A STING (stimulator of interferon genes) agonist GSK3996915 under investigation in early discovery for hepatitis B was orally dosed to a mouse model for understanding the parent drug distribution in liver, the target organ. Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) was used to quantify the distribution of GSK3996915 in liver collected from mice administered a single oral dose at 90 mg/kg. GSK3996915 was detected with a zonal distribution localized in the portal triad and highly concentrated in the main bile ducts, indicating clearance through biliary excretion. High spatial resolution imaging showed the distribution of the parent drug localized to the cellular populations in the sinusoids, including the Kupffer cells. Additionally, a series of drug-related metabolites were observed to be localized in the central zones of the liver. These results exemplify the potential of utilizing MALDI IMS for measuring not only quantitative drug distribution and target exposure but also drug metabolism and elimination in a single suite of experiments.

Chronic pain conditions affect nearly 20% of the population in the United States. Current medical interventions, such as opioid drugs, are effective at relieving pain but are accompanied by many undesirable side effects. This is one reason increased numbers of chronic pain patients have been turning to Cannabis for pain management. Cannabis contains many bioactive chemical compounds; however, current research looking into lesser-studied minor cannabinoids in Cannabis lacks uniformity between experimental groups and/or excludes female mice from investigation. This makes it challenging to draw conclusions between experiments done with different minor cannabinoid compounds between laboratories or parse out potential sex differences that could be present. We chose five minor cannabinoids found in lower quantities within Cannabis: cannabinol (CBN), cannabidivarin (CBDV), cannabigerol (CBG), 8-tetrahydrocannabinol (8-THC), and 9-tetrahydrocannabivarin (THCV). These compounds were then tested for their cannabimimetic and pain-relieving behaviors in a cannabinoid tetrad assay and a chemotherapy-induced peripheral neuropathy (CIPN) pain model in male and female CD-1 mice. We found that the minor cannabinoids we tested differed in the cannabimimetic behaviors evoked, as well as the extent. We found that CBN, CBG, and high-dose 8-THC evoked some tetrad behaviors in both sexes, while THCV and low-dose 8-THC exhibited cannabimimetic tetrad behaviors only in females. Only CBN efficaciously relieved CIPN pain, which contrasts with reports from other researchers. Together these findings provide further clarity to the pharmacology of minor cannabinoids and suggest further investigation into their mechanism and therapeutic potential.

People with sickle cell disease (SCD) often experience chronic pain as well as unpredictable episodes of acute pain, which significantly affects their quality of life and life expectancy. Current treatment strategies for SCD-associated pain primarily rely on opioid analgesics, which have limited efficacy and cause serious adverse effects. Cannabis has emerged as a potential alternative, yet its efficacy remains uncertain. In this study, we investigated the antinociceptive effects of ⁹-tetrahydrocannabinol (THC), cannabis’ intoxicating constituent, in male HbSS mice, which express >99% human sickle hemoglobin, and male HbAA mice, which express normal human hemoglobin A, as a control. Acute THC administration (0.1–3 mg/kg^–1, i.p.) dose-dependently reduced mechanical and cold hypersensitivity in human sickle hemoglobin (HbSS) but not human normal hemoglobin A (HbAA) mice. In the tail-flick assay, THC (1 and 3 mg/kg^–1, i.p.) produced substantial antinociceptive effects in HbSS mice. By contrast, THC (1 mg/kg^–1, i.p.) did not alter anxiety-like behavior (elevated plus maze) or long-term memory (24-hour novel object recognition). Subchronic THC treatment (1 and 3 mg/kg^–1, i.p.) provided sustained relief of mechanical hypersensitivity but led to tolerance in cold hypersensitivity in HbSS mice. Together, the findings identify THC as a possible therapeutic option for the management of chronic pain in SCD. Further research is warranted to elucidate its mechanism of action and possible interaction with other cannabis constituents.

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HARI SREENIVASAN: Let’s turn to the continuing fallout and reaction to the Harvey Weinstein story.

Yesterday, Weinstein resigned from the board of his production company following numerous revelations of sexual harassment and several allegations of assault.

