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The free-solvent-based (FSB) model and osmotic pressure were used to probe the ion binding and protein hydration for self-crowded bovine serum albumin in 0.15 M NaF, NaCl, NaI, and NaSCN solutions. All experiments were conducted with solutions at pH 7.4. The regressed results of the FSB model behavior to the measured osmotic pressure were excellent, albeit, the osmotic pressure data for NaSCN were noisy. The resulting ion binding and hydration were realistic values and the covariance of the two parameters was exceptionally low, providing substantial credibility to the FSB model. The results showed that the kosmotropic F- and neutral Cl- solutions generated significantly higher ion binding and protein hydration than the chaotropic solutions of I- and SCN-. Further, the ionic strength ratio and resulting hydration implied that the chaotropic solutions had substantially higher aggregation than the other salts investigated. Overall, the FSB model provides an additional, complementary tool to contribute to the analysis of crowded protein solutions relative to anions in the Hofmeister series as it can interrogate crowded solutions directly; something that is not possible with many measurement techniques.

A 70-year-old man was diagnosed with small- cell lung cancer (SCLC) of the left upper lobe, with a TNM classification of cT4N3M1b (PUL, OSS, BRA, HEP). A single asymptomatic brain metastasis 1 cm in diameter was also identified. The patient underwent four cycles of cis- platin plus irinotecan therapy, with a total effect of partial response. Complete remission of the brain metastasis was also achieved, and whole-brain radiation therapy (WBRT) was postponed at the request of the patient. Six months after diagnosis, he was admitted to our hospital with a major complaint of dizziness. Computed tomography showed enlargement of the primary lesion and multiple brain metastases. WBRT was started, but performance status did not improve. While undergoing WBRT, the patient complained of blurred vision. The ophthalmologist found a metastasis on the right iris by chance, although blurred vision was caused by detachment of the left retina. Two months later, the patient died of respiratory failure. Autopsy histologically confirmed the iris metastasis of SCLC. Cases of iris metastasis diagnosed before death are rarely reported. Iris metastases are estimated to account for 9 % of uveal metastases. This may suggest that many iris metastases have few clinical signs and are difficult to diagnose. Asymptomatic iris metastases, particularly among patients with SCLC, are thus likely to be underdi- agnosed. Ocular metastasis should be considered when a cancer patient complains of visual disturbance.

Purpose: To study the usefulness of intravitreal injection of triamcinolone acetonide and trypan blue for facilitating visualization and dissection of the posterior vitreous cortex and internal limiting membrane (ILM) during vitrectomy in idiopathic stage 3 macular holes. Methods: Pars plana vitrectomy was performed in 10 eyes of 10 patientswith idiopathic stage 3 macular holes. After core vitrectomy had been performed, triamcinolone acetonide was injected over the posterior pole. After separation of the visualized posterior vitreous cortex, trypan blue was injected over the macular area. Excised specimens were examined by electron microscopy. Results: Upon injection of triamcinolone acetonide, the posterior vitreous cortex and residual vitreous cortex could be visualized in all patients. The posterior vitreous cortex and residual vitreous cortex were completely removed. The ILM of the retina was stained faint blue and was successfully removed in all patients. Electron microscopy revealed that the triamcinolone-acetonide-visualized layer and the trypan-blue-stained layer had different histological features. No complications related to the use of triamcinolone acetonide and trypan blue were encountered. Conclusion: Double visualization of the posterior vitreous cortex and ILM using triamcinolone acetonide and trypan blue during vitrectomy may facilitate separation of the posterior vitreous cortex from the retina and removal of the ILM around the macular hole in patients withidiopathic stage 3 macular holes.

