Rapamycin Controls Lymphoproliferation and Reverses T-Cell Responses in a Patient with a Novel STIM1 Loss-of-Function Deletion.

PURPOSE: Deficiency of stromal interaction molecule 1 (STIM1) results in combined immunodeficiency accompanied by extra-immunological findings like enamel defects and myopathy. We here studied a patient with a STIM1 loss-of-function mutation who presented with severe lymphoproliferation. We sought to explore the efficacy of the mTOR inhibitor rapamycin in controlling disease manifestations and reversing aberrant T-cell subsets and functions, which has never been used previously in this disorder. METHODS: Clinical findings of the patient were collected over time. We performed immunological evaluations before and after initiation of rapamycin treatment, including detailed lymphocyte subset analyses, alterations in frequencies of circulating T follicular helper (cTFH) and regulatory T (Treg) cells and their subtypes as well as T cell activation and proliferation capacities. RESULTS: A novel homozygous exon 2 deletion in STIM1 was detected in a 3-year-old girl with severe lymphoproliferation, recurrent infections, myopathy, iris hypoplasia, and enamel hypoplasia. Lymphoproliferation was associated with severe T-cell infiltrates. The deletion resulted in a complete loss of protein expression, associated with a lack of store-operated calcium entry response, defective T-cell activation, proliferation, and cytokine production. Interestingly, patient blood contained fewer cTFH and increased circulating follicular regulatory (cTFR) cells. Abnormal skewing towards TH2-like responses in certain T-cell subpopulations like cTFH, non-cTFH memory T-helper, and Treg cells was associated with increased eosinophil numbers and serum IgE levels. Treatment with rapamycin controlled lymphoproliferation, improved T-cell activation and proliferation capacities, reversed T-cell responses, and repressed high IgE levels and eosinophilia. CONCLUSIONS: This study enhances our understanding of STIM1 deficiency by uncovering additional abnormal T-cell responses, and reveals for the first time the potential therapeutic utility of rapamycin for this disorder.

Factors affecting prognosis and need for anti-vascular endothelial growth factor injections in wet age-related macular degeneration.

PURPOSE: To understand factors affecting visual prognosis and the number of intravitreal antivascular endothelial growth factor (anti-VEGF) injections needed to stabilize wet age-related macular degeneration (AMD). METHODS: In this retrospective cohort, 119 treatment-naïve wet AMD patients were followed for two years. In patients with bilateral disease, the eye with worse best-corrected visual acuity (BCVA) or that received more intravitreal injections was recruited as the study eye. In all visits, BCVA was recorded, ophthalmological examination was performed including macular optical coherence tomography imaging. Twenty health status/lifestyle questions were asked to the patients via phone as potential risk factors. All patients received 3 loading doses of intravitreal bevacizumab injections and received repeat injections of aflibercept or ranibizumab when the eye had a new, active neovascular lesion. RESULTS: Patients who took regular micronutrition had similar visual outcome and injection numbers compared to the ones who did not. Patients with bilateral disease needed less intravitreal injections compared to unilateral AMD patients (p = 0.016) and women on hormone replacement therapy (HRT) required less injections compared to the women who were not (p = 0.024). Female patients had a mean gain of 2.7 letters while male patients lost 3.8 letters (p = 0.038). Wet AMD started at an earlier age in smokers (p = 0.002). Patients with a better education level presented earlier with better BCVA (p = 0.037). CONCLUSION: HRT and anti-VEGF injections to the fellow eye improved the prognosis of wet AMD, while male patients had slightly worse prognosis. Estrogen's protective effects and potential contribution in wet AMD needs further attention. Retrospectively registered: 2020/0622.

Endothelial loss following postoperative intracameral triamcinolone acetonide and subconjunctival dexamethasone injections.

