Electrospun poly(l-lactide-co-dl-lactide) nanofibrous scaffold as substrate for ex vivo limbal epithelial cell cultivation.

Ultrathin electrospun poly (l-lactide-co-dl-lactide) nanofibrous membranes coated with fibronectin were explored as scaffolds for the ex vivo cultivation of limbal epithelial cells (LECs) for the treatment of limbal stem cell deficiency. The developed scaffolds were compared with the "gold-standard" fibrin gel. The resulting membranes composed of nanofibers possessed a very low thickness of 4 µm and allowed very good optical transparency in the wet state. The fibronectin-coated nanofibrous scaffolds demonstrated LEC expansion and successful cultivation similar to that on fibrin gel. Unlike the regular cobblestone epithelial cell morphology on the fibrin gel, the nanofibrous scaffold presented a mostly irregular epithelial morphology with a shift to a mesenchymal phenotype, as confirmed by the upregulation of profibroblastic genes: ACTA2 (p = 0.023), FBLN1 (p < 0.001), and THY1 (p < 0.001). Both culture conditions revealed comparable expression of stem cell markers, including KLF4, ΔNp63α and ABCG2, emphasizing the promise of polylactide-based nanofibrous membranes for further investigations.

The Efficacy of Fibrin Sealant in Sutureless Deep Sclerectomy - Preliminary Outcomes.

PRCIS: Deep sclerectomy with fibrin adhesive can constitute a safe alternative to the classic procedure using sutures, providing non-allergenic, non-toxic and secure adhesion with no sign of aqueous humor outflow obstruction postoperatively. PURPOSE: To evaluate short- and medium-term postoperative results of deep sclerectomy (DS) with a fibrin sealant. PATIENTS AND METHODS: This prospective, noncomparative, interventional case-series involves 12 eyes of 12 patients with uncontrolled open-angle glaucoma (OAG) who underwent DS with Esnoper (Clip or V2000) implant between February 2021 and March 2022. A novel method of wound closure (sclera, Tenon´s fascia and conjunctiva) employing fibrin glue was used instead of classic sutures. Surgical outcomes assessed include: intraocular pressure (IOP) and glaucoma therapy (GT) reduction, best-corrected visual acuity (BCVA) changes and number of complications registered peri- and postoperatively. All measurements were performed preoperatively, as well as at 1 day, at 1 and 2 weeks, and at 1, 2, 3, 6, 9 and 12 months after surgery. RESULTS: The mean IOP decreased from 24.0±9.1 mmHg to 13.8±6.3 mmHg at one year postoperatively (P<0.001). Kaplan-Meier survival analysis revealed complete and qualified success rates of 83.3% and 91.7%. The mean GT decreased from 3.2±1.1 to 0.8±1.3 drugs 12 months after surgery (P<0.001). Nd:YAG goniopunture was performed in 2 eyes at 1 and 12 months postoperatively. No significant BCVA changes were registered. Perioperatively, we noted a trabeculo-descemet microperforation in one eye, transient hypotony in 5 eyes and mild hyphema in 2 eyes. CONCLUSIONS: Fibrin adhesive provided an effective closure in sutureless DS in the patients included in our study. This modification of classical DS may simplify the surgical technique, ensure secure wound adaptation, optimize healing, and lower the risk of inflammation and fibrosis postoperatively.

Chemical, Physical, and Biological Corneal Decellularization Methods: A Review of Literature.

The cornea is one of the most commonly transplanted tissues worldwide. It is used to restore vision when severe visual impairment or blindness occurs in patients with corneal diseases or after trauma. Due to the global shortage of healthy donor corneas, decellularized corneal tissue has significant potential as an alternative to corneal transplantation. It preserves the native and biological ultrastructure of the cornea and, therefore, represents the most promising scaffold. This article discusses different methods of corneal decellularization based on the current literature. We searched PubMed.gov for articles from January 2009 to December 2023 using the following keywords: corneal decellularization, decellularization methods, and corneal transplantation. Although several methods of decellularization of corneal tissue have been reported, a universal standardised protocol of corneal decellularization has not yet been introduced. In general, a combination of decellularization methods has been used for efficient decellularization while preserving the optimal properties of the corneal tissue.

