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The vast majority of malignant melanomas occur in the skin. These are associated with sunlight exposure and thought to be due to ultraviolet radiation. Melanomas in rare instances (1.3-1.4 percent) may originate from extracutaneous tissues such as the leptomeninges.(1) They have also been reported arising from gastrointestinal, respiratory and genitourinary mucosa. Sunlight is not an aetiologic factor with mucosal melanomas. Herein, we report an extremely rare example of malignant melanoma arising from the mucocutaneous junction of an ileostomy. Only one such case has been reported in the literature.(2)., (Copyright © 2012 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Background and Purpose: To evaluate the current AJCC/UICC staging system (7th edition) for nasopharyngeal carcinoma and to explore for future improvement., Materials and Methods: A total of 985 patients, initially staged with preceding 5-6th edition, were retrospectively re-staged with the 7th edition. All were assessed by magnetic resonance imaging, and all 945 non-disseminated patients were irradiated with conformal/intensity-modulated technique., Results: Staging factors by both the 5-6th edition and the 7th edition were strongly significance for important endpoints (p<0.001). Down-staging of the previous T2a to T1 and, stages IIA to I in the 7th edition was appropriate. However, the impacts on overall stage distribution and prognostication were minimal. Further down-staging of the current T2 to T1, N2 to N1, stages II to I, and merging of N3a and N3b, stages IVA and IVB were suggested. With the 7th edition, the 5-year disease-specific survival (DSS) was 100% for stage I, 95% for II, 90% for III, 67% for IVA, 68% for IVB and 18% for IVC. The corresponding DSS for the proposed stages I, II, III and IV were 95%, 86%, 67% and 18%, respectively., Conclusions: The changes introduced in the 7th edition were appropriate, but the magnitude of improvement was minimal. With improving results by modern management, further simplification of the staging system is suggested. The proposed system could lead to more accurate prognostication, further validation is warranted., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The rights to reproduce and found a family are recognized as basic human rights. Infertile couples should enjoy the same right to reproduce as those who have the ability to do so without assistance. Both positive and negative rights to access to assisted reproductive technologies are required in order to fully realize the reproductive rights. However, there is a limit to such a claim. The positive right of individuals to have state-funded assisted reproductive treatments has to be balanced against the provision of other societal goods and healthcare rationing. The negative right to acquire access to assisted reproductive technologies by individuals' own resources is also restrained. The barrier to such access is often of a moral nature, the standard of which depends on the values of the society.

A general procedure is proposed for the rapid development of a reversed-phase liquid chromatographic (RP-LC) separation that is "orthogonal" to a pre-existing ("primary") method for the RP-LC separation of a given sample. The procedure involves a change of the mobile-phase organic solvent (B-solvent), the replacement of the primary column by one of very different selectivity, and (only if necessary) a change in mobile phase pH or the use of a third column. Following the selection of the "orthogonal" B-solvent, column and mobile phase pH, further optimization of peak spacing and resolution can be achieved by varying separation temperature and either isocratic %B or gradient time. The relative "orthogonality" of the primary and "orthogonal" RP-LC methods is then evaluated from plots of retention for one method versus the other. The present procedure was used to develop "orthogonal" methods for nine routine RP-LC methods from six pharmaceutical analysis laboratories. The relative success of this approach can be judged from the results reported here.

The column selectivity parameters (H, S*, A, B and C) described in the preceding paper [L.R. Snyder, A. Maule, A. Heebsch, R. Cuellar, S. Paulson, J. Carrano, L. Wrisley C.C. Chan, N. Pearson, J.W. Dolan, J.J. Gilroy, J. Chromatogr. A 1057 (2004) 49-57] can be used to compare columns in terms of selectivity. A detailed procedure for such column comparisons is presented here, and evaluated by its use in finding suitable replacement columns for 12 different routine separations performed in five different pharmaceutical analysis laboratories.

Previous work has shown that the selectivity of reversed-phase columns for HPLC can be described by means of five column parameters: H (hydrophobicity), S* (steric resistance), A (hydrogen-bond acidity), B (hydrogen-bond basicity) and C (cation-exchange capacity). Values of H, S*, etc. can be determined by carrying out retention measurements for 18 test solutes under standardized conditions. The reproducibility of the latter procedure has been evaluated by comparison testing in four different laboratories and found acceptable. An alternative 10-solute test procedure which is more reproducible and convenient (but somewhat less accurate), requires only 2-3 h per column.

Typically, pharmacokinetic studies in mice require one animal per time point, thus resulting in differences due to dosing error, animal to animal variation and more importantly the euthanasia of a large number of animals. A method for the determination of pharmacokinetic data from serially bled mice to support early drug discovery is described. Sample analysis relies on liquid chromatography coupled with tandem mass spectrometry permitting robust and reproducible analysis requiring approximately 3 min per sample. Several parameters are discussed including the method of sample collection, preparation and analysis. The use of serially bled mice has lead to a remarkable reduction in animal usage and a corresponding reduction in compound required for such experiments. Using conventional methodology, a nine-point pharmacokinetic curve with four animals per time point would require 36 mice. With the method described below, only four mice in total are used and euthanasia is not required, permitting reuse after several weeks recovery and washout. Also, pharmacodynamic-pharmacokinetic correlation is possible and is demonstrated using a mouse model of diabetes.

When standing height (StndHt) cannot be assessed, arm span (AS) or sitting height (SitHt) has been used as surrogate variables for prediction of StndHt in adult caucasians and blacks. We examined (1) the relationship between StndHt, AS and SitHt among adult Chinese, Malays and Indians; and (2) whether anthropometry could explain the ethnic differences in lung volumes (as StndHt-adjusted lung volumes are known to differ significantly: Chinese > Malays > Indians). We recruited 1250 consecutive outpatients aged 20-90 years. Prediction equations of StndHt (with AS, SitHt, weight, age as predictors) for each subgroup of race and sex were formulated with multiple linear regressions. Equations with both AS and SitHt as predictors had the best goodness of fit (SEE = 2.37-2.85 cm, adjusted R2 = 0.67-0.87), as compared to equations with either AS (SEE = 3.00-3.91 cm, adjusted R2 = 0.58-0.80) or SitHt alone (SEE = 3.48-4.00 cm, adjusted R2 = 0.45-0.76). GLM general factorial analyses found that age- and weight-adjusted SitHt-to-StndHt ratios differed significantly among Chinese (0.539), Malays (0.529) and Indians (0.518). This paralleled the ethnic differences in lung volumes. The equations with both AS and SitHt as predictors provide the most accurate estimate of StndHt. Ethnic differences in upper body segment length may explain in part the lung volume differences.

