New records of Heliopora hiberniana from SE Asia and the Central Indian Ocean

Coral reefs are among the most diverse ecosystems on the planet. They provide spawning sites for fishes and habitat for a myriad of fauna and flora. They protect coastlines from waves and storms and have important socio-economic value. However, coral reefs, as we know them, are seriously threatened by globalization and climate change [1]. The widespread bleaching of scleractinian corals threatens to destabilize critical ecosystem functions such as reef-building [2], and a growing body of data indicates that coral reefs are being transformed [3,4]. Future reefs are predicted to be dominated by non-constructional taxa [5,6], and the retreat of scleractinians threatens to cripple coral reef ecosystem functioning and endanger the lives of the millions of people that rely on coral reefs for protection, income and nutrition [7]. To detect coral community responses to climate change, and to identify which species may perform critical functional roles on future reefs, accurate taxonomic and systematic information is needed

Stem Cell-Derived Photoreceptor Transplants Differentially Integrate Into Mouse Models of Cone-Rod Dystrophy

Citation: Santos-Ferreira T, Völkner M, Borsch O, et al. Stem cell-derived photoreceptor transplants differentially integrate into mouse models of cone-rod dystrophy. Invest Ophthalmol Vis Sci. 2016;57:3509-3520. DOI:10.1167/iovs.16-19087 PURPOSE. Preclinical studies on photoreceptor transplantation provided evidence for restoration of visual function with pluripotent stem cells considered as a potential source for sufficient amounts of donor material. Adequate preclinical models representing retinal disease conditions of potential future patients are needed for translation research. Here we compared transplant integration in mouse models with mild (prominin1-deficient; Prom1 METHODS. For photoreceptor transplant production, we combined the mouse embryonic stem cell retinal organoid system with rhodopsin-driven GFP cell labeling by recombinant adenoassociated virus (AAV). Organoid-derived photoreceptors were enriched by CD73-based magnetic-activated cell sorting (MACS) and transplanted subretinally into wild-type, Prom1 and Cpfl1/Rho À/À hosts. The survival, maturation, and synapse formation of donor cells was analyzed by immunohistochemistry. RESULTS. Retinal organoids yielded high photoreceptor numbers that were further MACSenriched to 85% purity. Grafted photoreceptors survived in the subretinal space of all mouse models. Some cells integrated into wild-type as well as Prom1 À/À mouse retinas and acquired a mature morphology, expressing rod and synaptic markers in close proximity to secondorder neurons. In contrast, in the novel Cpfl1/Rho À/À model with complete photoreceptor degeneration, transplants remained confined to the subretinal space, expressed rod-specific but only reduced synaptic markers, and did not acquire mature morphology. CONCLUSIONS. Comparison of photoreceptor grafts in preclinical models with incomplete or complete photoreceptor loss, showed differential transplant success with effective and impaired integration, respectively. Thus, Cpfl1/Rho À/À mice represent a potential benchmark model resembling patients with severe retinal degeneration to optimize photoreceptor replacement therapies

Outer Segment Formation of Transplanted Photoreceptor Precursor Cells

<div><p>Transplantation of photoreceptor precursor cells (PPCs) into the retina represents a promising treatment for cell replacement in blinding diseases characterized by photoreceptor loss. In preclinical studies, we and others demonstrated that grafted PPCs integrate into the host outer nuclear layer (ONL) and develop into mature photoreceptors. However, a key feature of light detecting photoreceptors, the outer segment (OS) with natively aligned disc membrane staples, has not been studied in detail following transplantation. Therefore, we used as donor cells PPCs isolated from neonatal double transgenic reporter mice in which OSs are selectively labeled by green fluorescent protein while cell bodies are highlighted by red fluorescent protein. PPCs were enriched using CD73-based magnetic associated cell sorting and subsequently transplanted into either adult wild-type or a model of autosomal-dominant retinal degeneration mice. Three weeks post-transplantation, donor photoreceptors were identified based on fluorescent-reporter expression and OS formation was monitored at light and electron microscopy levels. Donor cells that properly integrated into the host wild-type retina developed OSs with the formation of a connecting cilium and well-aligned disc membrane staples similar to the surrounding native cells of the host. Surprisingly, the majority of not-integrated PPCs that remained in the sub-retinal space also generated native-like OSs in wild-type mice and those affected by retinal degeneration. Moreover, they showed an improved photoreceptor maturation and OS formation by comparison to donor cells located on the vitreous side suggesting that environmental cues influence the PPC differentiation and maturation. We conclude that transplanted PPCs, whether integrated or not into the host ONL, are able to generate the cellular structure for effective light detection, a phenomenon observed in wild-type as well as in degenerated retinas. Given that patients suffering from retinitis pigmentosa lose almost all photoreceptors, our findings are of utmost importance for the development of cell-based therapies.</p> </div