Novel Biomaterials in Glaucoma Treatment.

Glaucoma is a significant cause of blindness worldwide, and its treatment remains challenging. The disease progressively leads to damage to the optic disc and thus loss of visual acuity and visual field. High intraocular pressure (IOP) is a common risk factor. There are three major methods to treat this disease: topical, laser, and surgical. None of these are completely satisfactory; therefore, alternatives using new biomaterials are being sought. Since biomaterial engineering has experienced significant growth in recent decades, its products are gradually being introduced to various branches of medicine, with the exception of ophthalmology. Biomaterials, such as glaucoma drainage implants, have been successfully used to treat glaucoma. There is significant ongoing research on biomaterials as drug delivery systems that could overcome the disadvantages of topical glaucoma treatment, such as poor intraocular penetration or frequent drug administration. This article summarizes the use of novel biomaterials for glaucoma treatment presented in the literature. The literature search was based on articles published in English on PubMed.gov, Cochranelibrary.com, and Scopus.com between 2018 and 2023 using the following term "biomaterials in glaucoma." A total of 103 published articles, including twenty-two reviews, were included. Fifty-nine articles were excluded on the basis of their titles and abstracts.

Scleral Grafts in Ophthalmic Surgery. A Review.

AIM: To summarize the history and current trends in the use of scleral grafts in ophthalmology. MATERIALS AND METHODS: We conducted a review of the literature through the MEDLINE and Cochrane Library databases. The search terms were "sclera", "graft", and "surgery". The search resulted in 1596 articles, of which we evaluated 192 as relevant. The relevant articles were sorted chronologically and according to the method of using scleral grafts, which enabled the development of a review article. RESULTS: The sclera has been routinely used in ophthalmology since the 1950s in many different indications. Some of these indications have become practically obsolete over time (for example, use in the surgical management of retinal detachment), but a large number still find application today (especially use in glaucoma or oculoplastic surgery, or as a patch for a defect in the sclera or cornea). CONCLUSION: Even though allogeneic sclera is currently used less frequently in ophthalmology compared to other tissue banking products and the range of its indications has partially narrowed, it remains a useful material due to its availability and properties.

Corneal stromal lenticule transplantation for the treatment of corneal ulcers.

PURPOSE: To evaluate the safety and efficacy of using corneal stromal lenticules (CSLs) obtained during refractive surgery Refractive Lenticule Extraction (ReLEx) with the Small Incision Lenticule Extraction (SMILE) procedure for the treatment of corneal ulcers. METHODS: This retrospective study included 12 eyes of 12 patients, 7 men and 5 women with varying degrees of corneal ulcer. The mean age was 64 ± 18 (range 34 to 95 years). The monitoring included corrected distance visual acuity (CDVA), slit-lamp biomicroscopy examination, a Seidel test, stability of the graft and anterior segment optical coherence tomography (AS-OCT) inspection. Patients were closely monitored for possible postoperative complications for at least 6 months. RESULTS: In 7/12 (58%) eyes, the corneal ulcer was successfully sealed with CSL and amniotic membrane (AM) without the need for any additional surgical intervention. In 3 eyes, penetrating keratoplasty (PK) was needed in addition to CSL transplantation and in 2 eyes the scleral patch was used to fully seal after CSL transplantation. During the follow-up period no signs of rejection or infection were detected in any patient. CONCLUSION: The use of CSLs from ReLEx SMILE may be considered as an alternative method for the treatment of corneal ulcers before a more extensive and definitive solution - PK - is used. Our preliminary findings suggest that properly performed CSL transplantation using cryopreserved lenticules is a safe and effective method to temporarily cover the corneal partial-thickness defect or even perforation.