Objective: Vascular endothelial growth factor (VEGF) is considered the growth factor that stimulates vasculogenesis and angiogenesis. Recent studies have demonstrated its role in regulating placental growth and invasion. Its expression can be upregulated by hypoxia. Intrauterine growth restriction (IUGR) is thought to be associated with inadequate placental perfusion, which might result from a failure in the development of the villous vascular network. Our present study was undertaken to examine the relationship between VEGF expression and IUGR in pregnancies with preserved umbilical artery end-diastolic flow., Methods: VEGF Expression was determined by immunohistochemical analysis of placentas from 17 pregnancies with normal infant birth weight and 17 pregnancies complicated by IUGR., Results: We found no significant differences in the expression of VEGF in villous syncytiotrophoblasts and intermediate trophoblasts in maternal decidua between IUGR and normal pregnancies. However, in both groups there was a strong correlation in the expression of VEGF with villous syncytiotrophoblasts and intermediate trophoblasts. In normal and IUGR pregnancies the infants' Apgar scores at birth were significantly correlated with VEGF staining in both syncytiotrophoblasts and intermediate trophoblasts (P < .05). A strong correlation also was found between cord hematocrit and VEGF staining in villous syncytiotrophoblasts (P < .05), but VEGF staining in intermediate trophoblasts was not correlated with cord hemoglobin or hematocrit., Conclusions: Our results suggest that VEGF acts in an autocrine and paracrine fashion in both normal and IUGR placentas, and its expression can have an effect on the well being of the infant at birth.

Fly ash from municipal solid waste (MSW) incinerators contains leachable metals, including potentially hazardous heavy metals. The metal content of the fly ash can be reduced by thermal treatment, which vaporizes the volatile metal compounds. After heat treatment of fly ash at 1000 degrees C for 3 h, less metal was able to be leached from the thermally treated ash than from the ash without thermal treatment. Al and Cr were the exceptions. These metals were more soluble in the ash that had been thermally treated. This paper focuses on the leaching behaviour of Al only. Both simple and sequential extraction leaching tests showed that the leachable Al for the heat-treated fly ash is about twice that of the untreated fly ash. The sequential test further revealed that (i) the majority of the leachable Al is associated with Fe-Mn oxides in the fly ash, and (ii) most of the unleachable Al resides in the silicate matrices of the heat-treated and untreated fly ash. Pure chemicals, Al(2)O(3), CaO and CaCl(2), simulating the relevant ingredients in the fly ash, were used for studying their reactions at 1000 degrees C. The aluminum compounds were identified by X-ray Diffraction (XRD). Two new chemical phases produced by the thermal treatment were identified; Ca(AlO(2))(2) and 12CaO.7Al(2)O(3). Their formation suggests a mechanism whereby thermal treatment of fly ash would produce more soluble Al.

Background/aims: Excessive formation of nitric oxide may mediate the generalized vasorelaxation and hyporesponsiveness to vasoconstrictors observed in portal hypertensive states. Endotoxin, released from the bowel and detoxified by the liver, could stimulate inducible nitric oxide synthase directly or indirectly via the cytokine cascade. This study investigated the effect of chronic intraperitoneal injection of polymyxin B, a neutralizing antagonist of endotoxin, on the hemodynamics of partially portal vein-ligated (PVL) rats., Methods: Concomitantly with endotoxin (600 EU) and dactinomycin (80 microg), polymyxin B (0.1 mg) or normal saline (N/S) was administered via an intraperitoneal route to male Sprague-Dawley rats. Twenty-four hours later, mean arterial pressure was determined. In PVL rats polymyxin B (0.1 mg in 5 cc N/S) or N/S was given intraperitoneally twice daily from 2 days prior to operation until 5 days (short-term) or 14 days (long-term) after the operation. Long-term polymyxin B- or N/S-treated sham-operated rats were included as controls. Hemodynamic studies with a thermodilution technique were performed at the end of treatment. Blood samples were collected from another series of PVL rats with long-term treatment to determine plasma levels of endotoxin and tumor necrosis factor-alpha. Plasma levels of endotoxin and tumor necrosis factor-alpha were measured by Limulus assay and the ELISA method, respectively., Results: With the dosage of 0.1 mg polymyxin B, hypotension in rats subjected to endotoxin and dactinomycin administration could be corrected (polymyxin B vs. placebo: 130.0+/-7.7 vs. 108.8+/-6.7 mm Hg, p<0.05). However, long-term or short-term treatment with the same dosage of polymyxin B failed to ameliorate the hyperdynamic circulation of PVL rats. In addition, long-term treatment with polymyxin B did not change systemic and portal hemodynamics in sham-operated rats. Plasma levels of endotoxin and tumor necrosis factor-alpha were comparable in PVL rats treated with long-term polymyxin B or N/S (p>0.05)., Conclusions: Our findings do not support the role of endotoxin in the hyperdynamic circulation of PVL rats.

