Search results for: complicated corneal ulceration
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 867

Search results for: complicated corneal ulceration

867 Complicated Corneal Ulceration in Cats: Clinical Diagnosis and Surgical Management of 80 Cases

Authors: Khaled M. Ali, Ayman A. Mostafa, Soliman M. Soliman

Abstract:

Objectives: To describe the most common clinical and endoscopic findings associated with complicated corneal ulcers in cats, and to determine the short-term outcomes after surgical treatment of these cats. Animals Eighteen client-owned cats of different breeds (52 females and 28 males), ranging in age from 3 months to 6 years, with corneal ulcers. Procedures: Cats were clinically evaluated to initially determine the concurrent corneal abnormalities. Endoscopic examination was performed to determine the anterior and posterior segments abnormalities. Superficial and deep stromal ulcers were treated using conjunctival flap. Corneal sequestrum was treated by partial keratectomy and conjunctival flap. Anterior synechia was treated via peripheral iridectomy and separation of the adhesion between the iris and the inner cornea. Symblepharon was treated by removal of the adhered conjunctival membrane from the cornea. Incurable endophthalmitis was treated surgically by extirpation. Short-term outcomes after surgical managements of selected corneal abnormalities were then assessed clinically and endoscopically. Results: Deep stromal ulcer with descemetocele, endophthalmitis, symblepharon, corneal sequestration and anterior synechia with secondary glaucoma and corneal scarring were the most common complications of corneal ulcer. FHV-1 was a common etiologic factor of corneal ulceration. Persistent corneal scars of varying shape and size developed in cats with deep stromal ulcer, anterior synechia, and corneal sequestration. Conclusions: Domestic shorthaired and Persian cats were the most predisposed breeds to FHV-1 infection and subsequent corneal ulceration. Immediate management of patients with corneal ulcer would prevent serious complications. No age or sex predisposition to complicated corneal ulceration in cats.

Keywords: cats, complicated corneal ulceration, clinical, endoscopic diagnosis, FHV-1

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866 Complications of Contact Lens-Associated Keratitis: A Refresher for Emergency Departments

Authors: S. Selman, T. Gout

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Microbial keratitis is a serious complication of contact lens wear that can be vision and eye-threatening. Diverse presentations relating to contact lens wear include dry corneal surface, corneal infiltrate, ulceration, scarring, and complete corneal melt leading to perforation. Contact lens wear is a major risk factor and, as such, is an important consideration in any patient presenting with a red eye in the primary care setting. This paper aims to provide an overview of the risk factors, common organisms, and spectrum of contact lens-associated keratitis (CLAK) complications. It will highlight some of the salient points relevant to the assessment and workup of patients suspected of CLAK in the emergency department based on the recent literature and therapeutic guidelines. An overview of the management principles will also be provided.

Keywords: microbial keratitis, corneal pathology, contact lens-associated complications, painful vision loss

Procedia PDF Downloads 109
865 Visual and Clinical Outcome in Patients with Corneal Lacerations

Authors: Avantika Verma

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In industrialized nations, corneal lacerations are one of the most common reason for hospitalization. This study was designed to study visual and clinical outcome in patients presenting with full thickness corneal lacerations in Indian population and to ascertain the impact of various preoperative and operative factors influencing prognosis after repair of corneal lacerations. Males in third decade with injuries at work with metallic objects were common. Lens damage, hyphema, vitreous hemorrhage, retinal detachment and endophthalmitis were seen. All the patients underwent primary repair within first 24 hours of presentation. At 3 months, 74.3% had a good visual outcome. About 5.7% of patients had no perception of light.In conclusion, various demographic and preoperative factors like age, time of presentation, vision at presentation, length of corneal wound, involvement of visual axis, associated ocular features like hyphaema, lenticular changes, vitreous haemorrhage and retinal detachment are significant prognostic indicators for final visual outcome.

Keywords: corneal laceration, corneal wound repair, injury, visual outcome

Procedia PDF Downloads 356
864 Outcomes of Combined Penetrating keratoplasty and Vitreo-Retinal Surgery in Management of Endophthalmitis with Obscured Corneal Clarity

Authors: Abhishek Dave, Manisha Singh

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Purpose: The study aims to evaluate the outcomes of combined Penetrating keratoplasty (PKP) and Vitreo-Retinal (VR) surgery in patients having endophthalmitis with poor corneal clarity. Methods: PKP with VR Surgery was performed in 43 eyes. This is a retrospective analysis of their preoperative, intraoperative and microbiological characteristics and anatomical and functional outcomes. Results: Corneal opacification was due to corneal ulcer in 30 (69.7%), graft infection in 8 (18.6%), bullous keratopathy in 4 and corneal scar in 1 eye. Postoperative visual acuity improved in 20 (46.5%), not changed in 14 (32.5%) and deteriorated in 9 eyes (20.9%). Poor anatomic success was seen in 15 (34.88%) eyes (9-phthisis bulbi, 6-eviscerated). Graft remained clear in 24 eyes (1 year). Microbiology revealed bacteria in 26, fungus in 14 and no growth in 3 eyes. Six out of 11 patients having poor vision in the fellow eye, too, achieved functional success. Conclusion: PKP with VR surgery is a complex but globe-salvaging procedure for poor prognosis eyes, which otherwise would need evisceration.

Keywords: penetrating keratoplasty, VR surgery, endophthalmitis, corneal ulcer

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863 Smart Coating for Enhanced Corneal Healing via Delivering Progranulin

Authors: Dan Yan, Yunuo Zhang, Yuhan Huang, Weijie Ouyang

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The cornea serves as a vital protective barrier for the eye; however, it is prone to injury and damage that can disrupt corneal epithelium and nerves, triggering inflammation. Therefore, understanding the biological effects and molecular mechanisms involved in corneal wound healing and identifying drugs targeting these pathways is crucial for researchers in this field. This study aimed to investigate the therapeutic potential of progranulin (PGRN) in treating corneal injuries. Our findings demonstrated that PGRN significantly enhanced corneal wound repair by accelerating corneal re-epithelialization and re-innervation. In vitro experiments with cultured epithelial cells and trigeminal ganglion cells further revealed that PGRN stimulated corneal epithelial cell proliferation and promoted axon growth in trigeminal ganglion cells. Through RNA-sequencing (RNA-seq) analysis and other experimental techniques, we discovered that PGRN exerted its healing effects by modulating the Wnt signaling pathway, which played a critical role in repairing epithelial cells and promoting axon regeneration in trigeminal neurons. Importantly, our study highlighted the anti-inflammatory properties of PGRN by inhibiting the NF-κB signaling pathway, leading to decreased infiltration of macrophages. In conclusion, our findings underscored the potential of PGRN in facilitating corneal wound healing by promoting corneal epithelial cell proliferation, trigeminal ganglion cell axon regeneration, and suppressing ocular inflammation. These results suggest that PGRN could potentially expedite the healing process and improve visual outcomes in patients with corneal injuries.

