Loss of aryl hydrocarbon receptor potentiates FoxM1 signaling to enhance self-renewal of colonic stem and progenitor cells.

The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that senses xenobiotics, diet, and gut microbial-derived metabolites, is increasingly recognized as a key regulator of intestinal biology. However, its effects on the function of colonic stem and progenitor cells remain largely unexplored. Here, we observed that inducible deletion of AhR in Lgr5+ stem cells increases the percentage of colonic stem cells and enhances organoid initiating capacity and growth of sorted stem and progenitor cells, while AhR activation has the opposite effect. Moreover, intestinal-specific AhR knockout increases basal stem cell and crypt injury-induced cell proliferation and promotes colon tumorigenesis in a preclinical colitis-associated tumor model by upregulating FoxM1 signaling. Mechanistically, AhR transcriptionally suppresses FoxM1 expression. Activation of AhR in human organoids recapitulates phenotypes observed in mice, such as reduction in the percentage of colonic stem cells, promotion of stem cell differentiation, and attenuation of FoxM1 signaling. These findings indicate that the AhR-FoxM1 axis, at least in part, mediates colonic stem/progenitor cell behavior.

Chlorogenic Acid and Quercetin in a Diet with Fermentable Fiber Influence Multiple Processes Involved in DSS-Induced Ulcerative Colitis but Do Not Reduce Injury.

Ulcerative colitis (UC) patients often avoid foods containing fermentable fibers as some can promote symptoms during active disease. Pectin has been identified as a more protective fermentable fiber, but little has been done to determine the interaction between pectin and bioactive compounds present in foods containing that fiber type. Quercetin and chlorogenic acid, two bioactives in stone fruits, may have anti-cancer, anti-oxidant, and anti-inflammatory properties. We hypothesized that quercetin and chlorogenic acid, in the presence of the fermentable fiber pectin, may suppress the expression of pro-inflammatory molecules, alter the luminal environment, and alter colonocyte proliferation, thereby protecting against recurring bouts of UC. Rats (n = 63) received one of three purified diets (control, 0.45% quercetin, 0.05% chlorogenic acid) containing 6% pectin for 3 weeks before exposure to dextran sodium sulfate (DSS, 3% for 48 h, 3x, 2 wk separation, n = 11/diet) in drinking water to initiate UC, or control (no DSS, n = 10/diet) treatments prior to termination at 9 weeks. DSS increased the fecal moisture content (p < 0.05) and SCFA concentrations (acetate, p < 0.05; butyrate, p < 0.05). Quercetin and chlorogenic acid diets maintained SLC5A8 (SCFA transporter) mRNA levels in DSS-treated rats at levels similar to those not exposed to DSS. DSS increased injury (p < 0.0001) and inflammation (p < 0.01) scores, with no differences noted due to diet. Compared to the control diet, chlorogenic acid decreased NF-κB activity in DSS-treated rats (p < 0.05). Quercetin and chlorogenic acid may contribute to the healthy regulation of NF-κB activation (via mRNA expression of IκΒα, Tollip, and IL-1). Quercetin enhanced injury-repair molecule FGF-2 expression (p < 0.01), but neither diet nor DSS treatment altered proliferation. Although quercetin and chlorogenic acid did not protect against overt indicators of injury and inflammation, or fecal SCFA concentrations, compared to the control diet, their influence on the expression of injury repair molecules, pro-inflammatory cytokines, SCFA transport proteins, and NF-κB inhibitory molecules suggests beneficial influences on major pathways involved in DSS-induced UC. Therefore, in healthy individuals or during periods of remission, quercetin and chlorogenic acid may promote a healthier colon, and may suppress some of the signaling involved in inflammation promotion during active disease.

Diffusible contrast-enhanced micro-CT improves visualization of stented vessels.