The dose of methacholine causing a 20% fall in forced expiratory volume in 1 sec (FEV1) from baseline (PD20) has been used as an index of asthma severity. The aim of this study was to determine if the percentage fall in forced vital capacity (FVC) from baseline at the PD20 (dFVC%) is an independent marker of asthma severity. We first retrospectively studied the dFVC% and PD20 obtained from 149 consecutive newly diagnosed asthmatics with a positive methacholine-challenge test (MCT). We then performed MCT on 20 normal subjects and 35 stable asthmatics. The 'milder' asthmatics (n = 20) and 'more severe' asthmatics (n = 15) were on regular inhaled corticosteroids: 200 micrograms or less and 800 micrograms or more daily, respectively. A dosimeter technique was used, and normal subjects were given a cumulative dose of 2400 micrograms. The PD20 and dFVC% were calculated using log-linear interpolation of the last two points. Student's unpaired t-tests and linear regression analyses were used for comparison and correlation of results. There was no significant correlation between dFVC% and PD20 among the 149 newly diagnosed asthmatics (r = 0.1), or among the 35 known stable asthmatics (r = 0.008). The more severe asthmatics had a larger dFVC% compared with the milder asthmatics (15.8% vs. 9.6%; P = 0.0005). In addition, inhaled corticosteroid usage correlated better with dFVC% (r = 0.56) than with PD20 (r = 0.36). The normal subjects had a mean fall in FVC of only 4.8%. The percentage fall in FVC at PD20 (dFVC%) may be a useful index of asthma severity which is independent of PD20. This index is potentially complementary to the PD20 in the assessment of asthma severity.

Apoptosis and cell proliferation are widely recognized to be important physiological processes which together maintain tissue homeostasis. The apoptotic and proliferative processes in 38 first trimester placentae were quantified using terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling (TUNEL) and an antibody against Ki-67 antigen, respectively. In 14 cases, the estimation of apoptotic index was repeated with sections stained with haematoxylin and eosin by identifying the morphological features of apoptosis. We found that the indices obtained by either method correlated well with each other although a lower rate was obtained with the haematoxylin and eosin stained sections. Apoptosis was found in clusters mainly in syncytiotrophoblasts in association with fibrinoid deposits while proliferating activity was limited to cytotrophoblasts and stromal cells. A significant inverse correlation was observed between the proliferative index and gestational age as well as the apoptotic index. While the extent of apoptosis decreased with advanced gestational age, the correlation was not statistically significant. These findings provide a potential explanation for villous remodelling during trophoblastic invasion in early pregnancy.

A total elemental analysis was performed on a municipal solid waste (MSW) fly ash sample, before and after it was treated at 1000 degrees C, to reveal the metal distribution between the volatile matter and the ash residue. Metals such as Pb, Zn, Cd, and to a lesser degree, Cr, Mn and Ni, were volatilized. Addition of chlorinating agents generally increased the volatility of certain elements. More acid resistant compounds were formed in the ash residue after the heat treatment using CaCl2 as a chlorinating agent. The efficiencies of volatilization of the metals, using Cl2 as a chlorinating agent, were generally higher compared with using CaCl2. However, CaCl2 was found to be a more selective chlorinating agent for volatilizing the heavy metals of concern, i.e., Pb, Cd, Zn and Cu. The efficiencies of volatilization of the recovered metals were approximately proportional to their standard free-energy changes (delta G(o) for the corresponding chlorination reactions.

Objective: Primary intraocular lymphoma, a non-Hodgkin's lymphoma, is a primary central nervous system lymphoma (PCNSL). Diagnosis is usually made by identifying malignant, large B lymphocytes in the vitreous, eye, brain, and cerebral spinal fluid; however, these cells are few, friable, and difficult to recognize. Recently, clonal heavy chain immunoglobulin (IgH) gene rearrangement and bcl-2 gene translocation have been reported in systemic B-cell lymphoma and are used for the detection of malignant cells and in making a diagnosis. The authors investigated the molecular changes in three eyes and a chorioretinal biopsy specimen of four patients with PCNSL., Design: Human tissue study., Materials: Five ocular specimens of PCNSL were collected., Intervention: The first patient had a diagnostic enucleation of the left eye. The second patient underwent diagnostic chorioretinal biopsy. In the third case, a pair of autopsied eyes with reactive lymphoplasmacytic infiltrates of a patient with acquired immune deficiency syndrome (AIDS) were studied. In the fourth case, an enucleated eye of a patient with AIDS-associated lymphoma was sampled., Main Outcome Measures: The bcl-2 and IgH genes of the lymphoma cells from routine, paraffin-embedded, formaldehyde-fixed, or frozen histologic tissue sections were analyzed using microdissection and polymerase chain reaction (PCR) technique., Results: Lymphoma cells obtained from the above four cases showed IgH rearrangement gene in the third framework of the VH region. Bcl-2-associated translocation also was detected in three cases (cases 1, 2, and 4)., Conclusion: Rearrangement of the IgH gene can serve as a molecular marker for PCNSL. Microdissection allows for procurement and analysis of specific, selected, minute cell populations that are obtained from histologic sections of the complex, heterogeneous tissue. Translocation of IgH and bcl-2, the apoptotic "survival" signal and proto-oncogene, could contribute to the pathogenesis of PCNSL. The combination of microdissection and PCR is a powerful tool for studies of small lesions and cell populations and for understanding disease mechanisms.

The purpose of this study was to compare the cancer risk with chloroform exposure during showering. The study concentrated on the three major metropolitan areas of Taiwan. Total exposure was measured based on a combination of ingestion, inhalation and skin absorption. A total of 137 tap water samples were taken from 26 locations within the Taipei (north), Taichung (central) and Kaohsiung (south) areas. Analysis of VOC compounds was performed according to the US EPA Method 524. Chloroform concentrations were highest in Kaohsiung (60.19 micrograms/l), followed by Taipei (18.83 micrograms/l) and Taichung (17.55 micrograms/l). Based on the two-resistance theory to volatilization in showers, when air flow rate is increased, chloroform concentrations in the air significantly decrease. A 10-min shower would result in chloroform exposure with a 3:4:3 ratio (ingestion, inhalation, skin absorption). However, that changes to 1:7:2 for a 20-min shower under the same conditions. The cancer risk was highest in Kaohsiung at 17.59 per million for a 10-min shower and 64.77 per million for a 20-min shower. The lowest cancer risk was found in Taichung at 4.99 and 11.50 per million for a 10- and 20-min shower, respectively. Although ingestion is commonly considered to be the primary source of exposure to chloroform from tap water, inhalation and skin absorption exposure concentrations were found to be even higher.