Keywords: cornea, wound healing, progranulin, corneal epithelial cells, trigeminal ganglion cells

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862 Viscoelastic Response of the Human Corneal Stroma Induced by Riboflavin/UVA Cross-Linking

Authors: C. Labate, M. P. De Santo, G. Lombardo, R. Barberi, M. Lombardo, N. M. Ziebarth

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In the past decades, the importance of corneal biomechanics in the normal and pathological functions of the eye has gained its credibility. In fact, the mechanical properties of biological tissues are essential to their physiological function. We are convinced that an improved understanding of the nanomechanics of corneal tissue is important to understand the basic molecular interactions between collagen fibrils. Ultimately, this information will help in the development of new techniques to cure ocular diseases and in the development of biomimetic materials. Therefore, nanotechnology techniques are powerful tools and, in particular, Atomic Force Microscopy has demonstrated its ability to reliably characterize the biomechanics of biological tissues either at the micro- or nano-level. In the last years, we have investigated the mechanical anisotropy of the human corneal stroma at both the tissue and molecular levels. In particular, we have focused on corneal cross-linking, an established procedure aimed at slowing down or halting the progression of the disease known as keratoconus. We have obtained the first evidence that riboflavin/UV-A corneal cross-linking induces both an increase of the elastic response and a decrease of the viscous response of the most anterior stroma at the scale of stromal molecular interactions.

Keywords: atomic force spectroscopy, corneal stroma, cross-linking, viscoelasticity

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861 Xenografts: Successful Penetrating Keratoplasty Between Two Species

Authors: Francisco Alvarado, Luz Ramírez

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Corneal diseases are one of the main causes of visual impairment and affect almost 4 million, and this study assesses the effects of deep anterior lamellar keratoplasty (DALK) with porcine corneal stroma and postoperative topical treatment with tacrolimus in patients with infectious keratitis. No patient was observed with clinical graft rejection. Among the cases: 2 were positive to fungal culture, 2 with Aspergillus and the other 8 cases were confirmed by bacteriological culture. Corneal diseases are one of the main causes of visual impairment and affect almost 4 million. This study assesses the effects of deep anterior lamellar keratoplasty (DALK) with porcine corneal stroma and postoperative topical treatment with tacrolimus in patients with infectious keratitis. Receiver bed diameters ranged from 7.00 to 9.00 mm. No incidents of Descemet's membrane perforation were observed during surgery. During the follow-up period, no corneal graft splitting, IOP increase, or intolerance to tacrolimus were observed. Deep anterior lamellar keratoplasty seems to be the best option to avoid xenograft rejection, and it could help new surgical techniques in humans.

Keywords: ophthalmology, cornea, corneal transplant, xenografts, surgical innovations

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860 Intrastromal Donor Limbal Segments Implantation as a Surgical Treatment of Progressive Keratoconus: Clinical and Functional Results

Authors: Mikhail Panes, Sergei Pozniak, Nikolai Pozniak

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Purpose: To evaluate the effectiveness of intrastromal donor limbal segments implantation for treatment of progressive keratoconus considering on main characteristics of corneal endothelial cells. Setting: Outpatient ophthalmic clinic. Methods: Twenty patients (20 eyes) with progressive keratoconus II-III of Amsler classification were recruited. The worst eye was treated with the transplantation of donor limbal segments in the recipient corneal stroma, while the fellow eye was left untreated as a control of functional and morphological changes. Furthermore, twenty patients (20 eyes) without progressive keratoconus was used as a control of corneal endothelial cells changes. All patients underwent a complete ocular examination including uncorrected and corrected distance visual acuity (UDVA, CDVA), slit lamp examination fundus examination, corneal topography and pachymetry, auto-keratometry, Anterior Segment Optical Coherence Tomography and Corneal Endothelial Specular Microscopy. Results: After two years, statistically significant improvement in the UDVA and CDVA (on the average on two lines for UDVA and three-four lines for CDVA) were noted. Besides corneal astigmatism decreased from 5.82 ± 2.64 to 1.92 ± 1.4 D. Moreover there were no statistically significant differences in the changes of mean spherical equivalent, keratometry and pachymetry indicators. It should be noted that after two years there were no significant differences in the changes of the number and form of corneal endothelial cells. It can be regarded as a process stabilization. In untreated control eyes, there was a general trend towards worsening of UDVA, CDVA and corneal thickness, while corneal astigmatism was increased. Conclusion: Intrastromal donor segments implantation is a safe technique for keratoconus treatment. Intrastromal donor segments implantation is an efficient procedure to stabilize and improve progressive keratoconus.

Keywords: corneal endothelial cells, intrastromal donor limbal segments, progressive keratoconus, surgical treatment of keratoconus

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859 Anterior Segment Optical Coherence Tomography Study of Cornea and Tear Film Parameters in Juvenile Systemic Lupus Erythematous Patients

Authors: Mohamed Salah El-Din Mahmoud, Ahmed Hamed, Asmaa Anwar Mohamed

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Purpose: To study the tear film parameters, total corneal thickness (CT), corneal epithelial thickness and, corneal power in Juvenile systemic lupus erythematosus (JSLE) patients compared to age-matched controls using anterior segment optical coherence tomography (AS-OCT). Methods: This was a cross-sectional study. Study participants were divided into 2 groups: Group A: 75 eyes of JSLE patients, Group B: 75 eyes of healthy controls. Tear meniscus height (TMH), tear meniscus depth (TMD), and tear meniscus area (TMA) were the lower tear meniscus parameters that were measured. The corneal power, CT, and epithelial thickness were all determined automatically. Results: In the JSLE group, the range of age was 10 to 15 years while the control group was 11 to 16 years. TMH, TMA, and TMD were 527.7±46.8, 0.059±0.015 and 343.3±59.9 respectively in JSLE group while 525.4±44.6, 0.058±0.011 and 340.6±58.0 respectively in control group without significant difference (p-value<0.001). The corneal power was 43.3±0.55 in the JSLE while 43.2±0.54 in the control group without significant difference (p-value= 0.407). CT was 551.1±13.5 in JSLE group while 551.2±15.3 in control group without significant difference (p-value= 0.982). Epithelial thickness was 52.66±1.35 in the JSLE group while 52.60±1.36 in the control group without significant difference (p-value= 0.765). Conclusion: We demonstrated no significant difference in tear meniscus dimensions, CT, epithelial thickness, and corneal power in the JSLE patients compared to age-matched controls using AS-OCT.

