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Lipopolysaccharides (LPS), also termed endotoxins, are the major component of the outer membrane of Gram-negative bacteria. In general, endotoxins in the intestine are considered harmless in healthy animals. However, different stressors, such as heat stress, can lead to a compromised gut barrier, resulting in endotoxin translocation. Chickens are considered to be less sensitive to the effects of LPS compared with other species, for example, humans, pigs, or calves, probably because of the lack of the functional-specific TRAM-TRIF signalling pathway (MyD88-independent). Therefore, six LPS preparations (three different strains with two different preparation methods each) were compared in murine macrophages and characterized according to their MyD88-dependent pathway activation. All tested LPS preparations induced a strong inflammatory response after 4 and 24 h on a murine macrophage cell line. However, there was a similar strong response in the gene expression profile as well as production of nitrite oxide and TNF-alpha from LPS of different strains and preparation methods. On the basis of the results of the in vitro study, one LPS preparation was chosen for the subsequent in vivo study with broilers to assess the effect of an oral LPS bolus (E. coli O55:B5 phenol extracted; 2 mg/kg b.w.) during heat stress conditions (10 h, 36 °C). The most pronounced effects were seen in broilers receiving the oral LPS bolus during heat stress conditions. The endotoxin activity in the intestine as well as the serum concentration of the 3-OH C14 (part of LPS) were increased. In addition, an increased expression of genes related to inflammation and stress response (e.g., IL-6, IL-1beta, HSP70) was observed, whereas the expression of genes associated with gut health (e.g., MUC2, FABP2) was decreased. To conclude, an increase of intestinal LPS combined with heat stress can pose a risk to animal health.
Nicole Reisinger; Caroline Emsenhuber; Barbara Doupovec; Elisabeth Mayer; Gerd Schatzmayr; Veronika Nagl; Bertrand Grenier. Endotoxin Translocation and Gut Inflammation Are Increased in Broiler Chickens Receiving an Oral Lipopolysaccharide (LPS) Bolus during Heat Stress. Toxins 2020, 12, 622 .
AMA StyleNicole Reisinger, Caroline Emsenhuber, Barbara Doupovec, Elisabeth Mayer, Gerd Schatzmayr, Veronika Nagl, Bertrand Grenier. Endotoxin Translocation and Gut Inflammation Are Increased in Broiler Chickens Receiving an Oral Lipopolysaccharide (LPS) Bolus during Heat Stress. Toxins. 2020; 12 (10):622.
Chicago/Turabian StyleNicole Reisinger; Caroline Emsenhuber; Barbara Doupovec; Elisabeth Mayer; Gerd Schatzmayr; Veronika Nagl; Bertrand Grenier. 2020. "Endotoxin Translocation and Gut Inflammation Are Increased in Broiler Chickens Receiving an Oral Lipopolysaccharide (LPS) Bolus during Heat Stress." Toxins 12, no. 10: 622.
The Fusarium mycotoxin deoxynivalenol (DON) contaminates animal feed worldwide. In vivo, DON modifies the cellular protein synthesis, thereby also affecting the immune system. However, the functional consequences of this are still ill-defined. In this study, peripheral blood mononuclear cells from healthy pigs were incubated with different DON concentrations in the presence of Concanavalin A (ConA), a plant-derived polyclonal T-cell stimulant. T-cell subsets were investigated for proliferation and expression of CD8α, CD27, and CD28, which are involved in activation and costimulation of porcine T cells. A clear decrease in proliferation of all ConA-stimulated major T-cell subsets (CD4+, CD8+, and γδ T cells) was observed in DON concentrations higher than 0.4 µM. This applied in particular to naïve CD4+ and CD8+ T cells. From 0.8 μM onwards, DON induced a reduction of CD8α (CD4+) and CD27 expression (CD4+ and CD8+ T cells). CD28 expression was diminished in CD4+ and CD8+ T cells at a concentration of 1.6 µM DON. None of these effects were observed with the DON-derivative deepoxy-deoxynivalenol (DOM-1) at 16 µM. These results indicate that DON reduces T-cell proliferation and the expression of molecules involved in T-cell activation, providing a molecular basis for some of the described immunosuppressive effects of DON.