OBJECTIVES: To compare endothelial toxicity and efficacy of two local steroid injections (intracameral triamcinolone acetonide and subconjunctival dexamethasone) in controlling postoperative inflammation following pars plana vitrectomy (PPV) combined with phacoemulsification cataract surgery. METHODS: This cohort included 54 patients that underwent combined surgery and received either intracameral triamcinolone acetonide injections (n = 27, IC-TA group) or subconjunctival dexamethasone (n = 27, Sc-Dex group) injections at the end of the surgery. All participants had at least 4 months or longer follow-up. A detailed ophthalmologic examination including intraocular pressure (IOP) measurement and specular microscopy was performed at every visit. RESULTS: Endothelial cell density (ECD) reduced significantly in IC-TA group postoperatively (2418 vs. 2249, p = 0.019), while it did not change significantly in Sc-Dex group (2541 vs. 2492, p = 0.247). Postoperative ECD was also significantly lower in IC-TA group compared to Sc-Dex group (p = 0.011). Preoperative and postoperative IOP values remained unchanged both in IC-TA and Sc-Dex groups (p = 0.424 and p = 0.523, respectively). However, 4 patients in IC-TA group and 5 patients in the Sc-Dex group needed glaucoma medications. The postoperative need for glaucoma medications was similar between the groups (p = 0.347). Postoperative inflammation was well controlled in both groups and none of the patients developed fibrin membrane or synechiae postoperatively. CONCLUSION: Both treatments were effective in controlling postoperative inflammation, but patients in IC-TA group experienced significantly higher endothelial loss. Sc-Dex injections are safer in terms of endothelial loss and preferable to control postoperative inflammation following complex intraocular surgeries.

Corneal neuropathic pain in irritable bowel syndrome: clinical findings and in vivo corneal confocal microscopy.

PURPOSE: Ocular discomfort is a common symptom in central sensitization syndromes. The aim of this study was to evaluate ocular surface discomfort and related corneal changes in patients with irritable bowel syndrome. METHODS: Twenty-nine patients with IBS (20 female, 9 male, mean age: 45.3 ± 10.1 years) and 37 healthy control subjects (25 female, 12 male, mean age: 44.95 ± 9.76 years) were included. A detailed ophthalmological examination was performed to all participants including tear break-up time (TBUT) and Schirmer test I with anesthetic (SIT). Ocular discomfort was evaluated using the ocular surface disease index (OSDI) questionnaire and corneal sensation was evaluated with Cochet-Bonnet esthesiometer. Corneal subbasal nerve plexus was evaluated with in vivo corneal confocal microscopy (IVCM). RESULTS: There was no significant difference between the groups for age, gender distribution, and visual acuity. OSDI scores were significantly higher (p = 0.008) and TBUT was significantly reduced in patients with IBS compared to controls (p = 0.001 for right eye, p = 0.014 for left eye). However, there was no significant difference in corneal touch sensation and SIT results between the groups. IVCM revealed that corneal nerve fiber density, corneal nerve branch density, and corneal nerve fiber length were significantly reduced in patients with IBS (p < 0.001, p < 0.001, and p = 0.023, respectively). CONCLUSION: Patients with IBS have increased dry eye-associated ocular surface complaints and nerve fiber loss in corneal subbasal nerve plexus. IBS should be remembered in the differential diagnosis, when there is discordance between the level of ocular surface discomfort and dry eye disease associated corneal findings.

Double-Organ Bias in Controlled Trials on Eyelid Ptosis.

PURPOSE: Proper statistical analysis is an essential element in the evaluation of clinical trial outcomes. We had the informal observation that double-organ bias was a neglected issue during the statistical analyses of clinical trials on eyelid ptosis. The aim of this study was to formally document the prevalence of this bias in these studies. METHODS: Clinical trials on eyelid ptosis, published in the last 20 years, were searched in PubMed with the terms; "((blepharoptosis) OR upper eyelid ptosis) OR eyelid ptosis" and with the filters "Randomized Controlled Trial (RCT), Clinical Trial, Humans." Two independent observers evaluated the articles for eligibility, field of the journal, field of the authors, presence of randomization, type of the study (surgical, medical), and statistical approach to double organ bias. RESULTS: The PubMed search yielded 101 articles and 23 of them met the above-mentioned criteria. In 3 articles, primary outcome measure was not related with the eyes. Among the remaining 20 articles, 14 (70%) had double-organ bias in the statistics. The bias was slightly less common in randomized trials (60% vs. 80%), but the difference was not statistically significant. Its prevalence was similar between ophthalmology journals and the rest (p = 0.64). Interestingly, the bias tended to be more in Q1 journals (87.5%) compared to Q2 and Q3 journals (58.3%) and median impact factor was higher in biased articles (1.82 vs. 1.29), but the differences weren't statistically significant (p = 0.32, p = 0.24). There was no difference between the last 2 decades (66.6% vs. 75%, p = 0.64). CONCLUSIONS: The prevalence of the double-organ bias was very high in published clinical trials on eyelid ptosis (70%) and even among RCTs (60%). The prevalence of the bias didn't prevent publication in higher impact factor journals and didn't change between journals from different disciplines or over time.