Keywords: tear film, ASOCT, JSLE, pachymetry, corneal thickness

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858 Ophthalmic Hashing Based Supervision of Glaucoma and Corneal Disorders Imposed on Deep Graphical Model

Authors: P. S. Jagadeesh Kumar, Yang Yung, Mingmin Pan, Xianpei Li, Wenli Hu

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Glaucoma is impelled by optic nerve mutilation habitually represented as cupping and visual field injury frequently with an arcuate pattern of mid-peripheral loss, subordinate to retinal ganglion cell damage and death. Glaucoma is the second foremost cause of blindness and the chief cause of permanent blindness worldwide. Consequently, all-embracing study into the analysis and empathy of glaucoma is happening to escort deep learning based neural network intrusions to deliberate this substantial optic neuropathy. This paper advances an ophthalmic hashing based supervision of glaucoma and corneal disorders preeminent on deep graphical model. Ophthalmic hashing is a newly proposed method extending the efficacy of visual hash-coding to predict glaucoma corneal disorder matching, which is the faster than the existing methods. Deep graphical model is proficient of learning interior explications of corneal disorders in satisfactory time to solve hard combinatoric incongruities using deep Boltzmann machines.

Keywords: corneal disorders, deep Boltzmann machines, deep graphical model, glaucoma, neural networks, ophthalmic hashing

Procedia PDF Downloads 249
857 Laser Corneoplastique™: A Refractive Surgery for Corneal Scars

Authors: Arun C. Gulani, Aaishwariya A. Gulani, Amanda Southall

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Background: Laser Corneoplastique™ as a least interventional, visually promising technique for patients with vision disability from corneal scars of varied causes has been retrospectively reviewed and proves to cause a paradigm shift in mindset and approach towards corneal scars as a Refractive surgery aiming for emmetropic, unaided vision of 20;/20 in most cases. Three decades of work on this technique has been compiled in this 15-year study. Subject and Methods: The objective of this study was to determine the success of Laser Corneoplastique™ surgery as a treatment of corneal scar cases. A survey of corneal scar cases caused by various medical histories that had undergone Laser Corneoplastique™ surgery over the past twenty years by a single surgeon Arun C. Gulani, M.D. were retrospectively reviewed. The details of each of the cases were retrieved from their medical records and analyzed. Each patient had been examined thoroughly at their preoperative appointments for stability of refraction and vision, depth of scar, pachymetry, topography, pattern of the scar and uncorrected and best corrected vision potential, which were all taken into account in the patients' treatment plans. Results: 64 eyes of 53 patients were investigated for scar etiology, keratometry, visual acuity, and complications. There were 25 different etiologies seen, with the most common being a Herpetic scar. The average visual acuity post-op was, on average, 20/23.55 (±7.05). Laser parameters used were depth and pulses. Overall, the mean Laser ablation depth was 30.67 (±19.05), ranging from 2 to 73 µm. Number of Laser pulses averaged 191.85 (±112.02). Conclusion: Refractive Laser Corneoplastique™ surgery, when practiced as an art, can address all levels of ametropia while reversing complex corneas and scars from refractive surgery complications back to 20/20 vision.

Keywords: corneal scar, refractive surgery, corneal transplant, laser corneoplastique

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856 Impact of Glaucoma Surgery on Corneal Endothelium

Authors: Majid Moshirfar, Kyle Margulies, Yasmyne C. Ronquillo, Phillip Hoopes

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A total of 66 articles were reviewed to compare glaucoma and its associated surgeries' effect on central corneal endothelium cell density (CECD). The paper reports the average reported central CECD loss at 3-, 6-, 12-, 24-, 36-, 48-, and 60-month post-operation for each glaucoma surgery. ALT, MLT, SLT, CS AGV, VC BGI, Hydrus + phaco, XEN gel + phaco, PRESERFLO, Dual iStent, or Trabectome had no significant impact on postoperative CECD compared to either preoperative CECD or control group CECD. The highest CECD loss was found to be EXPRESS-phaco, AC AGV, CS BGI, CS BGI, AC BGI, and AC BGI at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups, respectively. AC AGV, Trab + MMC, Trab, AC BGI, Trab + MMC, Cypass, and Cypass showed the smallest reduction of CECD at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups.

Keywords: glaucoma, corneal endothelium, cell density, surgery outcome

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855 Alternate Optical Coherence Tomography Technologies in Use for Corneal Diseases Diagnosis in Dogs and Cats

Authors: U. E. Mochalova, A. V. Demeneva, Shilkin A. G., J. Yu. Artiushina

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Objective. In medical ophthalmology OCT has been actively used in the last decade. It is a modern non-invasive method of high-precision hardware examination, which gives a detailed cross-sectional image of eye tissues structure with a high level of resolution, which provides in vivo morphological information at the microscopic level about corneal tissue, structures of the anterior segment, retina and optic nerve. The purpose of this study was to explore the possibility of using the OCT technology in complex ophthalmological examination in dogs and cats, to characterize the revealed pathological structural changes in corneal tissue in cats and dogs with some of the most common corneal diseases. Procedures. Optical coherence tomography of the cornea was performed in 112 animals: 68 dogs and 44 cats. In total, 224 eyes were examined. Pathologies of the organ of vision included: dystrophy and degeneration of the cornea, endothelial corneal dystrophy, dry eye syndrome, chronic superficial vascular keratitis, pigmented keratitis, corneal erosion, ulcerative stromal keratitis, corneal sequestration, chronic glaucoma and also postoperative period after performed keratoplasty. When performing OCT, we used certified medical devices: "Huvitz HOCT-1/1F», «Optovue iVue 80» and "SOCT Copernicus Revo (60)". Results. The results of a clinical study on the use of optical coherence tomography (OCT)of the cornea in cats and dogs, performed by the authors of the article in the complex diagnosis of keratopathies of variousorigins: endothelial corneal dystrophy, pigmented keratitis, chronic keratoconjunctivitis, chronic herpetic keratitis, ulcerative keratitis, traumatic corneal damage, sequestration of the cornea of cats, chronic keratitis, complicating the course of glaucoma. The characteristics of the OCT scans are givencorneas of cats and dogs that do not have corneal pathologies. OCT scans of various corneal pathologies in dogs and cats with a description of the revealed pathological changes are presented. Of great clinical interest are the data obtained during OCT of the cornea of animals undergoing keratoplasty operations using various forms of grafts. Conclusions. OCT makes it possible to assess the thickness and pathological structural changes of the corneal surface epithelium, corneal stroma and descemet membrane. We can measure them, determine the exact localization, and record pathological changes. Clinical observation of the dynamics of the pathological process in the cornea using OCT makes it possible to evaluate the effectiveness of drug treatment. In case of negative dynamics of corneal disease, it is necessary to determine the indications for surgical treatment (to assess the thickness of the cornea, the localization of its thinning zones, to characterize the depth and area of pathological changes). According to the OCT of the cornea, it is possible to choose the optimal surgical treatment for the patient, the technique and depth of optically constructive surgery (penetrating or anterior lamellar keratoplasty).; determine the depth and diameter of the planned microsurgical trepanation of corneal tissue, which will ensure good adaptation of the edges of the donor material.

