15-Hydroxyeicosatrienoic acid induces nasal congestion by changing vascular functions in mice.

BACKGROUND: Nasal congestion in allergic rhinitis (AR) is caused by vascular hyperpermeability and vascular relaxation of the nasal mucosa. We previously detected high levels of a lipoxygenation metabolite of dihomogammalinolenic acid, 15-hydroxy-8Z,11Z,13E-eicosatrienoic acid (15-HETrE) in the nasal lavage fluid of AR model mice. Here, we investigated the effects of 15-HETrE on vascular functions associated with nasal congestion. METHODS: We measured 15-HETrE levels in the nasal lavage fluid of ovalbumin-induced AR model mice and nasal discharge of patients with AR. We also assessed nasal congestion and vascular relaxation in mice. Vascular contractility was investigated using isolated mouse aortas. RESULTS: Five ovalbumin challenges increased 15-HETrE levels in AR model mice. 15-HETrE was also detected in patients who exhibiting AR-related symptoms. Intranasal administration of 15-HETrE elicited dyspnea-related behavior and decreased the nasal cavity volume in mice. Miles assay and whole-mount immunostaining revealed that 15-HETrE administration caused vascular hyperpermeability and relaxation of the nasal mucosa. Intravital imaging demonstrated that 15-HETrE relaxed the ear vessels that were precontracted via thromboxane receptor stimulation. Moreover, 15-HETrE dilated the isolated mouse aortas, and this effect was attenuated by K+ channel inhibitors and prostaglandin D2 (DP) and prostacyclin (IP) receptor antagonists. Additionally, vasodilatory effects of 15-HETrE were accompanied by an increase in intracellular cAMP levels. CONCLUSIONS: Our results indicate that 15-HETrE, whose levels are elevated in the nasal cavity upon AR, can be a novel lipid mediator that exacerbates nasal congestion. Moreover, it can stimulate DP and IP receptors and downstream K+ channels to dilate the nasal mucosal vasculature.

Marker-less tracking system for multiple mice using Mask R-CNN.

Although the appropriate evaluation of mouse behavior is crucial in pharmacological research, most current methods focus on single mouse behavior under light conditions, owing to the limitations of human observation and experimental tools. In this study, we aimed to develop a novel marker-less tracking method for multiple mice with top-view videos using deep-learning-based techniques. The following stepwise method was introduced: (i) detection of mouse contours, (ii) assignment of identifiers (IDs) to each mouse, and (iii) correction of mis-predictions. The behavior of C57BL/6 mice was recorded in an open-field arena, and the mouse contours were manually annotated for hundreds of frame images. Then, we trained the mask regional convolutional neural network (Mask R-CNN) with all annotated images. The mouse contours predicted by the trained model in each frame were assigned to IDs by calculating the similarities of every mouse pair between frames. After assigning IDs, correction steps were applied to remove the predictive errors semi-automatically. The established method could accurately predict two to four mice for first-look videos recorded under light conditions. The method could also be applied to videos recorded under dark conditions, extending our ability to accurately observe and analyze the sociality of nocturnal mice. This technology would enable a new approach to understand mouse sociality and advance the pharmacological research.

Interrogating the Osmotic Pressure of Self-Crowded Bovine Serum Albumin Solutions: Implications of Specific Monovalent Anion Effects Relative to the Hofmeister Series.

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.

Clinical features of IgG4-related dacryoadenitis.

BACKGROUND: To elucidate the clinical characteristics of IgG4-related dacryoadenitis. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Human immunodeficiency virus-related retinal microangiopathy and systemic cytomegalovirus disease association.

PURPOSE: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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 that developed over a 3-month observation period for IgG4-related dacryoadenitis and sialadenitis.

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.

Visual field deficit: a rare initial symptom of autoimmune pancreatitis.

An autoimmune pancreatitis (AIP) patient with metachronous and multiple extrapancreatic lesions is reported. Initial symptoms were proptosis, oculomotor deficits, and a visual field deficit of the left eye, and swelling of bilateral lacrimal glands. Swelling of the right salivary gland and elevated serum levels of hepatobiliary enzymes were detected. AIP associated with IgG4-related orbital pseudotumor, IgG4-related sclerosing dacryoadenitis and sialadenitis, and IgG4-related sclerosing cholangitis was diagnosed. All symptoms and lesions improved with steroid therapy. Although an orbital pseudotumor is a rare extrapancreatic lesion of AIP, we should know that AIP patients may describe unusual symptoms such as abnormal visual field.

Lacrimal gland function in autoimmune pancreatitis.

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 beta2 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.

Immunohistochemical observation of amniotic membrane patching on a corneal alkali burn in vivo.

PURPOSE: 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. METHODS: 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 alpha-smooth muscle actin (alpha-SMA), vimentin, MMP-1, MMP-2, MMP-9, and membrane-type1 (MT1)-MMP. Observation after Masson trichrome staining was also performed. RESULTS: In the AM group, alpha-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 alpha-SMA-positive cells were observed in the AM group. CONCLUSIONS: 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.

Trypan-blue- and endoillumination-assisted phacoemulsification in eyes with vitreous hemorrhage during combined cataract and vitreous surgery.

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.

Double visualization using triamcinolone acetonide and trypan blue during stage 3 macular hole surgery.

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 patients with 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 with idiopathic stage 3 macular holes.

Triamcinolone acetonide facilitates removal of the epiretinal membrane and separation of the residual vitreous cortex in highly myopic eyes with retinal detachment due to a macular hole.

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.