P37-A124†Smile bank - corneal stromal lenticules from relex smile - the new eye bank product.

PURPOSE: The aim of our presentation is to introduce future eye bank product - corneal stromal lenticule from living donors, which can be used for allotransplantation. METHODS: ReLEx (refractive lenticule extraction) SMILE (small incision lenticule extraction) is a common approach in laser eye surgery. It is minimally invasive and flap-free procedure. During this procedure part of corneal stroma (lenticule) is created by femtosecond laser and consequently removed through small incision. The lenticule is basically waste material of the ReLEx SMILE procedure. In the International Eye Bank of Prague, we decided to establish new protocol for lenticule withdrawal, storage and release for transplantation. RESULTS: All donors signed an informed consent, and their serum was tested for the presence of infectious diseases. After ReLEx SMILE procedure the lenticule was stored in container with cryopreservation solution and frozen in the eye bank using the same protocol for frozen amniotic membrane. After 6 months in -80°C tissues were defrosted and examined histologically, using conventional light histology staining and electron microscopy. CONCLUSION: We believe, that lenticule from living donor is a safe and effective tissue, that can be used for many indications and in particular situations represents good alternative to whole donor cornea and amniotic membrane.

The new future perspective in corneal tissue utilisation - methods of preparation and preservation.

PURPOSE: The goal of our study is to find an optimal approach to the preparation and preservation of corneal stromal tissue. We want to compare different methods of corneal stromal tissue creation and storage to optimize the efficacy of this process under the conditions of an eye bank. After we find the most suitable method to create a safe high quality product, we want to prove the possibility of using a single donor cornea for more than one patient. We would also like to verify the feasibility of making more corneal lenticules after the removal of a corneal endothelium for DMEK transplantation. METHODS: We provided morphological (histology, scanning electron microscope) and microbiological analysis in order to compare different methods of corneal lenticule and corneal stromal lamellae preparation and preservation. We also tested the surgical handling of the tissue to secure a safe manipulation of the tissue for clinical use. We compared two methods of corneal lenticule preparation: microkeratome dissection and femtosecond laser. As methods of preservation, we tested hypothermia, cryopreservation at -80 degrees Celsius in DMSO (dimethyl sulfoxide) and storage at room temperature with glycerol. Some intrastromal lenticules and lamellae in each group were previously irradiated with gamma radiation of 25 kGy (KiloGray). RESULTS: Corneal stromal lamellae prepared with a microkeratome have a smoother cut - side surface compared to lamellae prepared with a femtosecond laser. Femtosecond laser preparation caused more irregularities on the surface and we detected more conglomerates of the fibrils, while lamellae made with microkeratome had more sparse network. Using femtosecond laser, we were able to make more than five lenticules from a single donor cornea. Gamma irradiation led to damage of collagen fibrils in corneal stroma and a loss of their regular arrangement. Corneal tissue stored in glycerol showed collagen fibril aggregates and empty spaces between fibrils caused by dehydration. Cryopreserved tissue without previous gamma irradiation showed the most regular structure of the fibrils comparable to storage in hypothermia. CONCLUSION: Our results suggest that formation of a corneal lenticule lamellae by microkeratome results in smoother corneal lenticules, while being much cheaper than formation by femtosecond laser. Gamma irradiation of 25 kGy caused damage of the collagen fibres as well as their network arrangement, which correlated with loss of transparency and stiffer structure. These changes impair possible surgical utilisation of gamma irradiated corneas. Storage in glycerol at room temperature and cryopreservation had similar outcomes and we believe that both methods are appropriate and safe for further clinical use .

Refractive Outcome After Corneal Lenticule Implantation Ex Vivo Non-Human Study.