Refractive outcome of trocar-assisted sutureless scleral fixation with 3-piece intraocular lenses.

PURPOSE: Intraocular lens (IOL) implantation with a trocar-assisted sutureless scleral fixation technique is a relatively new IOL implantation approach for aphakic eyes. During this surgery, the intrascleral position of the haptics can change the location of the IOL optic and may alter the refractive outcome. This study aimed to evaluate the refractive outcome of this surgery. METHODS: The files of 22 patients who had undergone IOL implantation with the trocar-assisted sutureless scleral fixation technique were retrospectively reviewed, and the patients were invited for final examination. IOL power was calculated with optical biometry (Lenstar LS900). IOL power calculations were performed according to formulas designed for in-the-bag IOL implantation. The final refractive error was determined with an autorefractometer (Topcon KR-1/RM-1). RESULTS: This study included 14 patients (8 male, 6 female, mean age: 62.7 ± 18.7). There were no significant differences between the preoperative and postoperative corneal astigmatism values (p = 0.16). There were also no significant differences between the postoperative corneal and total astigmatism values (p = 0.44), confirming the absence of significant IOL tilt. The difference between the implanted IOLs and the calculated IOL power for emmetropia was 0.09 D ± 0.49 D (p = 0.52). Although emmetropia was targeted, the refractive outcome shifted to hyperopia (+ 0.85 D ± 1.15 D) and was significantly different from the refractive status calculated with optical biometry (p = 0.034). CONCLUSIONS: The intrascleral fixation of IOLs results in increased posterior positioning of the IOL optic and can induce approximately 0.85 D of hyperopia if proper adjustments are not performed during IOL power calculations.

Clinical outcome and endothelial loss following prepupillary and retropupillary implantation of iris claw intraocular lenses.

PURPOSE: Iris claw intraocular lenses (IOLs) were successfully used for the management of aphakia in patients that lack capsular support. The aim of this study was to compare the clinical outcome of prepupillary and retropupillary implantation of these IOLs. METHODS: The files of the 26 patients that had an iris claw IOL implantation between 2010 and 2020 were retrospectively reviewed. Detailed ophthalmological examination findings including corrected distance visual acuity (CDVA), intraocular pressure, endothelial cell counts, slit lamp and dilated fundus examination findings were specifically tabulated. Intraoperative and postoperative complications were also specifically recorded. RESULTS: There were 18 patients in the prepupillary implantation group and eight patients in the retropupillary implantation group. Age and gender distribution were similar between the groups. CDVA significantly increased in prepupillary and retropupillary implantation groups (p = 0.001 and p = 0.012, respectively). Median endothelial cell loss was 6.7% in prepupillary group and 7.2% in retropupillary group. The only intraoperative complication was iridodialysis (n = 1). Postoperative complications included retinal detachment (n = 1), cystoid macular edema (n = 2) and IOL tilt (n = 1). All of these complications occurred in the prepupillary implantation group. CONCLUSION: Prepupillary and retropupillary implantation of iris claw IOLs resulted with similar visual gain and endothelial loss rates in a follow-up time of 6 months. There was an insignificant trend toward a reduced complication rate following retropupillary implantation.

Complete blood count parameters and neutrophil-to-lymphocyte ratio values as markers for differentiation between systemic infectious and non-infectious uveitis.

