Toxicological and pharmacokinetic properties of sucralose-6-acetate and its parent sucralose: in vitro screening assays.

The purpose of this study was to determine the toxicological and pharmacokinetic properties of sucralose-6-acetate, a structural analog of the artificial sweetener sucralose. Sucralose-6-acetate is an intermediate and impurity in the manufacture of sucralose, and recent commercial sucralose samples were found to contain up to 0.67% sucralose-6-acetate. Studies in a rodent model found that sucralose-6-acetate is also present in fecal samples with levels up to 10% relative to sucralose which suggest that sucralose is also acetylated in the intestines. A MultiFlow® assay, a high-throughput genotoxicity screening tool, and a micronucleus (MN) test that detects cytogenetic damage both indicated that sucralose-6-acetate is genotoxic. The mechanism of action was classified as clastogenic (produces DNA strand breaks) using the MultiFlow® assay. The amount of sucralose-6-acetate in a single daily sucralose-sweetened drink might far exceed the threshold of toxicological concern for genotoxicity (TTCgenotox) of 0.15 µg/person/day. The RepliGut® System was employed to expose human intestinal epithelium to sucralose-6-acetate and sucralose, and an RNA-seq analysis was performed to determine gene expression induced by these exposures. Sucralose-6-acetate significantly increased the expression of genes associated with inflammation, oxidative stress, and cancer with greatest expression for the metallothionein 1 G gene (MT1G). Measurements of transepithelial electrical resistance (TEER) and permeability in human transverse colon epithelium indicated that sucralose-6-acetate and sucralose both impaired intestinal barrier integrity. Sucralose-6-acetate also inhibited two members of the cytochrome P450 family (CYP1A2 and CYP2C19). Overall, the toxicological and pharmacokinetic findings for sucralose-6-acetate raise significant health concerns regarding the safety and regulatory status of sucralose itself.

Revisited: Assessing the in vivo data on low/no-calorie sweeteners and the gut microbiota.

Over the last two decades, safety concerns about low/no-calorie sweeteners (LNCS) have been described in the archival scientific literature including elevated risk of metabolic syndrome, type 2 diabetes, excessive weight gain, cardiovascular disease, safety, and disruption of the gut microbiome. A recent review by Lobach, Roberts, and Roland in Food and Chemical Toxicology examined 17 research articles on modulation of gut bacteria by LNCS along with other selected publications. In the conclusions of their paper, they claim that LNCS 1) do not affect gut microbiota at use levels and 2) are safe at levels approved by regulatory agencies. Both of these claims are incorrect. The scientific literature on LNCS clearly indicates that it is inappropriate to draw generalized conclusions regarding effects on gut microbiota and safety issues for compounds that vary widely chemical structure and pharmacokinetics. Scientific studies on the sweetener sucralose, used here as a representative LNCS, indicate that this organochlorine compound unequivocally and irrefutably disrupts the gut microbiome at doses relevant to human use. Results of dozens of additional research publications added and reviewed here also raise significant and extensive concerns about the safety of sucralose for the human food supply.

Intestinal Metabolism and Bioaccumulation of Sucralose In Adipose Tissue In The Rat.

The aim of this study was to (1) determine if the organochlorine artificial sweetener sucralose is metabolized in rat intestine with repeated dosing and (2) examine whether sucralose might bioaccumulate in rat adipose tissue. Sucralose was administered to 10 rats by gavage daily for 40 days at an average dosage of 80.4 mg/kg/day. The dosages were within the range utilized in historical toxicology studies submitted for regulatory approval in North America, Europe, and Asia. Feces and urine were collected individually from each animal for every 24-hr period during the 40-day dosing period. Analysis of the urine and fecal extracts by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) revealed two new biotransformation products that have not previously been reported. These two metabolites are both acetylated forms of sucralose that are less polar and hence more lipophilic than sucralose itself. These metabolites were present in urine and feces throughout the sucralose dosing period and still detected at low levels in the urine 11 days after discontinuation of sucralose administration and 6 days after sucralose was no longer detected in the urine or feces. The finding of acetylated sucralose metabolites in urine and feces do not support early metabolism studies, on which regulatory approval was based, that claimed ingested sucralose is excreted unchanged (i.e. not metabolized). The historical metabolic studies apparently failed to detect these metabolites in part because investigators used a methanol fraction from feces for analysis along with thin layer chromatography and a low-resolution linear radioactivity analyzer. Further, sucralose was found in adipose tissue in rats two weeks after cessation of the 40-day feeding period even though this compound had disappeared from the urine and feces. Thus, depuration of sucralose which accumulated in fatty tissue requires an extended period of time after discontinuation of chemical ingestion. These new findings of metabolism of sucralose in the gastrointestinal tract (GIT) and its accumulation in adipose tissue were not part of the original regulatory decision process for this agent and indicate that it now may be time to revisit the safety and regulatory status of this organochlorine artificial sweetener.