FGF-3, originally named int-2, was discovered as an oncogene frequently activated in mammary carcinomas resulting from the chromosomal integration of the mouse mammary tumor virus (MMTV). Int-2 was later designated FGF-3 based on sequence homology with other members of the fibroblast growth factor (FGF) family. FGF-1 is the prototypical member of the FGF family, and is the only family member which activates all known FGF receptor isoforms. Transgenic mice expressing in the lens a form of FGF-1 engineered to be secreted show premature differentiation of the entire lens epithelium. In contrast, transgenic mice engineered to secrete FGF-2 in the lens do not undergo premature differentiation of the lens epithelium (C. M. Stolen et al., 1997, Development 124, 4009-4017). To further assess the roles of FGFs and FGF receptors in lens development, the alpha A-crystallin promoter was used to target expression of FGF-3 to the developing lens of transgenic mice. The expression of FGF-3 in the lens rapidly induced epithelial cells throughout the lens to elongate and to express fiber cell-specific proteins including MIP and beta-crystallins. This premature differentiation of the lens epithelium was followed by the degeneration of the entire lens. Since FGF-1 and FGF-3 can both activate one FGF receptor isoform (FGFR2 IIIb) that is not activated by FGF-2, these results suggest that activation of FGFR2 IIIb is sufficient to induce fiber cell differentiation throughout the lens epithelium in vivo. Furthermore, transgenic lens cells expressing FGF-3 were able to induce the differentiation of neighboring nontransgenic lens epithelial cells in chimeric mice. Expression of FGF-3 in the lens also resulted in developmental alterations of the eyelids, cornea, and retina, and in the most severely affected transgenic lines, the postnatal appearance of intraocular glandular structures.

The objective of this study is to analyze volatile organic compound (VOC) concentrations in Taiwan's drinking water supply. Focusing on Taiwan's three major metropolitan areas--Taipei, Taichung and Kaohsiung (in the north, middle and south, respectively)--171 samples were taken from tap water and 68 from boiled water. Tests showed VOC concentrations were highest in Kaohsiung. This is due to different water sources and methods of treatment. Except for bromoform, trihalomethane (THM) concentrations were highest. Detection rates of toluene and 1,2-dichloroethane were slightly higher than other VOC compounds. VOC concentrations decreased significantly after water was boiled. THMs had a removal rate from 61% to 82%. The authors conclude that the three metropolitan areas contain significantly different levels of VOCs and that boiling can significantly reduce the presence of VOCs. Other sources of pollution that contaminate drinking water such as industrial plants and gas stations must be further investigated.

Reported herewith are the results from an ongoing study of outdoor air pollution and the health of persons living in the communities in close proximity to petrochemical industrial complexes. To determine if there is an excess of adverse health outcomes in the population exposed to petrochemical industrial emissions, a health survey was undertaken in 1996 in this area and in one reference area which has no local industrial emissions. The subjects were 436 adults (30-64 years of age) living in the Sanwei area (exposed area) and 488 in Taicei (reference area). For several indicators of respiratory health, including cough, wheezing, and chronic bronchitis, the prevalence rates were not significantly different between the study and the control populations. Acute irritative symptoms (eye irritation, nausea, throat irritation, and chemical odor perception) were significantly more common in the exposed area, particularly perception of chemical odors (84.6% vs 2.1%). It is concluded that exposure to petrochemical air emissions may be associated with increased rates of acute irritative symptoms. Future studies are needed to identify the potential role of petrochemical industrial emissions (particularly volatile organic compounds) in the genesis of acute irritative symptoms in a nearby petrochemical industrial area.

Background: Bronchoalveolar lavage and transbronchial biopsy are often used for definitive diagnosis of lung rejection and infection in lung transplant recipients. Although protected specimen brushing is of value in nosocomial bacterial pneumonia, its role in lung transplant recipients had not been widely reported. The aim of the study is to review the diagnostic yield and therapeutic impact of flexible bronchoscopy with the use of a combination of bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy in lung transplant recipients., Methods: We reviewed flexible bronchoscopy data in 83 transplant recipients between February 1990 and March 1995. Only those with bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy were included in the analysis. There were 282 bronchoscopies performed for clinically suspected lung rejection or infection (clinical bronchoscopy) and 38 bronchoscopies for follow-up of a previously detected histologic abnormality (follow-up bronchoscopy)., Results: The total yields for rejection and infection for clinical and follow-up bronchoscopies were 67.4% and 58.9%, respectively. Acute rejection was detected with transbronchial biopsy in 26.2% and 34.2% of clinical and follow-up bronchoscopies, respectively. Cytomegalovirus pneumonitis was detected with transbronchial biopsy in 4.0% and 11.4% of clinical and follow-up bronchoscopies, respectively. Overall, bacteria was the most common cause of lower respiratory tract infection. When used together, protected specimen brushing and bronchoalveolar lavage were complementary techniques for detection of bacterial lower respiratory tract infection with a significantly higher proportion detected with protected specimen brushing ( > or = 10(3) colony forming units/ml) compared with bronchoalveolar lavage ( > or = 10(5) colony forming units/ml) (p < 0.001). Complications were hemorrhage (1.9%), pneumothorax (2.5%) and transient hypoxemia (10.5%). The results had an impact on management of rejection and infection in 57.8% of clinical and 39.5% of follow-up bronchoscopies., Conclusions: We conclude that bronchoscopy, with the use of a combination of bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy, is safe with a high diagnostic yield and therapeutic impact for treating lung transplant recipients.

We previously generated an animal model for the study of autoimmune diseases of the eye by targeting gamma interferon (gamma IFN) expression to the lens of transgenic mice. Here, we have studied the effect of constitutive lens expression of gamma IFN on eye development of these transgenic mice. By Day 18 of embryonic development, lens and retinal differentiation programs are completely disrupted; normal lens epithelia and fibers are replaced by balloon-like cells and retinal differentiation into inner and outer neuroblastic layers is already affected. The mRNA levels of gamma E- and/or gamma F-crystallin and MIP, markers of lens cell differentiation, are drastically reduced, while expression of ICSBP, a gamma IFN-inducible transcriptional factor, is induced in the alpha ACry-gamma IFN transgenic mouse eyes. Taken together, our results suggest that constitutive expression of gamma IFN and its induction and activation of gamma IFN-inducible transcriptional factors in the eye altered the developmental fate of cells destined to become lens fiber cells by altering the pattern of lens gene expression.

