Guidance for Anti-VEGF Intravitreal Injections During the COVID-19 Pandemic

The World Health Organization designated the outbreak of the novel coronavirus, COVID-19, as a pandemic on March 11, 2020, after its rapid spread to countries throughout the  world.1 There is a strong need to support the ophthalmic community to help guide decision- making during these unprecedented times, where infection control is of utmost concern and  disruption   to   regular   clinical   practice is highly likely. This is  particularly  important for patients with retinal diseases who are receiving intravitreal injections of anti-vascular endothelial growth factor (VEGF) agents.Older age (>65 years), living in a care facility  or nursing home, and having an underlying health condition are key risk factors for developing severe illness following infection with COVID-19,  and  are  also  common among patients with retinal disease. Health conditions  including  chronic   lung   disease  or moderate-to-severe asthma, serious cardiovascular conditions, diabetes, being immunocompromised, severe obesity, liver disease, and chronic kidney disease requiring dialysis are considered particularly high risk, especially when they are not well-controlled.Several organizations have produced general guidance for ophthalmologists on managing patients during the pandemic. The Vision Academy Steering Committee convened in March 2020 to review these relevant guidelines and  documents,  discuss  key  challenges,  and  develop  recommendations  specifically  for managing patients receiving intravitreal injections of anti-VEGF agents during the COVID-19 pandemic. Recommendations were systematically refined before being voted on by the Steering Committee for consensus

The safety of using anti-VEGF: Is there strength in numbers? Curtis LH, Hammill BG, Schulman KA, Cousins SW (2010) Risks of mortality, myocardial infarction, bleeding, and stroke associated with therapies for age-related macular degeneration. Arch Ophthalmol 128(10):1273-1279

published_or_final_versionSpringer Open Choice, 21 Feb 201

Welcome to the Ipad generation

published_or_final_versionSpringer Open Choice, 31 May 201

Graefe’s Archives for Clinical and Experimental Ophthalmology: the past and the future

published_or_final_versio

Remarkable macrolichens in the urban area of Aachen and the north-western Eifel Mountains

Es wird über Großflechtenfunde, speziell epiphytische Arten, aus dem Aachener Stadtgebiet und der nordwestlichen Eifel berichtet. Zu den folgenden Arten werden genauere Angaben zu Fundorten gemacht: Bryoria fuscescens, Flavoparmelia soredians, Hyperphyscia adglutinata, Hypotrachyna afrorevoluta, Hypotrachyna revoluta s. str., Melanohalea laciniatula (= Melanelia laciniatula), Parmelia submontana, Parmelina pastillifera, Physconia distorta, Physconia enteroxantha, Physconia perisidiosa, Punctelia borreri, Ramalina fastigiata, Sphaerophorus globosus, Tuckermanopsis chlorophylla (= Cetraria chlorophylla), Usnea dasypoga (= Usnea filipendula) und Xanthomendoza fallax (= Xanthoria fallax). Diese und weitere Arten werden anhand von Fotos dargestellt.The article reports on macrolichens, especially epiphytic species, from the urban area of Aachen and the northwestern Eifel Mountains. Localities of the following species are presented: Bryoria fuscescens, Flavoparmelia soredians, Hyperphyscia adglutinata, Hypotrachyna afrorevoluta, Hypotrachyna revoluta s. str., Melanohalea laciniatula (= Melanelia laciniatula), Parmelia submontana, Parmelina pastillifera, Physconia distorta, Physconia enteroxantha, Physconia perisidiosa, Punctelia borreri, Ramalina fastigiata, Sphaerophorus globosus, Tuckermanopsis chlorophylla (= Cetraria chlorophylla), Usnea dasypoga (= Usnea filipendula) and Xanthomendoza fallax (= Xanthoria fallax). These and further species are illustrated on the basis of pictures

Expression of connexin genes in the human retina

Background: Gap junction channels allow direct metabolically and electrical coupling between adjacent cells in various mammalian tissues. Each channel is composed of 12 protein subunits, termed connexins (Cx). In the mouse retina, Cx43 could be localized mostly between astroglial cells whereas expression of Cx36, Cx45 and Cx57 genes has been detected in different neuronal subtypes. In the human retina, however, the expression pattern of connexin genes is largely unknown. Methods: Northern blot hybridizations, RT-PCR as well as immunofluorescence analyses helped to explore at least partially the expression pattern of the following human connexin genes GJD2 (hCx36), GJC1 (hCx45), GJA9 (hCx59) and GJA10 (hCx62) in the human retina. Results: Here we report that Northern blot hybridization signals of the orthologuous hCx36 and hCx45 were found in human retinal RNA. Immunofluorescence signals for both connexins could be located in both inner and outer plexiform layer (IPL, OPL). Expression of a third connexin gene denoted as GJA10 (Cx62) was also detected after Northern blot hybridization in the human retina. Interestingly, its gene structure is similar to that of Gja10 (mCx57) being expressed in mouse horizontal cells. RT-PCR analysis suggested that an additional exon of about 25 kb further downstream, coding for 12 amino acid residues, is spliced to the nearly complete reading frame on exon2 of GJA10 (Cx62). Cx59 mRNA, however, with high sequence identity to zebrafish Cx55.5 was only weakly detectable by RT-PCR in cDNA of human retina. Conclusion: In contrast to the neuron-expressed connexin genes Gjd2 coding for mCx36, Gjc1 coding for mCx45 and Gja10 coding for mCx57 in the mouse, a subset of 4 connexin genes, including the unique GJA9 (Cx59) and GJA10 (Cx62), could be detected at least as transcript isoforms in the human retina. First immunofluorescence analyses revealed a staining pattern of hCx36 and hCx45 expression both in the IPL and OPL, partially reminiscent to that in the mouse, although additional post-mortem material is needed to further explore their sublamina-specific distribution. Appropriate antibodies against Cx59 and Cx62 protein will clarify expression of these proteins in future studies