More than three dozen women have said Weinstein harassed them. While Weinstein has admitted to behaving inappropriately, he has said he didn’t physically assault anyone.

One of those women is Katherine Kendall. She was a 23-year-old actress who met Weinstein in 1993. She alleges that he invited her to his apartment in New York, where, she says, he took off his clothes and asked for a massage.

As other actresses began coming forward about their painful experiences, she also went public with her own story.

She joins me now from Los Angeles.

First, thanks for joining us.

And I don’t want to relive something that’s painful for you, but you are taking a public stance on it.

For people who don’t know your story, what happened?

KATHERINE KENDALL, Actress/ Photographer: Well, I was you know, a young actress, and I had had a formal meeting at the Miramax office earlier that day.

And then, at the end of the meeting, which I thought went really well, he invited me to come to screenings. He said: “Welcome to the Miramax family. You know, come to premieres, screenings, et cetera. In fact, there’s one this afternoon. Would you like to come?”

And I said, “Sure.”

And I ended up going to see a movie with him. It ended up just being a movie, not a screening, but the film “Red Rock West.” And, you know, that’s right when I had this sort of sinking feeling that something wasn’t going right.

And then, after the movie, we walked for a few blocks. And he said he needed to go up to his apartment to get something, and would I just come with him real quick? And I sort of said no, and we went back and forth on that for a minute. It was sort of a negotiation with him always, trying to sort of stand my ground, but then be convinced it was OK.

I did go into his apartment. Once there, we talked for a long time about art and movies. And I felt like he was treating me like an intellect.

And I felt like the meeting was going really well, and sort of continued. I didn’t feel unsafe once I was in there. And, at one point, then, he got up to go to the bathroom. And he came back in a robe and asked me to give him a massage.

And I was extremely uncomfortable. And I was like, oh, God, no, I’m not comfortable with that. And we went back and forth on that.

And then he went back to the bathroom again, and came back this time completely naked. And, you know, that changed it entirely for me, too. It just took it to the next place. It was completely disorienting. And I was scared, you know? I was really scared.

And then it became sort of a cat-and-mouse game of, like, how am I going to get out of there?

And I’m — it’s hard to make sense of what someone is trying to do to you when they’re fully naked, and they’re…

HARI SREENIVASAN: Yes.

KATHERINE KENDALL: You know, I’m 105 pounds. He’s a large man standing between me and the door.

And, I mean, I felt very resolute, like, I will definitely get out of here somehow. But I’m not — I’m not sure — I’m not sure what’s going to happen here. You know, a lot was going through my head.

And he said, well, if you won’t give me a massage, will you at least show me your breasts? And it was just — you know, it was, all in all, an extremely humiliating experience for me.

And even though I got away, I felt like something had still — like something horrible had just happened to me.

HARI SREENIVASAN: You know, in the immediate aftermath, did you tell someone about it? Because you have said before that you felt ashamed…

KATHERINE KENDALL: I did.

HARI SREENIVASAN: … even though you were the victim.

KATHERINE KENDALL: I did.

It’s really interesting how that happens. And I think — you know, I’m older now, and I have done some work on myself. And I have learned that a lot of people feel that way.

It’s — it’s not — it wasn’t just me. But the just me feeling that this is my fault, this must have only happened to me, there’s something wrong with me, is so common when someone perpetrates against you.

HARI SREENIVASAN: What were the…

KATHERINE KENDALL: And I did. I told my mom.

And I told some good friends. But, you know, one of the things that happened was, I didn’t want them to tell anybody. You know, people wanted to help me, but they didn’t know how, and I didn’t want them to try too hard, because I didn’t want it to backlash.

I was scared. And I think that it’s important to remember that we don’t really come from a culture that supports women in talking about sexual harassment, in my — in my experience, that is. And, you know, I just haven’t felt like it was something I was going to get support on…

HARI SREENIVASAN: You know, how long…

KATHERINE KENDALL: … in the bigger picture.

HARI SREENIVASAN: Yes.

How long did this feeling last? Or, I guess, what are the longer-term ripple effects here? Did it shake your confidence in your abilities?

