Lipopolysaccharide (LPS) induced inflammatory changes to differentially expressed miRNAs of the host inflammatory response.

In veterinary medicine, inflammation in swine is evaluated principally by clinical signs. This method is often unreliable when assessing large animal populations because of inconsistent interpretations of clinical observations. This study examined whether changes in miRNA expression can predict the severity of the inflammatory response in swine after administration of Escherichia coli lipopolysaccharide (LPS). Whole blood from swine challenged with LPS at 0.125 µg/kg to 2.0 µg/kg body weight was collected at 0, 1, 3, and 8 h post LPS-challenge. Mature miRNAs were extracted from plasma and quantitative real-time-PCR (qRT-PCR) was used to evaluate the 84 most abundant swine miRNAs found in plasma. The miRNA changes in expression were assessed using the comparative CT Method (ΔΔCT method) for normalization with an exogenous control. The results revealed that expression of ssc-let-7e-5p, ssc-mir-22-3p, and ssc-miR-146a-5p were the most significantly changed miRNA over the time course. At 1 h post-LPS, ssc-let-7e-5p decreased as the LPS dosage levels increased from 0.125 to 1.0 µg/kg. Similarly, as the LPS doses increased from 0.125 to 0.5 µg/kg, ssc-miR-22-3p levels significantly decreased at 1 h post-LPS. In the 2.0 µg/kg LPS, ssc-miR-146a-5p levels increased between 0 and 3 h post-LPS; however, expression was downregulated with a 145 % decrease from 3 to 8 h. The three miRNA biomarkers suggest potentially useful surrogate endpoints for the evaluation of inflammatory and endotoxemia responses in swine.

Substance Use Disorders and Behavioral Addictions During the COVID-19 Pandemic and COVID-19-Related Restrictions.

COVID-19 was first identified in Wuhan, China in December of 2019 and appeared in the United States 1 month later. Between the onset of the pandemic and January 13, 2021, over 92 million people have tested positive for the virus and over 1.9 million people have died globally. Virtually every country in the world has been impacted by this virus. Beginning in March 2020, many U.S. state governments enforced a "quarantine" to respond to the growing health crisis. Citizens were required to remain at home; schools, restaurants, and non-essential businesses were forced to close, and large gatherings were prohibited. Americans' lives were transformed in a span of days as daily routines were interrupted and people were shuttered indoors. Mounting fear and unpredictability coupled with widespread unemployment and social isolation escalated anxiety and impacted the mental health of millions across the globe. Most (53%) U.S. adults reported that the coronavirus outbreak has had a negative impact on their mental health, including inducing or exacerbating use of alcohol, drugs, gambling and overeating as coping mechanisms. In this paper, we will examine substance use and addictive behaviors that have been used to manage the stress and uncertainty wrought by the COVID-19 pandemic. We review the changing treatment landscape as therapy pivoted online and telemedicine became the norm.

Molecular mechanism of action responsible for carrageenan-induced inflammatory response.

Carrageenan-induced inflammation has long been used as an in vivo model of local inflammation. We developed an in vitro model of inflammation using primary blood cells to characterize gene induction following carrageenan (λ-CGN) stimulation and identify the signal transduction pathway(s) through which λ-CGN worked, using swine whole blood cultures from Yorkshire barrows. Blood samples were divided into stimulated and unstimulated groups. Unstimulated blood was a control for λ-CGN treated cultures to delineate treatment effects from time-in-culture effects. All cultures were collected and separated into two fractions; supernatant for ELISA analyses and white blood cells for mRNA expression. Lambda (λ)-CGN induced MCP-1 at the proteomic and the genomic levels. Lambda-CGN increased IL-8 protein production but had no impact on serum amyloid A protein levels. Alveolar Macrophage-Derived Neutrophil Chemotactic Factor-II (AMCF-II), a swine-specific member of the IL8/GRO family, showed increased gene expression. TNF-α and IL-6 protein levels were not induced by λ-CGN stimulation. Stimulation of HEK-293 cells co-transfected with a single pattern recognition receptor and the secreted embryonic alkaline phosphatase (SEAP) read-out system demonstrated that λ-CGN signals through the TLR-2 and TLR-4 signal transduction pathways. Using silencing RNA to inhibit TLR6 expression in TLR2 transfected HEK-293 cells indicated that λ-CGN works through the TLR2/6 pathway. Silencing TLR6 expression in TLR4 transfected HEK-293 cells showed that λ-CGN stimulation of this cell line worked through a TLR4/6 heterodimer, as lipopolysaccharide (LPS) induced SEAP production through a TLR4 homodimer. These results demonstrate that although carrageenan can stimulate through TLR4 signaling pathways, it initiates an inflammatory response in these cells that differs from a typical endotoxin effect such as LPS stimulation, in terms of the pathways and gene products altered, suggesting that activation of TLR2/6 and TLR4/6 are the predominant pathways through which carrageenan induces inflammatory responses.