Influence of medications on taste and smell.

Medications frequently have chemosensory side effects that can adversely affect compliance with medical treatment regimens. Hundreds of drugs have been reported to induce unpleasant tastes and/or odors as well as altered chemosensations when administered alone or in combination with other medications. Some chemosensory complaints are due to the sensory properties of the drug itself such as aversive bitter and metallic tastes. However, most chemosensory side effects of drugs are due to alterations in the transduction pathways, biochemical targets, enzymes, and transporters by the offending medications. Studies of chemosensory perception in medicated older individuals have found that taste and smell loss is greatest for those consuming the largest number of prescription drugs. There are no standard treatments for drug-induced chemosensory disorders because each drug has unique biological effects. However, there are a few treatment options to ameliorate chemosensory alterations including addition of simulated flavors to food to compensate for losses and to override offending tastes and smells.

Sucralose, a synthetic organochlorine sweetener: overview of biological issues.

Sucralose is a synthetic organochlorine sweetener (OC) that is a common ingredient in the world's food supply. Sucralose interacts with chemosensors in the alimentary tract that play a role in sweet taste sensation and hormone secretion. In rats, sucralose ingestion was shown to increase the expression of the efflux transporter P-glycoprotein (P-gp) and two cytochrome P-450 (CYP) isozymes in the intestine. P-gp and CYP are key components of the presystemic detoxification system involved in first-pass drug metabolism. The effect of sucralose on first-pass drug metabolism in humans, however, has not yet been determined. In rats, sucralose alters the microbial composition in the gastrointestinal tract (GIT), with relatively greater reduction in beneficial bacteria. Although early studies asserted that sucralose passes through the GIT unchanged, subsequent analysis suggested that some of the ingested sweetener is metabolized in the GIT, as indicated by multiple peaks found in thin-layer radiochromatographic profiles of methanolic fecal extracts after oral sucralose administration. The identity and safety profile of these putative sucralose metabolites are not known at this time. Sucralose and one of its hydrolysis products were found to be mutagenic at elevated concentrations in several testing methods. Cooking with sucralose at high temperatures was reported to generate chloropropanols, a potentially toxic class of compounds. Both human and rodent studies demonstrated that sucralose may alter glucose, insulin, and glucagon-like peptide 1 (GLP-1) levels. Taken together, these findings indicate that sucralose is not a biologically inert compound.

Rationale for further medical and health research on high-potency sweeteners.

High-potency or artificial sweeteners have historically been considered inert compounds without physiological consequences other than taste sensations. However, recent data suggest that some of these sweeteners have biological effects that may impact human health. Furthermore, there are significant gaps in our current knowledge of the pharmacokinetics of these sweeteners, their potential for "sweetener-drug interactions" and their impact on appetite and body weight regulation. Nine research needs are described that address some of the major unknown issues associated with ingestion of high-potency sweeteners.

Effects of aging on the human taste system.

Losses in taste perception as well as distortions of gustatory function occur with greater frequency in older individuals, and these changes are exacerbated by certain medical conditions, pharmacologic interventions, radiation, and exposure to toxic chemicals. Medications, especially drug-drug interactions, are the most significant yet underappreciated contributors to taste disorders in the elderly. Taste disorders frequently occur when the fraction of an orally administered drug dose that reaches the systemic circulation is markedly increased due to inhibition (by a coadministered drug) of the efflux transporter P-glycoprotein and/or inhibition of the cytochrome P450 metabolism system, which normally limit systemic availability of drugs. The supratherapeutic plasma drug concentrations that result from drug-drug interactions can induce a taste from the blood side of taste cells (intravascular taste), alter taste cell biochemistry, and/or modify other aspects of the taste pathways.

Air pollution and odor in communities near industrial swine operations.