Eleni Vatzia; Alix Pierron; Armin Saalmüller; Elisabeth Mayer; Wilhelm Gerner. Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets. Toxins 2019, 11, 644 .
AMA StyleEleni Vatzia, Alix Pierron, Armin Saalmüller, Elisabeth Mayer, Wilhelm Gerner. Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets. Toxins. 2019; 11 (11):644.
Chicago/Turabian StyleEleni Vatzia; Alix Pierron; Armin Saalmüller; Elisabeth Mayer; Wilhelm Gerner. 2019. "Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets." Toxins 11, no. 11: 644.
Forages are important components of dairy cattle rations but might harbor a plethora of mycotoxins. Ruminants are considered to be less susceptible to the adverse health effects of mycotoxins, mainly because the ruminal microflora degrades certain mycotoxins. Yet, impairment of the ruminal degradation capacity or high ruminal stability of toxins can entail that the intestinal epithelium is exposed to significant mycotoxin amounts. The aims of our study were to assess i) the mycotoxin occurrence in maize silage and ii) the cytotoxicity of relevant mycotoxins on bovine intestinal cells. In total, 158 maize silage samples were collected from European dairy cattle farms. LC-MS/MS-based analysis of 61 mycotoxins revealed the presence of emerging mycotoxins (e.g. emodin, culmorin, enniatin B1, enniatin B, and beauvericin) in more than 70% of samples. Among the regulated mycotoxins, deoxynivalenol and zearalenone were most frequently detected (67.7%). Overall, 87% of maize silages contained more than five mycotoxins. Using an in vitro model with calf small intestinal epithelial cells B, the cytotoxicity of deoxynivalenol, nivalenol, fumonisin B1 and enniatin B was evaluated (0–200 µM). Absolute IC50 values varied in dependence of employed assay and were 1.2–3.6 µM, 0.8–1.0 µM, 8.6–18.3 µM, and 4.0–6.7 µM for deoxynivalenol, nivalenol, fumonisin B1, and enniatin B, respectively. Results highlight the potential relevance of mycotoxins for bovine gut health, a previously neglected target in ruminants.
Nicole Reisinger; Sonja Schürer-Waldheim; Elisabeth Mayer; Sandra Debevere; Gunther Antonissen; Michael Sulyok; Veronika Nagl. Mycotoxin Occurrence in Maize Silage—A Neglected Risk for Bovine Gut Health? Toxins 2019, 11, 577 .
AMA StyleNicole Reisinger, Sonja Schürer-Waldheim, Elisabeth Mayer, Sandra Debevere, Gunther Antonissen, Michael Sulyok, Veronika Nagl. Mycotoxin Occurrence in Maize Silage—A Neglected Risk for Bovine Gut Health? Toxins. 2019; 11 (10):577.
Chicago/Turabian StyleNicole Reisinger; Sonja Schürer-Waldheim; Elisabeth Mayer; Sandra Debevere; Gunther Antonissen; Michael Sulyok; Veronika Nagl. 2019. "Mycotoxin Occurrence in Maize Silage—A Neglected Risk for Bovine Gut Health?" Toxins 11, no. 10: 577.
Feed samples are frequently contaminated by a wide range of chemically diverse natural products, which can be determined using highly sensitive analytical techniques. Next to already well-investigated mycotoxins, unknown or unregulated fungal secondary metabolites have also been found, some of which at significant concentrations. In our study, 1141 pig feed samples were analyzed for more than 800 secondary fungal metabolites using the same LC-MS/MS method and ranked according to their prevalence. Effects on the viability of the 28 most relevant were tested on an intestinal porcine epithelial cell line (IPEC-J2). The most frequently occurring compounds were determined as being cyclo-(L-Pro-L-Tyr), moniliformin, and enniatin B, followed by enniatin B1, aurofusarin, culmorin, and enniatin A1. The main mycotoxins, deoxynivalenol and zearalenone, were found only at ranks 8 and 10. Regarding cytotoxicity, apicidin, gliotoxin, bikaverin, and beauvericin led to lower IC50 values, between 0.52 and 2.43 µM, compared to deoxynivalenol (IC50 = 2.55 µM). Significant cytotoxic effects were also seen for the group of enniatins, which occurred in up to 82.2% of the feed samples. Our study gives an overall insight into the amount of fungal secondary metabolites found in pig feed samples compared to their cytotoxic effects in vitro.
