Mitochondria as a Platform for Dictating the Cell Fate of Cultured Human Corneal Endothelial Cells

PurposeAiming to clarify the role of mitochondria in cell fate decision of cultured human corneal endothelial cell (cHCEC) subpopulations.MethodsThe mitochondrial respiratory ability were examined with Mito stress and Mito fuel flex test assays using an extracellular flux analyzer (XFe24; Agilent Technologies; Santa Clara, CA) for human corneal endothelium tissues, mature cHCECs and a variety of cell state transitioned cHCECs. Tricarboxylic acid cycle and acetyl-coenzyme A–related enzymes was analyzed by proteomics for cell lysates using liquid chromatography–tandem mass spectrometry for cHCEC subpopulations.ResultsThe maximum oxygen consumption rate was found to become stable depending on the maturation of cHCECs. In the Mito stress tests, culture supplements, epidermal growth factor, SB203580, and SB431543 significantly repressed oxygen consumption rate, whereas a Rho-associated protein kinase inhibitor Y-27632 increased. Tricarboxylic acid cycle and mitochondria acetyl-coenzyme A–related enzymes were selectively upregulated in mature cHCECs, but not in cell state transitioned cHCECs. The maximum oxygen consumption rate was found to be higher in healthy human corneal endothelium tissues than those with deeply reduced cell density. An upregulated tricarboxylic acid cycle was linked with metabolic rewiring converting cHCECs to acquire the mitochondria-dependent oxidative phenotype.ConclusionsMitochondrial metabolic intermediates and energy metabolism are tightly linked to the endothelial cell fate and function. These findings will help us to standardize a protocol for endothelial cell injection.     

Early stage gastric MALT lymphoma with high-grade component cured by Helicobacter pylori eradication

A 68-year-old woman was diagnosed with gastric lymphoma of the mucosa-associated lymphoid tissue (MALT) type with a high-grade component. Surgical treatment was recommended because of the presence of the high-grade component, but she refused surgery. As an alternative, she received Helicobacter pylori eradication treatment, which successfully induced regression of the lymphoma. She shows no sign of recurrence endoscopically and histologically, as of 29 months after the eradication treatment. Moreover, the B-cell monoclonality and Helicobacter pylori infection demonstrated at diagnosis has disappeared. This is one of the rare cases of gastric lymphoma of the MALT type with a high-grade component cured by Helicobacter pylori eradication alone. Received: October 7, 1999 / Accepted: May 26, 2000     

A successfully treated case of infective endocarditis due to Candida tropicalis

A 34-year-old man, a heavy drinker, was admitted with a high fever and hematuria two months previously. Surgery was performed for acute sever pancreatitis and postoperatively antibiotics were administered with intravenous hyperalimentation. After discharge he was readmitted and infective endocarditis was strongly suspected because of high fever, hematuria, Osler's nodes, Janeway's lesions, splinter hemorrhages and mitral regurgitation. Penicillin G in combination with Gentamycine therapy was started on the first hospital day. On the second hospital day, blood culture revealed Candida tropicalis so Miconazole therapy was commenced. On the forth hospital day, he underwent surgery for replacement of a mitral prosthesis with a prosthetic valve because he had embolus in the radial artery. Despite intensive antifungal therapy, he showed no improvement in clinical symptoms. Then we changed the antifungal drug from Miconazole to Amphotericin B and 5-fluorocytosine. On the 109th hospital day, his clinical symptoms improved. Antifungal therapy was halted and at present 10 months later, he is healthy.     

Wide-field contact specular microscopy analysis of corneal endothelium post trabeculectomy

Graefe's Archive for Clinical and Experimental Ophthalmology - This study investigates the possible role of the filtration bleb in the continuous decrease in corneal endothelial cell (CEC)...     

Identification of a novel HLA-B allele, HLA-B*5904

The new human leukocyte antigen (HLA) class I allele, HLA-B*5904 was identified in Japanese individual. HLA-B*5904 differs from HLA-B*5901 by two non-synonymous nucleotide exchanges at codon 163 (ACG to CTG).     

Integrin β4 is a major target antigen in pure ocular mucous membrane pemphigoid

Background

Previous studies of ocular mucous membrane pemphigoid (OMMP) have identified several components of the basement membrane zone to be autoantigens, including integrin β4. However, there are no extensive or definitive reported studies that address this, particularly in pure OMMP.

Objectives

To clarify the major autoantigens in pure OMMP.

Materials and methods

In this study, we examined sera from 43 pure OMMP patients for both IgG and IgA antibodies using newly developed immunoblotting analyses with a hemidesmosome-rich fraction and various recombinant proteins of integrin α6β4, in addition to our routine immune-serological tests.

Results

Using a hemidesmosome-rich fraction, sera from patients with pure OMMP demonstrated reactivity of IgG and/or IgA antibodies to integrin β4, BP180 and laminin-332. The reactivity of pure OMMP sera to integrin β4 was further confirmed by immunoblotting using integrin β4 recombinant proteins. Using concentrated supernatant of HaCaT cells, only one serum sample showed positive IgG and IgA reactivity to LAD-1, the ectodomain of BP180. None of the pure OMMP sera reacted with any autoantigens on immunoblotting using normal human epidermal or dermal extracts, or purified human laminin-332.

Conclusions

Integrin β4 was considered to be the major and specific autoantigen for pureOMMP. The new methods established in this study are useful for detection of various autoantigens, particularly integrin β4.

     

Effect of metalloproteinase inhibitor on corneal cytokine expression after alkali injury.

PURPOSE: Interleukin (IL)-1alpha and IL-6 levels in the cornea are greatly elevated during the early stages after an alkali burn in mice. The authors investigated the effect of synthetic inhibitor of matrix metalloproteinase (SIMP) on the expression of inflammatory cytokines in alkali-burned murine corneas and evaluated the clinical appearance of the eyes. METHODS: After 0.5N NaOH-alkali burns to 400 corneas of ICR mice, 200 received 400 microg/ml of SIMP topically 4 times a day while 200 corneas were similarly treated with vehicle only. At days 4, 7 and 14 after injury, each cornea was assigned a clinical score for corneal opacity, corneal epithelial defect, hyphema and cataract. Extracts of injured corneas in each group were then assayed for cytokine production using ELISA systems for IL-1alpha, IL-1beta, IL-6 and tumor necrosis factor-alpha (TNF-alpha). RESULTS: The levels of IL-1alpha, IL-1beta and IL-6 were significantly lower in the SIMP-treated group than in the vehicle-treated group 7 days after the burn. However, levels of these cytokines were similar in the SIMP and non-SIMP groups at days 4 and 14. Levels of TNF-alpha did not differ between both groups at any postinjury time. In the SIMP-treated corneas, there was less opacification and hyphema formation and epithelial regeneration was faster. CONCLUSIONS: Topical application of SIMP in alkali-burned murine corneas reduced the expression of IL-1alpha, IL-1beta, and IL-6 and lessened the severity of the injury.