BACKGROUND: Odors can affect health and quality of life. Industrialized animal agriculture creates odorant compounds that are components of a mixture of agents that could trigger symptoms reported by neighbors of livestock operations. OBJECTIVE: We quantified swine odor episodes reported by neighbors and the relationships of these episodes with environmental measurements. METHODS: Between September 2003 and September 2005, 101 nonsmoking volunteers living within 1.5 mi of industrial swine operations in 16 neighborhoods in eastern North Carolina completed twice-daily odor diaries for approximately 2 weeks. Meteorological conditions, hydrogen sulfide, and particulate matter 6.75 miles per hour. The odds of reporting a change in daily activities due to odor increased 62% for each unit increase in average odor during the prior 12 hr (t-value = 7.17). CONCLUSIONS: This study indicates that malodor from swine operations is commonly present in these communities and that the odors reported by neighbors are related to objective environmental measurements and interruption of activities of daily life.

Splenda alters gut microflora and increases intestinal p-glycoprotein and cytochrome p-450 in male rats.

Splenda is comprised of the high-potency artificial sweetener sucralose (1.1%) and the fillers maltodextrin and glucose. Splenda was administered by oral gavage at 100, 300, 500, or 1000 mg/kg to male Sprague-Dawley rats for 12-wk, during which fecal samples were collected weekly for bacterial analysis and measurement of fecal pH. After 12-wk, half of the animals from each treatment group were sacrificed to determine the intestinal expression of the membrane efflux transporter P-glycoprotein (P-gp) and the cytochrome P-450 (CYP) metabolism system by Western blot. The remaining animals were allowed to recover for an additional 12-wk, and further assessments of fecal microflora, fecal pH, and expression of P-gp and CYP were determined. At the end of the 12-wk treatment period, the numbers of total anaerobes, bifidobacteria, lactobacilli, Bacteroides, clostridia, and total aerobic bacteria were significantly decreased; however, there was no significant treatment effect on enterobacteria. Splenda also increased fecal pH and enhanced the expression of P-gp by 2.43-fold, CYP3A4 by 2.51-fold, and CYP2D1 by 3.49-fold. Following the 12-wk recovery period, only the total anaerobes and bifidobacteria remained significantly depressed, whereas pH values, P-gp, and CYP3A4 and CYP2D1 remained elevated. These changes occurred at Splenda dosages that contained sucralose at 1.1-11 mg/kg (the US FDA Acceptable Daily Intake for sucralose is 5 mg/kg). Evidence indicates that a 12-wk administration of Splenda exerted numerous adverse effects, including (1) reduction in beneficial fecal microflora, (2) increased fecal pH, and (3) enhanced expression levels of P-gp, CYP3A4, and CYP2D1, which are known to limit the bioavailability of orally administered drugs.

Dispersion modeling to compare alternative technologies for odor remediation at swine facilities.

The effectiveness of 18 alternative technologies for reducing odor dispersion at and beyond the boundary of swine facilities was assessed in conjunction with an initiative sponsored through agreements between the Attorney General of North Carolina and Smithfield Foods, Premium Standard Farms, and Frontline Farmers. The trajectory and spatial distribution of odor emitted at each facility were modeled at 200 and 400 m downwind from each site under two meteorological conditions (daytime and nighttime) using a Eulerian-Lagrangian model. To predict the dispersion of odor downwind, the geographical area containing the odorant sources at each facility was partitioned into 10-m2 grids on the basis of satellite photographs and architectural drawings. Relative odorant concentrations were assigned to each grid point on the basis of intensity measurements made by the trained odor panel at each facility using a 9-point rating scale. The results of the modeling indicated that odor did not extend significantly beyond 400 m downwind of any of the test sites during the daytime when the layer of air above the earth's surface is usually turbulent. However, modeling indicated that odor from all full-scale farms extended beyond 400 m onto neighboring property in the evenings when deep surface cooling through long-wave radiation to space produces a stable (nocturnal) boundary layer. The results also indicated that swine housing, independent of waste management type, plays a significant role in odor downwind, as do odor sources of moderate to moderately high intensity that emanate from a large surface area such as a lagoon. Human odor assessments were utilized for modeling rather than instrument measurements of volatile organic compounds (VOCs), hydrogen sulfide, ammonia, or particulates less than 10 microm in diameter (PM10) because these physical measurements obtained simultaneously with human panel ratings were not found to accurately predict human odor intensity in the field.

Critical illness and changes in sensory perception.