PURPOSE: To compare complete blood count (CBC) parameters and neutrophil/lymphocyte ratio (NLR) between patients with infectious uveitis (IU) and those with non-infectious uveitis (NIU) during the first acute uveitis attack (AUA). METHODS: The records of 119 patients admitted with the first AUA between 2016 and 2019 and whose diagnosis was unknown at the time of admission were retrospectively reviewed. The patients were divided into two groups, IU and NIU according to diagnoses after ocular and systemic workup. The IU group was also divided into subgroups as uveitis associated with local ocular infections and systemic infections. The complete blood count and associated indices of patients calculated from samples taken during the attack were compared between the groups. RESULTS: A total of 60 NIU cases (mean age: 43.5 ± 11.6 years) and 59 IU cases (43.3 ± 14.7 years) were examined. Twenty-six of the NIU cases were female and 34 were male, while 32 of the IU cases were female and 27 were male. The localization of uveitis was similar in the IU and NIU groups (anterior: 11 vs. 18, intermediate: 3 vs. 6, posterior: 28 vs. 14, panuveitis: 17 vs. 22). The NLR values were significantly increased in patients with IU compared to those with NIU (p = 0.047). When the NLR is compared between NIU and subgroups of IU, this value was only found significantly increased in uveitis due to a systemic infection (n = 12) (p < 0.001). The mean white blood cell (WBC) count (p < 0.001) and neutrophil values (p < 0.001) were also observed significantly higher in the uveitis associated with systemic infectious diseases when compared with uveitis associated with local ocular infections and NIU group. CONCLUSION: In the current study, NLR values are compared with autoimmune uveitis and systemic infectious uveitis for the first time in patients with AUA. This ratio has been found significantly higher in uveitis associated with systemic infections. The use of CBC parameters and indices, especially the NLR, may be instrumental in assessing patients presenting with first AUA, particularly when there is no clear explanation or underlying cause.

Patterns of Altered Corneal Sensation in Patients With Chronic Migraine.

OBJECTIVE: Migraine is a central sensitization disease characterized by recurrent unilateral headache. The aim of this study was to evaluate corneal sensitivity and associated findings in patients with migraine. METHODS: In this study, we included 58 patients with migraine (52 female and 6 male, mean age: 32.98±8.51 years) and 30 healthy control subjects (27 females and 3 males, mean age: 35.29±8.00 years). Only patients who did not have any eye disease were included in this study. Corneal sensation was evaluated with Cochet-Bonnet esthesiometer in five different regions of the cornea. RESULTS: There was no significant difference between the groups for age, sex, and visual acuity. Increased mean sensitivity of the nasal cornea in the patients with migraine was found (55.0 vs. 53.75 mm, P=0.020). There was no significant difference in mean sensitivity for central (57.5 vs. 55 mm, P=0.163), temporal (57.5 vs. 55 mm, P=0.066), superior (52.5 vs. 52.5 mm, P=0.551), and inferior cornea (48.75 vs. 47.5 mm, P=0.818), or mean overall sensation (54.0 vs. 52.75, P=0.14), although there was a trend toward increased sensitivity in the patients with migraine. Among the subgroup of patients with unilateral migraine, mean overall sensation (54.0 vs. 53.0 mm, P=0.049) and temporal sensation (57.5 vs. 55.0 mm, P=0.043) were increased on the affected side. CONCLUSION: In this study, we have demonstrated that patients with migraine have increased corneal sensation, especially in the cornea of the affected side. This finding supports the idea that corneal sensation is altered in patients with migraine.

Efficacy and safety of low-concentration, bisulphite-containing, intracameral epinephrine and topical atropine treatments for the prevention of intraoperative floppy iris syndrome.