BACKGROUND/AIMS—The Arthus type allergic reaction is characterised by inflammatory cell infiltration and marked neovascularisation in the cornea. During the healing stages, inflammatory cells and newly formed microvessels gradually disappear. The aim was to establish whether apoptosis affected the regression of inflammatory cells and newly formed microvessels, in order to define more clearly the cellular mechanisms involved in the pathobiology of corneal diseases. METHODS—Albino male rabbits were injected subcutaneously with 5 mg/ml bovine serum albumin (BSA) incorporated in Freund's complete adjuvant twice weekly. Under the anaesthesia, 30 µl of a 0.5 mg/ml BSA solution was injected into the central corneal stroma to induce an Arthus type allergic reaction. The injured corneas were collected at various time points ranging from 3 to 20 days. Apoptotic cells were identified by both light microscopy using in situ TdT-dUTP nick end labelling (TUNEL) method and electron microscopy. RESULTS—With increasing time after induction of the Arthus reaction, marked neovascularisation and infiltrated inflammatory cells such as polymorphonuclear cells (PMNs) and plasma cells were observed in the cornea. Thereafter, the inflammatory cells and newly formed microvessels gradually disappeared. Coincidently, the numbers of microvessel endothelial cells and infiltrated inflammatory cells undergoing apoptosis were increased. Apoptotic bodies were taken up by macrophages, PMNs, as well as myofibroblasts derived presumably from transformation of migrated keratocytes. CONCLUSIONS—These data demonstrate that regression of the cellular infiltrates and microvessel endothelial cells associated with the Arthus reaction in the cornea occurs via apoptosis. This finding adds insights into the cellular mechanisms regulating the pathobiology of corneal diseases.

To elucidate the clinical characteristics of IgG4-related dacryoadenitis. Clinical features, laboratory findings, radiological findings, associated diseases, treatment, and prognosis were prospectively examined in 12 patients (seven men, five women; mean age, 60.9 ± 15.1 years) with IgG4-related dacryoadenitis. In addition to eyelid swelling, other ophthalmologic symptoms were observed in seven patients, including diplopia (n = 4), ptosis (n = 2), visual field disturbance (n = 2), eye pain (n = 2), decrease of visual acuity (n = 2), eye-movement disturbance (n = 1), dry eye (n = 1), corneal ulcer (n = 1), and epiphora (n = 1). Swelling of the lacrimal glands was bilateral in half of the patients. Other IgG4-related diseases were present in nine patients, including sialadenitis (n = 5), autoimmune pancreatitis (n = 4), retroperitoneal fibrosis (n = 2), and lymphadenopathy (n = 8). Serum IgG4 levels were significantly higher in patients with other IgG4-related disease (1070 ± 813 mg/dl) than in those without (197 ± 59 mg/dl, p = 0.017). Allergic histories and elevated serum IgE levels were each detected in six patients. Eight patients showed inflammatory extension beyond the lacrimal gland, such as thickened rectus muscle (n = 6), inflammation of the optic nerve (n = 2), and retrobulbar inflammation (n = 3). Steroid therapy was effective in seven patients, but dacryoadenitis relapsed in two patients with markedly higher serum IgG4 levels and autoimmune pancreatitis. IgG4-related dacryoadenitis showed various ophthalmologic symptoms due to extensive inflammation beyond the lacrimal gland, frequent association with other IgG4-related disease or allergic phenomena, and steroid responsiveness.

To determine whether there is a significant association between human immunodeficiency virus (HIV)-related retinal microangiopathy and systemic cytomegalovirus (CMV) disease in HIV-infected patients. Participants in this single-center, cross-sectional, retrospective study were 383 HIV-infected patients assessed for ocular manifestations before the beginning of antiretroviral therapy. The presence of HIV-related retinal microangiopathy, the presence of systemic CMV disease, laboratory data, and demographic information were determined by referring to medical records. The significance of any association between HIV-related retinal microangiopathy and systemic CMV disease was determined by use of the Chi-squared test and by multivariate analysis. HIV-related retinal microangiopathy was present in 85 patients, and was significantly associated with systemic CMV disease both by use of the Chi-squared test (P = 0.006) and by multivariate analysis (P = 0.045, odds ratio 2.03, 95 % confidence interval 1.02–4.06 adjusted for CD4+ cell count and plasma HIV-RNA level). These findings indicate that microangiopathy may be involved in the development of CMV disease in HIV-infected patients. Thus, detection of the presence of HIV-related retinal microangiopathy is important in the management of HIV-infected patients.