PURPOSE: To evaluate changes in corneal refractive parameters after implantation of a stromal lenticule of different thickness. We assume that the refractive outcome depends on the optical power of the used lenticule. METHODS: We conducted an ex-vivo non-human study on 33 normotonic porcine eyeballs divided into two groups, for 4D and 8D human lenticule implantation. Corneal stromal lenticules were obtained as a by-product from a laser procedure ReLEx SMILE. We evaluated corneal refractive parameters measured on Oculus Pentacam© device before and immediately after the intrastromal lenticule implantation. RESULTS: There was no statistically significant difference in corneal refractive parameters between the eyeball groups before lenticule implantation. In both groups, the intrastromal implantation in the depth of 300um led to a significant increase of central corneal pachymetry and corneal anterior steepening. In the 4D group the average central corneal pachymetry increased from 903 ± 124.59 to 1230 ± 148.99 (p = 0.0022) and in 8D group from 733.35 ± 69.60 to 1109 ± 161.64 (p = 0.0008). Induced changes in other studied parameters were not statistically significant, Kmax changed from 45.57 ± 2.78 to 72.07 ± 16.83 (p = 0.0094) and Km front from 40.72 ± 1.60 to 48.87 ± 5.83 (p = 0.0037) in 4D group and in the 8D group average Kmax increased from 42.22 ± 1.54 to 62.95 ± 12.67 (p = 0.0001) and K2 front 40.46 ± 1.64 to 51.51 ± 9.63 (p = 0.0037). There were no significant differences in refractive changes between the 4D and 8D groups after lenticule implantation. CONCLUSION: Intrastromal corneal lenticule implantation induces changes in corneal refractive parameters. In both groups, the implantation induced a significant increase of an anterior corneal steepening without any significant influence on posterior corneal flattening. Corneal lenticule implantation did not lead to any significant change of corneal astigmatism. However, in order to have more precise data for future clinical applications we need to continue with the experiments and verify the results on human corneas.

Primary Posterior Continuous Circular Capsulorhexis without the Use of Ophthalmic Viscosurgical Device.

Performing primary posterior capsulorhexis (PPCCC) during cataract surgery is an effective prevention of secondary cataracts. It is important not to damage the anterior vitreous membrane to minimize the possible complications and adverse side effects of this step of the surgery. Most authors use a viscoelastic material to protect the anterior vitreous membrane, injected into the space between the posterior lens capsule and the vitreous membrane through a small hole in the posterior capsule. The creation of posterior capsulorhexis using 25G forceps under an implanted intraocular lens while continuously filling the anterior chamber with irrigation fluid from an irrigation cannula, without the use of a viscoelastic material is, in our opinion, a safe, fast and cheap technique of PPCCC. This technique could theoretically lead to a lower incidence of postoperative complications such as higher postoperative intraocular pressure or anterior segment inflammatory response.

Efficacy and Safety of Deep Sclerectomy With the Esnoper Clip Implant for Uncontrolled Primary Open Angle Glaucoma: A 1 Year Prospective Study.

PRCIS: Deep sclerectomy (DS) with the Esnoper Clip drainage implant in patients with uncontrolled primary open angle glaucoma (POAG) achieved a complete success rate of 87.2% at the 1-year follow-up. PURPOSE: To investigate the efficacy and safety of DS followed by Esnoper Clip implantation in patients with uncontrolled POAG. MATERIALS AND METHODS: In a prospective, consecutive, interventional study, we investigated 39 eyes of 35 patients with uncontrolled POAG who underwent DS with Esnoper Clip implantation. Complete ophthalmologic examinations including corrected visual acuity and intraocular pressure (IOP), were performed preoperatively, and at 1 day, at 1 week as well as at 1, 3, 6, 9, and 12 months postoperatively. Moreover, any goniopunctures and glaucoma medications required postoperatively were noted. RESULTS: The mean preoperative IOP was 20.8±5.2 mm Hg and it decreased to 13.9±3.1 mm Hg at 1 year postoperatively ( P <0.001). The number of glaucoma medications decreased from 2.9±0.7 preoperatively to 0.3±0.8 after 1 year ( P <0.001). The complete success rate (IOP≤21 mm Hg without glaucoma medication) and the qualified success rate (IOP ≤21 mm Hg with or without glaucoma medication) were 87.2% and 94.9%, respectively. Goniopuncture was performed in 33.3% of cases. No significant corrected visual acuity changes were registered at the final follow-up. Perioperative complications consisted of 3 micro-perforations of the trabeculo-descemet membrane. Postoperative complications included: hyphema (6 eyes), hypotony (6 eyes), shallow anterior chamber (3 eyes), choroidal detachment (4 eyes)-all of which were resolved without surgical intervention during the first postoperative month-and conjunctival dehiscence, which required resuture (2 eyes). CONCLUSION: Deep sclerectomy with the Esnoper Clip implant was safe and effectively lowered IOP in patients with uncontrolled POAG.