Impairments of sensory perception that occur during a period of critical care can seriously impact on health and nutritional status, activities of daily living, independence, quality of life and the possibility of recovery. It is emphasized from the outset that sensory losses in critically-ill patients may or may not be related to their current medical condition. The present paper provides an overview of all five senses (vision, hearing, taste, smell and touch) and describes the factors that contribute to sensory losses in critically-ill patients, including medications, medical conditions and treatments and the process of aging itself. Cancer and stroke are two critical illnesses in which profound sensory decrements often occur. Many sensory complaints in patients with cancer are related to alteration in sensory signals caused by damage to the sensory receptors. However, some complaints, such as taste aversions in patients with cancer, are not related to altered sensory physiology per se but to learned aversions that arise during the noxious effects of radiotherapy and chemotherapy. The paper also reviews a study in which the sensory performance (of all five senses) was compared in three groups of elderly subjects: (1) patients who had undergone coronary artery bypass surgery; (2) patients with cardiovascular conditions but with no history of surgery; (3) healthy non-medicated age-matched controls. Performance of patients who had undergone coronary artery bypass surgery was worse than that for the other two groups, with taste and smell losses greater than for the other senses. The study demonstrates that critical illness (e.g. coronary artery bypass surgery) can exacerbate sensory losses in an older cohort.

Symptomatic effects of exposure to diluted air sampled from a swine confinement atmosphere on healthy human subjects.

Aerial emissions from a swine house at North Carolina State University's field laboratory were diluted to a level that could occur at varying distances downwind from a confined animal feeding operation (CAFO) both within and beyond the property line, and these emissions were delivered to an environmental exposure chamber. The study design consisted of two 1-hr sessions, one in which 48 healthy human adult volunteers were exposed to diluted swine air and another in which they were exposed to clean air (control). Objective measures of blood pressure, temperature, heart rate, respiratory rate, lung function, nasal inflammation, secretory immunity, mood, attention, and memory were correlated with objective measures of air quality. Ratings of perceived (self-reported) health symptoms were also obtained. The mean levels of airborne constituents in the swine air condition were hydrogen sulfide (24 ppb), ammonia (817 ppb), total suspended particulates (0.0241 mg/m3), endotoxin (7.40 endotoxin units/m3), and odor (57 times above odor threshold). No statistical differences on objective measures of physical symptoms, mood, or attention resulted from the 1-hr exposure to swine emissions in the environmental chamber when compared with clean air for healthy human volunteers. However, subjects were 4.1 (p = 0.001) times more likely to report headaches, 6.1 (p = 0.004) times more likely to report eye irritation, and 7.8 (p = 0.014) times more likely to report nausea in the swine air (experimental) condition than in the control condition. These results indicate that short-term exposure in an environmental chamber to malodorous emissions from a swine house at levels expected downwind can induce clinically important symptoms in healthy human volunteers.

Science of odor as a potential health issue.

Historically, unpleasant odors have been considered warning signs or indicators of potential risks to human health but not necessarily direct triggers of health effects. However, citizen complaints to public health agencies suggest that odors may not simply serve as a warning of potential risks but that odor sensations themselves may cause health symptoms. Mal-odors emitted from large animal production facilities and wastewater treatment plants, for example, elicit complaints of eye, nose, and throat irritation, headache, nausea, diarrhea, hoarseness, sore throat, cough, chest tightness, nasal congestion, palpitations, shortness of breath, stress, drowsiness, and alterations in mood. There are at least three mechanisms by which ambient odors may produce health symptoms. First, symptoms can be induced by exposure to odorants (compounds with odor properties) at levels that also cause irritation or other toxicological effects. That is, irritation--rather than the odor--is the cause of the health symptoms, and odor (the sensation) simply serves as an exposure marker. Second, health symptoms from odorants at non-irritant concentrations can be due to innate (genetically coded) or learned aversions. Third, symptoms may be due to a co-pollutant (such as endotoxin) that is part of an odorant mixture. Objective biomarkers of health symptoms must be obtained, however, to determine if health complaints constitute health effects. One industry that is receiving much attention, worldwide, related to this subject is concentrated animal production agriculture. Sustainability of this industry will likely necessitate the development of new technologies to mitigate odorous aerial emissions. Examples of such "environmentally superior technologies" (EST) developed under the initiative sponsored through agreements between the Attorney General of North Carolina and Smithfield Foods and Premium Standard Farms are described.

Potential health effects of odor from animal operations, wastewater treatment, and recycling of byproducts.