Purpose: To study the immunopathology of experimental melanin-protein-induced uveitis in the Lewis rat., Methods: Rats were immunized with bovine ocular melanin protein. The kinetics of experimental melanin-protein-induced uveitis was studied by clinical examination and immunopathology. Cellular and humoral responses were evaluated by lymphocyte proliferation, delayed-type hypersensitivity, and agglutination. After clinical disease subsided, recurrent experimental uveitis was induced with a low-dose footpad injection of lipopolysaccharide., Results: Experimental melanin-protein-induced uveitis was characterized by bilateral uveal infiltration mainly with lymphocytes and monocytes. Delayed-type hypersensitivity, lymphocyte proliferation, and agglutination to bovine ocular melanin protein were positive. Expressions of major histocompatibility complex class II and intercellular adhesion molecule-1 were observed before ocular infiltration. The predominantly infiltrating cells were CD4+ lymphocytes. Experimental melanin-protein-induced uveitis subsided within 1 month, spontaneously recurred within 1 week in approximately one quarter of the rats, and was inducible in most rats with 5 micrograms of lipopolysaccharide confirmed by histopathology., Conclusions: Experimental melanin-protein-induced uveitis is a T-cell-mediated autoimmune uveitis, resembling noninfectious recurrent iridocyclitis and choroiditis in humans.

S-antigen (S-Ag), a major protein on the retinal photoreceptor cell, can induce experimental autoimmune uveitis (EAU). We have previously detected S-Ag and its mRNA in irises of chronic uveitis patients receiving long-term steroid therapy. To further investigate the effect of steroid therapy on ocular tissue, we compared levels of S-Ag and its mRNA in EAU rats with and without steroid treatment. EAU was induced in 72 Lewis rats by two footpad injections of S-Ag. The rats were then treated with Depo-Medrol or phosphate-buffered saline by intramuscular injection. Eyes were collected at different time points. S-Ag mRNA was detected in the lens epithelium, trabecular meshwork, iris, and ciliary body of 47.2% of the steroid-treated group vs 22.2% of the controls. S-antigen was also detected in 30.6% of the eyes of the treated group vs 16.7% of the controls. S-Ag and its mRNA was found only in the retina of normal rats. These results support the hypothesis that prolonged corticosteroid therapy in uveitic patients might enhance the expression of S-Ag in nonretinal ocular tissues and might contribute to the ocular side effects of corticosteroid.

Rat prostaglandin endoperoxidase synthase-2 (PGHS-2) cDNA was cloned from rat calvarial osteoblasts total RNA by RT-PCR. The primary sequence of rat PGHS-2 had 98% and 92% identity to the mouse and human enzymes, respectively. Transfection of the rat PGHS-2 cDNA into COS 7 cells, followed by the addition of 20 microM arachidonic acid, resulted in a dramatic increase in PGE2 released from these cells. The amount of PGE2 produced was comparable to that obtained from cells similarly transfected with human PGHS-1 cDNA. In the rat paw carrageenin-oedema inflammatory model, the injected paw had elevated levels of PGHS-2 mRNA compared to the control paw. In a rat pyrexia model, injection of the pyrogen lipopolysaccharide, resulted in elevated levels of PGHS-2 mRNA in the brain. These results suggest that PGHS-2 expression plays a role both in inflammation and fever.

Background: Intraocular lymphoma is associated with significant morbidity and mortality, but early diagnosis and treatment may improve prognosis., Methods: The diagnostic features of 12 cases of intraocular lymphoma diagnosed at the National Eye Institute between 1984 and 1992 were retrospectively reviewed., Results: A pathologic diagnosis of large B-cell lymphoma was made on vitrectomy specimens in ten patients, cerebral spinal fluid in one, and on an enucleation specimen in one. The mean time from onset of symptoms to diagnosis was 21.4 months (range, 1-66 months). All 12 patients were given a final diagnosis of non-Hodgkin's lymphoma of the central nervous system (NHL-CNS), based on the epidemiology, pathology, and clinical course of their tumors. Although an initial vitrectomy was negative for malignant cells in three of ten patients, a repeat vitrectomy specimen subsequently showed intraocular lymphoma. Results of examination of the cerebrospinal fluid (CSF) showed malignant cells in 5 of 11 patients, although malignant cells were only identified after repeat examination of additional samples of CSF in three of these patients. Malignant cells often are difficult to identify, and an experienced cytopathologist was critical in making the correct diagnosis. In addition, corticosteroids are lympholytic to the lymphoma cells, and they appeared to decrease the viability of tumor cells obtained in samples of vitreous and CSF., Conclusion: The prompt, appropriate handling of specimens and review by an experienced cytopathologist are critical to the diagnosis of intraocular lymphoma. Malignant cells often are present in the cerebral spinal fluid at the time that ocular lymphoma is diagnosed. Nevertheless, multiple vitrectomies and lumbar punctures may be necessary before the correct diagnosis is made.

Background: The ideal management of intraocular inflammation may require a tissue diagnosis. Diagnostic vitrectomy is a well-established method for acquiring such tissue. However, in some patients, the diagnostic pathology is limited to the choroid and retina and vitrectomy may yield no useful information. In such cases, a more aggressive surgical approach to obtain tissue may be useful., Methods: The authors performed chorioretinal biopsies on seven patients with progressive chorioretinal lesions of unknown etiology. Indications for biopsy included: (1) macular-threatening lesions unresponsive to therapy, (2) suspicion of malignancy, or (3) suspicion of an infectious etiology. In all cases, it was expected that the results would alter therapy or other aspects of clinical care. Preoperative visual acuity was 20/200 or worse in each eye biopsied with one or more peripheral chorioretinal lesions present. Biopsy specimens were divided into three parts and submitted for light and electron microscopy, immunohistochemistry, and tissue culture., Results: On the basis of the biopsy findings, a diagnosis of multifocal choroiditis and subretinal fibrosis was rendered in three eyes, sarcoidosis in two eyes, and viral retinitis in two eyes. Therapy was changed in five patients. Final visual acuity was unchanged or improved in five eyes. Complications included the progression of lens opacity in all eyes and the development of phthisis in one eye that was extensively diseased preoperatively., Conclusion: Chorioretinal biopsy may provide useful information for determining the diagnosis and guiding the subsequent management of patients with progressive chorioretinal lesions of unknown etiology.