Effect of the position of the corneal lamella on the frequency of its detachment.

AIM: To determine the influence of gaps [places where neither the donor's nor the recipient's Descemet's membrane (DM) is present] and overlaps (places where the recipient's DM is covered by the donor's DM) on the frequency of postoperative detachment of DM endothelial keratoplasty (DMEK) lamellae. METHODS: Totally 64 eyes of 64 patients with Fuchs' endothelial dystrophy or bullous keratopathy indicated for DMEK were randomly divided in two groups. The diameter of the implanted DMEK lamella was the same in both groups (8 mm), but we changed the diameter of the removed recipient DM. In the first group (32 eyes), the circular area was approximately 8.5 mm (gaps); in the second group (32 eyes), the diameter was 7.5 mm (overlaps). Postoperatively we noted all cases of detachment visible on the slit lamp and these cases we indicated for rebubbling. We also measured the uncorrected distance visual acuity (UDVA) as well as corrected distance visual acuity (CDVA) in decimal and postoperative endothelial cell density (ECD). The minimum follow-up time was 6mo. RESULTS: The number of rebubbling procedures in the entire group of patients was 13, i.e., 20.3%, with 6 eyes (18.7%) in the gap group, and 7 eyes (21.9%) in the overlap group. Lamella replacement (re-DMEK) was required in 3 (gap group) and 2 patients (overlap group), respectively. The difference between the groups was statistically insignificant. The UDVA was 0.54±0.21 in the gap group and 0.58±0.24 in the overlap group. The CDVA was 0.74±0.22 and 0.80±0.16, respectively. ECD was 1920±491 and 2149±570 cells/mm2. The small differences between both groups were not statistically significant. CONCLUSION: We do not notice any difference in the group of patients with overlaps or gaps of DM. The presence of small areas of gaps or overlaps does not affect the frequency of detachment of the DMEK lamellae.

Interleukin-13 increases the stemness of limbal epithelial stem cells cultures.

This study aimed to determine the effect of interleukin-13 (IL13) on the stemness, differentiation, proliferation, clonogenicity, and morphology of cultured limbal epithelial cells (LECs). Human limbal explants were used to culture LECs up to the second passage (P0-P2) with or without IL13 (IL13+ and IL13-, respectively). Cells were analyzed by qPCR (for the expression of ΔNp63α, BMI-1, keratin (K) 3, K7, K12, K14, K17, mucin 4, and MKI67) and immunofluorescence staining for p63α. The clonogenic ability was determined by colony-forming assay (CFA), and their metabolic activity was measured by WST-1 assay. The results of the CFA showed a significantly increased clonogenic ability in P1 and P2 cultures when LECs were cultured with IL13. In addition, the expression of putative stem cell markers (ΔNp63α, K14, and K17) was significantly higher in all IL13+ cultures compared to IL13-. Similarly, immunofluorescence analysis showed a significantly higher percentage of p63α positive cells in P2 cultures with IL13 than without it. LECs cultures without IL13 lost their cuboidal morphology with a high nucleocytoplasmic ratio after P1. The use of IL13 also led to significantly higher proliferation in P2, which can be reflected by a higher ability to reach confluence in P2 cultures. On the other hand, IL13 had no effect on corneal epithelial cell differentiation (K3 and K12 expression), and the expression of the conjunctival marker K7 significantly increased in all IL13+ cultures compared to the respective cell culture without IL13. This study showed that IL13 enhanced the stemness of LECs by increasing the clonogenicity and the expression of putative stem cell markers of LECs while maintaining their stem cell morphology. We established IL13 as a culture supplement for LESCs, which increases their stemness potential in culture, even after the second passage, and may lead to the greater success of LESCs transplantation in patients with LSCD.

