference in the number of fibroblasts between ABC-treated implants and bFGF-treated and control implants was also statistically significant 2 weeks after implantation.At the concentration of bFGF of 1 microg/10 microL, no acceleration of tissue ingrowth into porous high-density polyethylene implants was noted. However, when porous high-density polyethylene implants were treated with ABC, the implants were invaded to a greater degree by soft tissue, particularly in the early postoperative period (first 2 weeks). Bioactive substances associated with the coagulation and platelet cascades present in the ABC may be responsible for this accelerated incorporation of the porous implant and may have clinical implications. Arch Facial Plast Surg. 2000;2:27-33

Objective To assess the histologic behavior and clinical efficacy of autologous collagen dispersion (Autologen) in augmenting human dermis. Subjects Adult patients of the Facial Plastic Surgery Clinic at The New York Eye and Ear Infirmary who were undergoing facial aesthetic surgery with skin excision. Methods Five patients were injected intradermally with Autologen in one postauricular area and bovine cross-linked collagen (Zyplast) on the contralateral side. Patients were examined clinically for signs of infection, skin necrosis, or implant rejection/allergy 2, 4, and 12 weeks postinjection. Impressions and photographs of all implant sites were taken at all follow-up visits. Biopsy specimens of each implant were taken 4 and 12 weeks after injection and examined histologically for signs of integration, rejection, and resorption. Results All implants were well tolerated. No identifiable differences were noted in the clinical persistence of Zyplast vs Autologen. Histologically, there was more variability in the degree of fibroblast infiltration of Autologen vs Zyplast deposits. Conclusions Our trial suggests that autologous collagen dispersion may represent a viable alternative to bovine collagen. Clinical persistence and histologic behavior of Autologen appear to be at least as favorable as those of Zyplast, and Autologen obviates the need for allergy testing and eliminates the possibility of disease transmission.

Combining degenerate oligonucleotide-primed PCR (DOP-PCR) with comparative genomic hybridization (CGH) has made it possible to analyze genomic changes in single cells. Although DOP-PCR-CGH methodology has been reported, the reproducibility of the method has been uncertain. We have developed a reproducible DOP-PCR-CGH protocol by systematically evaluating different labeling methods (including nick translation, PCR incorporation, and random-primed labeling) and different hybridization mixtures (including amplified test DNA vs. amplified reference DNA, termed homo-hybridization; and amplified test DNA vs. unamplified reference DNA or vice versa, termed hetero-hybridization). We have analyzed DNA samples obtained from 16 tissue sources including fresh/frozen normal and tumor samples, formalin fixed and paraffin embedded tumor tissue, and tumor cell lines by using differently labeled probes and hybridization combinations, and we calculated the corresponding rate (CR) of DOP-PCR-CGH with standard CGH. We found that homo-hybridization produced reproducible results with high CRs as compared to standard CGH (91–100% CR, mean 97%); In contrast, hetero-hybridization failed to generate reproducible hybridization with low CRs (57–97% CR, mean 80%; χ2 = 1245.8, P < 0.0001), high background, uneven hybridization, and false deletions or amplifications. In addition, our improved DOP-PCR protocol raised the amplification efficiency at least five times as compared to previously reported protocols, allowing for the detection of genomic imbalances in as little as 12.5 pg of starting DNA. In conclusion, the DOP-PCR-CHG homo-hybridization method, especially when combined with labeling by nick translation, is reliable and reproducible. The method can be used in screening for genomic imbalances using minute amounts of tumor DNA, thereby facilitating CGH application. Genes Chromosomes Cancer 28:395–403, 2000. © 2000 Wiley-Liss, Inc.

