Orally Delivered Connexin43 Hemichannel Blocker, Tonabersat, Inhibits Vascular Breakdown and Inflammasome Activation in a Mouse Model of Diabetic Retinopathy.

Diabetic retinopathy (DR), a microvascular complication of diabetes, is associated with pronounced inflammation arising from the activation of a nucleotide-binding and oligomerization domain-like receptor (NLR) protein 3 (NLRP3) inflammasome. Cell culture models have shown that a connexin43 hemichannel blocker can prevent inflammasome activation in DR. The aim of this study was to evaluate the ocular safety and efficacy of tonabersat, an orally bioavailable connexin43 hemichannel blocker, to protect against DR signs in an inflammatory non-obese diabetic (NOD) DR mouse model. For retina safety studies, tonabersat was applied to retinal pigment epithelial (ARPE-19) cells or given orally to control NOD mice in the absence of any other stimuli. For efficacy studies, either tonabersat or a vehicle was given orally to the inflammatory NOD mouse model two hours before an intravitreal injection of pro-inflammatory cytokines, interleukin-1 beta, and tumour necrosis factor-alpha. Fundus and optical coherence tomography images were acquired at the baseline as well as at 2- and 7-day timepoints to assess microvascular abnormalities and sub-retinal fluid accumulation. Retinal inflammation and inflammasome activation were also assessed using immunohistochemistry. Tonabersat did not have any effect on ARPE-19 cells or control NOD mouse retinas in the absence of other stimuli. However, the tonabersat treatment in the inflammatory NOD mice significantly reduced macrovascular abnormalities, hyperreflective foci, sub-retinal fluid accumulation, vascular leak, inflammation, and inflammasome activation. These findings suggest that tonabersat may be a safe and effective treatment for DR.

Evaluation of the Effectiveness of Aeration and Chlorination during Washing to Reduce E. coli O157:H7, Salmonella enterica, and L. innocua on Cucumbers and Bell Peppers.

The attachment strength of bacteria to surfaces can affect the efficacy of sanitizers during washing. This study aimed to determine the effectiveness of chlorination and aeration in the removal of pathogens from the surface of produce. Cucumbers and bell peppers were inoculated with Listeria innocua, Escherichia coli O157:H7, or Salmonella enterica; afterwards, the produce was washed with or without chlorinated water (100 ppm) for 3 min in combination with or without aeration. Cucumbers washed with chlorinated water, with or without aeration, presented significant reductions of L. innocua (3.65 log CFU/cm2 and 1.13 log CFU/cm2, respectively) (p < 0.05). Similarly, bell peppers washed in chlorinated water with aeration (1.91 log CFU/g) and without aeration (2.49 log CFU/g) presented significant reductions of L. innocua. A significant reduction of L. innocua was observed on bell peppers washed with non-chlorinated water with aeration (2.49 log CFU/g) (p < 0.05). Non-chlorinated water was also effective in significantly reducing the level of Salmonella enterica (p < 0.05) on cucumbers and bell peppers. Washing with chlorinated water with aeration reduced Salmonella enterica levels from 4.45 log CFU/cm2 on cucumbers to below the detectable limit (0.16 log CFU/cm2). The highest reduction of Salmonella enterica from bell peppers occurred after washing with chlorinated water with aeration (2.48 log CFU/g). E. coli O157:H7, L. innocua, and Salmonella enterica levels present in non-chlorinated water after washing contaminated produce with or without aeration were significantly greater than those in chlorinated water (p < 0.05). After treatment, the population levels of all pathogens in chlorinated water with or without aeration were below the detectable limit for bell peppers (<1.10 log CFU/mL) and cucumbers (<1.20 log CFU/mL). Using chlorine in combination with forced aeration during washing minimizes cross-contamination of bacterial pathogens.