KATHERINE KENDALL: I think it did. I think it did. I think it did.

I think it made me feel like, wow, you know, that was a wash. He wasn’t interested at all in what I had to say, or, you know, he didn’t see any talent there or intellect there. He was assessing the situation the whole time for something else.

And I think that — that did hurt. You know, I wish it didn’t.

HARI SREENIVASAN: Yes.

KATHERINE KENDALL: But he had produced so many movies that I thought were wonderful. And it was — it’s hard when someone has made art that you love, and how do you stay attached to liking their art, but feeling conflicted about them?

And, yes, I think it does have long-term effects. I think you tuck it away. And then, for me, also, I realized that it came back when I would see his name or see him in person. I would start to sort of tremble all over again.

I mean, I wouldn’t think about him on a daily basis or anything for years, and then I would see him, and I would think, oh, I don’t feel well. I got to get out of here.

HARI SREENIVASAN: Right.

KATHERINE KENDALL: You know, it would bring up so much emotion.

And the most recent one was the woman in New York, the Italian model. I felt so, so enraged when I saw what happened there, and that they sort of — the police had him, and that then he got away. And then she was being dragged through the press as somebody who just, you know, wanted a payout, et cetera.

HARI SREENIVASAN: You know, in the wake of that, there was — a friend of yours had tweeted, “At some point, all the women who have been afraid to speak out about Harvey Weinstein are going to have to hold hands and jump.” This was back in 2015.

And from your Twitter account, you said, “Agreed.”

It seemed like you almost had the opportunity to come forward.

What made you want to come forward now? Has this become a turning point in the industry?

KATHERINE KENDALL: This is a turning point. It’s a turning point.

There are so many times when I thought about it, and then felt like — there were times when I thought about it and said, well, I have nothing to lose, I will just do it. And then I thought, I — I just didn’t have the strength or the courage yet.

And I think somebody like Jodi Kantor doing the story for The New York Times, the fact that she thought it was a story at all was startling to me and made me feel like, wow, something is going to be done.

And I knew she had told me — I mean, they were looking for women that this had happened to, because they’d been hearing rumors for so long that it happened to so many people. And she had told me other people were coming out.

And I thought, I can’t — when I watched Rose McGowan or any of the other actresses come forward, I just — or Ashley Judd — I just thought, they look strong to me, and I don’t want to be the one that stays silent.

HARI SREENIVASAN: Well, Katherine Kendall…

KATHERINE KENDALL: I want to stand beside them.

HARI SREENIVASAN: Katherine Kendall, thank you very much for speaking with us.

And, hopefully, there are other people that are empowered by you coming forward.

KATHERINE KENDALL: I hope so. Thank you.

The post Escaping Harvey Weinstein was a ‘cat-and-mouse game,’ says Katherine Kendall appeared first on PBS NewsHour.

A lithium-ion battery made from three droplets of hydrogel is the smallest soft battery of its kind – and it could be used in biocompatible and biodegradable implants

The Alloy Origins Core is a tenkeyless mechanical keyboard made of airplane-grade aluminum, which prompts the question: Why don't they make the whole plane out of keyboard?

A lithium-ion battery made from three droplets of hydrogel is the smallest soft battery of its kind – and it could be used in biocompatible and biodegradable implants

ASUS has introduced the Wireless Mouse MD102 in India, offering a blend of durability and wireless dependability in an ergonomic design. Available in Dark Grey and White, this stylish mouse is priced at Rs 1799 and will be available from September

The future of investing online is here.