Clinicopathologic correlation of a subconjunctival foreign body using ultrasound biomicroscopy and anterior segment ocular coherence tomography

PURPOSE To characterize the histopathology of recurrent Meesmann's corneal epithelial dystrophy after penetrating keratoplasty. METHODS Postmortem examination by light and electron microscopy of the eyes of an 84-year-old patient with Meesmann's dystrophy who underwent a penetrating keratoplasty in the right eye at age 74 years and a lamellar keratoplasty in the left eye at age 51 years. RESULTS In the right eye, the characteristic features of Meesmann's dystrophy were demonstrated in both the donor and recipient corneas. The pathologic findings were limited to the corneal epithelium and included increased thickness, architectural disorganization, loss of cell polarity, increased amounts of intracellular glycogen, presence of intraepithelial microcysts containing degenerated cells, and in some cells, the presence of an electron-dense fibrillogranular material associated with disrupted cytoplasmic filaments. In the left eye, the corneal findings were consistent with but not specific for Meesmann's dystrophy. These included architectural disorganization, loss of cell polarity, presence of intraepithelial microcysts, and irregular thickening of the basement membrane in the donor cornea. CONCLUSION Meesmann's corneal epithelial dystrophy is demonstrated to recur after penetrating keratoplasty. This finding suggests that the abnormalities that lead to the disease are localized to the corneal epithelial cells and not in the stroma, as previously proposed.

The effect of TGF-beta2 on growth of uveal melanocytes in vitro was studied and the dose-dependent inhibitory effect of TGF-beta2 was compared with the known concentration of TGF-beta2 in aqueous humor. Uveal melanocytes were isolated and cultured with medium supplemented with cAMP elevating agents and basic fibroblast growth factor. The uveal melanocytes were plated into multi-well plates. After 24 hr, TGF-beta2 was added to the medium in various concentrations. After 5 days, the cells were detached, counted and compared to the controls. The effect of TGF-beta2 on DNA synthesis (as evaluated by uptake of bromodeoxyuridine) were also tested. TGF-beta2 inhibited growth and DNA synthesis of cultured uveal melanocytes in a dose-dependent manner at concentrations from 0.03-10.0 ng ml-1. The growth-inhibition of TGF-beta2 was present even in serum-free medium. TGF-beta2 had little or no effect on melanogenesis of cultured uveal melanocytes. The serum used for cultivation did not contain active TGF-beta1 or TGF-beta2 as measured by immunoassay. The known amount of active TGF-beta2 in aqueous humor (0.2-0.4 ng ml-1) is sufficient to inhibit the growth of uveal melanocytes. It indicates that TGF-beta2 is a potent growth inhibit factor of uveal melanocytes and may play an important role in maintaining the non-proliferative, relatively quiescence status of uveal melanocytes in vivo.

BACKGROUND Inverted papilloma (IP) of the paranasal sinus is a benign neoplastic condition that can be associated with squamous cell carcinoma (SCC). To understand the etiology of the disease better, paranasal sinus tumor specimens were examined for alterations in either p53 protein expression or genomic DNA sequence, and for infection by human papilloma virus (HPV). METHODS Tumor specimens were categorized as follows: benign, nondysplastic IP; IP with dysplasia; SCC arising within IP; or SCC without IP. Sections of each tumor specimen were stained for p53 protein overexpression, and mutations in exons 5-9 of the p53 gene were determined in DNA purified from all tumor samples. HPV infection was screened by degenerate polymerase chain reaction (PCR) amplification and typed by multiplex PCR and direct DNA sequencing of PCR-amplified HPV sequences. RESULTS Altered p53, either in genetic sequence or protein overexpression, was observed in 0 of 7 benign, nondysplastic IP specimens. A significantly higher p53 alteration incidence was observed for IP specimens exhibiting dysplasia (57%; P < 0.05) and IP specimens that were associated with SCC (75%; P < 0.025). HPV sequences were detected in 9 of 24 (38%) tumor specimens, 78% of which were of the oncogenic HPV16 strain. A significantly higher incidence (P < 0.05) of HPV infection was observed in IP tumors exhibiting dysplasia or containing SCC than in nondysplastic IPs. None of the p53-mutated tumors were infected with oncogenic HPV16. CONCLUSIONS These data suggest that p53 alterations and/or HPV infection are associated predominantly with IPs exhibiting evidence of dysplasia or IPs associated with SCC, but not in nondysplastic, benign IPs. In addition, an inverse correlation may exist between oncogenic HPV infection and p53 alterations in paranasal sinus tumors. The authors postulate that patients with IPs containing altered p53 may be at increased risk for SCC of the paranasal sinus. Cancer 1997; 79:1320-8. © 1997 American Cancer Society.