Autoimmune pancreatitis (AIP) that developed in a short period in a patient with IgG4-related dacryoadenitis and sialadenitis is reported. A 67-year-old man was diagnosed as having IgG4-related dacryoadenitis and sialadenitis histologically, serologically, and radiologically. At that time, the pancreas was normal on computed tomographic scanning, magnetic resonance imaging, and 18F-fluorodeoxyglucose positron emission tomography. However, AIP occurred in the pancreatic body and tail during the 3-month period of observation of the initial diseases. All IgG4-related lesions improved promptly after steroid therapy. This report emphasizes that, since IgG4-related disease can develop in other organs within a short period in a patient with IgG4-related disease, periodic imaging follow-up is necessary.

Objective Autoimmune pancreatitis (AIP) may be a pancreatic lesion of IgG4-related systemic disease. Lacrimal gland swelling is a rare extrapancreatic lesion of AIP. The aim of the present study was to investigate lacrimal gland function in AIP patients, and to determine changes after steroid therapy. Patients and Methods Schirmer's test and sialochemistry were done prospectively in 11 AIP patients. These tests were also performed after steroid therapy in 7 patients. Results Dysfunction of tear secretion was found in at least one eye in 7 (64%) patients. The average lower level in both eyes was 4.3±1.5 mm in the 7 patients with lacrimal gland dysfunction, which was significantly lower than the 8.2±2.4 mm in patients with normal lacrimal gland function (p=0.005). There were no significant differences between the two groups in age at diagnosis of AIP, sex ratio, and the presence of swelling of the lacrimal glands and the salivary glands. Although there was no significant difference, mean serum IgG4 levels and mean salivary Na+ and β2 microglobulin levels were lower in patients with normal lacrimal gland function. After steroid therapy, lacrimal gland function improved in 3 of 5 patients with impaired lacrimal gland function, though the degree of improvement was not marked compared to the improvement of salivary gland function. Conclusion Lacrimal gland function was frequently impaired in AIP patients, even when no lacrimal gland swelling was observed clinically. Lacrimal gland function impairment appears to be similar to impairment of salivary gland function in AIP patients.

To investigate by immunohistochemical observation the effects of amniotic membrane (AM) patching on myofibroblastic differentiation and matrix metalloproteinase (MMP) expression in the corneal stroma after an alkali burn in vivo. A corneal alkali burn was made by placing a circular piece of filter paper containing 1 N NaOH on the central cornea of rabbits. Burning was done unilaterally in each rabbit. Immediately after the wounding, in the AM group, AM was sutured onto the cornea and removed on day 1. Rabbits with no AM patching were controls. On day 14, corneas were excised and immunohistochemical observation was carried out using antibodies against α-smooth muscle actin (α-SMA), vimentin, MMP-1, MMP-2, MMP-9, and membrane-type1 (MT1)-MMP. Observation after Masson trichrome staining was also performed. In the AM group, α-SMA positive cells were noticeably fewer, and MMP-2, MMP-9, and MT1-MMP expression was clearly inhibited. Also, collagen fibers were more regularly arranged than in control eyes. The more proximate the cells were to the epithelial side, the fewer α-SMA-positive cells were observed in the AM group. AM patching suppressed myofibroblastic differentiation and MMP expression in the stroma after an alkali burn. An inhibition gradient suggests that AM may release unknown soluble factors possessing some antiscarring capability. Jpn J Ophthalmol 2007;51:3–9 © Japanese Ophthalmological Society 2007