In this report, we showcase diffusible iodine-based contrast-enhanced computed tomography (DICE-CT) as a method for improving soft tissue visualization and reducing beam hardening artifact within a stented vessel. This technique is commonly used in our pathology lab to image soft tissue specimens with dense metal implants and to ensure reliable morphological analysis through clear delineation of tissue structures. For this report, a porcine right coronary artery with an implanted metal stent was scanned using both conventional and DICE-CT methods. Upon reconstruction, DICE-CT produced less beam hardening artifact in comparison to traditional micro-CT; furthermore, DICE-CT produced results with morphometric similarity to histology. Accordingly, these differences illustrated the clear advantage of using DICE-CT over conventional micro-CT when imaging soft tissue specimens with dense metal implants.

Role of tumor necrosis factor alpha in disease using a mouse model of Shiga toxin-mediated renal damage.

Mice have been extensively employed as an animal model of renal damage caused by Shiga toxins. In this study, we examined the role of the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) in the development of toxin-mediated renal disease in mice. Mice pretreated with TNF-alpha and challenged with Shiga toxin type 1 (Stx1) showed increased survival compared to that of mice treated with Stx1 alone. Conversely, mice treated with Stx1 before TNF-alpha administration succumbed more quickly than mice given Stx1 alone. Increased lethality in mice treated with Stx1 followed by TNF-alpha was associated with evidence of glomerular damage and the loss of renal function. No differences in renal histopathology were noted between animals treated with Stx1 alone and the TNF-alpha pretreatment group, although we noted a sparing of renal function when TNF-alpha was administered before toxin. Compared to that of treatment with Stx1 alone, treatment with TNF-alpha after toxin altered the renal cytokine profile so that the expression of proinflammatory cytokines TNF-alpha and interleukin-1beta (IL-1beta) increased, and the expression of the anti-inflammatory cytokine IL-10 decreased. Increased lethality in mice treated with Stx1 followed by TNF-alpha was associated with higher numbers of dUTP-biotin nick end labeling-positive renal tubule cells, suggesting that increased lethality involved enhanced apoptosis. These data suggest that the early administration of TNF-alpha is a candidate interventional strategy blocking disease progression, while TNF-alpha production after intoxication exacerbates disease.

Intrauterine infusion of latency-associated peptide (LAP) during early porcine pregnancy affects conceptus elongation and placental size.

In the pig, transforming growth factor beta (TGFB), TGFB receptors (TGFBRs), and integrins are present during the peri-implantation period. Latency-associated peptide (LAP), a part of latent TGFB, can bind to integrin heterodimers via its Arg-Gly-Asp (RGD) sequence; therefore, ligand-receptor interactions between TGFB and TGFBRs, along with LAP and integrin heterodimers, may be functional in mediating events supporting conceptus elongation and attachment. With the use of surgically implantable osmotic pumps, we were able to maintain pregnancy with the aim of mechanistically altering in vivo receptor-ligand interactions involving TGFB with TGFBRs and LAP with integrins during porcine pregnancy. Day 9 pregnant gilts received intrauterine infusions of LAP-RGD, a recombinant mutant of LAP (LAP-RGE), or vehicle control and were ovariohysterectomized on Day 13 or 24 of pregnancy. We hypothesized that intrauterine infusion of LAP-RGD would decrease downstream signaling of TGFB while increasing LAP-integrin interactions and that net effect would enhance conceptus survival and attachment early in the peri-implantation period but possibly increase the chance of abnormal placentation later in pregnancy. Additionally, we hypothesized that infusion of LAP-RGE would disrupt TGFB signals but not alter integrin signaling, and thus the net result would be decreased conceptus survival and abnormal development. Unexpectedly, LAP-RGD intrauterine infusions resulted in a reduction of conceptus elongation, whereas infusions of LAP-RGE permitted implantation and placentation but resulted in larger fetal weight, allantois length, and allantoic fluid volume. Results suggest TGFB and integrins are contributing factors in the regulation of conceptus elongation and placental and fetal size.

Dietary polyunsaturated fatty acids modulate resistance to Mycobacterium tuberculosis in guinea pigs.