Keywords: optical coherence tomography, corneal sequestration, optical coherence tomography of the cornea, corneal transplantation, cat, dog

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854 Treatment of Drug-Induced Oral Ulceration with Hyaluronic Acid Gel: A Case Report

Authors: Meltem Koray, Arda Ozgon, Duygu Ofluoglu, Mehmet Yaltirik

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Oral ulcerations can be seen as a side effect of different drugs. These ulcers usually appear within a few weeks following drug treatment. In most of cases, these ulcers resist to conventional treatments, such as anesthetics, antiseptics, anti-inflammatory agents, cauterization, topical tetracycline and corticosteroid treatment. The diagnosis is usually difficult, especially in patients receiving multiple drug therapies. Hyaluronan or hyaluronic acid (HA) is a biomaterial that has been introduced as an alternative approach to enhance wound healing and also used for oral ulcer treatment. The aim of this report is to present the treatment of drug-induced oral ulceration on maxillary mucosa with HA gel. 60-year-old male patient was referred to Department of Oral and Maxillofacial Surgery complaining of oral ulcerations during few weeks. He had received chemotherapy and radiotherapy in 2014 with the diagnosis of nasopharyngeal carcinoma, and he has accompanying systemic diseases such as; cardiological, neurological diseases and gout. He is medicated with Escitalopram (Cipralex® 20mg), Quetiapine (Seroquel® 100mg), Mirtazapine (Zestat® 15mg), Acetylsalicylic acid (Coraspin® 100mg), Ramipril-hydrochlorothiazide (Delix® 2.5mg), Theophylline anhydrous (Teokap Sr® 200mg), Colchicine (Colchicum Dispert® 0.5mg), Spironolactone (Aldactone® 100mg), Levothyroxine sodium (Levotiron® 50mg). He had painful oral ulceration on the right side of maxillary mucosa. The diagnosis was 'drug-induced oral ulceration' and HA oral gel (Aftamed® Oral gel) was prescribed 3 times a day for 2 weeks. Complete healing was achieved within 3 weeks without any side effect and discomfort. We suggest that HA oral gel is a potentially useful local drug which can be an alternative for management of drug-induced oral ulcerations.

Keywords: drug-induced, hyaluronic acid, oral ulceration, maxillary mucosa

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853 The Morphological Changes of POV in Diabetic Patients and Its Correlation with Changes in Corneal Epithelium, Corneal Nerve, and the Fundus in Using Vivo Confocal Microscopy

Authors: Ji Jiazheng, Wang Jingrao, Jin Xin, Zhang Hong

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Diabetes mellitus is a metabolic disease characterized by high blood sugar. A long-standing hyperglycemic state can lead to various tissue damage. Diabetic retinopathy is the most common and widely studied ocular complication and has become the leading cause of blindness in my country. At the same time, diabetes has profound clinically relevant effects on the cornea, leading to keratopathy and vision-threatening. The cornea is an avascular tissue and is sensitive to hyperglycemia, Keratopathy caused by diabetes is usually chronic, they are called diabetic keratopathy or diabetic neurotrophic keratopathy, leading to several diabetic corneal complications including delayed epithelial wound healing, recurrent erosions, neuropathy, loss of sensitivity. Corneal stem cell dysfunction in diabetic patients as an important influencing factor of diabetic keratopathy. The consequences of this condition are often underestimated. The limbus is located between the cornea and the sclera tissue. The limbal stroma consists of a series of radial elevations with fibrovascular centers known as palisades of Vogt (POV). Previous studies have shown that palisades of Vogt (POV), as the main site of limbal stem cells, plays an important role in the homeostasis of the corneal epithelium. Therefore, POV plays a vital role in the healing of corneal epithelial surgery and postoperative evaluation. IVCM can observe the condition of the corneal epithelium at the cellular level. It has profound significance and guidance for the evaluation of limbal and limbal stem cells. We have previously observed structural changes in POV in HSK and HZO patients on IVCM. At present, there have been reports involving limbal stem cell dysfunction in diabetic patients, but the specific pathogenesis is still unclear. However, there are no studies on POV morphological changes in patients with DM. Therefore, we performed statistics and compared the correlation between POV morphological changes and corneal epithelial basal cell density, corneal nerves, and length of disease in DM patients and normal humans using IVCM studies. At the same time, fundoscopy was used to observe the correlation between the thickness of RNFL and the thickness of GCC and POV in diabetic patients. And to observe the correlation between SVD, DVD and POV for research.

Keywords: confocal microscopy, fundus, limbal stem cells, diabetes

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852 Treatment Outcome Of Corneal Ulcers Using Levofloxacin Hydrate 1.5% Ophthalmic Solution And Adjuvant Oral Ciprofloxacin, A Treatment Strategy Applicable To Primary Healthcare

Authors: Celine Shi Ying Lee, Jong Jian Lee

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Background: Infectious keratitis is one of the leading causes of blindness worldwide. Prompt treatment with effective medication will control the infection early, preventing corneal scarring and visual loss. fluoroquinolones ophthalmic medication is used because of its broad-spectrum properties, potency, good intraocular penetration, and low toxicity. The study aims to evaluate the treatment outcome of corneal ulcers using Levofloxacin 1.5% ophthalmic solution (LVFX) with adjuvant oral ciprofloxacin when indicated and apply this treatment strategy in primary health care as first-line treatment. Methods: Patients with infective corneal ulcer treated in an eye center were recruited. Inclusion criteria includes Corneal infection consistent with bacterial keratitis, single or multiple small corneal ulcers. Treatment regime: LVFX hourly for the first 2 days, 2 hourly from the 3rd day, and 3 hourly on the 5th day of review. Adjuvant oral ciprofloxacin 500mg BD was administered for 5 days if there were multiple corneal ulcers or when the location of the cornea ulcer was central or paracentral. Results: 47 subjects were recruited. There were 16 (34%) males and 31 (66%) females. 40 subjects (85%) were contact lens (CL) related to corneal ulcer, and 7 subjects (15%) were non-contact lens related. 42 subjects (89%) presented with one ulcer, of which 20 of them (48%) needed adjuvant therapy. 5 subjects presented with 2 or 3 ulcers, of which 3 needed adjuvant therapy. A total of 23 subjects (49%) was given adjuvant therapy (oral ciprofloxacin 500mg BD for 5 days).21 of them (91%) were CL related. All subjects recovered fully, and the average duration of treatment was 3.7 days, with 49% of the subjects resolved on the 3rd day, 38% on the 5thday of and 13% on the 7thday. All subjects showed symptoms of relief of pain, light-sensitivity, and redness on the 3rd day with full visual recovery post-treatment. No adverse drug reactions were recorded. Conclusion: Our treatment regime demonstrated good clinical outcome as first-line treatment for corneal ulcers. A corneal ulcer is a common eye condition in Singapore, mainly due to CL wear. Pseudomonas aeruginosa is the most frequent and potentially sight-threatening pathogen involved in CL related corneal ulcer. Coagulase-negative Staphylococci, Staphylococcus aureus, and Streptococcus Pneumoniae were seen in non-CL users. All these bacteria exhibit good sensitivity rates to ciprofloxacin and levofloxacin. It is therefore logical in our study to use LVFX Eyedrops and adjuvant ciprofloxacin oral antibiotics when indicated as first line treatment for most corneal ulcers. Our study of patients, both CL related and non-CL related, have shown good clinical response and full recovery using the above treatment strategy. There was also a full restoration of visual acuity in all the patients. Eye-trained primary Healthcare practitioners can consider adopting this treatment strategy as first line treatment in patients with corneal ulcers. This is relevant during the COVID pandemic, where hospitals are overwhelmed with patients and in regions with limited access to specialist eye care. This strategy would enable early treatment with better clinical outcome.

