Aqueous-deficient dry eye disease: Preferred practice pattern guidelines on clinical approach, diagnosis, and management

Dry eye disease (DED) is a broad term that includes a diverse group of clinical disorders. Aqueous-deficient dry eye (ADDE), a subtype of DED, is characterized by decreased tear production by the lacrimal gland. It can be seen in up to one-third of individuals with DED and can be comorbid with a systemic autoimmune process or occur secondary to an environmental insult. Since ADDE can be a source of long-term suffering and severe visual impairment, early identification and adequate treatment are imperative. Multiple etiologies can underlie ADDE, and it is critical to identify the underlying cause to not only improve the ocular health but also to improve the overall quality of life and well-being of affected individuals. This review discusses the various etiologies of ADDE, highlights a pathophysiology-based approach for evaluating underlying contributors, outlines various diagnostic tests, and reviews treatment options. We present the current standards and discuss ongoing research in this field. Through this review, we propose a treatment algorithm that would be useful for an ophthalmologist in diagnosing and managing individuals with ADDE.

Systemic investigations in dry eye disease

The incidence of dry eye disease has increased manifold in the past few years with more patients presenting with these complaints to our clinics every day. In the more severe forms of disease, it is important to evaluate for any systemic association which could be driving the disease such as in Sjogren’s syndrome. Understanding the possible varied etiopathogenesis and knowing when to evaluate, form an important part of treating this condition effectively. In addition, it is sometimes confusing as to which investigations to order and how to prognosticate the disease in these situations. This article simplifies this into an algorithmic approach with insights from the ocular and systemic point of view

Formation of FADD amyloid fiber and its role in immune signaling in Drosophila melanogaster / 生物工程学报

In this research, we studied the formation of Drosophila melanogaster FADD (Fas-associated death domain-containing protein) amyloid fiber and its influence on signal transduction in IMD (Immune deficiency) signaling pathway to better understand the regulation mechanism of Drosophila innate immune signaling pathway, which will provide reference for the immune regulation in other species. First, we purified dFADD protein expressed in Escherichia coli and performed Sulfur flavin T binding and transmission electron microscopy to identify the dFADD amyloid fibers formed in vitro. Then we investigated the formation of dFADD polymers in S2 cells using SDD-AGE and confocal microscope. We also constructed dFADD mutants to find out which domain is essential to fiber formation and its effect on IMD signal transduction. Our results revealed that dFADD could be polymerized to form amyloid fiber polymers in vitro and inside the cells. Formation of fibers relies on DED (Death-effector domain) domain of dFADD, since DED domain-deleted mutant existed as a monomer. Dual luciferase reporter assay showed that intact DED domain was required for the induction of downstream antimicrobial peptides, indicating that fiber formation was the key to IMD signal transduction. Our study revealed the role of dFADD in mediating the cascade between IMD and Dredd in the IMD signaling pathway by forming amyloid fibers, suggesting an evolutionarily conserved regulatory mechanism of innate immune signaling pathway.

Comparative analysis of efficacy and adverse effects of tacrolimus vs cyclosporine in chronic dry eye disease: a tertiary care hospital based study

Background: Dry eye disease (DED) is the very common and underdiagnosed ocular condition affecting vision, quality of life, and the outcomes of cataract or refractive surgery. Dry eye disease (DED), either alone or in combination with other conditions, is a frequent cause of ocular irritation that leads the patients to seek ophthalmic care. Due to a wide variety of presenting symptoms, it is often unrecognized and this causes great frustration of the patient and treating physician. While these symptoms often improve with appropriate treatment, usually in majority of the cases the disease may not be curable. Aim of the study was to compare the efficacy of tacrolimus and cyclosporine in dry eye disease.Methods: This was a single centred, 3 months prospective study. Patients with unilateral or bilateral dry eye disease and an ocular surface disease index score >12, atleast one eye with schirmer score <5mm and TBUT <10 s were enrolled in the study. The enrolled patients were randomly divided into two groups, twenty-five patients in Group 1 and twenty three patients in Group 2 completed the follow up. Group 1(n =25) who received 0.03% tacrolimus eye ointment twice daily for consecutive 3 months and Group 2 (n= 23) received 0.05% cyclosporine eye drops twice daily for consecutive 3 months the primary efficacy outcome was Schirmer score after 3 months. The secondary outcomes were TBUT and adverse effects.Results: After 3 months, both the treatment groups showed significant improvement in mean Schirmer score (p<0.001) and mean TBUT (p <0.0001). However, on comparing both the groups, mean Schirmer score and mean TBUT, results were comparable. No patient discontinued treatment because of minor ocular adverse effects.Conclusions: Dry eye patients demonstrated improvement in Schirmer score and TBUT after 3 months of treatment with tacrolimus 0.03% ointment and cyclosporine 0.05% eye drops.