Although a robust inflammatory response is needed to combat infection, this response must ultimately be terminated to prevent chronic inflammation. One mechanism that terminates inflammatory signaling is the production of alternative mRNA splice forms in the Toll-like receptor (TLR) signaling pathway. Whereas most genes in the TLR pathway encode positive mediators of inflammatory signaling, several, including that encoding the MyD88 signaling adaptor, also produce alternative spliced mRNA isoforms that encode dominant-negative inhibitors of the response. Production of these negatively acting alternatively spliced isoforms is induced by stimulation with the TLR4 agonist lipopolysaccharide (LPS); thus, this alternative pre-mRNA splicing represents a negative feedback loop that terminates TLR signaling and prevents chronic inflammation. In the current study, we investigated the mechanisms regulating the LPS-induced alternative pre-mRNA splicing of the MyD88 transcript in murine macrophages. We found that 1) the induction of the alternatively spliced MyD88 form is due to alternative pre-mRNA splicing and not caused by another RNA regulatory mechanism, 2) MyD88 splicing is regulated by both the MyD88- and TRIF-dependent arms of the TLR signaling pathway, 3) MyD88 splicing is regulated by the NF-κB transcription factor, and 4) NF-κB likely regulates MyD88 alternative pre-mRNA splicing per se rather than regulating splicing indirectly by altering MyD88 transcription. We conclude that alternative splicing of MyD88 may provide a sensitive mechanism that ensures robust termination of inflammation for tissue repair and restoration of normal tissue homeostasis once an infection is controlled.

Scientists propose dropping poison to kill mice on the Farallon Islands, but there's concern that wildlife and water quality will suffer.

New York City's Feral Cat Initiative pairs feral cats that need a range with humans who want a defense against rodents.

The mouse deer. Or the pig deer. Or the little goat. Whatever you call the chevrotain, this is a truly distinct looking (and tiny!) ungulate.

Vacationers are becoming wary of theme parks as the swine flu outbreak worsens.

Researchers say the mutant mice study could increase our understanding of perception systems.

Today, Mouseflow, a fast-growing SaaS platform for website analytics, welcomes Mikkel Wakefield as the company's new CEO.

Mouseflow today announced the launch of Live Heatmaps. The Live Heatmap is going to change the way businesses are using traditional web heat maps. Live Heatmaps has been on its way for the past nine (9) months.

Robert Iger, CEO of Disney.

Using CSS3 to produce click gallery with permanent images to rival jQuery/javascript.

A CSS and javascript responsive swipe action gallery with momentum scrolling. For touch screen, trackpad and mouse.

We present a new mouse cursor designed to facilitate the use of the mouse by people with peripheral vision loss. The pointer consists of a collection of converging straight lines covering the whole screen and following the position of the mouse cursor. We measured its positive effects with a group of participants with peripheral vision loss of different kinds and we found that it can reduce by a factor of 7 the time required to complete a targeting task using the mouse. Using eye tracking, we show that this system makes it possible to initiate the movement towards the target without having to precisely locate the mouse pointer. Using Fitts' Law, we compare these performances with those of full visual field users in order to understand the relation between the accuracy of the estimated mouse cursor position and the index of performance obtained with our tool.

A technique is provided for post selection location of a mouse pointer icon in a display screen of a computing device. A software tool receives input of the post selection location for the mouse pointer icon. The post selection location defines a default location to move the mouse pointer icon in response to a window action taken on a window displayed in the display screen. In response to the window action in which the mouse pointer icon is initially displayed at a selection location corresponding to the window action, the mouse pointer icon is moved to the post selection location such that the mouse pointer icon is displayed at the post selection location in the display screen.

HE may have made his first on-screen appearance more than ninety years ago, but in 1989 he showed his happy face at Marks and Spencer, Southampton.

In this podcast, The Evil Chocolate Cookie gives us a demonstration of the Tap Strap 2, a single handed all-in-one wearable keyboard, mouse & air gesture controller.

Tap is a wearable keyboard and mouse that enables you to type, mouse and control any Bluetooth enabled device, using any surface and in any position. Tap has accelerometers built into each finger ring, which register which fingers you are tapping and sending the associated letter, number, symbol, or macro to a paired Bluetooth device.

Tap is the only ergonomic solution that allows you to type in any position that is comfortable to your specific body without being bound to a desk. Tap’s design takes stress off your median nerve, allowing for hours of typing without strain. Tap also allows you to write without hunching forward, which causes posture problems as well as neck, shoulder and back pain.

Learning to Tap takes about 2 Hours – which is approximately 10 times faster than it takes to learn to QWERTY touch-typing. You learn to Tap using finger combinations, not key locations, making it usable in a variety of situations where you could not use a QWERTY. The fastest way to learn is by playing our free TapGenius Learning Game, which takes you through the Tap Alphabet using a series of fast, engaging challenges.