Keywords: corneal ulcer, levofloxacin hydrate, treatment strategy, ciprofloxacin

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851 Study into the Interactions of Primary Limbal Epithelial Stem Cells and HTCEPI Using Tissue Engineered Cornea

Authors: Masoud Sakhinia, Sajjad Ahmad

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Introduction: Though knowledge of the compositional makeup and structure of the limbal niche has progressed exponentially during the past decade, much is yet to be understood. Identifying the precise profile and role of the stromal makeup which spans the ocular surface may inform researchers of the most optimum conditions needed to effectively expand LESCs in vitro, whilst preserving their differentiation status and phenotype. Limbal fibroblasts, as opposed to corneal fibroblasts are thought to form an important component of the microenvironment where LESCs reside. Methods: The corneal stroma was tissue engineered in vitro using both limbal and corneal fibroblasts embedded within a tissue engineered 3D collagen matrix. The effect of these two different fibroblasts on LESCs and hTCEpi corneal epithelial cell line were then subsequently determined using phase contrast microscopy, histolological analysis and PCR for specific stem cell markers. The study aimed to develop an in vitro model which could be used to determine whether limbal, as opposed to corneal fibroblasts, maintained the stem cell phenotype of LESCs and hTCEpi cell line. Results: Tissue culture analysis was inconclusive and required further quantitative analysis for remarks on cell proliferation within the varying stroma. Histological analysis of the tissue-engineered cornea showed a comparable structure to that of the human cornea, though with limited epithelial stratification. PCR results for epithelial cell markers of cells cultured on limbal fibroblasts showed reduced expression of CK3, a negative marker for LESC’s, whilst also exhibiting a relatively low expression level of P63, a marker for undifferentiated LESCs. Conclusion: We have shown the potential for the construction of a tissue engineered human cornea using a 3D collagen matrix and described some preliminary results in the analysis of the effects of varying stroma consisting of limbal and corneal fibroblasts, respectively, on the proliferation of stem cell phenotype of primary LESCs and hTCEpi corneal epithelial cells. Although no definitive marker exists to conclusively illustrate the presence of LESCs, the combination of positive and negative stem cell markers in our study were inconclusive. Though it is less traslational to the human corneal model, the use of conditioned medium from that of limbal and corneal fibroblasts may provide a more simple avenue. Moreover, combinations of extracellular matrices could be used as a surrogate in these culture models.

Keywords: cornea, Limbal Stem Cells, tissue engineering, PCR

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850 The First Report of Aberrant Corneal Occlusion in Rabbit in Iran

Authors: Bahador Bardshiri, Omid Moradi, Amir Komeilian, Nima Panahifar

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Formation of a conjunctival membrane over the corneal surface is a condition unique to rabbits that has been labeled aberrant corneal occlusion or pseudopterygium. In the summer of 2013, a five years old male Standard Chinchilla rabbit were presented to Karaj Central Veterinary hospital and the owner complained that his rabbit shows degrees of blindness and there were opacities on both eyes of the presented rabbit. Ophthalmic examination of the affected eyes revealed a conjunctival fold stretching over the cornea of both eyes. The fold originated from limbus and it was vascularized and centrally thickened. There were no attachments to the corneal epithelium and the fold could be easily lifted. Surgery was performed under general anesthesia. The conjunctival fold was incised centrifugally up to its attachment at the limbus and the lid margin using small scissors. The central rim of the segment was then replaced to its normal position in the fornix and fixed with mattress sutures (7/0) passing through outside skin. After the surgery, eye drops containing dexamethasone, gentamicin and polymixin were applied twice daily up to 3 weeks. Within the observation period (8 months) no recurrence was noted. "Pseudo" in the term pseudopterygium refers to the fact that the conjunctival membrane is not adhering to the underlying cornea, but growing over it. In rare cases, the membrane may be loosely attached to the cornea, but can be easily separated without causing damage. It can cover only a small part of the cornea with an annular peripheral opacification of the cornea, or cover it almost fully, leading to blindness. Ethiopathogenesis remains unclear and recurrence of the problem is very likely. The surgical technique that used here decreases probability of recurrence of conjunctival fold.

Keywords: rabbit, cornea, aberrant corneal occlusion, pseudopterygium

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849 Modified Model for UV-Laser Corneal Ablation

Authors: Salah Hassab Elnaby, Omnia Hamdy, Aziza Ahmed Hassan, Salwa Abdelkawi, Ibrahim Abdelhalim

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Laser corneal reshaping has been proposed as a successful treatment of many refraction disorders. However, some physical and chemical demonstrations of the laser effect upon interaction with the corneal tissue are still not fully explained. Therefore, different computational and mathematical models have been implemented to predict the depth of the ablated channel and calculate the ablation threshold and the local temperature rise. In the current paper, we present a modified model that aims to answer some of the open questions about the ablation threshold, the ablation rate, and the physical and chemical mechanisms of that action. The proposed model consists of three parts. The first part deals with possible photochemical reactions between the incident photons and various components of the cornea (collagen, water, etc.). Such photochemical reactions may end by photo-ablation or just the electronic excitation of molecules. Then a chemical reaction is responsible for the ablation threshold. Finally, another chemical reaction produces fragments that can be cleared out. The model takes into account all processes at the same time with different probabilities. Moreover, the effect of applying different laser wavelengths that have been studied before, namely the common excimer laser (193-nm) and the solid state lasers (213-nm & 266-nm), has been investigated. Despite the success and ubiquity of the ArF laser, the presented results reveal that a carefully designed 213-nm laser gives the same results with lower operational drawbacks. Moreover, the use of mode locked laser could also decrease the risk of heat generation and diffusion.

