Deletion of thioredoxin-interacting protein preserves retinal neuronal function by preventing inflammation and vascular injury

BACKGROUND AND PURPOSE

Retinal neurodegeneration is an early and critical event in several diseases associated with blindness. Clinically, therapies that target neurodegeneration fail. We aimed to elucidate the multiple roles by which thioredoxin-interacting protein (TXNIP) contributes to initial and sustained retinal neurodegeneration.

EXPERIMENTAL APPROACH

Neurotoxicity was induced by intravitreal injection of NMDA into wild-type (WT) and TXNIP-knockout (TKO) mice. The expression of apoptotic and inflammatory markers was assessed by immunohistochemistry, elisa and Western blot. Microvascular degeneration was assessed by periodic acid-Schiff and haematoxylin staining and retinal function by electroretinogram.

KEY RESULTS

NMDA induced early (1 day) and significant retinal PARP activation, a threefold increase in TUNEL-positive nuclei and 40% neuronal loss in ganglion cell layer (GCL); and vascular permeability in WT but not TKO mice. NMDA induced glial activation, expression of TNF-α and IL-1β that co-localized with Müller cells in WT but not TKO mice. In parallel, NMDA triggered the expression of NOD-like receptor protein (NLRP3), activation of caspase-1, and release of IL-1β and TNF-α in primary WT but not TKO Müller cultures. After 14 days, NMDA induced 1.9-fold microvascular degeneration, 60% neuronal loss in GCL and increased TUNEL-labelled cells in the GCL and inner nuclear layer in WT but not TKO mice. Electroretinogram analysis showed more significant reductions in b-wave amplitudes in WT than in TKO mice.

CONCLUSION AND IMPLICATIONS

Targeting TXNIP expression prevented early retinal ganglion cell death, glial activation, retinal inflammation and secondary neuro/microvascular degeneration and preserved retinal function. TXNIP is a promising new therapeutic target for retinal neurodegenerative diseases.     

Dye Labelling of Nucleosides

Dye‐labelled nucleosides were obtained in 30–79% (average 45%) yields by treating N‐(4‐arylazobenzoyl)‐1H‐benzotriazoles 3a–b with appropriate nucleosides. Similarly, 3a–b afforded dye‐labelled threoninol conjugates in 55–89% (average 67%) yields. All novel products were characterized by NMR and elemental analysis.     

Attenuation of bleomycin-induced lung fibrosis in rats by mesna

Lung fibrosis is a common side effect of the chemotherapeutic agent, bleomycin. Current evidence suggests that reactive oxygen species may play a key role in the development of lung fibrosis. The present study examined the effect of mesna on bleomycin-induced lung fibrosis in rats. Animals were divided into three groups: (1) saline control group; (2) Bleomycin group in which rats were injected with bleomycin (15 mg/kg, i.p.) three times a week for four weeks; (3) Bleomycin and mesna group, in which mesna was given to rats (180 mg/kg/day, i.p.) a week prior to bleomycin and daily during bleomycin injections for 4 weeks until the end of the treatment. Bleomycin treatment resulted in a pronounced fall in the average body weight of animals. Bleomycin-induced pulmonary injury and lung fibrosis was indicated by increased lung hydroxyproline content, and elevated nitric oxide synthase, myeoloperoxidase, platelet activating factor, and tumor necrosis factor-alpha in lung tissues. On the other hand, bleomycin induced a reduction in reduced glutathione concentration and angiotensin converting enzyme activity in lung tissues. Moreover, bleomycin-induced severe histological changes in lung tissues revealed as lymphocytes and neutrophils infiltration, increased collagen deposition and fibrosis. Co-administration of bleomycin and mesna reduced bleomycin-induced weight loss and attenuated lung injury as evaluated by the significant reduction in hydroxyproline content, nitric oxide synthase activity, and concentrations of myeoloperoxidase, platelet activating factor, and tumor necrosis factor-alpha in lung tissues. Furthermore, mesna ameliorated bleomycin-induced reduction in reduced glutathione concentration and angiotensin activity in lung tissues. Finally, histological evidence supported the ability of mesna to attenuate bleomycin-induced lung fibrosis and consolidation. Thus, the findings of the present study provide evidence that mesna may serve as a novel target for potential therapeutic treatment of lung fibrosis.     

