Chronic Alcohol Exposure Promotes Cancer Stemness and Glycolysis in Oral/Oropharyngeal Squamous Cell Carcinoma Cell Lines by Activating NFAT Signaling.

Alcohol consumption is associated with an increased risk of several cancers, including oral/oropharyngeal squamous cell carcinoma (OSCC). Alcohol also enhances the progression and aggressiveness of existing cancers; however, its underlying molecular mechanism remains elusive. Especially, the local carcinogenic effects of alcohol on OSCC in closest contact with ingestion of alcohol are poorly understood. We demonstrated that chronic ethanol exposure to OSCC increased cancer stem cell (CSC) populations and their stemness features, including self-renewal capacity, expression of stem cell markers, ALDH activity, and migration ability. The ethanol exposure also led to a significant increase in aerobic glycolysis. Moreover, increased aerobic glycolytic activity was required to support the stemness phenotype of ethanol-exposed OSCC, suggesting a molecular coupling between cancer stemness and metabolic reprogramming. We further demonstrated that chronic ethanol exposure activated NFAT (nuclear factor of activated T cells) signaling in OSCC. Functional studies revealed that pharmacological and genetic inhibition of NFAT suppressed CSC phenotype and aerobic glycolysis in ethanol-exposed OSCC. Collectively, chronic ethanol exposure promotes cancer stemness and aerobic glycolysis via activation of NFAT signaling. Our study provides a novel insight into the roles of cancer stemness and metabolic reprogramming in the molecular mechanism of alcohol-mediated carcinogenesis.

Near-lifespan longitudinal tracking of brain microvascular morphology, topology, and flow in male mice.

In age-related neurodegenerative diseases, pathology often develops slowly across the lifespan. As one example, in diseases such as Alzheimer's, vascular decline is believed to onset decades ahead of symptomology. However, challenges inherent in current microscopic methods make longitudinal tracking of such vascular decline difficult. Here, we describe a suite of methods for measuring brain vascular dynamics and anatomy in mice for over seven months in the same field of view. This approach is enabled by advances in optical coherence tomography (OCT) and image processing algorithms including deep learning. These integrated methods enabled us to simultaneously monitor distinct vascular properties spanning morphology, topology, and function of the microvasculature across all scales: large pial vessels, penetrating cortical vessels, and capillaries. We have demonstrated this technical capability in wild-type and 3xTg male mice. The capability will allow comprehensive and longitudinal study of a broad range of progressive vascular diseases, and normal aging, in key model systems.

Effect of Sodium Polyphosphates on Growth of Listeria monocytogenes.

Evaluation of sodium polyphosphates (SPP), multifunctional food additives, indicated that only the higher polymers sodaphos, hexaphos, and glass H (average chain length = 6, 13, and 21, respectively) significantly inhibited growth of Listeria monocytogenes . The effect of the three compounds (0-2%) on growth of L. monocytogenes was studied in brain heart infusion + 0.3% glucose medium, pH 6.0, at 28, 19, 10, and 5°C. The organism grew well under all test conditions in the absence of SPP. Hexaphos and glass H were considerably more inhibitory than sodaphos. The most pronounced effect of SPP was on lag times, which increased with increasing SPP concentration and decreasing temperature. At 10°C, addition of 0.3% hexaphos or glass H increased lag time from 22 h to 197 and 186 h, respectively, and no growth was observed after 40 d in the presence of 2.0% of these compounds. Addition of 2.0% NaCl increased the inhibitory effect of SPP but had little effect on growth in SPP-free media. Results suggest that high molecular weight SPP may be useful in controlling the growth of L. monocytogenes , particularly at low temperatures and in combination with NaCl.

Effect of Sodium Nitrite on Growth of Shigella flexneri.

A partial factorial design study of the effect of NaNO2 (0, 100, 200, 1000 ppm) in combination with NaCl (0.5, 2.5, 4.0%), pH (7.5, 6.5, 5.5), and temperature (37, 28, 19°C) on growth of Shigella flexneri is reported. Experiments were done aerobically in brain-heart infusion medium, using an inoculum of 1 × 103 CFU/ml. Growth curves were fitted from plate count data by the Gompertz equation; exponential growth rates, lag times, generation times, and maximum populations were derived for all variable combinations. In the absence of nitrite, the organism grew well under all test conditions at 37 and 28°C but did not grow at 19°C at pH 5.5 nor at pH 7.5 with 4% NaCl. Nitrite did not affect growth in media of pH 7.5 at 37 and 28°C. At pH 6.5 growth was inhibited by 1000 ppm NaNO2. The organism failed to grow at 19°C at all nitrite levels in the presence of 2.5 or 4.0% NaCl. The inhibitory effect of nitrite was much greater in media of pH 5.5 and increased with increasing salt levels. More inhibition was apparent at 28 than at 37°C. While lack of growth was used as a paradigm of the effect of nitrite on S. flexneri , nitrite also increased the lag and generation times and decreased the exponential growth rate. Results indicated that NaNO2 in combinations with low temperature, low pH, and high salt content can effectively inhibit the growth of S. flexneri .

Effect of Sodium Chloride, pH and Temperature on Growth of Shigella flexneri.

A systematic study of the effect of sodium chloride (0.5, 2.0, 3.5, 5.0%), pH (7.5, 6.5, 5.5), and temperature (37, 28, 19, 10°C) on growth of Shigella flexneri is reported for the first time, using a factorial design. Experiments were done using Brain-Heart Infusion media inoculated to contain 1 × 103 cfu/ml and incubated on rotary shakers (150 rpm). Growth curves were plotted from the experimental data by means of the Gompertz equation, and growth rates, lag times, generation times, and maximum populations were derived for all variable combinations. Results indicated that the three variables interact to affect the growth of S. flexneri , and combinations of low temperature, low pH, and high sodium chloride contents are strongly inhibitory.