Barbara Novak; Valentina Rainer; Michael Sulyok; Dietmar Haltrich; Gerd Schatzmayr; Elisabeth Mayer. Twenty-Eight Fungal Secondary Metabolites Detected in Pig Feed Samples: Their Occurrence, Relevance and Cytotoxic Effects In Vitro. Toxins 2019, 11, 537 .
AMA StyleBarbara Novak, Valentina Rainer, Michael Sulyok, Dietmar Haltrich, Gerd Schatzmayr, Elisabeth Mayer. Twenty-Eight Fungal Secondary Metabolites Detected in Pig Feed Samples: Their Occurrence, Relevance and Cytotoxic Effects In Vitro. Toxins. 2019; 11 (9):537.
Chicago/Turabian StyleBarbara Novak; Valentina Rainer; Michael Sulyok; Dietmar Haltrich; Gerd Schatzmayr; Elisabeth Mayer. 2019. "Twenty-Eight Fungal Secondary Metabolites Detected in Pig Feed Samples: Their Occurrence, Relevance and Cytotoxic Effects In Vitro." Toxins 11, no. 9: 537.
Deoxynivalenol (DON) is one of the most prevalent mycotoxins, contaminating cereals and cereal-derived products. Its derivative deepoxy-deoxynivalenol (DOM-1) is produced by certain bacteria, which either occur naturally or are supplemented in feed additive. DON-induced impairments in protein synthesis are particularly problematic for highly proliferating immune cells. This study provides the first comparison of the effects of DON and DOM-1 on the concanavalin A-induced proliferation of porcine, chicken, and bovine peripheral blood mononuclear cells (PBMCs). Therefore, isolated PBMCs were treated with DON (0.01–3.37 µM) and DOM-1 (1.39–357 µM) separately, and proliferation was measured using a bromodeoxyuridine (BrdU) assay. Although pigs are considered highly sensitive to DON, the present study revealed a substantially higher sensitivity of bovine (IC50 = 0.314 µM) PBMCs compared to chicken (IC50 = 0.691 µM) and porcine (IC50 = 0.693 µM) PBMCs. Analyses on the proliferation of bovine T-cell subsets showed that all major subsets, namely, CD4+, CD8β+, and γδ T cells, were affected to a similar extent. In contrast, DOM-1 did not affect bovine PBMCs, but reduced the proliferation of chicken and porcine PBMCs at the highest tested concentration (357 µM). Results confirm the necessity of feed additives containing DON-to-DOM-1-transforming bacteria and highlights species-specific differences in the DON sensitivity of immune cells.
Barbara Novak; Eleni Vatzia; Alexandra Springler; Alix Pierron; Wilhelm Gerner; Nicole Reisinger; Sabine Hessenberger; Gerd Schatzmayr; Elisabeth Mayer. Bovine Peripheral Blood Mononuclear Cells Are More Sensitive to Deoxynivalenol Than Those Derived from Poultry and Swine. Toxins 2018, 10, 152 .
AMA StyleBarbara Novak, Eleni Vatzia, Alexandra Springler, Alix Pierron, Wilhelm Gerner, Nicole Reisinger, Sabine Hessenberger, Gerd Schatzmayr, Elisabeth Mayer. Bovine Peripheral Blood Mononuclear Cells Are More Sensitive to Deoxynivalenol Than Those Derived from Poultry and Swine. Toxins. 2018; 10 (4):152.
Chicago/Turabian StyleBarbara Novak; Eleni Vatzia; Alexandra Springler; Alix Pierron; Wilhelm Gerner; Nicole Reisinger; Sabine Hessenberger; Gerd Schatzmayr; Elisabeth Mayer. 2018. "Bovine Peripheral Blood Mononuclear Cells Are More Sensitive to Deoxynivalenol Than Those Derived from Poultry and Swine." Toxins 10, no. 4: 152.