Simultaneous Presbyopia and Astigmatism Correction with a Novel Trifocal Toric Intraocular Lens-A One-Year Follow-Up.

The current investigation evaluates the efficiency of the trifocal toric Liberty 677MTY intraocular lens (IOL) in correcting preoperative corneal astigmatism in cataract patients demanding spectacle independence. The retrospective evaluation included 28 eyes of 15 patients with preoperative corneal astigmatism of at least 1.0 Dioptre (D). All patients were followed up for one year postoperatively. Residual refractive errors and visual acuities at multiple distances were measured. Binocular visual acuity and contrast sensitivity defocus curves were plotted. Visual functions and patient satisfaction were assessed. The efficiency of astigmatism correction was determined using the vector analysis method. The mean spherical equivalent refraction (SEQ) improved from 2.72 ± 1.62 D to 0.10 ± 0.48 D. The cylindric refraction decreased from 1.18 ± 0.45 D to 0.16 ± 0.31 D. Vector analysis proved efficient astigmatism correction with a centroid of 0.10 ± 0.34 D at 161°. Ninety-two percent of eyes resulted within 0.5 D from the target refraction. Visual acuities were 0.1 logMAR or better from +1.0 to -3.5 D defocus values. Visual tasks could be performed without major difficulties. Our patients were highly satisfied. Refractive and visual outcomes with the investigated presbyopia-correcting toric IOL are predictable and the lens provides excellent trifocal vision.

Preservation of corneal stromal lenticule: review.

Corneal stromal lenticule is a part of corneal stroma, which can be created by manual dissection, by femtosecond laser from the donor cornea, but chiefly it is a waste product of a refractive procedure ReLex SMILE (Small Incision Lenticule Extraction). Corneal lenticule has a huge potential in corneal surgery. In recent years, many studies have been published to show the possibility to use this tissue to treat corneal defects, as well as in refractive surgery. Thanks to the quantity of lenticules which arise every day during SMILE operations, this tissue is much more accessible than any other kind of corneal tissue. According to the experience with lenticule implantation in animal models, or even human patients, lenticule implantation is considered safe, reversible method, which is not associated with immune rejection or other severe complications. However, the crucial step before the process of lenticule implantation, is proper preservation of this tissue. Donor corneal tissue containing endothelium is usually preserved in hypothermia and then usable maximally for two weeks. Newer methods such as organ culture storage and use of a sterile cornea prolong the time of usability of the tissue. The possibilities for corneal lenticule storage are theoretically wider thanks to the fact, that we do not need to preserve fragile cellular structures. Besides the storage in hypothermia, other preserving methods such as cryopreservation and storage after decellularization have been tested. This review aimed to examine the current literature that describes possible methods of corneal lenticule preservation. A comprehensive search was created based on articles published in English on PubMed.gov, Cochranelibrary.com and Scopus.com using following keywords: corneal lenticule preservation, corneal lenticule storage, cold storage corneal lenticule, corneal lenticule cryopreservation till 2020.

Comparison of Clinical Outcomes, Visual Quality and Visual Function of Two Presbyopia-Correcting Intraocular Lenses Made from the Same Material, but with Different Design and Optics.