PURPOSE: The aim of this study was to evaluate the efficacy and safety of topical atropine and topical atropine combined with intracameral low-concentration, bisulphite-containing epinephrine treatment for the prophylaxis of intraoperative floppy iris syndrome. MATERIALS AND METHODS: Seventy-two eyes of 55 male patients who were treated with alpha-adrenergic antagonist medications for benign prostatic hyperplasia were included in this study. Standard premedication with topical cyclopentolate, phenylephrine, tropicamide and ketorolac was applied to all of the patients. In 22 cases no further prophylactic method was used (Group-NP), while in 29 cases topical atropine drops was instilled 12 h and 30 min before surgery (Group-A) and in 21 cases 1:16 000 epinephrine was injected to the anterior chamber at the beginning of surgery (Group-EA) in addition to topical atropine prophylaxis. RESULTS: In Group-NP, 62.8% of the cases developed IFIS, while development of IFIS was significantly lower in Group-A (17.2%, p = 0.0004) and Group-EA (9.5%, p < 0.0001). Posterior capsule rupture was observed in two cases (9.1%) in Group-NP, in one case (3.4%) in Group-A and was not observed in Group-EA. There was no statistically significant difference between the groups for the development of surgical complications. We did not observe any adverse events or significant endothelial cell loss (p = 0.462). CONCLUSIONS: Our results indicate that preoperative use of topical atropine reduces the incidence of IFIS. Use of low-concentration, bisulphite-containing epinephrine is more effective in the prevention of IFIS and does not cause preservative related endothelial damage. This prophylaxis may be preferred when preservative free epinephrine is not available.

Influence of Apical Clearance on Mini-Scleral Lens Settling, Clinical Performance, and Corneal Thickness Changes.

PURPOSE: The purpose of this work was to define the timing and magnitude of scleral contact lens settling, the factors influencing settling, and to examine whether the amount of apical clearance has an impact on clinical performance or is associated with hypoxia-induced corneal changes. METHODS: Eleven patients (22 eyes) with keratoconus were fitted with a mini-scleral lens (Esclera, Brazil). Three different lenses with successively greater sagittal depths were applied to achieve 3 levels of initial apical clearance: 100 to 200 µm (low), 200 to 300 µm (medium), and greater than 300 µm (high). Corneal apical clearance was measured at 15 min, 1, 2, 4, 6, and 8 hr with optical coherence tomography (OCT). Central corneal thickness was measured with OCT and Pentacam, in the morning and immediately after removal of the contact lens. RESULTS: Mean settling was 26.8±18.8 µm (42.7%) at 1 hr, 39.5±26.5 µm (62.9%) at 2 hr, 50.7±31.6 µm (80.8%) at 4 hr, 57.4±34.6 µm (91.4%) at 6 hr, and 62.8±38.4 µm (100%) at 8 hr. Settling rate was significantly lower in low apical clearance group (P=0.01). The smaller diameter lenses settled more (P=0.03). There was a slight statistically significant increase of 1.3% in central corneal thickness measured with OCT (P=0.03). Central corneal thickness measured with Pentacam at three locations increased slightly (P=0.001). Settling showed significant intrasubject and intersubject variations. The amount of corneal swelling and comfort scores did not differ significantly according to apical clearance. CONCLUSIONS: The average amount of settling was 62.8 µm after 8 hr, 80% of which occurred during the first 4 hr. Slight corneal swelling (1.3%) occurred after 8 hr of wear.

Orbital myositis associated with celiac disease.

A 26-year-old female patient presented with redness, lid edema on the right eye and diplopia on left gaze. There was a minimal limitation of adduction of the right eye, accompanied with pain and diplopia. Orbital magnetic resonance imaging confirmed the diagnosis of orbital myositis. Anti-endomysial immunoglobulin (Ig) A, anti-endomysial IgG, anti-gliadin IgA and anti-tissue transglutaminase IgG antibodies were positive, while other tests for autoimmune diseases were unremarkable. The patient stated that her diarrhea had been relieved by a gluten-free diet. Methylprednisolone therapy (1 mg/kg) was started and on her next visit her complaints were relieved. Later, methotrexate (15 mg/week) was added to the therapy as the patient became steroid-dependent, and she has been attack-free for the last 3 months under methotrexate and low-dose methylprednisolone (4 mg/day) treatment. Associations between orbital myositis and various autoimmune diseases have previously been reported. We report here the first case of associated orbital myositis and celiac disease.

Autologous-Fibrin Induction: A Novel Approach for Sutureless Sclerotomy Closure in Pars Plana Vitrectomy.