Autoimmune T lymphocytes can be used under appropriate conditions to induce resistance to the specific autoimmune disease that they usually produce. This practice, termed T cell vaccination, was found to be effective with the injection of a low (subpathogenic) number of autoaggressive T line lymphocytes. We report here that T cell vaccination produced marked resistance to the expression of experimental autoimmune uveoretinitis (EAU) in Lewis rats. In addition, vaccination led to the appearance of lymphoid cells in the vaccinated rats that demonstrated proliferative responses against idiotypic and ergotypic specificities of the injected T cells. This is the first report demonstrating the effector T lymphocytes specific for ocular antigens may be used as agents to modulate immunopathogenic responses responsible for EAU.

Diagnostic vitrectomy is often performed because of suspected infection or malignancy. Giemsa, Gram, and Papanicolaou stains are used routinely to identify the components in the vitreous. Immunocytochemical staining of cellular components of vitreous specimens has the potential to significantly increase the amount of useful information that can be gained from histopathologic study. Vitreous specimens from 14 patients undergoing diagnostic or therapeutic vitrectomy for infection, suspected primary intraocular lymphoma, or uveitis were examined by immunocytochemical staining using monoclonal antibodies specific for leukocyte subclass antigens and immunoglobulin. The three classes of disorders showed characteristic patterns of staining, which were useful in confirming microbiologic and clinical diagnoses. Infections showed more pronounced neutrophils and macrophages, primary intraocular lymphomas demonstrated light chain restriction of the malignant B lymphocytes, and uveitis was characterized by the predominance of T lymphocytes. The routine use of immunocytochemical staining is recommended to characterize cellular infiltrates and increase the diagnostic yield from vitrectomy specimens.

Two patients who had similar clinical presentations of bilateral multiple chorioretinal lesions and needed a correct diagnosis underwent chorioretinal biopsy. The biopsy from one patient demonstrated mainly a B cell infiltrate in choroidal and subretinal nodules, while the biopsy from the second patient showed mainly macrophages in the retina. These findings directed the therapeutic approach taken in each patient. Although chorioretinal biopsy is an invasive procedure with the potential for serious complications, the resultant finding may aid in the diagnosis and guide the subsequent management of certain patients presenting with serious ocular findings of undefined etiology.

We examined the action of a chimeric protein, IL-2-PE40, on the development of a T cell-mediated disease of the central nervous system with numerous similarities to multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). EAE is caused by IL-2 receptor-bearing T cells specific for myelin basic protein (BP). We report here that the treatment of Lewis rats with IL-2-PE40 delayed and shortened the course of EAE induced by BP in adjuvant and dramatically prevented EAE mediated by anti-myelin basic protein T line cells. The absence of paralytic signs, the absence of cell infiltration in the central nervous system, and the abatement of cellular immunity to myelin basic protein in the treated rats are direct consequences of the specific mechanism of action of IL-2-PE40. Our data support the notion that IL-2-PE40 may be efficient as an immunosuppressive agent for those disorders in which activated T cells play a crucial role.

A 38-year-old male homosexual with the new Acquired Immunodeficiency Syndrome (AIDS) had biopsy proven Kaposi's sarcoma of the right palpebral conjunctiva, extraocular mucocutaneous areas, and lymph nodes. Histologically, the palpebral tumor was characterized by atypical spindle cell proliferation and multiple slit-like vessels. Staining for Factor VIII-related antigen was positive in the cytoplasm of some tumor cells. Electron microscopy disclosed Weibel-Palade bodies in cells lining scattered slit-like vascular channels. The palpebral tumor mass was relatively nonadherent to its surrounding tissues and thus its simple excision was noteworthy; despite the prominent vascularity of the tumor, minimal bleeding was associated with its surgical resection. Any patient with AIDS should have all mucosal surfaces routinely examined for the presence of Kaposi's sarcoma lesions.

Forty-eight patients with the clinical diagnosis of central retinal vein occlusion and ten surgically enulceated eyes with central retinal vein occlusion documented by histopathologic examination were studied and reviewed. Retinal neovascularization developed in only one of the ten eyes. Rubeosis iridis occurred in 14% (7) of the clinical cases and in all of the histopathologic cases. Ten eyes with neovascular glaucoma, enucleated following central retinal vein occlusion, showed absence of retinal endothelial cells. The absence of retinal capillary endothelial cells is significant in explaining the rare occurrence of retinal neovascularization following central retinal vein occlusion. However, the small number of cases may reflect a biased result. Further studies of more cases over a longer period are required to reach the final conclusion.

The kinetic changes of T-lymphocyte subtypes in the eyes of Lewis rats which developed experimental autoimmune uveitis (EAU) were studied. Lymphoid cells were the majority of the ocular inflammatory cells in different stages of the disease. As EAU proceeds, there is an increase in the relative number of OX-8 (suppressor/cytotoxic) T cells, and the decrease of W3/25 (helper/inducer) T cells. These findings were also observed in the animals with EAU and partially treated with corticosteroids or cyclosporine. The kinetics may reflect the role of immunoregulation of the inflammatory response in autoimmune diseases.

Different patterns of iron deposition in the epithelium in 96 of 120 corneal buttons that had superficial irregularities of different causes, as examined after staining grossly with Prussian blue by Gomori's method were detected. These observations were confirmed by light microscopy in 86 cases. The deposition was either diffuse or it formed a linear pattern within the epithelium adjacent to elevated areas of corneal scarring. The authors believe that the tear film is the source of iron in these cases, and that relative pooling of tears on an irregular surface is the mechanism by which iron lines are formed.