This semi-prospective, parallel, comparative investigation evaluated the clinical outcomes and quality of vision (contrast sensitivity, visual function, dysphotopsia, spectacle use, overall satisfaction) after mono- or bilateral implantation of two presbyopia-correcting intraocular lenses (IOL)-the Liberty® 677MY or the AT LISA® tri 839M-in 50 eyes of 25 cataract patients. Clinical outcomes were assessed 3 and 12 months postoperatively. Eighty-nine percent of eyes implanted with the Liberty IOL and 59% of eyes implanted with the AT LISA IOL achieved a refractive outcome ±0.5 diopters of the target (emmetropia). Refractive outcomes were stable with both lenses. The proportions of eyes with 20/20 uncorrected distance visual acuity (UDVA) and 20/20 uncorrected near visual acuity (UNVA) were higher in the Liberty group than in the AT LISA group (UDVA: 56% vs. 41%; UNVA: 83% vs. 66%). Optical quality assessment results were comparable for the two IOLs. Superior photopic contrast sensitivity was found with the Liberty lens. The rate of Nd:YAG capsulotomy at the 12-month follow-up was 16.7% in the Liberty group and 40.6% for the AT LISA IOL. Considering that both lenses are made from the same material, we propose that the noted differences in clinical outcomes may derive from differences in design and optical surface between the two IOLs.

Evaluation of Astigmatism-Correcting Efficiency and Rotational Stability after Cataract Surgery with a Double-Loop Haptic Toric Intraocular Lens: A 1-Year Follow-Up.

OBJECTIVES: The aim of this study was to assess the clinical outcomes, predictability of results, efficiency of astigmatism correction, and rotational stability of the Bi-Flex 677TAY (Medicontur Medical Engineering Ltd., Zsámbék, Hungary) monofocal toric intraocular lens (IOL) designed for cataract patients with astigmatism. METHODS: The IOLs were implanted either mono- or binocularly, following routine cataract surgery. Visual and refractive outcomes, as well as off-axis rotation were assessed throughout a 1-year follow-up period. All clinical data for this work were collected retrospectively. Vector analysis based on the Alpins method was performed to assess the efficiency of astigmatism correction. RESULTS: No complications or adverse events occurred during surgery or the follow-up period. IOL implantation brought 88% of eyes into the ±0.50 D, and 100% into the ± 1.00 D range compared to the target spherical equivalent refraction, emmetropia. Astigmatism correction brought similar results: 94% of eyes had a residual cylindrical error of not higher than ±0.50 D, and 97% were within ±1.00 D. Vector analysis resulted in a correction index of 0.96 and a difference vector of 0.17. Both refractive and visual outcomes showed long-term stability. During the 12-month follow-up period, no eyes had a rotation of >5°. Absolute rotation after 1 year was 1.42 ± 1.89° (median = 0°), while signed rotation was 1.06 ± 2.12° (median = 0°). CONCLUSION: The Bi-Flex 677TAY monofocal toric IOL, designed by Medicontur Medical Engineering Ltd., represents an efficient and safe solution for cataract patients with astigmatism. Clinical and refractive outcomes are predictable, and rotational stability ensures long-term visual comfort.

Long-term Results of a Combined Procedure of Cataract Surgery and Descemet Membrane Endothelial Keratoplasty With Stromal Rim.