Age-related macular degeneration associated polymorphism rs10490924 in ARMS2 results in deficiency of a complement activator

Background Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. Methods Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM). Biolayer interferometry defined protein interactions. Furthermore, endogenous ARMS2 gene expression was studied in human blood derived monocytes and in human induced pluripotent stem cell- derived microglia (iPSdM) by PCR and LSM. The ARMS2 protein was localized in human genotyped retinal sections and in purified monocytes derived from AMD patients without the ARMS2 risk variant by LSM. ARMS2 expression in monocytes under oxidative stress was determined by Western blot analysis. Results Here, we demonstrate for the first time that ARMS2 functions as surface complement regulator. Recombinant ARMS2 binds to human apoptotic and necrotic cells and initiates complement activation by recruiting the complement activator properdin. ARMS2-properdin complexes augment C3b surface opsonization for phagocytosis. We also demonstrate for the first time expression of ARMS2 in human monocytes especially under oxidative stress and in microglia cells of the human retina. The ARMS2 protein is absent in monocytes and also in microglia cells, derived from patients homozygous for the ARMS2 AMD risk variant (rs10490924). Conclusions ARMS2 is likely involved in complement- mediated clearance of cellular debris. As AMD patients present with accumulated proteins and lipids on Bruch’s membrane, ARMS2 protein deficiency due to the genetic risk variant might be involved in drusen formation

CD171- and GD2-specific CAR-T cells potently target retinoblastoma cells in preclinical in vitro testing

BACKGROUND: Chimeric antigen receptor (CAR)-based T cell therapy is in early clinical trials to target the neuroectodermal tumor, neuroblastoma. No preclinical or clinical efficacy data are available for retinoblastoma to date. Whereas unilateral intraocular retinoblastoma is cured by enucleation of the eye, infiltration of the optic nerve indicates potential diffuse scattering and tumor spread leading to a major therapeutic challenge. CAR-T cell therapy could improve the currently limited therapeutic strategies for metastasized retinoblastoma by simultaneously killing both primary tumor and metastasizing malignant cells and by reducing chemotherapy-related late effects. METHODS: CD171 and GD2 expression was flow cytometrically analyzed in 11 retinoblastoma cell lines. CD171 expression and T cell infiltration (CD3+) was immunohistochemically assessed in retrospectively collected primary retinoblastomas. The efficacy of CAR-T cells targeting the CD171 and GD2 tumor-associated antigens was preclinically tested against three antigen-expressing retinoblastoma cell lines. CAR-T cell activation and exhaustion were assessed by cytokine release assays and flow cytometric detection of cell surface markers, and killing ability was assessed in cytotoxic assays. CAR constructs harboring different extracellular spacer lengths (short/long) and intracellular co-stimulatory domains (CD28/4-1BB) were compared to select the most potent constructs. RESULTS: All retinoblastoma cell lines investigated expressed CD171 and GD2. CD171 was expressed in 15/30 primary retinoblastomas. Retinoblastoma cell encounter strongly activated both CD171-specific and GD2-specific CAR-T cells. Targeting either CD171 or GD2 effectively killed all retinoblastoma cell lines examined. Similar activation and killing ability for either target was achieved by all CAR constructs irrespective of the length of the extracellular spacers and the co-stimulatory domain. Cell lines differentially lost tumor antigen expression upon CAR-T cell encounter, with CD171 being completely lost by all tested cell lines and GD2 further down-regulated in cell lines expressing low GD2 levels before CAR-T cell challenge. Alternating the CAR-T cell target in sequential challenges enhanced retinoblastoma cell killing. CONCLUSION: Both CD171 and GD2 are effective targets on human retinoblastoma cell lines, and CAR-T cell therapy is highly effective against retinoblastoma in vitro. Targeting of two different antigens by sequential CAR-T cell applications enhanced tumor cell killing and preempted tumor antigen loss in preclinical testing