The mouse clicked in Indiana, but the gun that fired the shot heard 'round the Internet sits on a game preserve in Texas. Now, 14 states and an influential congressman want to ban online hunting. -Robert MacMillan

Sometimes the people who make the biggest contributions to our culture can be pretty strange characters.

Adhesive mouse traps which use glue to slowly inflict fatal injuries are still being sold at discount shops throughout Australia and are harming native animals, according to an animal rights group.

A West Australian man has captured on camera what may be one of the first documented cases of a redback spider capturing a mouse.

Here's your first look at Disney's new masks that will benefit families in vulnerable communities.

Colorways & Toeboxes Live from L.A.

Colorways & Toeboxes Live from L.A.

Assembled α-synuclein in nerve cells and glial cells is the defining pathological feature of neurodegenerative diseases called synucleinopathies. Seeds of α-synuclein can induce the assembly of monomeric protein. Here, we used sucrose gradient centrifugation and transiently transfected HEK 293T cells to identify the species of α-synuclein from the brains of homozygous, symptomatic mice transgenic for human mutant A53T α-synuclein (line M83) that seed aggregation. The most potent fractions contained Sarkosyl-insoluble assemblies enriched in filaments. We also analyzed six cases of idiopathic Parkinson's disease (PD), one case of familial PD, and six cases of multiple system atrophy (MSA) for their ability to induce α-synuclein aggregation. The MSA samples were more potent than those of idiopathic PD in seeding aggregation. We found that following sucrose gradient centrifugation, the most seed-competent fractions from PD and MSA brains are those that contain Sarkosyl-insoluble α-synuclein. The fractions differed between PD and MSA, consistent with the presence of distinct conformers of assembled α-synuclein in these different samples. We conclude that α-synuclein filaments are the main driving force for amplification and propagation of pathology in synucleinopathies.

The plasmas of diabetic or uremic patients and of those receiving peritoneal dialysis treatment have increased levels of the glucose-derived dicarbonyl metabolites like methylglyoxal (MGO), glyoxal (GO), and 3-deoxyglucosone (3-DG). The elevated dicarbonyl levels can contribute to the development of painful neuropathies. Here, we used stimulated immunoreactive Calcitonin Gene–Related Peptide (iCGRP) release as a measure of nociceptor activation, and we found that each dicarbonyl metabolite induces a concentration-, TRPA1-, and Ca2+-dependent iCGRP release. MGO, GO, and 3-DG were about equally potent in the millimolar range. We hypothesized that another dicarbonyl, 3,4-dideoxyglucosone-3-ene (3,4-DGE), which is present in peritoneal dialysis (PD) solutions after heat sterilization, activates nociceptors. We also showed that at body temperatures 3,4-DGE is formed from 3-DG and that concentrations of 3,4-DGE in the micromolar range effectively induced iCGRP release from isolated murine skin. In a novel preparation of the isolated parietal peritoneum PD fluid or 3,4-DGE alone, at concentrations found in PD solutions, stimulated iCGRP release. We also tested whether inflammatory tissue conditions synergize with dicarbonyls to induce iCGRP release from isolated skin. Application of MGO together with bradykinin or prostaglandin E2 resulted in an overadditive effect on iCGRP release, whereas MGO applied at a pH of 5.2 resulted in reduced release, probably due to an MGO-mediated inhibition of transient receptor potential (TRP) V1 receptors. These results indicate that several reactive dicarbonyls activate nociceptors and potentiate inflammatory mediators. Our findings underline the roles of dicarbonyls and TRPA1 receptors in causing pain during diabetes or renal disease.