Apoptosis of Renal Cell Carcinoma Cells by Expression of FADD (Fas-Associated Death Domain) / 대한비뇨기과학회지

PURPOSE: The Fas-associated death domain (FADD) constitutes a novel protein that specifically associates with the cytoplasmic death domain of Fas, and induces apoptosis. FADD is composed of a death effector domain (DED) and a death domain. In this study, we evaluated the in vivo antitumor effect of the FADD, or FADD-DED, gene in renal cell carcinoma cells, using a plasmid vector expressing the human FADD and FADD-DED genes. MATERIALS AND METHODS: The cDNA of the human FADD and FADD-DED genes were amplified by RT-PCR, and cloned to the pCR(R) 3.1. The expressions of the cloned FADD and FADD-DED (pCR(R)3.1-FADD and pCR3.1-FADD-DED) were observed by Western blot analysis. The efficacy of the growth inhibition by the cloned FADD and FADD-DED genes was tested, in vitro, on A498 and Caki-1 human renal cell carcinoma cell lines using the MTT assay. To evaluate the apoptosis, DNA fragmentation and caspase-3 assays were performed. RESULTS: Expressions of the FADD protein, and the FADD-DED, of the transfected A498 and Caki-1 cells had increased by 48 and 24 hours, respectively, compared with the control cell lines. The cytotoxicity of the pCR3.1-FADD and pCR3.1-FADD-DED on the A498 and Caki-1 cells significantly increased compared to the empty vector. The increased cytotoxicity of the FADD- or FADD-DED-transfected cell lines was associated with enhanced apoptosis, as assessed by DNA fragmentation and caspase-3 assays. CONCLUSIONS: Our results showed that the cloned FADD or FADD-DED expression plasmid vector efficiently inhibited the growth of A498 and Caki-1 human renal cell carcinoma cell lines. These data suggest that the exogenous FADD or FADD-DED expressions may have therapeutic applications in renal cell carcinomas.

Organotypic Culture of HaCaT cells: Use of Dermal Substrate that Combines de-epidermized Dermis with Fibroblast-populated Collagen Matrix

BACKGROUND: The immortalized human keratinocyte line, HaCaT cells have been widely used as substitutes for normal epidermal keratinocytes. Recently, reconstruction of a skin equivalent using HaCaT cells showed a multilayered epithelium,but somewhat different tissue architecture as compared with normal epidermis. OBJECTIVE: In this study, using HaCaT cells we tried to reconstruct an epidermis resembling more closely to normal epidermis than the previous results. MATERIALS AND METHODS: HaCaT cells were cultured in air-liquid interface on a recently developed dermal substrated in our laboratory, de-epidermized dermis (DED) raised on fibroblast-populated collagen matrix and the result was compared with those on DED or fibroblast-populated collagen matrix alone. RESULTS: HaCaT cells on the new dermal substrate formed a multilayered epithelium with rete ridges, showing rather orderly cellular organization compared with those on fibroblast-populated collagen matrix. However, horny and granular layers were not observed contrary to normal epidermis. Immunohistochemical studies revealed that differentiation markers such as keratin 1, keratin 6 and involucrin showed the similar pattern to those in HaCaT cells cultured on fibroblast-populated collagen matrix. Markers of terminal differentiation, loricrin and filaggrin were not expressed contrary to normal epidermis. CONCLUSION: These results suggest that organotypic culture HaCaT cells on the dermal substrate combines DED with fivroblast-populated collagen matrix results in incomplete differentiation of HaCaT cells contrary to normal keratinocytes.