Chemokine Receptor Expression Pattern Correlates to Progression of Conjunctival Melanocytic Lesions

Purpose: Chemokines play a role in the progression and metastatic spread of both cutaneous and uveal melanomas. The aim of this study was to examine the prognostic value of expression of chemokine receptors CCR7, CXCR4, and CCR10 in conjunctival melanocytic lesions. Methods: In total, 44 conjunctival nevi, 21 cases of primary acquired melanosis (PAM) with atypia and 35 conjunctival melanomas, were included. After immunohistochemical staining for CCR7, CXCR4, and CCR10 the immunoreactive score (IRS) was determined. The findings were correlated for association with melanoma and development of metastasis. For mechanistic evaluation, we used a mouse melanoma metastasis model using two human conjunctival melanoma cell lines, CM2005.1 and CRMM1. Results: All tested chemokines showed a significantly higher expression in conjunctival melanoma than conjunctival nevi. There was a statistically significant difference between the IRS in nevi and PAM with atypia for nuclear IRS in CCR10 (P = 0.03) and both nuclear and cytoplasmic IRS in CXCR4 (P < 0.01 and P = 0.03, respectively); this was also true evaluating the groups PAM with atypia and melanoma all together (P < 0.01). Furthermore, a trend for lower IRS was seen in cases of melanoma without metastasis, with a suggestive pattern of a higher IRS in cases that did develop metastases, supported for CXCR4 using the mouse melanoma metastasis model. Conclusions: Expression of specific chemokines changes during the progression and metastatic spread of conjunctival melanocytic lesions. Differential chemokine profiles may hold prognostic value for patients with conjunctival melanomas and might be considered as a therapeutic target

Soluble Guanylate Cyclase α1–Deficient Mice: A Novel Murine Model for Primary Open Angle Glaucoma

Primary open angle glaucoma (POAG) is a leading cause of blindness worldwide. The molecular signaling involved in the pathogenesis of POAG remains unknown. Here, we report that mice lacking the $α_1$ subunit of the nitric oxide receptor soluble guanylate cyclase represent a novel and translatable animal model of POAG, characterized by thinning of the retinal nerve fiber layer and loss of optic nerve axons in the context of an open iridocorneal angle. The optic neuropathy associated with soluble guanylate cyclase $α_1$–deficiency was accompanied by modestly increased intraocular pressure and retinal vascular dysfunction. Moreover, data from a candidate gene association study suggests that a variant in the locus containing the genes encoding for the $α_1$ and $β_1$ subunits of soluble guanylate cyclase is associated with POAG in patients presenting with initial paracentral vision loss, a disease subtype thought to be associated with vascular dysregulation. These findings provide new insights into the pathogenesis and genetics of POAG and suggest new therapeutic strategies for POAG

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Liver Injury and Acute Liver Failure after Bariatric Surgery: An Overview of Potential Injury Mechanisms

The obesity epidemic has caused a surge in the use of bariatric surgery. Although surgery-induced weight loss is an effective treatment of nonalcoholic fatty liver disease, it may precipitate severe hepatic complications under certain circumstances. Acute liver injury (ALI) and acute liver failure (ALF) following bariatric surgery have been reported in several case series. Although rare, ALI and ALF tend to emerge several months after bariatric surgery. If so, it can result in prolonged hospitalization, may necessitate liver transplantation, and in some cases prove fatal. However, little is known about the risk factors for developing ALI or ALF after bariatric surgery and the mechanisms of liver damage in this context are poorly defined. This review provides an account of the available data on ALI and ALF caused by bariatric surgery, with emphasis on potential injury mechanisms and the outcomes of liver transplantation for ALF after bariatric surgery

Liver Injury and Acute Liver Failure after Bariatric Surgery: An Overview of Potential Injury Mechanisms

The obesity epidemic has caused a surge in the use of bariatric surgery. Although surgery-induced weight loss is an effective treatment of nonalcoholic fatty liver disease, it may precipitate severe hepatic complications under certain circumstances. Acute liver injury (ALI) and acute liver failure (ALF) following bariatric surgery have been reported in several case series. Although rare, ALI and ALF tend to emerge several months after bariatric surgery. If so, it can result in prolonged hospitalization, may necessitate liver transplantation, and in some cases prove fatal. However, little is known about the risk factors for developing ALI or ALF after bariatric surgery and the mechanisms of liver damage in this context are poorly defined. This review provides an account of the available data on ALI and ALF caused by bariatric surgery, with emphasis on potential injury mechanisms and the outcomes of liver transplantation for ALF after bariatric surgery

Liver Injury and Acute Liver Failure after Bariatric Surgery: An Overview of Potential Injury Mechanisms

The obesity epidemic has caused a surge in the use of bariatric surgery. Although surgery-induced weight loss is an effective treatment of nonalcoholic fatty liver disease, it may precipitate severe hepatic complications under certain circumstances. Acute liver injury (ALI) and acute liver failure (ALF) following bariatric surgery have been reported in several case series. Although rare, ALI and ALF tend to emerge several months after bariatric surgery. If so, it can result in prolonged hospitalization, may necessitate liver transplantation, and in some cases prove fatal. However, little is known about the risk factors for developing ALI or ALF after bariatric surgery and the mechanisms of liver damage in this context are poorly defined. This review provides an account of the available data on ALI and ALF caused by bariatric surgery, with emphasis on potential injury mechanisms and the outcomes of liver transplantation for ALF after bariatric surgery

HLA class I antigen expression in conjunctival melanoma is not associated with PD-L1/PD-1 status

