TACSTD2 in gelatinous drop-like corneal dystrophy: variant functional analysis and expression in the cornea after limbal stem cell transplantation.

Gelatinous drop-like corneal dystrophy (GDLD) is a rare autosomal recessive eye disease. GDLD is characterized by the loss of barrier function in corneal epithelial cells (CECs) and amyloid deposition due to pathogenic variants in the TACSTD2 gene. Limbal stem cell transplantation (LSCT) has been suggested as an effective therapeutic alternative for patients with GDLD. However, despite LSCT, amyloid deposition recurs in some patients. The pathogenesis of recurrence is poorly studied. We present the case of a patient with GDLD. Genetic analysis revealed a homozygous deletion, NM_002353.3:c.653del, in the TACSTD2 gene. Functional analysis in a cell model system revealed the loss of the transmembrane domain and subcellular protein mislocalization. The patient with GDLD underwent direct allogeneic LSCT with epithelial debridement followed by deep anterior lamellar keratoplasty 10 months later due to amyloid deposition and deterioration of vision. Taken together, the results of transcriptome analysis and immunofluorescence staining of post-LSCT corneal sample with amyloid deposits obtained during keratoplasty demonstrated complete restoration of wild-type TACSTD2 expression, indicating that donor CECs replaced host CECs. Our study provides experimental evidence that amyloid deposition can recur after LSCT despite complete restoration of wild-type TACSTD2 expression.

Three-quarter DMEK in eyes with glaucoma draining devices to avoid secondary graft failure.

PURPOSE: To present a case series of a modified three-quarter Descemet's membrane endothelial keratoplasty (3/4-DMEK) technique to treat pseudophakic bullous keratopathy in the presence of a glaucoma drainage device (GDD) tube in the anterior chamber by reducing the risk of donor endothelial damage due to absence of donor endothelial cells overlying the GGD tube area. METHODS: In this prospective case series, four eyes of three patients with stable glaucoma underwent 3/4-DMEK surgery for pseudophakic bullous keratopathy after GDD insertion. The patients were followed up to 24 ± 2.5 months postoperatively. RESULTS: No intraoperative or postoperative complications were noted. The average central endothelial cell density (ECD) was 1093 ± 74 cells/mm2 at 12 months postoperatively, corresponding to an ECD decrease of 58 (±6)% as compared to preoperative values. Average best-corrected visual acuity increased from finger counting before surgery to 20/60 (logMar 0.5) at 12 months after 3/4-DMEK and remained stable up to 24 months postoperatively. All corneas remained clear at the last available follow-up. CONCLUSION: This case series demonstrates the technical feasibility of 3/4-DMEK in eyes with pseudophakic bullous keratopathy in the presence of a GDD tube. The absence of a donor DM and donor cells above the silicone tube excludes direct tube contact with the graft. Longer term studies are needed to show the effect of this modified graft pattern and dimensions on transplant survival.

Two-Dimensional Autofocus Technique Based on Spatial Frequency Domain Fragmentation.

Existing two-dimensional (2-D) autofocus algorithms, exploiting the known structure of 2-D phase error, lack the error estimation accuracy in the case of a low signal-tonoise ratio (SNR) due to the non-exhaustive use of the echo signal. In this paper, we propose a novel 2-D autofocus algorithm, thoroughly utilizing all the available data and therefore achieving superior estimation performance. Via analytical study, we show that the partial derivative of the 2-D error with respect to the azimuth frequency is approximable as a function of single argument, after appropriate change of variable. The established property enables a scheme for the azimuth phase error (APE) measurement, where the polar formatted data are fragmented along the range frequency and then aligned along the azimuth frequency, in order to equalize phase gradients in different fragments. On the one hand, such a scheme avoids the necessity to divide the full-aperture signal into subapertures, while on the other hand, it involves the whole signal support. Improved accuracy of the resulting estimate is achieved through the joint inter-fragment estimation of the APE gradient. The proposed algorithm, based on the mentioned scheme, was validated via computer simulations. The conducted experiments confirmed its preference against the existing techniques. The preference is particularly distinct for low SNR imagery.

The Impact of the Complexing Cation on the Sensitivity of the Collisional-Induced Dissociation Spectra to Fatty Acid Position for a Set of YXY/YYX-type Triglycerides.

RATIONAL: The development of an automated platform for the positional analysis of triglycerides based on electrospray tandem mass spectrometry continues to be pursued. This work compares the positional sensitivities of the collisional-induced dissociation spectra for a representative set of YXY/YYX triglycerides using ammonium, silver, sodium and lithium as complexing agents. METHODS: A set of triglycerides were synthesized and analyzed by electrospray tandem mass spectrometry using an ion trap mass spectrometer. Using different salt additives, the product ion spectra of the corresponding parent ions for twelve systems of the form YXY/YYX, where Y and X represent C16:0 , C18:1(c-9), C18:2(cc-9,12) and C20:4(cccc-5,8,11,14) , were collected. The data was used to prepare two-point calibration plots for each of the twelve positional isomer systems using each of the four complexing agents. RESULTS: The positional sensitivities for all twelve positional isomer systems were robust for both the sodium and lithium TAG adducts. The CID data for both the sodium and lithium TAG adducts are much less sensitive to the degree of unsaturation and double bond position of the fatty acids constituents than the CID data for the ammonium adducts. CONCLUSION: Using sodium or lithium TAG adducts may be advantageous for the development of an accurate predictive model for performing positional analysis of complex TAG mixtures based on electrospray tandem mass spectrometry. Ammonium adducts are likely complicated by the ability of the ammonium ion to provide extra stability to some parent ions through hydrogen bond-like interactions.