Vitreal-located transplanted photoreceptor precursor cells develop disturbed outer segment-like structures with misaligned disc membranes.

<p>CLEM analysis on resin sections allowed examination of vitreally-transplanted rhoEGFP–positive photoreceptor precursor cell aggregates. Cell nuclei are highlighted by DAPI staining (blue). A representative example is shown with fluorescence light microscopic- (A) and transmission electron microscopic-illumination (A′). The donor cells in A and A′ are shown at higher magnification in the respective panels B and B′. RhoEGFP-enriched structures (A, B; green, arrows) next to the cell bodies suggest the formation of OSs, and three of these (C, D and E) are displayed in the corresponding panels at a high-resolution level by TEM-analysis. These structures appear either as rhoEGFP-enriched cell debris (C) or rhoEGFP–positive OS-like structures with an abnormal organization/integrity including misaligned disc membranes (D, E). Additionally, mislocation of rhoEGFP to the plasma membrane of the soma was observed (F). Scale bars: A, A′: 20 µm, B, B′: 10 µm, C: 1 µm, D–F: 500 nm.</p

Differences in outer segment formation between sub-retinal and vitreal located transplanted photoreceptor precursor cells.

<p>CD73-enriched photoreceptor precursor cells (PPC) isolated from double-transgenic (rhoEGFP, actinDsRed) reporter animals were simultaneously transplanted into the sub-retinal (SRS) and vitreous (VS) space of 4-weeks old P347S hosts (A; B is the same image as A but with increased green fluorescence channel saturation level). Cell nuclei are highlighted with DAPI staining (blue). Donor cells (magenta) were detected in the SRS (e.g. boxed area in A) and VS (A and B, arrow) showing survival of transplanted cells in both locations. Whereas sub-retinally injected PPCs showed strong EGFP expression (green; e.g. boxed area in A), vitreally injected donor cells appeared almost devoid of EGFP signal (A, arrow). However, by increasing the saturation level of the green fluorescence signal channel in image A, faint EGFP signal could be detected in the vitreally located donor cells (B; B′ is a separate image with adjusted shutter time showing a magnification of the boxed area in B) demonstrating that donor cells located in the SRS had higher rhoEGFP expression levels than those in the VS (compare different cell populations in each individual panel A and B). Moreover, almost every single sub-retinal located donor PPC showed rhoEGFP labeled OSs (A′), in contrast to those located in the VS that generated only in few cases OS-like protrusions (B′, arrows). ONL, outer nuclear layer; INL, inner nuclear layer. Scale bars: 50 µm.</p

Transplantation of rhoEGFP–positive photoreceptors into the degenerated retina of heterozygous P347S mice.