PURPOSE: This study presents a novel sutureless closure approach for sclerotomies following pars plana vitrectomy (PPV) and assesses its efficacy and safety. METHODS: A total of 142 eyes were included in the study. PPV procedures were performed using 23-gauge (23 G) or 25-gauge (25 G) systems. Preoperative characteristics, intraoperative findings, and postoperative outcomes were documented. RESULTS: The cohort included 80 males and 62 females (mean age: 60.4 ± 12 years), primarily undergoing surgery for retinal detachment (59%). Among the patients, 87% underwent 25 G PPV (35% three-port, 52% four-port), while 13% underwent 23 G PPV (12% three-port, 1% four-port). Gas tamponade was administered in all cases, with perfluoropropane used in 45.7% of instances, sulfur hexafluoride in 29.5%, and air in 24.6%. Spontaneous closure was observed in 9.4% (47 of 501) of sclerotomies, autologous-fibrin induction approach successfully closed 75.8% (380 of 501) of the sclerotomies (83.7% of leaking sclerotomies) and 14.7% (74 of 501) of sclerotomies needed sutures. Visual acuity improved postoperatively, and first-day hypotony rate was 6.3%. Importantly, no serious complications such as choroidal detachment or endophthalmitis were observed during the postoperative period. CONCLUSION: The autologous-fibrin induction offers a simple, cost-efficient, and reliable approach for sutureless sclerotomy closure in PPV, with promising outcomes.

Clinical characteristics of pediatric noninfectious uveitis and risk factors for severe disease: a single-center study.

OBJECTIVES: We aimed to present the demographic, clinical, laboratory, and treatment data of children with non-infectious uveitis and to evaluate the risk factors for the development of complications and the need for biological treatment. METHOD: Patients diagnosed with non-infectious uveitis in childhood and followed up for at least 1 year were included in the study. Demographic data, including age, gender, age at diagnosis, uveitis in first-degree relatives, and rheumatologic diseases, were obtained retrospectively from medical records. The presence of complications or the need for biologic therapy was considered a composite outcome suggesting severe disease. RESULTS: The study included 123 patients (female: n = 59, 48%). The mean age at diagnosis was 14.89 ± 4.86 years. Uveitis was symptomatic in 104 patients (84.6%). Approximately one-quarter of the patients had at least one rheumatic disease (n = 35, 28.5%), the most common being juvenile idiopathic arthritis. Thirty-three patients (26.8%) had anti-nuclear antibody positivity. Biologic agents were needed in 60 patients (48.8%). Complications developed in 14 patients (11.4%). Early age at disease onset (aOR, 0.875; 95% C.I. 0.795-0.965, p = 0.007) and female gender (aOR, 2.99; 95% C.I. 1.439-6.248, p = 0.003) were significantly associated with the need for biologic treatment, while Behçet's disease (BD) was strongly associated with uveitis-related complications (aOR, 14.133; 95% C.I. 2.765-72.231, p = 0.001). CONCLUSION: We suggest that among pediatric patients with non-infectious uveitis, females, those with an early age of disease onset, and those with BD need to be closely monitored due to a significantly increased risk of severe disease. Key Points • Limited data exist on the clinical course of non-infectious uveitis in children and the associated risk factors for severe disease. • Our study reveals that nearly a quarter of pediatric patients with non-infectious uveitis also have a rheumatic disease. • Among pediatric patients diagnosed with non-infectious uveitis, we observed an increased risk of severe disease in those with an earlier onset age, in female patients, and in those diagnosed with Behçet's disease.

Bone marrow-derived CMPs and GMPs represent highly functional proangiogenic cells: implications for ischemic cardiovascular disease.

Clinical studies using bone marrow-derived proangiogenic cells (PACs) have demonstrated modest improvements of function and/or perfusion of ischemic myocardium or skeletal muscle. Because the identities of these PACs and their functional ability to promote neovascularization remain poorly understood, it is possible that a subset of robust PACs exists but is obscured by the heterogeneous nature of this cell population. Herein, we found that common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs) preferentially differentiate into PACs compared with megakaryocyte-erythrocyte progenitors, hematopoietic stem cells, and common lymphoid progenitors. In vivo hindlimb ischemia studies and Matrigel plug assays verified the enhanced neovascularization properties uniquely associated with PACs derived from CMPs and GMPs. Taken together, these observations identify CMPs and GMPs as key bone marrow progenitors for optimal PAC function in vitro and in vivo and provide a foundation for novel therapeutic approaches to modulate angiogenesis.