Keywords: UV lasers, mathematical model, corneal ablation, photochemical ablation

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848 Light and Scanning Electron Microscopic Studies on Corneal Ontogeny in Buffalo

Authors: M. P. S. Tomar, Neelam Bansal

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Histomorphological, histochemical and scanning electron microscopic observations were recorded in developing cornea of buffalo fetuses. The samples from fetal cornea were collected in appropriate fixative from slaughter house and Veterinary Clinics, GADVASU, Ludhiana. The microscopic slides were stained for detailed histomorphological and histochemical studies. The scanning electron microscopic studies were performed at Electron microscopy & Nanobiology Lab, PAU Ludhiana. In present study, it was observed that, in 36 days (d) fetus, the corneal epithelium was well marked single layered structure which was placed on stroma mesenchyme. Cornea appeared as the continuation of developing sclera. The thickness of cornea and its epithelium increased as well as the epithelium started becoming double layered in 47d fetus at corneo-scleral junction. The corneal thickness in this stage suddenly increased thus easily distinguished from developing sclera. The separation of corneal endothelium from stroma was evident as a single layered epithelium. The stroma possessed numerous fibroblasts in 49d stage eye. Descemet’s membrane was appeared at 52d stage. The limbus area was separated by a depression from the developing cornea in 61d stage. In 65d stage, the Bowman’s layer was more developed. Fibroblasts were arranged parallel to each other as well as parallel to the surface of developing cornea in superficial layers. These fibroblasts and fibers were arranged in wavy pattern in the center of stroma. Corneal epithelium started to be stratified as a double layered epithelium was present in this age of fetal eye. In group II (>120 Days), the corneal epithelium was stratified towards a well marked irido-corneal angle. The stromal fibroblasts followed a complete parallel arrangement in its entire thickness. In full term fetuses, a well developed cornea was observed. It was a fibrous layer which had five distinct layers. From outside to inwards were described as the outer most layer was the 7-8 layered corneal epithelial, subepithelial basement membrane (Bowman’s membrane), substantia propria or stroma, posterior limiting membrane (Descemet’s membrane) and the posterior epithelium (corneal endothelium). The corneal thickness and connective tissue elements were continued to be increased. It was 121.39 + 3.73µ at 36d stage which increased to 518.47 + 4.98 µ in group III fetuses. In fetal life, the basement membrane of corneal epithelium and endothelium depicted strong to intense periodic Acid Schiff’s (PAS) reaction. At the irido-corneal angle, the endothelium of blood vessels was also positive for PAS activity. However, cornea was found mild positive for alcian blue reaction. The developing cornea showed strong reaction for basic proteins in outer epithelium and the inner endothelium layers. Under low magnification scanning electron microscope, cornea showed two types of cells viz. light cells and dark cells. The light cells were smaller in size and had less number of microvilli in their surface than in the dark cells. Despite these surface differences between light and dark cells, the corneal surface showed the same general pattern of microvilli studding all exposed surfaces out to the cell margin. which were long (with variable height), slight tortuous slender and possessed a micro villus shaft with a very prominent knob.

Keywords: buffalo, cornea, eye, fetus, ontogeny, scanning electron microscopy

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847 Comparison of Corneal Curvature Measurements Conducted with Tomey AO-2000® and the Current Standard Biometer IOL Master®

Authors: Mohd Radzi Hilmi, Khairidzan Mohd Kamal, Che Azemin Mohd Zulfaezal, Ariffin Azrin Esmady

Abstract:

Purpose: Corneal curvature (CC) is an important anterior segment parameter. This study compared CC measurements conducted with two optical devices in phakic eyes. Methods: Sixty phakic eyes of 30 patients were enrolled in this study. CC was measured three times with the optical biometer and topography-keratometer Tomey AO-2000 (Tomey Corporation, Nagoya, Japan), then with the standard partial optical coherence interferometry (PCI) IOL Master (Carl Zeiss Meditec, Dublin, CA) and data were statistically analysed. Results: The measurements resulted in a mean CC of 43.86 ± 1.57 D with Tomey AO-2000 and 43.84 ± 1.55 D with IOL Master. Distribution of data is normal, and no significance difference in CC values was detected (P = 0.952) between the two devices. Correlation between CC measurements was highly significant (r = 0. 99; P < 0.0001). The mean difference of CC values between devices was 0.017 D and 95% limit of agreement was -0.088 to 0.12. Duration taken for measurements with the standard biometer IOL Master was longer (55.17 ± 2.24 seconds) than with Tomey AO-2000 (39.88 ± 2.38 seconds) in automatic mode. Duration of manual measurement with Tomey AO-2000 in manual mode was the shortest (28.57 ± 2.71 seconds). Conclusion: In phakic eyes, CC measured with Tomey AO-2000 and IOL Master showed similar values, and high correlation was observed between these two devices. This shows that both devices can be used interchangeably. Tomey AO-2000 is better in terms of faster to operate and has its own topography systems.

Keywords: corneal topography, corneal curvature, IOL Master, Tomey AO2000

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846 Antioxydant Properties and Gastroprotective Effect of Rosa canina Aqueous Extract against Alcohol-Induced Ulceration and Oxidative Stress in Rat Model

Authors: H. Sebai, M. A. Jabria, D. Wannes, H. Tounsi, L. Marzouki

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We aimed in the present study to investigate the protective effects of Tunisian Rosa canina aqueous extract (RCAE) against ethanol-induced gastric ulceration and oxidative stress in a rat model. In this respect, adult male Wistar rats were used and divided into six groups of ten each: control, EtOH, EtOH plus various doses of RCAE, EtOH plus famotidine and EtOH + gallic acid. Phytochemical and biochemical analysis were performed using colorimetric methods. We found that RCAE is rich in total polyphenols, total flavonoids, and condensed tannins, and exhibited an importance in vitro antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. In vivo, the results showed that oral administration of EtOH caused macroscopic and histological changes in gastric mucosa. These injuries are accompanied by an oxidative stress status as assessed by an increase of lipid peroxidation as well as a decrease of antioxidant enzyme activities such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Alcohol intoxication also induced intracellular mediators deregulation as assessed by an increase of hydrogen peroxide (H2O2), calcium and free iron levels in gastric mucosa. More, importantly, RCAE pretreatment reversed all macroscopic, histological and biochemical changes induced by EtOH administration. In conclusion, we suggest that RCAE has potent protective effects on acute ethanol-induced gastric ulceration related in part in part its antioxidant properties and its opposite effect on intracellular mediators. Indeed, Rosa canina can be offered as a food additive to protect against alcohol consumption-induced gastric and oxidative damage.

Keywords: alcohol, antioxidant properties, food additive, gastric ulceration, rat model, Rosa canina

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845 Utility of Routine Colonoscopy in Acute Diverticulitis

Authors: Naya Masood, Russell Hodgson, Mark Tacey

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Purpose: Patients with acute diverticulitis (AD) have an increased risk of underlying colorectal cancer (CRC); however, those with uncomplicated AD may have the same underlying population risk. This study informs on an Australian AD population who were not routinely offered colonoscopic follow-up. Methods: A 2-year (July 2016 – June 2018) retrospective study of patients admitted with CT-confirmed acute diverticulitis was conducted. CT findings were categorised as ‘complicated’ and ‘uncomplicated’ and were correlated with the detection of cancer in subsequent colonoscopy or follow-up. Results: 67.7% (n=292) of 431 patients were seen to have had complicated AD on an abdominopelvic CT scan. Patients were complicated most commonly due to bowel wall thickening reported on CT (90.4%), perforation (20.2%), or an abscess (12%). Follow-up colonoscopic evaluation was conducted in 52.9% (n=228) of total cases of AD, out of which 156 suffered complicated AD and the rest uncomplicated. None of the uncomplicated AD patients in our cohort were found to have CRC. Of those with complicated AD, six were found to have CRC. Conclusion: The only CRC diagnoses were made in patients with complicated AD. Despite available evidence, a significant proportion of uncomplicated AD patients were still undergoing colonoscopy. There is scope to further safely decrease the number of colonoscopies performed in AD patients.