• Background: Adenocarcinoma of the retinal pigment epithelium (RPE) is an uncommon intraocular tumor which has been rarely if ever diagnosed prior to enucleation. Our review of the literature suggests that when a melanotic and tuberous-shaped tumor presents in a woman with signs of uveitis one should consider the diagnosis of adenocarcinoma of the RPE. • Methods: We report a case in which a 4.9-mm-tall, mushroom-shaped, moderately reflective melanotic tumor found to be present in a 50-year-old woman. Other findings included a 12x10-mm dense transillumination shadow and 270 deg of posterior iris synechiae. With a presumptive diagnosis of uveal melanoma the patient was treated with palladium-103 plaque radiotherapy. Within 6 months she underwent enucleation due to uncontrollable ocular pain and progressive tumor growth. Histopathologic evaluation revealed an adenocarcinoma of the (RPE). • Results: Two months after radiotherapy the eye developed acute angle-closure glaucoma, secondary retinal detachment, and a painful scleritis (a result similar to that of Greer, who treated an intraocular adenoma with radiation). Our diagnosis of adenocarcinoma of the RPE was made after enucleation. • Conclusion: Adenocarcinomas of the RPE are rarely if ever diagnosed on clinical evaluation, should be suspected in women with a melanotic intraocular tumor and uveitis, and may respond poorly to ophthalmic plaque radiotherapy. All reported cases presenting without extrascleral extension have not metastasized.

The cytotoxic effects of zeaxanthin on two human uveal melanoma cell lines (SP6.5 and C918) and related signaling pathways were studied and compared to effects on normal ocular cells (uveal melanocytes, retinal pigment epithelial cells, and scleral fibroblasts). MTT assay revealed that zeaxanthin reduced the cell viability of melanoma cells in a dose-dependent manner (10, 30, and 100 μM), with IC50at 40.8 and 28.7 μM in SP6.5 and C918 cell lines, respectively. Zeaxanthin did not affect the viability of normal ocular cells even at the highest levels tested (300 μM), suggesting that zeaxanthin has a selectively cytotoxic effect on melanoma cells. Zeaxanthin induced apoptosis in melanoma cells as indicated by annexin V and ethidium III flow cytometry. Western blot analysis demonstrated that zeaxanthin decreased the expression of antiapoptotic proteins (Bcl-2 and Bcl-xL) and increased the expression of proapoptotic proteins (Bak and Bax) in zeaxanthin-treated melanoma cells. Zeaxanthin increased mitochondrial permeability as determined by JC-1 fluorescein study. Zeaxanthin also increased the level of cytosol cytochrome c and caspase-9 and -3 activities, but not caspase-8, as measured by ELISA assay or colorimetric assay. All of these findings indicate that the intrinsic (mitochondrial) pathway is involved in zeaxanthin-induced apoptosis in uveal melanoma cells.

BACKGROUND: We studied the histopathology of the stromal wound healing response in the cat cornea following intrastromal photorefractive keratectomy (IPRK) with the Nd: YLF picosecond laser. METHODS: Intrastromal PRK was performed in the anterior stroma of cat corneas with the Nd:YLF picosecond laser. The cats were sacrificed at predetermined intervals ranging from immediately to 6 months postoperatively. Effects of the laser treatment on the epithelium, Bowman's layer, stroma, and the endothelium were evaluated using light and scanning electron microscopy. No anti-inflammatory agents were used. RESULTS: Intrastromal PRK resulted in no perceptible damage to the corneal epithelium or Bowman's layer either acutely or at 6 months. The corneal stroma showed multiple cavitations immediately after intrastromal PRK, which collapsed over several hours, followed by thinning of the cornea over 2 weeks. At 1 month, the stromal collagen was abnormal with surrounding hyperceUularity. The endothelium showed no injury, acutely or at 6 months. No thermal effects on stromal collagen were observed at 6 months, and disruption of the lamellar pattern was not apparent after the cavitation bubbles were reabsorbed. CONCLUSION: Intrastromal PRK can effectively remove stromal tissue without acute damage to the adjacent lamellae, epithelium, or endothelium. There is a transient cellular wound healing response associated with a transient stromal collagen abnormality at 2 weeks to 1 month, which was not apparent 2 months after the procedure. [J Refract Surg. 1995;11:442-447.]