PURPOSE. Antitumor T cells need expression of HLA class I molecules but can be inhibited by ligands such as programmed death ligand 1 (PD-L1). We determined expression and regulation of these molecules in human conjunctival melanoma (CM) samples, cell lines, and murine xenografts. METHODS. Immunofluorescence staining was performed to examine the expression of HLA-A, HLA-B/C, and β-2-microglobulin (B2M) in 23 primary CM samples. HLA class I expression was compared with clinicopathologic characteristics, the presence of tumor-infiltrating leukocytes, and PD-L1/PD-1 status. The effect of interferon γ (IFN-γ) on HLA class I expression was tested on three CM cell lines using quantitative PCR and flow cytometry. Furthermore, HLA class I expression was determined in CM cell line-derived murine xenografts. RESULTS. One third of tumors had positive HLA-A, HLA-B/C, and B2M expression. A positive expression was especially seen in thin and epibulbar tumors but was not associated with recurrences. HLA class I expression was correlated with M2 macrophage density and tended to associate with CD8+ T-cell density but was independent of PD-L1 or PD-1 expression. IFN-γ upregulated HLA class I expression and genes involved in HLA transcription and transportation on CM cell lines. Murine xenografts showed a comparable HLA class I expression as their respective cell lines. CONCLUSIONS. Our data indicate that subsets of CM have positive HLA class I expression, and HLA class I and PD-L1/PD-1 are expressed independently. When one considers immunotherapy, one should also analyze HLA class I expression, whose downregulation can limit the efficacy of T cell-mediated therapies

Retinal vascular dysfunction in sGCα₁^−/− mice. A

<p>: Representative trace depicting the diameter in one segment of a retinal arteriole in a wild-type (WT) mouse before, during (arrow), and after injection of the NO-donor compound sodium nitroprusside. Dashed lines indicate the diameter before and after sodium nitroprusside injection. <b>B</b>: Quantitative analysis of the change in diameter (double arrow in fig. 6A) induced by injection of 0.8 mg/kg sodium nitroprusside in WT and sGCα₁^−/− mice. <i>n</i> = 5 mice (3–4 arterioles per mouse were assessed, see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060156#pone.0060156.s003" target="_blank">fig. S3</a>). *<i>P</i> = 4.1×10⁻³.</p

Localization of sGC α₁ and β₁ subunits in the human and murine eye.

<p>Panels <b>A–C</b> depict tissue sections from human eyes. Panels <b>D–F</b> depict tissue sections from mouse eyes. <b>A</b>: Ciliary muscle (CM), stained for α-smooth muscle actin (red), sGCα₁ (green, <b>upper panel</b>), or sGCβ₁ (green, <b>lower panel</b>). Both sGCα₁ and sGCβ₁ co-localized with α-smooth muscle actin in CM (yellow in merged images). Scale bars: 100 μm. <b>B</b>: An arteriole in the iris (IA) and an arteriole in the retina (RA) were stained for α-smooth muscle actin (red), sGCα₁ (green, <b>upper panels</b>), or sGCβ₁ (green, <b>lower panels</b>). Both sGCα₁ and sGCβ₁ co-localized with α-smooth muscle actin in the smooth muscle cell layer of arterioles in the iris and retina (yellow in merged images). ONL: outer nuclear layer, INL: inner nuclear layer. Scale bars: 20 μm. <b>C</b>: sGCα₁ (<b>left panel</b>) and sGCβ₁ (<b>right panel</b>) expression was detected histologically in the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL, white arrow) of the retina. sGCα₁ and sGCβ₁ are visualized by green fluorescence. Scale bars: 20 μm. <b>D</b>: Adjacent sections of a wild-type (WT) murine eye were stained for α-smooth muscle actin (green, <b>left panel</b>) or sGCα₁ (red, <b>right panel</b>). sGCα₁ co-localized with α-smooth muscle actin in ciliary muscle (CM) and in arterioles in the ciliary body (CA). The iridocorneal angle is indicated. Scale bars: 50 μm. <b>E</b>: Adjacent sections of a WT murine eye were stained for α-smooth muscle actin (green, <b>left panel</b>) or sGCα₁ (red, <b>right panel</b>). sGCα₁ co-localized with α-smooth muscle actin in retinal arterioles (RA). Scale bars: 50 μm. <b>F</b>: sGCα₁ (<b>left panel</b>) and sGCβ₁ (<b>right panel</b>) expression was detected histologically in the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL, white arrow) of the mouse retina. sGCα₁ is visualized by brown peroxidase stain and sGCβ₁ is visualized by green fluorescence. Scale bars: 20 μm.</p

Association between <i>GUCY1A3/GUCY1B3</i> single nucleotide polymorphisms (SNPs) and POAG in the Glaucoma Gene and Environment (GLAUGEN) study.

<p>Most significant single nucleotide polymorphisms (SNPs) stratified by gender and type of visual field (VF) loss. 51 SNPs were analyzed within the GUCY1A3/GUCY1B3 locus and 50 kb upstream and downstream of the region. A Bonferroni correction of 3.3×10⁻⁴, correcting for analyzing 51 SNPs and 3 subgroups, was applied to define statistical significance. The top SNP (rs11722059) reached significance in women with paracentral visual field (VF) loss (P value of 3.1×10⁻⁴, bold and italicized). MA: minor allele; OR: multivariable odds ratio associated with each minor allele dose, generated by multiple logistic regression analyses adjusting for age, gender, study site, DNA source, DNA extraction method, and three eigenvectors; CI: confidence interval; Chr: chromosome; N: number.</p