Deoxynivalenol (DON), a trichothecene produced by various Fusarium species, is one of the most prevalent food- and feed-associated mycotoxins. The effects of DON and deepoxy-deoxynivalenol (DOM-1) were assessed in five different cell lines from different tissues and species starting from the first line of defense, the trout gill (RTgill-W1) and pig intestinal cells (IPEC-1 and IPEC-J2) over immune cells, as second line of defense (mouse macrophages RAW 264.7) to human liver cells (HepG2). Viability was assessed with a WST-1 assay, except for RTgill-W1, where a neutral red (NR) and sulforhodamine B (SRB) assay was performed. Additionally, more sensitive parameters, such as interleukin-, nitric oxide (NO)-, and albumin-release were determined. Viability was affected by DON at concentrations starting at 10 μmol/L (RTgill-W1), 0.9 μmol/L (IPEC-1), 3.5 μmol/L (IPEC-J2), and 0.9 μmol/L (HepG2), whereas DOM-1 did not have such an effect. Additionally, NO was decreased (0.84 μmol/L DON), whereas interleukin (IL)-6 was increased (0.42 μmol/L DON) in lipopolysaccharide (LPS)-stimulated DON-, but not DOM-1-treated RAW cells. Tumor necrosis factor (TNF)-α release, however, was not affected. Interestingly, albumin secretion of HepG2 cells was decreased by both DON and DOM-1 but at a much higher concentration for DOM-1 (228 versus 0.9 μmol/L for DON). 98.9% of DOM-1 was retrieved by liquid chromatography tandem mass spectrometry at the end of the experiment, proving its stability. In this study, IL-6 was the most sensitive parameter, followed by NO and albumin release and viability for HepG2 and IPEC-1.
Elisabeth Mayer; Barbara Novak; Alexandra Springler; Heidi Schwartz-Zimmermann; Veronika Nagl; Nicole Reisinger; Sabine Hessenberger; Gerd Schatzmayr. Effects of deoxynivalenol (DON) and its microbial biotransformation product deepoxy-deoxynivalenol (DOM-1) on a trout, pig, mouse, and human cell line. Mycotoxin Research 2017, 33, 297 -308.
AMA StyleElisabeth Mayer, Barbara Novak, Alexandra Springler, Heidi Schwartz-Zimmermann, Veronika Nagl, Nicole Reisinger, Sabine Hessenberger, Gerd Schatzmayr. Effects of deoxynivalenol (DON) and its microbial biotransformation product deepoxy-deoxynivalenol (DOM-1) on a trout, pig, mouse, and human cell line. Mycotoxin Research. 2017; 33 (4):297-308.
Chicago/Turabian StyleElisabeth Mayer; Barbara Novak; Alexandra Springler; Heidi Schwartz-Zimmermann; Veronika Nagl; Nicole Reisinger; Sabine Hessenberger; Gerd Schatzmayr. 2017. "Effects of deoxynivalenol (DON) and its microbial biotransformation product deepoxy-deoxynivalenol (DOM-1) on a trout, pig, mouse, and human cell line." Mycotoxin Research 33, no. 4: 297-308.
The human, animal and plant pathogen Fusarium, which contaminates agricultural commodities worldwide, produces numerous secondary metabolites. An example is the thoroughly-investigated deoxynivalenol (DON), which severely impairs gastrointestinal barrier integrity. However, to date, the toxicological profile of other Fusarium-derived metabolites, such as enniatins, beauvericin, moniliformin, apicidin, aurofusarin, rubrofusarin, equisetin and bikaverin, are poorly characterized. Thus we examined their effects—as metabolites alone and as metabolites in combination with DON—on the intestinal barrier function of differentiated intestinal porcine epithelial cells (IPEC-J2) over 72 h. Transepithelial electrical resistance (TEER) was measured at 24-h intervals, followed by evaluation of cell viability using neutral red (NR) assay. Enniatins A, A1, B and B1, apicidin, aurofusarin and beauvericin significantly reduced TEER. Moniliformin, equisetin, bikaverin and rubrofusarin had no effect on TEER. In the case of apicidin, aurofusarin and beauvericin, TEER reductions were further substantiated by the addition of otherwise no-effect DON concentrations. In all cases, viability was unaffected, confirming that TEER reductions were not due to compromised viability. Considering the prevalence of mycotoxin contamination and the diseases associated with intestinal barrier disruption, consumption of contaminated food or feed may have substantial health implications.