Purpose We report a case of a choroidal metastasis from an adenoid cystic carcinoma of the lung. Design Interventional case report. Methods A 40-year old man, 9 months' status postresection of a bronchial adenoid cystic carcinoma, was diagnosed by clinical evaluation and fine-needle aspiration biopsy and treated with palladium-103 ophthalmic plaque brachytherapy. Results This unusual patient with uveal metastasis from adenoid cystic carcinoma presented with decreased vision in the right eye and a diaphanous amelanotic choroidal tumor in the superotemporal macula. Ultrasound revealed a dome-shaped tumor that measured 4.0 mm in apical height and 11 × 10 mm in basal diameter. Fluorescein angiography revealed a double circulation and late intense subretinal fluorescence. Plaque brachytherapy provided local control and preservation of the eye. Conclusion Bronchogenic adenoid cystic carcinoma can metastasize to the choroid.

Unprecedented recent advances in the molecular genetics of cutaneous malignancies have markedly improved our ability to diagnose, treat, and counsel patients with skin tumors. This review provides an update on molecular genetics of periocular cutaneous basal cell carcinoma, squamous cell carcinoma, sebaceous carcinoma, Merkel cell carcinoma, and malignant melanoma and describes how the knowledge of molecular genetics is translated into clinical practice. A literature search of peer-reviewed and indexed publications from 1965 to 2012 using the PubMed search engine was performed. Key terms included: molecular genetics, eyelid, basal cell carcinoma, squamous cell carcinoma, sebaceous adenoma, sebaceous epithelioma, sebaceoma, sebaceous carcinoma, Merkel cell carcinoma, and melanoma. Seminal articles prior to 1965 were selected from primary sources and reviews from the initial search. Articles were chosen based on pertinence to clinical, genetic, and therapeutic topics reviewed in this manuscript. We reviewed the literature regarding the advances in molecular genetics of cutaneous basal cell carcinoma, squamous cell carcinoma, sebaceous neoplasia, Merkel cell carcinoma, and malignant melanoma, and possible future directions towards diagnosing and treating cutaneous tumors at the genetic level. Cell culture experiments, animal models, and molecular genetic studies on the patients’ tumor tissues helped to elucidate genetic aberrations in these lesions. Cell culture experiments, animal studies and, ultimately, clinical trials provided means to test and develop novel therapeutic strategies, namely targeted therapy directed at specific molecular genetic defects. While remarkable progress has been made in this process, the complexity of the molecular genetics of skin tumors makes complete elucidation of the genetic mechanisms and the search for ideal therapies challenging. The recent studies focusing on molecular genetics of cutaneous malignancies show promising results, thereby improving our ability to diagnose, treat and counsel patients with these lesions. Future studies will hopefully help unravel further molecular mechanisms involved in cutaneous neoplasia and provide insights into novel preventative and therapeutic modalities.

Resting mature human B cells undergo a dynamic process of clonal expansion, followed by clonal contraction, during an in vitro response to surrogate C3d-coated Ag and innate immune system cytokines, IL-4 and BAFF. In this study, we explore the mechanism for clonal contraction through following the time- and division-influenced expression of several pro- and anti-apoptotic proteins within CFSE-labeled cultures. Several findings, involving both human and mouse B cells, show that a mitochondria-dependent apoptotic pathway involving p53 contributes to the high activation-induced cell death (AICD) susceptibility of replicating blasts. Activated B cell clones exhibit elevated p53 protein and elevated mRNA/protein of proapoptotic molecules known to be under direct p53 transcriptional control, Bax, Bad, Puma, Bid, and procaspase 6, accompanied by reduced anti-apoptotic Bcl-2. Under these conditions, Bim levels were not increased. The finding that full-length Bid protein significantly declines in AICD-susceptible replicating blasts, whereas Bid mRNA does not, suggests that Bid is actively cleaved to short-lived, proapoptotic truncated Bid. AICD was diminished, albeit not eliminated, by p53 small interfering RNA transfection, genetic deletion of p53, or Bcl-2 overexpression. DNA damage is a likely trigger for p53-dependent AICD because susceptible lymphoblasts expressed significantly elevated levels of both phosphorylated ataxia telangiectasia mutated-Ser1980 and phospho-H2AX-Ser139. Deficiency in activation-induced cytosine deaminase diminishes but does not ablate murine B cell AICD, indicating that activation-induced cytosine deaminase-induced DNA damage is only in part responsible. Evidence for p53-influenced AICD during this route of T cell-independent clonal expansion raises the possibility that progeny bearing p53 mutations might undergo positive selection in peripherally inflamed tissues with elevated levels of IL-4 and BAFF.