Qin Liu
Aug 1, 2007; 6:1299-1317
Research

Although a robust inflammatory response is needed to combat infection, this response must ultimately be terminated to prevent chronic inflammation. One mechanism that terminates inflammatory signaling is the production of alternative mRNA splice forms in the Toll-like receptor (TLR) signaling pathway. Whereas most genes in the TLR pathway encode positive mediators of inflammatory signaling, several, including that encoding the MyD88 signaling adaptor, also produce alternative spliced mRNA isoforms that encode dominant-negative inhibitors of the response. Production of these negatively acting alternatively spliced isoforms is induced by stimulation with the TLR4 agonist lipopolysaccharide (LPS); thus, this alternative pre-mRNA splicing represents a negative feedback loop that terminates TLR signaling and prevents chronic inflammation. In the current study, we investigated the mechanisms regulating the LPS-induced alternative pre-mRNA splicing of the MyD88 transcript in murine macrophages. We found that 1) the induction of the alternatively spliced MyD88 form is due to alternative pre-mRNA splicing and not caused by another RNA regulatory mechanism, 2) MyD88 splicing is regulated by both the MyD88- and TRIF-dependent arms of the TLR signaling pathway, 3) MyD88 splicing is regulated by the NF-κB transcription factor, and 4) NF-κB likely regulates MyD88 alternative pre-mRNA splicing per se rather than regulating splicing indirectly by altering MyD88 transcription. We conclude that alternative splicing of MyD88 may provide a sensitive mechanism that ensures robust termination of inflammation for tissue repair and restoration of normal tissue homeostasis once an infection is controlled.

The DOCK-D (dedicator of cytokinesis D) family proteins are atypical guanine nucleotide exchange factors that regulate Rho GTPase activity. The family consists of Zizimin1 (DOCK9), Zizimin2 (DOCK11), and Zizimin3 (DOCK10). Functions of the DOCK-D family proteins are presently not well-explored, and the role of the DOCK-D family in neuroinflammation is unknown. In this study, we generated three mouse lines in which DOCK9 (DOCK9−/−), DOCK10 (DOCK10−/−), or DOCK11 (DOCK11−/−) had been deleted and examined the phenotypic effects of these gene deletions in MOG35–55 peptide-induced experimental autoimmune encephalomyelitis, an animal model of the neuroinflammatory disorder multiple sclerosis. We found that all the gene knockout lines were healthy and viable. The only phenotype observed under normal conditions was a slightly smaller proportion of B cells in splenocytes in DOCK10−/− mice than in the other mouse lines. We also found that the migration ability of macrophages is impaired in DOCK10−/− and DOCK11−/− mice and that the severity of experimental autoimmune encephalomyelitis was ameliorated only in DOCK10−/− mice. No apparent phenotype was observed for DOCK9−/− mice. Further investigations indicated that lipopolysaccharide stimulation up-regulates DOCK10 expression in microglia and that microglial migration is decreased in DOCK10−/− mice. Up-regulation of C–C motif chemokine ligand 2 (CCL2) expression induced by activation of Toll-like receptor 4 or 9 signaling was reduced in DOCK10−/− astrocytes compared with WT astrocytes. Taken together, our findings suggest that DOCK10 plays a role in innate immunity and neuroinflammation and might represent a potential therapeutic target for managing multiple sclerosis.

Assembled α-synuclein in nerve cells and glial cells is the defining pathological feature of neurodegenerative diseases called synucleinopathies. Seeds of α-synuclein can induce the assembly of monomeric protein. Here, we used sucrose gradient centrifugation and transiently transfected HEK 293T cells to identify the species of α-synuclein from the brains of homozygous, symptomatic mice transgenic for human mutant A53T α-synuclein (line M83) that seed aggregation. The most potent fractions contained Sarkosyl-insoluble assemblies enriched in filaments. We also analyzed six cases of idiopathic Parkinson's disease (PD), one case of familial PD, and six cases of multiple system atrophy (MSA) for their ability to induce α-synuclein aggregation. The MSA samples were more potent than those of idiopathic PD in seeding aggregation. We found that following sucrose gradient centrifugation, the most seed-competent fractions from PD and MSA brains are those that contain Sarkosyl-insoluble α-synuclein. The fractions differed between PD and MSA, consistent with the presence of distinct conformers of assembled α-synuclein in these different samples. We conclude that α-synuclein filaments are the main driving force for amplification and propagation of pathology in synucleinopathies.