Alexandra Springler; Galina-Jacqueline Vrubel; Elisabeth Mayer; Gerd Schatzmayr; Barbara Novak. Effect of Fusarium-Derived Metabolites on the Barrier Integrity of Differentiated Intestinal Porcine Epithelial Cells (IPEC-J2). Toxins 2016, 8, 345 .
AMA StyleAlexandra Springler, Galina-Jacqueline Vrubel, Elisabeth Mayer, Gerd Schatzmayr, Barbara Novak. Effect of Fusarium-Derived Metabolites on the Barrier Integrity of Differentiated Intestinal Porcine Epithelial Cells (IPEC-J2). Toxins. 2016; 8 (11):345.
Chicago/Turabian StyleAlexandra Springler; Galina-Jacqueline Vrubel; Elisabeth Mayer; Gerd Schatzmayr; Barbara Novak. 2016. "Effect of Fusarium-Derived Metabolites on the Barrier Integrity of Differentiated Intestinal Porcine Epithelial Cells (IPEC-J2)." Toxins 8, no. 11: 345.
The mycotoxin deoxynivalenol (DON) contaminates agricultural commodities worldwide, posing health threats to humans and animals. Associated with DON are derivatives, such as deepoxy-deoxynivalenol (DOM-1), produced by enzymatic transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Using differentiated porcine intestinal epithelial cells (IPEC-J2), we provide the first multi-parameter comparative cytotoxicity analysis of DON and DOM-1, based on the parallel evaluation of lysosomal activity, total protein content, membrane integrity, mitochondrial metabolism and ATP synthesis. The study investigated the ability of DON and—for the first time of its metabolite DOM-1—to induce apoptosis, mitogen-activated protein kinase (MAPK) signalling, oxidative events and alterations of mitochondrial structure in porcine intestinal epithelial cells (IECs). The degree of DON toxicity strongly varied, depending on the cytotoxicity parameter evaluated. DON compromised viability according to the parameters of lysosomal activity, total protein content and membrane integrity, but increased viability according to assays based on mitochondrial metabolism and ATP synthesis. DON induced expression of cleaved caspase-3 (maximum induction 3.9-fold) and MAPK p38 and p42/p44 (maximum induction 2.51- and 2.30-fold, respectively). DON altered mitochondrial morphology, but did not increase intracellular ROS. DOM-1-treated IPEC-J2 remained unaffected at equimolar concentrations in all assays, thereby confirming the safety of feed additives using DON- to DOM-1-transforming bacteria. The study additionally highlights that an extensive multi-parameter analysis significantly contributes to the quality of in vitro data.
Alexandra Springler; Sabine Hessenberger; Nicole Reisinger; Corinna Kern; Veronika Nagl; Gerd Schatzmayr; Elisabeth Mayer. Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects. Mycotoxin Research 2016, 33, 25 -37.
AMA StyleAlexandra Springler, Sabine Hessenberger, Nicole Reisinger, Corinna Kern, Veronika Nagl, Gerd Schatzmayr, Elisabeth Mayer. Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects. Mycotoxin Research. 2016; 33 (1):25-37.
Chicago/Turabian StyleAlexandra Springler; Sabine Hessenberger; Nicole Reisinger; Corinna Kern; Veronika Nagl; Gerd Schatzmayr; Elisabeth Mayer. 2016. "Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects." Mycotoxin Research 33, no. 1: 25-37.