Ocular fluids and sera from 12 onchocerciasis patients and nine age-matched controls living in Tamale, Ghana, were examined for the presence of anti-retinal autoantibodies by the indirect immunoperoxidase technique. Antibodies directed against autoantigens of the inner retina (nerve fiber, ganglion cell, and Müller cell) were found in the sera of 10 of 12 patients, but in only three of nine controls (P less than 0.003). Autoimmune antibodies against the outer segment of the photoreceptor were noted in 7 of 12 patients, in contrast to only one of nine controls (P less than 0.02). Findings with the ocular fluids generally mirrored those with the sera. These autoantibodies could not be absorbed by conventional techniques using either retinal S-antigen (S-Ag) or the interphotoreceptor retinoid binding protein, an observation suggesting that other ocular antigens are involved. Such anti-retinal antibodies, especially those directed against the inner retina, may play a significant role in the pathogenesis of the retinal degeneration and optic atrophy that occur as a consequence of onchocerciasis.

Forty-six eyes in 26 patients with active, intermediate, and/or posterior uveitis with decreased visual acuity were evaluated with the laser interferometer before treatment with systemic cyclosporine. Pretreatment visual acuity (ETDRS), pretreatment laser interferometric visual acuity, and the best visual acuity obtained while under treatment were compared. Eighty-six percent of patients in whom the laser interferometer predicted an improvement of three or more lines did improve. In contrast, 52% of patients in whom no improvement was predicted improved three or more lines with therapy. There was a moderate correlation (R = 0.59) (P less than 0.001) between the number of lines of improvement predicted and the number of lines of improvement obtained. If the laser visual acuity is three or more lines better than the eye chart acuity, the prognosis for vision improvement after cyclosporine therapy is good, even in those patients with cystoid macula edema.

Standardization of observations is recognized as fundamental to clinical research. The methodology for the evaluation of anterior segment inflammatory disease has become well accepted, while the ocular inflammatory standardization of the posterior segment has not been so well described or accepted. A system for the evaluation of vitreal inflammatory activity in patients with intermediate and posterior uveitis is presented. A series of photographs representing various degrees of fundus vitreal haze is depicted. The observer examines the eye with an indirect ophthalmoscope, then chooses the photograph which most closely simulates what is being seen. This technique is rapid and its reproducibility is helpful in standardizing clinical observations.

Ocular tissue from six patients with a clinical diagnosis of sympathetic ophthalmia (SO) was examined using immunohistochemical techniques. All patients presented with a history of bilateral panuveitis after penetrating ocular injury or multiple intraocular surgeries and clinical features of SO. In four cases, classic histopathological features of SO were observed, including granulomatous uveal tract infiltration and subretinal pigment epithelium (RPE) collections of inflammatory cells (Dalen-Fuchs nodules). Bone marrow derived monocytes were the major cellular components in these granulomas. In two cases, histopathology failed to demonstrate typical Dalen-Fuchs nodules or granulomas in the choroid. However, the choroidal infiltrates were composed primarily of T-helper and B lymphocytes, without macrophages or epithelioid cells. The eyes examined in this report indicate that a varied spectrum of immunopathological and histopathological findings may occur in clinically diagnosed SO.

The local effects of synthetic leukotrienes (LT) were examined in the skin of the anesthetized pig. Blood flow was measured noninvasively with the use of a laser-Doppler flow meter and changes in vascular permeability were measured using technetium-labeled human serum albumin as a marker for extravasation. LTB4 and the peptidolipid leukotrienes, LTC4, LTD4, LTE4, LTF4, induced vasodilator responses when injected intradermally at a dose of 1 ng. The vasodilator effects of LTB4 and LTF4 were comparable in magnitude to those of prostaglandin E2 (PGE2) and histamine and persisted over a wide dose range. Vascular permeability was induced by histamine, PGE2, and LTB4 but not by the other leukotrienes. The effects of LTB4 were significantly increased in the presence of PGE2. Leukotrienes appear not to produce their effects through the generation of prostaglandins as neither the vasodilator nor the permeability-enhancing effects were affected by treatment with indomethacin. The present investigation demonstrates that the pig is the first animal model to be described which reflects the potent vasodilator actions of leukotrienes in human skin. The porcine skin may thus be a useful model in the study of human skin diseases.

Epidermal proliferation was studied in the guinea-pig ear in vivo using the incorporation of [3H]thymidine into DNA as a measure of cell proliferation. Twenty-four-hour pretreatment of the skin topically with nanomolar amounts of either leukotriene B4 (LTB4), 12-hydroxyeicosatetraenoic acid, ionophore A23187, or the epidermal mitogen 12-O-tetradecanoylphorbol-13-acetate induced dose-dependent increases in epidermal proliferation. The stimulatory effect of LTB4 was stereospecific since a number of biologically inactive LTB4 stereoisomers had no effect.

A chorioretinal biopsy was obtained from a patient with subretinal fibrosis and chronic uveitis. The biopsy specimen was analyzed by conventional light and electron microscopy and by immunohistochemical methods. These demonstrated a markedly inflamed choroid with a predominance of B cells and plasma cells. Complement and IgG were deposited above Bruch's membrane. The subretinal fibrosis tissue contained islands of cells with retinal pigment epithelial characteristics; however, some of these cells demonstrated surface determinants of Müller cells. There were no circulating antiretinal antibodies detected nor could virus be cultured from the biopsy specimen. The clinical appearance of this entity is distinctive and consists of a chronic posterior uveitis with transient initial yellow-white deep retinal lesions and the later development of large white subretinal fibrotic areas. The EOG is markedly decreased. We hypothesize that the immunologic events involved in this entity include antibody production with immunoglobulin and complement deposition leading to retinal and retinal pigment epithelial destruction and a subretinal fibrous proliferation.