The plasmas of diabetic or uremic patients and of those receiving peritoneal dialysis treatment have increased levels of the glucose-derived dicarbonyl metabolites like methylglyoxal (MGO), glyoxal (GO), and 3-deoxyglucosone (3-DG). The elevated dicarbonyl levels can contribute to the development of painful neuropathies. Here, we used stimulated immunoreactive Calcitonin Gene–Related Peptide (iCGRP) release as a measure of nociceptor activation, and we found that each dicarbonyl metabolite induces a concentration-, TRPA1-, and Ca2+-dependent iCGRP release. MGO, GO, and 3-DG were about equally potent in the millimolar range. We hypothesized that another dicarbonyl, 3,4-dideoxyglucosone-3-ene (3,4-DGE), which is present in peritoneal dialysis (PD) solutions after heat sterilization, activates nociceptors. We also showed that at body temperatures 3,4-DGE is formed from 3-DG and that concentrations of 3,4-DGE in the micromolar range effectively induced iCGRP release from isolated murine skin. In a novel preparation of the isolated parietal peritoneum PD fluid or 3,4-DGE alone, at concentrations found in PD solutions, stimulated iCGRP release. We also tested whether inflammatory tissue conditions synergize with dicarbonyls to induce iCGRP release from isolated skin. Application of MGO together with bradykinin or prostaglandin E2 resulted in an overadditive effect on iCGRP release, whereas MGO applied at a pH of 5.2 resulted in reduced release, probably due to an MGO-mediated inhibition of transient receptor potential (TRP) V1 receptors. These results indicate that several reactive dicarbonyls activate nociceptors and potentiate inflammatory mediators. Our findings underline the roles of dicarbonyls and TRPA1 receptors in causing pain during diabetes or renal disease.

Fatty acid transport protein 2 (FATP2) is highly expressed in the liver, small intestine, and kidney, where it functions in both the transport of exogenous long-chain fatty acids and the activation of very-long-chain fatty acids. Here, using a murine model, we investigated the phenotypic impacts of deleting FATP2, followed by a transcriptomic analysis using unbiased RNA-Seq to identify concomitant changes in the liver transcriptome. WT and FATP2-null (Fatp2−/−) mice (5 weeks) were maintained on a standard chow diet for 6 weeks. The Fatp2−/− mice had reduced weight gain, lowered serum triglyceride, and increased serum cholesterol levels and attenuated dietary fatty acid absorption. Transcriptomic analysis of the liver revealed 258 differentially expressed genes in male Fatp2−/− mice and a total of 91 in female Fatp2−/− mice. These genes mapped to the following gene ontology categories: fatty acid degradation, peroxisome biogenesis, fatty acid synthesis, and retinol and arachidonic acid metabolism. Targeted RT-quantitative PCR verified the altered expression of selected genes. Of note, most of the genes with increased expression were known to be regulated by peroxisome proliferator–activated receptor α (PPARα), suggesting that FATP2 activity is linked to a PPARα-specific proximal ligand. Targeted metabolomic experiments in the Fatp2−/− liver revealed increases of total C16:0, C16:1, and C18:1 fatty acids; increases in lipoxin A4 and prostaglandin J2; and a decrease in 20-hydroxyeicosatetraenoic acid. We conclude that the expression of FATP2 in the liver broadly affects the metabolic landscape through PPARα, indicating that FATP2 provides an important role in liver lipid metabolism through its transport or activation activities.