Schwannoma is a proliferation of neoplastic Schwann cells. Whereas schwannomas of the head and neck region are common, intraocular tissues are rarely affected. Uveal schwannoma has been aptly called a "pseudomelanoma", reflecting the difficulty in its clinical distinction from uveal malignant melanoma. Most of our current knowledge on intraocular schwannoma is limited to case reports, short case series, and non-comprehensive literature reviews. Three isolated reports of uveal schwannoma with extrascleral extension exist in literature, but the prognostic significance of this growth pattern is unknown. We present a patient with choroidal schwannoma with extrascleral extension and review 46 previously reported cases of uveal schwannomas to delineate clinical and pathologic characteristics of these intraocular tumors with a specific emphasis on schwannoma with extraocular extension.

Hepatocyte growth factor (HGF) is a pleiotropic growth factor that is mainly expressed in mesenchymal cells. MET (mesenchymal–epithelial transition factor) is a membrane receptor that binds HGF. The receptors for HGF (MET) are primarily found in epithelial cells and several stromal cells. Activation of MET by HGF promotes migration, mitosis, and survival of various cells. HGF protects various cells from oxidative stress-induced apoptosis mainly via the phosphorylation of phosphoinositide 3-kinase/Akt pathway. HGF also plays a role in embryogenesis, tissue repair, and angiogenesis. HGF levels in the ocular fluids are elevated in various ocular diseases related to cell proliferation and angiogenesis. HGF protects retinal pigment epithelial (RPE) cells from hydrogen peroxide-induced apoptosis by inhibition of the mitochondrial apoptotic pathway. In ceramide- and glutathione depletion-induced apoptosis of RPE cells, studies have also demonstrated that HGF can protect RPE cells in these oxidative stress models. These studies suggest that HGF is a natural protective factor for RPE cells and plays an autocrine role protecting RPE cells against oxidative stress.

Objective To evaluate the histological changes induced in human skin by injection of autologous platelet-rich fibrin matrix (PRFM). Methods Four healthy adult volunteers were included in the study. Platelet-rich fibrin matrix was prepared from 9 mL of autologous blood using a proprietary system (Selphyl; Aesthetic Factors, Wayne, New Jersey) and injected into the deep dermis and immediate subdermis of the upper arms of subjects. Full-thickness skin biopsy specimens were taken from the treated areas over a 10-week period, and the specimens were processed for histological evaluation. Results Findings from histological examination supported the clinical observation of soft-tissue augmentation. As early as 7 days after treatment, activated fibroblasts and new collagen deposition were noted and continued to be evident throughout the course of the study. Development of new blood vessels was noted by 19 days; also at this time, intradermal collections of adipocytes and stimulation of subdermal adipocytes were noted. These findings became more pronounced over the duration of the study, although the fibroblastic response became much less pronounced. No abnormal mitotic figures were observed at any point, and a very mild chronic inflammatory response was noted only at the earliest time points of the study. Conclusions Injection of PRFM into the deep dermis and subdermis of the skin stimulates a number of cellular changes that can be harnessed for use. Coupled with prior in vitro and in vivo studies, we now have a much clearer picture of the cellular effects of PRFM and its potential uses in facial plastic surgery. Further work is planned to more clearly elucidate the potential role of PRFM in aesthetic and reconstructive surgery. Trial Registration clinicaltrials.gov Identifier: NCT00956020