Complaints of health symptoms from ambient odors have become more frequent in communities with confined animal facilities, wastewater treatment plants, and biosolids recycling operations. The most frequently reported health complaints include eye, nose, and throat irritation, headache, nausea, diarrhea, hoarseness, sore throat, cough, chest tightness, nasal congestion, palpitations, shortness of breath, stress, drowsiness, and alterations in mood. Typically, these symptoms occur at the time of exposure and remit after a short period of time. However, for sensitive individuals such as asthmatic patients, exposure to odors may induce health symptoms that persist for longer periods of time as well as aggravate existing medical conditions. A workshop was held at Duke University on April 16-17, 1998 cosponsored by Duke University, the Environmental Protection Agency (EPA). and National Institute on Deafness and Other Communication Disorders (NIDCD) to assess the current state of knowledge regarding the health effects of ambient odors. This report summarizes the conclusions from the Workshop regarding the potential mechanisms responsible for health symptoms from ambient odors. Methods for validation of health symptoms, presence of odor, and efficacy of odor management techniques are described as well.

Odor from industrial hog farming operations and mucosal immune function in neighbors.

The authors evaluated whether exposure to malodor from industrial hog farming operations has a psychophysiologically mediated immunosuppressive effect on secretory immunoglobulin A (sIgA) in neighbors. Fifteen adults living within 2.4 km (1.5 mi) of at least one hog farming operation rated odor intensity on a 9-point scale and provided saliva samples twice daily for two weeks. The authors used hierarchical regression to model the association between reported odor and sIgA; study participants were their own controls. The natural log of slgA concentration and secretion rate declined, on average, 0.058 (0.032) and 0.116 (0.103), respectively, for each incremental 1-unit increase in reported odor from 4 to 9, adjusted for time of day, suggesting reduced levels of sIgA in response to moderate or high odor. Findings support the hypothesized immunosuppressive effect of malodor on mucosal immunity and provide preliminary data useful in understanding health effects related to malodor from industrial hog farming operations.

Effect of repeated presentation on sweetness intensity of binary and ternary mixtures of sweeteners.

The purpose of the present study was to determine the effect of repeated presentation of the same sweet stimulus on sweetness intensity ratings. The sweet stimuli tested in this study were binary and ternary blends of 14 sweeteners that varied widely in chemical structure. A trained panel evaluated the sweetness intensity over four sips of a given mixture presented at 30 s intervals. The individual components in the binary sweetener combinations were intensity-anchored with 5% sucrose, while the individual sweeteners in the ternary mixtures were intensity-anchored with 3% sucrose (according to formulae developed previously). Each self-mixture was also evaluated (e.g. acesulfame-K-acesulfame-K). The main finding of this study was that mixtures consisting of two or three different sweeteners exhibited less reduction in sweetness intensity over four repeated sips than a single sweetener at an equivalent sweetness level. Furthermore, ternary combinations tended to be slightly more effective than binary combinations at lessening the effect of repeated exposure to a given sweet stimulus. These findings suggest that the decline in sweetness intensity experienced over repeated exposure to a sweet stimulus could be reduced by the blending of sweeteners.

Aspartame: review of safety.

Over 20 years have elapsed since aspartame was approved by regulatory agencies as a sweetener and flavor enhancer. The safety of aspartame and its metabolic constituents was established through extensive toxicology studies in laboratory animals, using much greater doses than people could possibly consume. Its safety was further confirmed through studies in several human subpopulations, including healthy infants, children, adolescents, and adults; obese individuals; diabetics; lactating women; and individuals heterozygous (PKUH) for the genetic disease phenylketonuria (PKU) who have a decreased ability to metabolize the essential amino acid, phenylalanine. Several scientific issues continued to be raised after approval, largely as a concern for theoretical toxicity from its metabolic components--the amino acids, aspartate and phenylalanine, and methanol--even though dietary exposure to these components is much greater than from aspartame. Nonetheless, additional research, including evaluations of possible associations between aspartame and headaches, seizures, behavior, cognition, and mood as well as allergic-type reactions and use by potentially sensitive subpopulations, has continued after approval. These findings are reviewed here. The safety testing of aspartame has gone well beyond that required to evaluate the safety of a food additive. When all the research on aspartame, including evaluations in both the premarketing and postmarketing periods, is examined as a whole, it is clear that aspartame is safe, and there are no unresolved questions regarding its safety under conditions of intended use.