Keywords: acute diverticulitis, colonoscopy, colorectal cancer, advanced adenoma, complicated diverticulitis

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844 Psychiatric Symptoms in Keratoconus: Analyzing Anxiety and Depression in Affected Patients

Authors: Nida Amin, Fahad Tanveer, Hina Shabbir, Ayesha Saeed, Attiqa Riaz

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The gradual progression of corneal disorder keratoconus significantly impairs eyesight and quality of life, increasing the likelihood of depression. Using the Hospital Anxiety and Depression Scale (HADS) at the AL-Ibrahim Eye Hospital in Karachi, this study aimed to evaluate the occurrence of depression and anxiety symptoms in patients with keratoconus and to suggest better treatment. A descriptive-analytical study was conducted at Al-Ibrahim Eye Hospital Karachi from March to April 2022, and patients diagnosed with symptomatic keratoconus were recruited using a non-probability convenient sampling technique. After obtaining written informed consent from patients, keratoconus severity was assessed using visual acuity and corneal topography. Symptoms of anxiety and depression were assessed using the Hospital Anxiety and Depression (HADS) Scale. The data were analyzed using SPSS version 20.0. Spearman correlation coefficient. Of the 108 participants, 60 (56%) were female and 48 (44%) were male. Using the HADS scale, 44 (40.7%) patients were classified as normal with a HADS score of (0-7), 23 (21.3%) as borderline with a HADS score of (8-10) and 41 (38%) patients were diagnosed with anxiety and depression with a HADS score of (11-21). Depression and anxiety are highly prevalent among patients in advanced stages of the disease.

Keywords: cornea, keratoconus, anxiety, depression, corneal topography, mental health

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843 In vitro Modeling of Aniridia-Related Keratopathy by the Use of Crispr/Cas9 on Limbal Epithelial Cells and Rescue

Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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842 Corneal Confocal Microscopy As a Surrogate Marker of Neuronal Pathology In Schizophrenia

Authors: Peter W. Woodruff, Georgios Ponirakis, Reem Ibrahim, Amani Ahmed, Hoda Gad, Ioannis N. Petropoulos, Adnan Khan, Ahmed Elsotouhy, Surjith Vattoth, Mahmoud K. M. Alshawwaf, Mohamed Adil Shah Khoodoruth, Marwan Ramadan, Anjushri Bhagat, James Currie, Ziyad Mahfoud, Hanadi Al Hamad, Ahmed Own, Peter Haddad, Majid Alabdulla, Rayaz A. Malik

Abstract:

Introduction:- We aimed to test the hypothesis that, using corneal confocal microscopy (a non-invasive method for assessing corneal nerve fibre integrity), patients with schizophrenia would show neuronal abnormalities compared with healthy participants. Schizophrenia is a neurodevelopmental and progressive neurodegenerative disease, for which there are no validated biomarkers. Corneal confocal microscopy (CCM) is a non-invasive ophthalmic imaging biomarker that can be used to detect neuronal abnormalities in neuropsychiatric syndromes. Methods:- Patients with schizophrenia (DSM-V criteria) without other causes of peripheral neuropathy and healthy controls underwent CCM, vibration perception threshold (VPT) and sudomotor function testing. The diagnostic accuracy of CCM in distinguishing patients from controls was assessed using the area under the curve (AUC) of the Receiver Operating Characterstics (ROC) curve. Findings:- Participants with schizophrenia (n=17) and controls (n=38) with comparable age (35.7±8.5 vs 35.6±12.2, P=0.96) were recruited. Patients with schizophrenia had significantly higher body weight (93.9±25.5 vs 77.1±10.1, P=0.02), lower Low Density Lipoproteins (2.6±1.0 vs 3.4±0.7, P=0.02), but comparable systolic and diastolic blood pressure, HbA1c, total cholesterol, triglycerides and High Density Lipoproteins were comparable with control participants. Patients with schizophrenia had significantly lower corneal nerve fiber density (CNFD, fibers/mm2) (23.5±7.8 vs 35.6±6.5, p<0.0001), branch density (CNBD, branches/mm2) (34.4±26.9 vs 98.1±30.6, p<0.0001), and fiber length (CNFL, mm/mm2) (14.3±4.7 vs 24.2±3.9, p<0.0001) but no difference in VPT (6.1±3.1 vs 4.5±2.8, p=0.12) and electrochemical skin conductance (61.0±24.0 vs 68.9±12.3, p=0.23) compared with controls. The diagnostic accuracy of CNFD, CNBD and CNFL to distinguish patients with schizophrenia from healthy controls were, according to the AUC, (95% CI): 87.0% (76.8-98.2), 93.2% (84.2-102.3), 93.2% (84.4-102.1), respectively. Conclusion:- In conclusion, CCM can be used to help identify neuronal changes and has a high diagnostic accuracy to distinguish subjects with schizophrenia from healthy controls.

Keywords:

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841 Portable Palpation Probe for Diabetic Foot Ulceration Monitoring

Authors: Bummo Ahn

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Palpation is widely used to measure soft tissue firmness or stiffness in the living condition in order to apply detection, diagnosis, and treatment of tumors, scar tissue, abnormal muscle tone, or muscle spasticity. Since these methods are subjective and depend on the proficiency level, it is concluded that there are other diagnoses depending on the condition of the experts and the results are not objective. The mechanical property obtained by using the elasticity of the tissue is important to calculate a predictive variable for monitoring abnormal tissues. If the mechanical load such as reaction force on the foot increases in the same region under the same conditions, the mechanical property of the tissue is changed. Therefore, objective diagnosis is possible not only for experts but also for patients using this quantitative information. Furthermore, the portable system also allows non-experts to easily diagnose at home, not in hospitals or institutions. In this paper, we introduce a portable palpation system that can be used to measure the mechanical properties of human tissue, which can be applied to monitor diabetic foot ulceration patients with measuring the mechanical property change of foot tissue. The system was designed to be smaller and portable in comparison with the conventional palpation systems. It is consists of the probe, the force sensor, linear actuator, micro control unit, the display module, battery, and housing. Using this system, we performed validation experiments by applying different palpations (3 and 5 mm) to soft tissue (silicone rubber) and measured reaction forces. In addition, we estimated the elastic moduli of the soft tissue against different palpations and compare the estimated elastic moduli that show similar value even if the palpation depths are different.