Purpose To report 2 patients with progressive complex choristomas and to review the literature on this subject. Design Interventional case reports. Methods Clinical and pathologic correlation was performed on 2 patients with progressive epibulbar choristomas. PubMed database was searched to identify all the previously reported cases of progressive epibulbar choristomas (using key words choristoma, dermoid, growth, progression, and evolution). Results Growth of the epibulbar choristomas was noted in infancy in 1 patient with oculoectodermal syndrome and in puberty in another otherwise healthy patient. Both lesions were identified histopathologically as complex choristomas. In addition to the characteristic choristomatous tissues, both lesions demonstrated increased vascularity, inflammatory infiltrate, and fibroblast proliferation within myxomatous stroma. Review of the literature identified 4 patients with progressive complex choristomas, 1 of whom demonstrated histopathologic findings similar to those of the 2 cases reported here. Conclusions Epibulbar choristomas rarely enlarge, likely secondary to reactive changes within the tissue manifested by increased vascularity, inflammatory cell infiltration, and fibroblast proliferation with deposition of myxomatous stroma.

To report an immunocompetent patient with Nocardia exalbida endogenous endophthalmitis.Case report.Clinical-pathologic correlation and microbiologic evaluation were performed on an enucleated eye.A 56-year-old man presented with rapidly progressive vision loss associated with a posterior choroidal mass and serous retinal detachment. Pathologic evaluation of the enucleated eye demonstrated endogenous endophthalmitis. Nocardia exalbida was identified microbiologically. Systemic workup failed to demonstrate definite foci of systemic infection or evidence of immunocompromise. Review of literature failed to identify previously reported cases of Nocardia exalbida endophthalmitis.Nocardia can rarely cause isolated endogenous endophthalmitis in immunocompetent patients, which can contribute to a delay in diagnosis and vision loss. Endogenous Nocardia endophthalmitis typically occurs in immunocompromised patients with disseminated nocardiosis. Isolated endogenous Nocardia endophthalmitis in immunocompetent patients is rare. We describe isolated endogenous intraocular infection caused by Nocardia exalbida, a novel species, not previously associated with endophthalmitis.

Recently, we reported finding that circulating melatonin levels in age-related macular degeneration patients were significantly lower than those in age-matched controls. The purpose of this study was to investigate the hypothesis that melatonin deficiency may play a role in the oxidative damage of the retinal pigment epithelium (RPE) by testing the protective effect of melatonin and its receptor antagonist on RPE cells exposed to H(2)O(2) damage.Cultured human RPE cells were subjected to oxidative stress induced by 0.5 mM H(2)O(2). Cell viability was measured using the microculture tetrazoline test (MTT) assay. Cells were pretreated with or without melatonin for 24 h. Luzindole (50 μM), a melatonin membrane-receptor antagonist, was added to the culture 1 h before melatonin to distinguish direct antioxidant effects from indirect receptor-dependent effects. All tests were performed in triplicate.H(2)O(2) at 0.5 mM decreased cell viability to 20% of control levels. Melatonin showed dose-dependent protective effects on RPE cells against H(2)O(2). Cell viability of RPE cells pretreated with 10(-10), 10(-8), 10(-6), and 10(-4) M melatonin for 24 h was 130%, 160%, 187%, and 230% of cells treated with H(2)O(2) alone (all p0.05). Using cells cultured without H(2)O(2) as the control, cell viability of cells treated with H(2)O(2) after pretreatment with 10(-10)-10(-4) M melatonin was still significantly lower than that of the controls, suggesting that melatonin significantly decreased but did not completely abolish the in vitro cytotoxic effects of H(2)O(2). Luzindole completely blocked melatonin's protective effects at low concentrations of melatonin (10(-10)-10(-8) M) but not at high concentrations (10(-6)-10(-4) M).Melatonin has a partial protective effect on RPE cells against H(2)O(2) damage across a wide range of concentrations (10(-10)-10(-4) M). This protective effect occurs through the activation of melatonin membrane receptors at low concentrations (10(-10)-10(-8) M) and through both the direct antioxidant and indirect receptor activation effects at high concentrations (10(-6)-10(-4) M).