It is well established that the nutritional status of the host affects resistance to disease. The impact of dietary lipids on experimental pulmonary infection with mycobacteria has not been investigated. Therefore, the purpose of this study was to determine the role of dietary (n-3) and (n-6) fatty acids on immunity and resistance to aerosol infection with virulent Mycobacterium tuberculosis in guinea pigs. Weanling guinea pigs were fed purified, isocaloric diets differing only in lipid source, and the effects of diet on specific immune cell functions were evaluated after 3 or 6 wk. Dietary (n-3) fatty acid consumption reduced in vivo skin test and in vitro lympho-proliferative responses (P < 0.05) relative to (n-6) fatty acid consumption. The effect of diet on resistance to mycobacterial infection was assessed by enumerating viable mycobacteria in the lungs and spleens of guinea pigs infected with virulent M. tuberculosis by the aerosol route. (n-3) Fatty acid-fed guinea pigs had more bacteria in the lungs compared with (n-6) fatty acid-fed guinea pigs at 3 (P < 0.05) and 6 wk postinfection (P < 0.01). These data document the immunomodulatory effects of (n-3) fatty acid consumption in the context of tuberculosis resistance. The loss of antigen-specific T-cell functions in addition to impaired resistance to mycobacterial disease suggests a susceptible phenotype in (n-3) fatty acid-fed guinea pigs.

Intravascular hemolysis associated with severe cutaneous burn injuries in five horses.

Five horses were evaluated because of severe cutaneous burn injuries following a barn fire. Gross hemolysis and morphologic changes in RBCs consistent with oxidative damage were detected in all of the horses. Of these horses, 4 became azotemic. The overall goals of treatment included wound care, correction of dehydration and provision of diuresis, control of inflammation, pain management, and prophylaxis against sepsis. After treatment, 2 horses survived and were discharged from the hospital. Red blood cell damage and hemolysis following cutaneous burn injury have been investigated in other species and appear to be a result of the release of oxygen radicals from complement-activated neutrophils. Early intervention with aggressive fluid therapy is recommended in the treatment of human burn patients and is likely to be of benefit in horses with burn injuries; a beneficial role of free radical scavengers and xanthine oxidase inhibitors has also been suggested.

Oil Red O-positive lipid in peritoneal fluid from a horse with a rectal tear.

A 4-year-old Quarter Horse mare was presented to the Texas A&M University Veterinary Medical Teaching Hospital for evaluation of a rectal tear. On initial evaluation, rectal palpation and colonoscopy revealed a grade IIIb rectal tear. Analysis of peritoneal fluid revealed a modified transudate. Preliminary supportive care included fluid therapy and mineral oil administration via nasogastric tube. Approximately 48 hours after presentation, a second abdominocentesis was performed, and cytologic examination of the fluid revealed a marked suppurative exudate. Round clear nonrefractile material observed within neutrophils and macrophages and in the background stained bright pink to red with Oil Red O, confirming the material as lipid, likely from leakage of mineral oil through the rectal tear. The condition of the mare deteriorated and euthanasia was elected due to the poor prognosis. At necropsy, gross and histologic findings included peritoneal effusion and a full-thickness rectal tear with transmural necrotizing pyogranulomatous colitis and fibrinous peritonitis. To the authors' knowledge, this is the first reported case of Oil Red O-positive lipid vacuoles in the peritoneal fluid of a horse from presumed leakage of mineral oil through a transmural rectal perforation. The frequency of this occurrence in horses is unknown, but it is important for cytopathologists to be familiar with the appearance and significance of lipid-type droplets in phagocytic cells in cytologic fluid analysis specimens.

An improved ferritin assay for canine sera.

An improved serum ferritin assay for canine serum has been developed. It uses two monoclonal antibodies in a sandwich arrangement. Serum ferritin can be determined on undiluted canine sera with this assay. The recovery of ferritin added to canine serum ranged from 98 to 106%, the within-assay coefficient of variability was 3.3 to 4.5%, and the assay-to-assay variability was 9.8 to 10.2%. Serum ferritin from 61 apparently healthy dogs had a geometric mean of 252 ng/ml, with a range of 80 ng/ml to 800 ng/ml.