Keywords: palpation probe, portable, diabetic foot ulceration, monitoring, mechanical property

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840 Pluripotent Stem Cells as Therapeutic Tools for Limbal Stem Cell Deficiencies and Drug Testing

Authors: Aberdam Edith, Sangari Linda, Petit Isabelle, Aberdam Daniel

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Background and Rationale: Transparent avascularised cornea is essential for normal vision and depends on limbal stem cells (LSC) that reside between the cornea and the conjunctiva. Ocular burns or injuries may destroy the limbus, causing limbal stem cell deficiency (LSCD). The cornea becomes vascularised by invaded conjunctival cells, the stroma is scarring, resulting in corneal opacity and loss of vision. Grafted autologous limbus or cultivated autologous LCS can restore the vision, unless the two eyes are affected. Alternative cellular sources have been tested in the last decades, including oral mucosa or hair follicle epithelial cells. However, only partial success has been achieved by the use of these cells since they were not able to uniformly commit into corneal epithelial cells. Human pluripotent stem cells (iPSC) display both unlimited growth capacity and ability to differentiate into any cell type. Our goal was to design a standardized and reproducible protocol to produce transplantable autologous LSC from patients through cell reprogramming technology. Methodology: First, keratinocyte primary culture was established from a small number of plucked hair follicles of healthy donors. The resulting epithelial cells were reprogrammed into induced pluripotent stem cells (iPSCs) and further differentiate into corneal epithelial cells (CEC), according to a robust protocol that recapitulates the main step of corneal embryonic development. qRT-PCR analysis and immunofluorescent staining during the course of differentiation confirm the expression of stage specific markers of corneal embryonic lineage. First appear ectodermal progenitor-specific cytokeratins K8/K18, followed at day 7 by limbal-specific PAX6, TP63 and cytokeratins K5/K14. At day 15, K3/K12+-corneal cells are present. To amplify the iPSC-derived LSC (named COiPSC), intact small epithelial colonies were detached and cultivated in limbal cell-specific medium. In that culture conditions, the COiPSC can be frozen and thaw at any passage, while retaining their corneal characteristics for at least eight passages. To evaluate the potential of COiPSC as an alternative ocular toxicity model, COiPSC were treated at passage P0 to P4 with increasing amounts of SDS and Benzalkonium. Cell proliferation and apoptosis of treated cells was compared to LSC and the SV40-immortalized human corneal epithelial cell line (HCE) routinely used by cosmetological industrials. Of note, HCE are more resistant to toxicity than LSC. At P0, COiPSC were systematically more resistant to chemical toxicity than LSC and even to HCE. Remarkably, this behavior changed with passage since COiPSC at P2 became identical to LSC and thus closer to physiology than HCE. Comparative transcriptome analysis confirmed that COiPSC from P2 are similar to a mixture of LSC and CEC. Finally, by organotypic reconstitution assay, we demonstrated the ability of COiPSC to produce a 3D corneal epithelium on a stromal equivalent made of keratocytes. Conclusion: COiPSC could become valuable for two main applications: (1) an alternative robust tool to perform, in a reproducible and physiological manner, toxicity assays for cosmetic products and pharmacological tests of drugs. (2). COiPSC could become an alternative autologous source for cornea transplantation for LSCD.

Keywords: Limbal stem cell deficiency, iPSC, cornea, limbal stem cells

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839 The Role of Pulmonary Resection in Complicated Primary Pediatric Pulmonary Tuberculosis: An Evidence-Based Case Report

Authors: Hendra Wibowo, Suprayitno Wardoyo, Dhama Shinta

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Introduction: Pediatric pulmonary tuberculosis (TB) incidence was increasing, with many undetected cases. In complicated TB, treatment should consist of returning pulmonary function, preventing further complications, and eliminating bacteria. Complicated TB management was still controversial, and surgery was one of the treatments that should be evaluated in accordance with its role in the treatment of complicated TB. Method: This study was an evidence-based case report. The database used for the literature search were Cochrane, Medline, Proquest, and ScienceDirect. Keywords for the search were ‘primary pulmonary tuberculosis’, ‘surgery’, ‘lung resection’, and ‘children’. Inclusion criteria were studies in English or Indonesian, with children under 18 years old as subject, and full-text articles available. The assessment was done according to Oxford Centre for evidence-based medicine 2011. Results: Six cohort studies were analyzed. Surgery was indicated for patients with complicated TB that were unresponsive towards treatment. It should be noted that the experiments were done before the standard WHO antituberculosis therapy was applied; thus, the result may be different from the current application. Conclusion: Currently, there was no guideline on pulmonary resection. However, surgery yielded better mortality and morbidity in children with complicated pulmonary TB.

Keywords: pediatric, pulmonary, surgery, therapy, tuberculosis

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838 Photoactivated Chromophore for Keratitis-Cross Linking Window Absorption Alone versus Combined Pack-CXL Window Absorption and Standard Anti-microbial Therapy for Treatment of Infectious Keratitis: A Prospective Study

Authors: Mohammed M. Mahdy Tawfeek

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Objective: The aim of this work is to compare the outcome of photoactivated chromophore for keratitis-cross linking (PACK-CXL) window absorption (WA) alone with combined PACK-CXL WA and standard anti-microbial therapy (SAT) for the treatment of infectious keratitis. Patients and Methods: This is a randomized prospective comparative clinical trial. Thirty eyes with clinically suspected infectious keratitis were randomly assigned into two equal groups of 15 eyes each: Group (A) was treated by PACK-CXL WA alone and group (B) was treated by PACK-CXL WA combined with SAT. Identification of organisms was made by lab study before treatment. Corneal healing was evaluated by corneal examination and anterior segment OCT (AS-OCT). Written informed consent was obtained from all participants and the study was approved by the research ethics committee of the Faculty of Medicine, Zagazig University. The work has been carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) for studies involving humans. Results: Complete healing and resolution (Successful treatment) were observed in 10 eyes (66.7%) of a group (A) and 14 eyes (93.3%) of group (B) and failure was observed in 5 eyes (33.3%) of a group (A) and one eye (6.67%) of group (B). They were statistically significant (P =0.042 and 0.003) in a comparison between both groups regarding success and failure of treatment, respectively. Complete corneal healing was reported in the third month postoperatively in 10 eyes (66.7%) of group (A) and 14 eyes (93.3%) of group (B). Complications were absent in 12 patients (80%) in group (A) and 14 patients (93.3%) of group (B); however, perforation and impending perforation were found in 3 patients of group (A) and only one patient of group (B). Conclusion: PACK-CXL is a promising, non-invasive treatment option for infectious keratitis, especially when performed with the window absorption (WA) technique, either alone or combined with SAT. It has a synergistic effect with a standard antimicrobial treatment that gives good outcome results in the treatment of infectious keratitis. Also, it avoids the antibiotics resistance that has become rapidly spreading worldwide.

Keywords: corneal cross linking, infectious keratitis, PACK-CXL, window absorption

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