<p>Low and high magnification views (A, A′) of a degenerated retina from a heterozygous P347S mouse, 3 weeks post-transplantation of CD73-enriched PPCs isolated from double-transgenic (rhoEGFP, actinDsRed) reporter animals. Cell nuclei are stained with DAPI (blue). Note, that transplanted cells (cell body, magenta; outer segment, green) are found in a sheet-like structure in the sub-retinal space (SRS), i.e. above the host outer nuclear layer (ONL), which is reduced to only 1–2 cell rows. Electron microscopy analyses of rhoEGFP–positive structures reveal the formation of ultra-structurally normal OSs containing morphologically native disc membrane staples (A″, A′″). In light adapted animals (B–B′″), transducin (B″, white and B′″, magenta) is expressed in OSs of transplanted, sub-retinal PPCs at low levels (B′ and B″, arrows), whereas high levels of transducin could be detected in IS-like structures next to rhoEGFP-positive OSs (B′ and B″, arrowhead). In contrast, dark adapted animals show significantly higher levels of transducin in rhoEGFP-positive OSs (C′ and C″, arrows), indicated also by white overlay staining in C′″. This illustrates, that sub-retinal PPCs show the native translocation of transducin under different light conditions, which is a prerequisite for functionality. INL, inner nuclear layer. Scale bars: A: 50 µm, A′: 10 µm, A″, A′″: 500 nm, B–B′″ and C–C′″: 10 µm.</p

Transplanted photoreceptors remaining in the sub-retinal space generate outer segments.

<p>Upon transplantation into an adult wild-type mouse (A), many CD73-enriched photoreceptor precursor cells (PPC) isolated from double-transgenic (rhoEGFP, actinDsRed) reporter animals remain in the sub-retinal space (SRS) at the top of the host outer nuclear layer (ONL, dashed line) and develop outer segment (OS)-like structures that are labeled with rhoEGFP (green, arrows). The cell body of transplanted cells is observed by DsRed-expression (magenta) whereas cell nuclei are stained with DAPI (blue). An electron microscopy overview (B) of transplanted PPCs found in the SRS demonstrates that several of them (asterisks) lay above the host photoreceptor OSs, which are delimitated by a dashed line. Magnifications of the boxed areas in panel B reveal that the non-integrated PPCs developed as well organized disc membrane staples, which are positive for rhoEGFP (B′, B″, black dots). Scale bars: A: 10 µm, B: 5 µm, B′ and B″: 2 µm.</p

Stem Cell-Derived Photoreceptor Transplants Differentially Integrate Into Mouse Models of Cone-Rod Dystrophy

Preclinical studies on photoreceptor transplantation provided evidence for restoration of visual function with pluripotent stem cells considered as a potential source for sufficient amounts of donor material. Adequate preclinical models representing retinal disease conditions of potential future patients are needed for translation research. Here we compared transplant integration in mouse models with mild (prominin1-deficient; Prom1-/-) or severe (cone photoreceptor function loss 1/rhodopsin-deficient double-mutant; Cpfl1/Rho-/-) cone-rod degeneration.status: publishe

Evaluation of serum Mucorales PCR for the diagnosis of Mucormycoses: The MODIMUCOR prospective trial

International audienceBackground Early diagnosis and prompt initiation of specific antifungal treatment is essential for improving the prognosis of mucormycosis. We aimed to assess the performance of serum Mucorales quantitative PCR (qPCR) for the early diagnosis and follow-up of mucormycosis.Methods We prospectively enrolled 232 patients with suspicion of invasive mold disease, evaluated using standard imaging and mycological procedures. Thirteen additional patients with proven or probable mucormycosis were included to analyze DNA load kinetics. Serum samples were collected twice-a-week for Mucorales qPCR tests targeting the Mucorales species Lichtheimia, Rhizomucor and Mucor/Rhizopus.Results The sensitivity was 85·2%, specificity 89·8%, and positive and negative likelihood ratios 8·3 and 0·17, respectively in this prospective study. The first Mucorales qPCR-positive serum was observed a median of four days (IQR, 0-9) before sampling of the first mycological or histological positive specimen and a median of one day (IQR, (-2)-6) before the first imaging was performed. Negativity of Mucorales qPCR within seven days after liposomal-amphotericin B initiation was associated with an 85% lower 30-day mortality rate (adjusted hazard Ratio = 0·15, 95%CI [0·03-0·73], p = 0·02).Conclusion Our study argues for the inclusion of qPCR for the detection of circulating Mucorales DNA for mucormycosis diagnosis and follow-up after treatment initiation. Positive results should be added to the criteria for the consensual definitions from the European Organization for the Research and Treatment of Cancer/ Mycoses Study Group Education and Research Consortium (EORTC/MSGERC), as already done for Aspergillus PCR