Risk assessment induced by knapsack or conventional motor sprayer on pesticides applicators and farm workers in cotton season

Fakous Province in the Sharkia Governorate constitutes one of the largest agricultural areas in Egypt. The majority of people in this province rely on subsistence agriculture. In the cotton cultivation season the continuous application of pesticides is commonly used to increase agricultural productivity, using different types of spraying equipment. In this study a cohort of 210 intensive agricultural pesticide applicators and farm workers from Sawada and Akyad Elkepplia villages in Fakous Province were assessed according to the type of spraying equipment they used. Conventional motor (300l/feddan or 0.42ha) and knapsack motor sprayer (20l/feddan) were commonly used by farmers. Contamination on applicators was detected on head, body (thorax/abdomen) and legs at different percentages according to the spraying tools. The recorded results revealed that contamination with pesticides due to knapsack motor sprayers 0.76% on head, 4.8% on body and 5.8% on legs; however, conventional motor sprayers induce contamination of 3.6% on head, 23.7% on body and 29.1% on legs. Several criteria for estimating pesticide contamination by previous delivery systems were used. The most important reference biomarker was serum acetyl cholinesterase (AchE) depression. Sprayers showed changes in serum glucose levels as well as reduced erythrocytic glutathione levels (GSH). However, an increase in both total serum protein and albumin was recorded also, alongside elevation in lipid peroxidation biomarker malondialdehyde (MDA). Changes in serum biochemistry regarding enzymes reflecting cytotoxicity were also recorded, such as inhibition of alanine aminotransferase (ALT) and glutathione-S-transferase (GST). An increase in aspartate aminotransferase (AST) and glutathione reductase (GR) was observed particularly in conventional motor sprayers. Changes in enzymes activities found in this study are linked to the adverse health effects related to chronic pesticide toxicity that may lead to pathophysiological diseases, cancer or neurodegenerative disorders, which warrants further investigation.     

Preparation and first evaluation of [(18)F]FE@SUPPY: a new PET tracer for the adenosine A(3) receptor

INTRODUCTION: Changes of the adenosine A(3) receptor subtype (A3AR) expression have been shown in a variety of pathologies, especially neurological and affective disorders, cardiac diseases and oncological and inflammation processes. Recently, 5-(2-fluoroethyl) 2,4-diethyl-3-(ethylsulfanylcarbonyl)-6-phenylpyridine-5-carboxylate (FE@SUPPY) was presented as a high-affinity ligand for the A3AR with good selectivity. Our aims were the development of a suitable labeling precursor, the establishment of a reliable radiosynthesis for the fluorine-18-labeled analogue [(18)F]FE@SUPPY and a first evaluation of [(18)F]FE@SUPPY in rats. METHODS: [(18)F]FE@SUPPY was prepared in a feasible and reliable manner by radiofluorination of the corresponding tosylated precursor. Biodistribution was carried out in rats, and organs were removed and counted. Autoradiography was performed on rat brain slices in the presence or absence of 2-Cl-IB-MECA. RESULTS: Overall yields and radiochemical purity were sufficient for further preclinical and clinical applications. The uptake pattern of [(18)F]FE@SUPPY found in rats mainly followed the described mRNA distribution pattern of the A3AR. Specific uptake in brain was demonstrated by blocking with a selective A3AR agonist. CONCLUSION: We conclude that [(18)F]FE@SUPPY has the potential to serve as the first positron emission tomography tracer for the A3AR.     

Small Intestinal Bacterial Overgrowth in Nonalcoholic Steatohepatitis: Association with Toll-Like Receptor 4 Expression and Plasma Levels of Interleukin 8

Background  

Experimental and clinical studies suggest an association between small intestinal bacterial overgrowth (SIBO) and nonalcoholic steatohepatitis (NASH). Liver injury and fibrosis could be related to exposure to bacterial products of intestinal origin and, most notably, endotoxin, including lipopolysaccharide (LPS).