Deoxynivalenol (DON), produced by the plant pathogens Fusarium graminearum and Fusarium culmorum, is one of the most common mycotoxins, contaminating cereal and cereal-derived products. Although worldwide contamination of food and feed poses health threats to humans and animals, pigs are particularly susceptible to this mycotoxin. DON derivatives, such as deepoxy-deoxynivalenol (DOM-1), are produced by bacterial transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Intestinal epithelial cells are the initial barrier against these food- and feed-borne toxins. The present study confirms DON-induced activation of MAPK p44/42 and inhibition of p44/42 by MAPK-inhibitor U0126 monoethanolate. Influence of DON and DOM-1 on transepithelial electrical resistance (TEER), viability and expression of seven tight junction proteins (TJ), as well as the potential of U0126 to counteract DON-induced effects, was assessed. While DOM-1 showed no effect, DON significantly reduced TEER of differentiated IPEC-J2 and decreased expression of claudin-1 and -3, while leaving claudin-4; ZO-1, -2, and -3 and occludin unaffected. Inhibition of p44/42 counteracted DON-induced TEER decrease and restored claudin-3, but not claudin-1 expression. Therefore, effects of DON on TEER and claudin-3 are at least partially p44/42 mediated, while effects on viability and claudin-1 are likely mediated via alternative pathways.
Alexandra Springler; Sabine Hessenberger; Gerd Schatzmayr; Elisabeth Mayer. Early Activation of MAPK p44/42 Is Partially Involved in DON-Induced Disruption of the Intestinal Barrier Function and Tight Junction Network. Toxins 2016, 8, 264 .
AMA StyleAlexandra Springler, Sabine Hessenberger, Gerd Schatzmayr, Elisabeth Mayer. Early Activation of MAPK p44/42 Is Partially Involved in DON-Induced Disruption of the Intestinal Barrier Function and Tight Junction Network. Toxins. 2016; 8 (9):264.
Chicago/Turabian StyleAlexandra Springler; Sabine Hessenberger; Gerd Schatzmayr; Elisabeth Mayer. 2016. "Early Activation of MAPK p44/42 Is Partially Involved in DON-Induced Disruption of the Intestinal Barrier Function and Tight Junction Network." Toxins 8, no. 9: 264.
One of the most important hoof diseases is laminitis. Yet, the pathology of laminitis is not fully understood. Different bacterial toxins, e.g. endotoxins or exotoxins, seem to play an important role. Additionally, ingestion of mycotoxins, toxic secondary metabolites of fungi, might contribute to the onset of laminitis. In this respect, fumonsins are of special interest since horses are regarded as species most susceptible to this group of mycotoxins. The aim of our study was to investigate the influence of fumonisin B1 (FB1) on primary isolated epidermal and dermal hoof cells, as well as on the lamellar tissue integrity and sphingolipid metabolism of hoof explants in vitro. There was no effect of FB1 at any concentration on dermal or epidermal cells. However, FB1 significantly reduced the separation force of explants after 24 h of incubation. The Sa/So ratio was significantly increased in supernatants of explants incubated with FB1 (2.5–10 µg/mL) after 24 h. Observed effects on Sa/So ratio were linked to significantly increased sphinganine concentrations. Our study showed that FB1 impairs the sphingolipid metabolism of explants and reduces lamellar integrity at non-cytotoxic concentrations. FB1 might, therefore, affect hoof health. Further in vitro and in vivo studies are necessary to elucidate the effects of FB1 on the equine hoof in more detail.
Nicole Reisinger; Ilse Dohnal; Veronika Nagl; Simone Schaumberger; Gerd Schatzmayr; Elisabeth Mayer. Fumonisin B1 (FB1) Induces Lamellar Separation and Alters Sphingolipid Metabolism of In Vitro Cultured Hoof Explants. Toxins 2016, 8, 89 .
AMA StyleNicole Reisinger, Ilse Dohnal, Veronika Nagl, Simone Schaumberger, Gerd Schatzmayr, Elisabeth Mayer. Fumonisin B1 (FB1) Induces Lamellar Separation and Alters Sphingolipid Metabolism of In Vitro Cultured Hoof Explants. Toxins. 2016; 8 (4):89.
Chicago/Turabian StyleNicole Reisinger; Ilse Dohnal; Veronika Nagl; Simone Schaumberger; Gerd Schatzmayr; Elisabeth Mayer. 2016. "Fumonisin B1 (FB1) Induces Lamellar Separation and Alters Sphingolipid Metabolism of In Vitro Cultured Hoof Explants." Toxins 8, no. 4: 89.