Purpose: To study the safety and usefulness of trypan blue application and conventional endoillumination for enhancing visualization during phacoemulsification and foldable intraocular lens implantation in cases of cataract with vitreous hemorrhage. Methods: Ten eyes of 10 patients with cataract and coexisting vitreous hemorrhage underwent combined trypan-blue- and endoillumination-assisted phacoemulsification and foldable intraocular lens implantation with vitrectomy. In a two-step surgery, after staining the anterior capsule with trypan blue, continuous circular capsulorhexis was performed; then, phacoemulsification was performed using conventional endoillumination. Specular microscopy was performed before and 3 months after surgery on eyes that underwent trypan-blue- and endoillumination-assisted phacoemulsification and on control eyes that underwent standard phacoemulsification to evaluate the toxic effect of trypan blue on corneal endothelial cells. Results: Trypan blue application improved visualization of the anterior capsule, and a complete capsulorhexis could be performed in all eyes. In all 10 eyes, endoillumination was required to visualize the lens structure, and phacoemulsification could be accomplished. Specular microscopy revealed that there was no significant difference in the level of corneal endothelial cell loss between the trypan-blue- and endoillumination-assisted phacoemulsification group and the control group. Conclusion: The described double visualization technique may assist phacoemulsification in selected cases of cataract with vitreous hemorrhage.

Purpose: To study the usefulness of intravitreal triamcinolone acetonide injection during vitrectomy in highly myopic eyes with retinal detachment due to a macular hole. Methods: Pars plana vitrectomy was performed in 6 patients with retinal detachment resulting from a highly myopic eye with a macular hole. After separation of the posterior hyaloid and removal of any visible epiretinal membrane, triamcinolone acetonide was injected over the posterior pole. Excised specimens were evaluated by transmission electron microscopy. Results: Upon injection of triamcinolone acetonide, the entire epiretinal membrane and residual vitreous cortex could be visualized in all patients. The epiretinal membrane and residual posterior vitreous cortex were completely removed. Successful reattachment was performed without retinal damage in all cases. Electron microscopy revealed a cellular epiretinal membrane within a collagenous matrix lining the smooth internal surface of the internal limiting membrane. No complications related to the use of triamcinolone acetonide were encountered. Conclusion: Intraoperative visualization of the epiretinal membrane and residual posterior vitreous cortex with triamcinolone acetonide was found to be a useful adjunct to vitrectomy. Using triamcinolone acetonide during vitrectomy may facilitate both removal of the epiretinal membrane around the macular hole and separation of the residual vitreous cortex from the retina in highly myopic eyes with retinal detachment.

The fine structure of the intestinal epithelium in the swell fish (Spheroides stictonotus) was studied by electron microscopy in materials fixed in buffered osmic acid and embedded in Epon epoxy resin. Three types of cells can be distinguished in the epithelium.1. Columnar cells with a striated border are most numerous and common throughout the entire midgut. The luminal surface of the columnar epithelial cell is tightly packed with regular microvilli. In a few places the intermicrovillous plasma membranes are invaginated into the terminal web, giving rise to numerous furrows or a variety of vesicles in the apical cytoplasm. The lateral plasma membranes do not show complex interdigitations between neighboring cells, but desmosomes occur here sporadically. Particular concentration of mitochondria is observed both in the apical and basal cytoplasm of the columnar epithelial cell.2. The most striking feature in the columnar epithelial cells is the occurrence of endoplasmic lamellar structures which are found mainly infra-nuclear regions. These structures are composed of two very regular, parallel membranes which are separted by a distance of about 250A. From this characteristic appearance, they are readily distinguished from the usual endoplasmic reticulum. In cross sections, the cavity of these lamellae often shows continuity with the intercellular space. These membranes are never seen associated with RNP particles. These lamellae are independent of each other, being never anastmosed.3. Typical goblet cells, which are basically the same in structure as those found in other vertebrates, are encountered among columnar epithelial cells.4. A few of granular cells are occasionally observed singly in the epithelium. These cells contain large granules bounded by a single membrane, aboundant glycogen particles, and a variety of vesicles and tubules. Since these cells do not show any desmosomes between adjacent epithelia, and a similar type of cell is recognized in the lamina propria mucosae, they may be migratory in nature.5. The present study demonstrates the fine structure of the intestinal epithelium in a marine teleost. Of particular interest is the existence of lamellar structures which have been reported so far only in fresh water fishes. From the observations in fresh water teleosts and in the present study, it is strongly suggested that these lamellar structures are in general characteristic of the fish intestine, even though not in all species of fishes. These structures are assumed to be involved in the mechanism of concentration of salts or of water transport, probably the latter.