TGF-ß1 signaling and Krüppel-like factor 10 regulate bone marrow-derived proangiogenic cell differentiation, function, and neovascularization.

Emerging evidence demonstrates that proangiogenic cells (PACs) originate from the BM and are capable of being recruited to sites of ischemic injury where they contribute to neovascularization. We previously determined that among hematopoietic progenitor stem cells, common myeloid progenitors (CMPs) and granulocyte-macrophage progenitor cells (GMPs) differentiate into PACs and possess robust angiogenic activity under ischemic conditions. Herein, we report that a TGF-ß1-responsive Krüppel- like factor, KLF10, is strongly expressed in PACs derived from CMPs and GMPs, ∼ 60-fold higher than in progenitors lacking PAC markers. KLF10(-/-) mice present with marked defects in PAC differentiation, function, TGF-ß responsiveness, and impaired blood flow recovery after hindlimb ischemia, an effect rescued by wild-type PACs, but not KLF10(-/-) PACs. Overexpression studies revealed that KLF10 could rescue PAC formation from TGF-ß1(+/-) CMPs and GMPs. Mechanistically, KLF10 targets the VEGFR2 promoter in PACs which may underlie the observed effects. These findings may be clinically relevant because KLF10 expression was also found to be significantly reduced in PACs from patients with peripheral artery disease. Collectively, these observations identify TGF-ß1 signaling and KLF10 as key regulators of functional PACs derived from CMPs and GMPs and may provide a therapeutic target during cardiovascular ischemic states.

Darbepoetin α ameliorates neuronal damage in a rat model of acute ethanol intoxication.

OBJECTIVE: Acute ethanol intoxication has been shown to cause oxidative damage in many organ systems including the brain. Erythropoietin has antioxidant effects and prevents neuronal damage in the animal model of ischemic brain injury. In this study, we aimed to investigate the effects of darbepoetin alpha, an analog of erythropoietin with a longer half-life and higher in vivo activity, on ethanol-induced acute brain injury. METHODS: Forty-eight Wistar albino rats were allocated to four groups. The first group received ethanol treatment (E), the second group was treated with ethanol and darbepoetin (ED), the third group received only saline treatment (S), and the fourth group received both saline and darbepoetin treatment (SD). Plasma S100-ß and neuron-specific enolase (NSE) levels were measured. Histopathological evaluation of the brains was performed. RESULTS: The plasma S100-ß and NSE levels were significantly lower in group ED compared with group E. In group E, we have observed focal red-neuron formation at the granular layer of the dentate gyrus. We did not observe any histopathological changes in the other groups (ED, S, and SD). CONCLUSION: Our findings suggest that darbepoetin alpha has neuroprotective effect in acute ethanol intoxication, possibly through its antioxidant effect.

Long-term follow-up of choroidal changes following COVID-19 infection: analysis of choroidal thickness and choroidal vascularity index.

OBJECTIVE: To investigate subclinical choroidal involvement in patients with systemic coronavirus disease 2019 (COVID-19) infection and evaluate its long-term course. MATERIALS AND METHODS: This prospective, longitudinal study included 32 eyes of 16 COVID-19 patients and 34 eyes of 17 age-matched healthy control subjects. All the participants had a detailed ophthalmologic assessment, including visual acuity assessment, slit-lamp examination, and indirect ophthalmoscopy. Enhanced depth optical coherence tomography imaging of the posterior pole and peripapillary region was performed in the early (days 15-40) and late (ninth month) postinfectious periods. Choroidal vascularity index (CVI) was calculated using ImageJ software (National Institutes of Health, Bethesda, Md.). RESULTS: None of the patients had any examination finding associated with the ocular involvement of COVID-19. Subfoveal choroidal thickness (SFCT) decreased significantly in the early postinfectious period compared with the healthy control individuals (p = 0.045). SFCT increased significantly in the late postinfectious period compared with the early period (p = 0.002), and the difference between patients and control individuals became statistically insignificant (p = 0.362). There was a similar trend for the peripapillary choroidal thickness measurements. CVI remained unchanged (p = 0.721) despite the significant decrease in SFCT and total choroidal area (p = 0.042), indicating that this decrease occurred both in choroidal stroma and in blood vessels. CVI remained unchanged in the late postinfectious period (p = 0.575) compared with the early period, indicating that recovery occurred in the entire choroidal tissue. CONCLUSION: This study demonstrates that choroidal thickness was reduced in all measured areas and that this decrease affected all choroidal layers. This choroidopathy was reversible and recovered in the ninth postinfectious month.