To analyze the role of the retinal vascular endothelial cells in the development of experimental autoimmune uveitis (EAU), we studied the presence of Ia antigen and FN in retinal vessels of Lewis rats immunized with retinal S antigen. Immunopathologic studies were performed on frozen tissues obtained during various stages of the disease. Our results show that Ia antigen was not present in the normal rat retina, and there was very little FN present in a few retinal vessels. One to two days prior to the histologic and clinical onset of EAU, FN was found to be increased in the retinal vessels. Ia antigen was found to be present in the retinal vessels coincident with the first signs of cellular infiltration. During the stage of maximal cellular infiltration, FN was present diffusely throughout the retina, as well as in the subretinal space, and Ia antigen was found diffusely in the cellular infiltrate. Therefore, FN and Ia antigen reflect the immunomodulation of vascular endothelial cells in EAU, which may be very important in the pathogenesis of retinal S antigen-induced uveitis. Two possible mechanisms for the role of the activation of the retinal vascular endothelium in the development of retinal inflammation in uveitis are discussed.

Forty patients with acquired immunodeficiency syndrome (AIDS) were examined for ocular abnormalities. Twenty of these patients died and the eyes were obtained for culture and histopathologic examination. These patients have multiple opportunistic infections and neoplasms as the result of a severe depression of cellular immunity. Fifty percent of all patients with AIDS and 75% of the autopsy group have ocular signs attributable to AIDS. Ocular findings were confined to four major categories: cytomegalovirus retinitis (10 patients), retinal cotton wool spots (11 patients), conjunctival Kaposi's sarcoma (2 patients) and neuro-ophthalmic motility abnormalities (3 patients). Cytomegalovirus retinitis was a significant cause of visual loss. Seven of 40 autopsy eyes had hand motion or worse visual acuity prior to the patients' death because of CMV retinitis. This necrotic retinitis showed minimal inflammation and progressed to involve the entire retina in three to six months, resulting in a gliotic retinal membrane. Therapy with antiviral agents was not effective. Recognizing the ocular signs of AIDS may facilitate the diagnosis. The ophthalmologist also has a major role in the observation of progression or regression of these ocular manifestations, and can assist in the evaluation of therapy in patients with AIDS.

S-antigen is a well-characterized retinal protein that is highly pathogenic for the induction of experimental autoimmune uveitis (EAU), a severe inflammatory disease of the eye and the pineal gland. EAU was observed following the immunization of Lewis rats with various doses (50 to 200 micrograms) of a small synthetic peptide, peptide N (22 amino acids in length), which corresponds to amino acid positions 281 to 302 in bovine S-antigen. Peptide N consistently induced an EAU that was identical to the disease caused by native S-antigen. Clinically, the disease that developed in the eye was characterized by iris and pericorneal hyperemia, followed by inflammatory exudates in the anterior chamber and vitreous. Histopathologically, a severe inflammatory response was observed that resulted in the complete destruction of the photoreceptor cell layer of the retina. In addition, animals with ocular inflammatory disease had an associated pinealitis characterized by a lymphocytic infiltration of the pineal gland. Furthermore, draining lymph node cells of rats immunized with peptide N showed strong in vitro proliferative responses toward peptide N as measured by [3H]thymidine uptake. Our results indicate that several synthetic peptides, which correspond to the amino acid sequence of bovine S-antigen, are capable of inducing an EAU and, as such, suggest that multiple uveitopathogenic sites may be present in the molecule.

An immunohistological study using monoclonal antibodies directed at specific membrane antigens of various inflammatory cells was carried out in order to evaluate the identity and topographic localization of the immuno-competent cells in an enucleated eye from a 6-year-old black patient with a three-month history of sympathetic ophthalmia. Correlative light and transmission electron microscopic examination of serial sections was also performed. The data demonstrated that the predominant cells within the choroidal infiltrate were T-lymphocytes (Leu 1+). T-cell subset analysis disclosed that most of these cells harbored specific antigenic determinants of the helper phenotype (Leu 3a+). A smaller proportion of the T cells demonstrated the specific determinants of the suppressor subtype (Leu 2a+). The helper/suppressor ratio varied slightly and ranged in most areas of the choroid between 3:1 and 4:1. Additionally, approximately 15% of the infiltrating lymphocytes harbored the Leu 14+ determinant specific for B cells. The latter were located in the outer choroid adjacent to the sclera. Very few natural killer (NK) cells (Leu 7+) were identified throughout the choroid. The granulomatous foci in the choroid were composed mainly of epithelioid cells and histiocytes expressing the OKM1+ and M221+ antigenic determinants on their membranes and demonstrating a high cytoplasmic nonspecific esterase activity (ANAE+). Within the Dalen-Fuchs nodules, similar to the choroidal nodules, there was a predominance of histiocytes and epithelioid cells (OKM1+, M221+, ANAE+), a few T-helper cells (Leu 1+, Leu3a+) and some OKM1-, M221- cells whose origin could not be determined. These findings were corroborated by electron microscopic observations of serial sections. Careful light and electron microscopic studies disclosed breaks in Bruch's membrane underlying some of Dalen-Fuchs nodules. In our opinion, these observations may be interpreted as the demonstration that Dalen-Fuchs' nodules and the choroidal granulomatous foci could be formed by identical cells of similar function and origin.

The more than coincidental occurrence of phacoanaphylatic endophthalmitis (PE) in sympathetic ophthalmia, and the similarity of the dissimilarity between Vogt-Koyanagi-Harada (VKH) disease and sympathetic ophthalmia have been well described both clinically and histopathologically. The etiology and pathogenesis of these three diseases are still not fully understood. Identifying and distinctive characteristics among them include the history of ocular trauma in sympathetic ophthalmia, rupture of the lens capsule in phacoanaphylatic endophthalmitis, and involvement of the skin, ear, and central nervous system in VKH disease. A T-cell-mediated immune reaction to ocular antigens seems to play a major role in sympathetic ophthalmia and VKH disease. A B-cell-related Arthus reaction to lens antigen seems to be the principle mechanism of PE. Thus, these three diseases may represent a spectrum of uveitis. At one end is the delayed-type hypersensitivity disease of sympathetic ophthalmia, whereas at the opposite end is the immune complex disease of PE, with VKH disease in the middle of this uveitic spectrum.