Fabry disease is a heritable lipid disorder caused by the low activity of α-galactosidase A and characterized by the systemic accumulation of globotriaosylceramide (Gb3). Recent studies have reported a structural heterogeneity of Gb3 in Fabry disease, including Gb3 isoforms with different fatty acids and Gb3 analogs with modifications on the sphingosine moiety. However, Gb3 assays are often performed only on the selected Gb3 isoforms. To precisely determine the total Gb3 concentration, here we established two methods for determining both Gb3 isoforms and analogs. One was the deacylation method, involving Gb3 treatment with sphingolipid ceramide N-deacylase, followed by an assay of the deacylated products, globotriaosylsphingosine (lyso-Gb3) and its analogs, by ultra-performance LC coupled to tandem MS (UPLC-MS/MS). The other method was a direct assay established in the present study for 37 Gb3 isoforms and analogs/isoforms by UPLC-MS/MS. Gb3s from the organs of symptomatic animals of a Fabry disease mouse model were mainly Gb3 isoforms and two Gb3 analogs, such as Gb3(+18) containing the lyso-Gb3(+18) moiety and Gb3(−2) containing the lyso-Gb3(−2) moiety. The total concentrations and Gb3 analog distributions determined by the two methods were comparable. Gb3(+18) levels were high in the kidneys (24% of total Gb3) and the liver (13%), and we observed Gb3(−2) in the heart (10%) and the kidneys (5%). These results indicate organ-specific expression of Gb3 analogs, insights that may lead to a deeper understanding of the pathophysiology of Fabry disease.

Although a robust inflammatory response is needed to combat infection, this response must ultimately be terminated to prevent chronic inflammation. One mechanism that terminates inflammatory signaling is the production of alternative mRNA splice forms in the Toll-like receptor (TLR) signaling pathway. Whereas most genes in the TLR pathway encode positive mediators of inflammatory signaling, several, including that encoding the MyD88 signaling adaptor, also produce alternative spliced mRNA isoforms that encode dominant-negative inhibitors of the response. Production of these negatively acting alternatively spliced isoforms is induced by stimulation with the TLR4 agonist lipopolysaccharide (LPS); thus, this alternative pre-mRNA splicing represents a negative feedback loop that terminates TLR signaling and prevents chronic inflammation. In the current study, we investigated the mechanisms regulating the LPS-induced alternative pre-mRNA splicing of the MyD88 transcript in murine macrophages. We found that 1) the induction of the alternatively spliced MyD88 form is due to alternative pre-mRNA splicing and not caused by another RNA regulatory mechanism, 2) MyD88 splicing is regulated by both the MyD88- and TRIF-dependent arms of the TLR signaling pathway, 3) MyD88 splicing is regulated by the NF-κB transcription factor, and 4) NF-κB likely regulates MyD88 alternative pre-mRNA splicing per se rather than regulating splicing indirectly by altering MyD88 transcription. We conclude that alternative splicing of MyD88 may provide a sensitive mechanism that ensures robust termination of inflammation for tissue repair and restoration of normal tissue homeostasis once an infection is controlled.

Fatty acid transport protein 2 (FATP2) is highly expressed in the liver, small intestine, and kidney, where it functions in both the transport of exogenous long-chain fatty acids and the activation of very-long-chain fatty acids. Here, using a murine model, we investigated the phenotypic impacts of deleting FATP2, followed by a transcriptomic analysis using unbiased RNA-Seq to identify concomitant changes in the liver transcriptome. WT and FATP2-null (Fatp2−/−) mice (5 weeks) were maintained on a standard chow diet for 6 weeks. The Fatp2−/− mice had reduced weight gain, lowered serum triglyceride, and increased serum cholesterol levels and attenuated dietary fatty acid absorption. Transcriptomic analysis of the liver revealed 258 differentially expressed genes in male Fatp2−/− mice and a total of 91 in female Fatp2−/− mice. These genes mapped to the following gene ontology categories: fatty acid degradation, peroxisome biogenesis, fatty acid synthesis, and retinol and arachidonic acid metabolism. Targeted RT-quantitative PCR verified the altered expression of selected genes. Of note, most of the genes with increased expression were known to be regulated by peroxisome proliferator–activated receptor α (PPARα), suggesting that FATP2 activity is linked to a PPARα-specific proximal ligand. Targeted metabolomic experiments in the Fatp2−/− liver revealed increases of total C16:0, C16:1, and C18:1 fatty acids; increases in lipoxin A4 and prostaglandin J2; and a decrease in 20-hydroxyeicosatetraenoic acid. We conclude that the expression of FATP2 in the liver broadly affects the metabolic landscape through PPARα, indicating that FATP2 provides an important role in liver lipid metabolism through its transport or activation activities.