Visual Outcomes, Endothelial Loss, and Complications Following Iris-Claw Intraocular Lens Implantation and Trocar-Assisted Sutureless Scleral Fixation of Intraocular Lenses.

Objectives: Iris-claw intraocular lens (IC-IOL) implantation and sutureless scleral fixation of intraocular lenses (SSF-IOL) are two commonly preferred surgical approaches for the management of aphakic patients without sufficient capsular support. The aim of this study was to compare the outcomes of IC-IOL implantation and trocar-assisted SSF-IOL implantation. Methods: The medical records of secondary IOL implantation patients were retrospectively reviewed. All patients had a detailed ophthalmological examination, including LogMAR best-corrected distance visual acuity (CDVA), intraocular pressure (IOP), and endothelial cell density (ECD) preoperatively and postoperatively. SPSS 21.0 software was used for the statistical analysis. Results: There were 15 patients in the IC-IOL group and 12 patients in the SSF-IOL group. Age and gender distributions were similar between the groups (p=0.456 and p=0.398, respectively). Similarly, patients in both groups had similar CDVA preoperatively and postoperatively (p=0.51, p=0.48, respectively). Both IC-IOL and SSF-IOL implantation significantly increased CDVA (p=0.001 and p=0.005, respectively). IOP remained unchanged in both groups. However, ECD reduced significantly following both IC-IOL and SSF-IOL implantation (p=0.001 and p=0.005, respectively) and trocar-assisted SSF-IOL implantation resulted in significantly more endothelial loss compared to IC-IOL implantation (439.5±89 vs. 164.4±53, p=0.013). Conclusion: Both surgical approaches increased CDVA significantly and at similar levels. However, trocar-assisted SSF-IOL implantation resulted in significantly more endothelial loss compared to IC-IOL implantation. None of the patients developed bullous keratopathy, but this difference should be kept in mind, especially in patients with critically low ECD.

Mediastinal lymph node removal modulates natural killer cell exhaustion in patients with non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death globally. In this study, the effect of complete removal of mediastinal lymph nodes by video-assisted mediastinoscopic lymphadenectomy (VAMLA) on natural killer (NK) cell phenotype and functions in patients with NSCLC was evaluated. The study included 21 NSCLC patients (cIA-IVA) undergoing VAMLA staging and 33 healthy controls. Mononuclear cells were isolated from peripheral blood of all participants and mediastinal lymph nodes of the patients. NK cells were analyzed by flow cytometry to define NK subsets, expressions of PD-1, CTLA-4, activating/inhibitory receptors, granzyme A, and CD107a. The plasma levels of soluble PD-1, PDL-1, and CTLA-4 were measured by ELISA. Mediastinal lymph nodes of NSCLC patients had increased ratios of exhausted NK cells, increased expression of PD-1 and IL-10, and impaired cytotoxicity. Mediastinal lymph nodes removal increased CD56dimCD16bright cytotoxic effector phenotype and reduced exhausted NK cells. PD-1+ NK cells were significantly more abundant in patients' blood, and VAMLA significantly reduced their ratio as well. The ratio of IL-10 secreting regulatory NK cells was also reduced after VAMLA. Blood NK cells had increased cytotoxic functions and spontaneous IFN-γ secretion, and these NK cell functions were also recovered by VAMLA. Mediastinal lymph node removal reversed NK cell exhaustion, reduced regulatory NK cells, and improved antitumoral functions of NK cells. Tumor-draining lymph nodes may contribute to tumor evasion from antitumoral immune responses. The role of their removal needs to be further studied both to better understand this mechanism and as a potential immunotherapeutic approach.