PURPOSE: To evaluate the long-term results of combining cataract surgery, intraocular lens (IOL) implantation, and Descemet membrane endothelial keratoplasty with peripheral stromal support (DMEK-S). METHODS: The outcomes of 107 eyes of 37 patients who had undergone a combination of cataract surgery, IOL implantation, and DMEK-S between October 2007 and February 2015 were retrospectively evaluated. The average follow-up duration was 18.2 ± 18.3 months (range, 6-84 months). Changes in uncorrected and corrected distance visual acuity, spherical equivalent, and corneal endothelial cell density were analyzed. Intraoperative and postoperative complications were also recorded. RESULTS: The mean logMAR uncorrected distance visual acuity improved from 0.92 ± 0.60 preoperatively to 0.33 ± 0.33, 0.28 ± 0.25, 0.24 ± 0.20, and 0.27 ± 0.27 at 6, 12, 24, and 36 months postoperatively (P < 0.001), respectively. The mean logMAR corrected distance visual acuity improved from 0.62 ± 0.65 preoperatively to 0.17 ± 0.29, 0.13 ± 0.21, 0.07 ± 0.16, and 0.11 ± 0.23 at 6, 12, 24, and 36 months postoperatively (P < 0.001), respectively. The mean endothelial cell density decreases were 50.7% ± 21.1%, 50.8% ± 19.8%, 60.7% ± 16.6%, 62.2% ± 11.8%, and 61.8% ± 20.0% at 6, 12, 24, 36, and 48 months after surgery, respectively. Similar trends, although with slightly better visual outcomes, were found in the subgroup of patients without other eye diseases. The mean final deviation from intended spherical equivalent was 0.77 ± 1.19 D, representing a hyperopic shift. CONCLUSIONS: A combined procedure of cataract surgery, IOL implantation, and DMEK-S is an effective method of treatment in patients with combined corneal disease and cataract, maintaining visual rehabilitation in the long term. Future developments should be performed to minimize the incidence of complications.

Discontinuous transcription of ribosomal DNA in human cells.

Numerous studies show that various genes in all kinds of organisms are transcribed discontinuously, i.e. in short bursts or pulses with periods of inactivity between them. But it remains unclear whether ribosomal DNA (rDNA), represented by multiple copies in every cell, is also expressed in such manner. In this work, we synchronized the pol I activity in the populations of tumour derived as well as normal human cells by cold block and release. Our experiments with 5-fluorouridine (FU) and BrUTP confirmed that the nucleolar transcription can be efficiently and reversibly arrested at +4°C. Then using special software for analysis of the microscopic images, we measured the intensity of transcription signal (incorporated FU) in the nucleoli at different time points after the release. We found that the ribosomal genes in the human cells are transcribed discontinuously with periods ranging from 45 min to 75 min. Our data indicate that the dynamics of rDNA transcription follows the undulating pattern, in which the bursts are alternated by periods of rare transcription events.

The utility of massively parallel sequencing for posterior polymorphous corneal dystrophy type 3 molecular diagnosis.

The aim of this study was to identify the molecular genetic cause of disease in posterior polymorphous corneal dystrophy (PPCD) probands of diverse origin and to assess the utility of massively parallel sequencing in the detection of ZEB1 mutations. We investigated a total of 12 families (five British, four Czech, one Slovak and two Swiss). Ten novel and two recurrent disease-causing mutations in ZEB1, were identified in probands by Sanger (n = 5), exome (n = 4) and genome (n = 3) sequencing. Sanger sequencing was used to confirm the mutations detected by massively parallel sequencing, and to perform segregation analysis. Genome sequencing revealed that one proband harboured a novel ∼0.34 Mb heterozygous de novo deletion spanning exons 1-7 and part of exon 8. Transcript analysis confirmed that the ZEB1 transcript is detectable in blood-derived RNA samples and that the disease-associated variant c.482-2A>G leads to aberrant pre-mRNA splicing. De novo mutations, which are a feature of PPCD3, were found in the current study with an incidence rate of at least 16.6%. In general, massively parallel sequencing is a time-efficient way to detect PPCD3-associated mutations and, importantly, genome sequencing enables the identification of full or partial heterozygous ZEB1 deletions that can evade detection by both Sanger and exome sequencing. These findings contribute to our understanding of PPCD3, for which currently, 49 pathogenic variants have been identified, all of which are predicted to be null alleles.