A novel in situ permeation system and its utility in cancer tissue ablation.

Focal ablation therapy is an emerging treatment modality for localized cancer lesions. It is an attractive strategy for inhibiting tumor progression and preventing morbidity associated with open surgery. As for intratissue drug delivery systems for use in local therapy, the convection-enhanced delivery (CED) of liquid drugs has been utilized, particularly for the treatment of malignant brain tumors. Although the conventional CED system is useful for providing drug/vehicle-based local therapy, there are several reported disadvantages in terms of the ability to control the extent of drug diffusion. We herein developed and validated a novel in situ permeation (ISP)-MW-1 system for achieving intratissue drug diffusion. The ISP system includes a perfusion catheter connected to an injector and aspirator, which enables intratissue perfusion of the solute diluted in the vehicle in the tip-inserted cavity. We subsequently evaluated the utility of the ISP-MW-1 system for in situ permeation in a subcutaneous tumor model in hamsters. Dehydrated ethanol, saline and 50% acetic acid were evaluated as the vehicle, and methylene blue was used as a dissolved substance for evaluating the diffusion of the agent. As a result, almost all of the tumor tissue within the capsule (tumor size: ~3 cm) was permeated with the dehydrated ethanol and 50% acetic acid and partially with the saline. We further demonstrated that ISP treatment with 50% acetic acid completely ablated the subcutaneous tumors in all of the treated hamsters (n=3). Therefore, the ISP-MW-1 system is a promising approach for controlling the intratissue diffusion of therapeutic agents and for providing local ablation therapy for cancer lesions. We believe that this system may be applicable to a broad range of medicinal and industrial fields, such as regenerative medicine, drug delivery systems, biochemistry and material technologies as well as cancer therapy.

[(4-Hydroxyl-benzo[4,5]thieno[3,2-c]pyridine-3-carbonyl)-amino]-acetic acid derivatives; HIF prolyl 4-hydroxylase inhibitors as oral erythropoietin secretagogues.

A new series of PHD (HIF prolyl 4-hydroxylase) inhibitors was designed based on the X-ray co-crystal structure of FG-2216. Using a lead generation process, a series of [(4-Hydroxyl-benzo[4,5]thieno[3,2-c]pyridine-3-carbonyl)-amino]-acetic acid derivatives was developed as potent PHD2 inhibitors. This class of compounds also showed the ability to stabilize HIF-α, to stimulate EPO secretion in in vitro studies, and to increase hematocrit, red blood cell count, and hemoglobin levels in an animal efficacy study.

Development and validation of a RP-HPLC method for the quantification of sparfloxacin in pharmaceutical dosage forms

Un modo rápido y sensible de cromatografía líquida en fase reversa de alta resolución ha sido desarrollado para el análisis cuantitativo de parfloxacino en preparaciones farmacéuticas. El método fue validado de acuerdo a las normas de precisión, especificidad y linealidad aportadas por la FDA, ICH y USP. El método fue desarrollado utilizando la fase móvil compuesta por solución acuosa de ácido acético al 1% y acetonitrilo en la proporción 71:29% (v/v) bajo una velocidad de flujo de 0.7 ml/min sobre una columna de sílica c-8 a temperatura ambiente. La recuperación fue de más del 97% para cada muestra adicionada de parfloxacino lo que demuestra la precisión del protocolo. La precisión intradía e interdiario fue inferior al límite máximo permitido (RSD ≤ 2.0), de acuerdo a la FDA. El método mostró una respuesta linear con un coeficiente de correlación del 0.998. Por lo tanto, el método fue preciso, reproducible, sensible y rápido, por lo que puede ser utilizado para el análisis de formulaciones de sparfloxacino (AU)

A randomized comparison of pharmacokinetics of a single vaginal dose of dry misoprostol or misoprostol moistened with normal saline or with acetic acid.

BACKGROUND: The pharmacokinetics of vaginal misoprostol as a dry tablet or as a tablet moistened with normal saline or with acetic acid were studied. METHODS: For this study, 42 women requesting termination of pregnancy at gestational age of <12 weeks were recruited and received 400 µg vaginal misoprostol tablets. They were randomized into three groups: (i) dry tablets, (ii) tablets moistened with 3 ml of normal saline and (iii) tablets moistened with 3 ml of 5% acetic acid. Venous blood samples were taken at 0, 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330 and 360 min after misoprostol administration. Misoprostol acid (MPA) was determined in serum samples using gas chromatography/tandem mass spectrometry. RESULTS: The serum peak MPA concentration (C(max)) was significantly higher and the time-to-peak concentration (T(max)) was significantly shorter in the normal saline and acetic acid groups, when compared with the dry tablet group. Both areas under the curve at 240 and 360 min (AUC(240) and AUC(360)) of the normal saline and acetic acid groups were also significantly greater than that of the dry tablet group. The coefficients of variation in C(max) and T(max) were highest in the normal saline group, while that of AUC(240) and AUC(360) were highest in the dry tablet group. The C(max) was significantly higher in subjects in the dry tablet group with vaginal pH < 5 than in those with pH 5. There were no significant differences in other pharmacokinetic parameters between subjects with vaginal pH < 5 and those with vaginal pH 5 in all three groups. CONCLUSIONS: Vaginal misoprostol tablets moistened with normal saline or 5% acetic acid achieved better absorption than the dry tablet. The use of vaginal misoprostol tablets moistened with normal saline or 5% acetic acid would potentially improve the clinical efficacy of misoprostol. HKClinicalTrials.com registration: HKCTR-821.

Bioavailability of acetate from two vinegar supplements: capsule and drink.

The bioavailability of acetate in various vinegar supplements, e.g. as capsules and drinks, remains unclear. Thus, we conducted a cross-over clinical study in 30 healthy subjects. After an overnight fast, subjects received each test sample in a randomised sequence: 9 vinegar capsules (containing 750 mg acetic acid in total) with 150 mL of water, 100 mL of vinegar drink (containing 750 mg acetic acid), and 150 mL of water as reference. Blood samples were collected before (defined as 0 min), at 15, 30, 45, 60, 90, 120 and 180 min after each test sample intake. In the vinegar drink group, serum acetate concentration increased immediately after intake, peaked at 15 min and returned to baseline at 90 min. That in the vinegar capsule group rose slowly, peaked at 30 min and returned to baseline at 120 min. The peak values in both groups exceeded 200 µmol/L, the physiologically active concentration confirmed by in vitro experiment. In the reference group, levels remained constant throughout the 180-min period. The amount of absorbed acetate from the vinegar capsule group and the drink group was evaluated by the difference value of the area under the serum acetate concentration-time curve (AUC) between in each vinegar group and in the reference group (expressed as AUC(capsule-ref) and AUC(drink-ref ), respectively). AUC(capsule-ref) was about 80% of AUC(drink-ref ), but there was no significant difference between them.

Reduction in cholesterol synthesis in response to serum starvation in lymphoblasts of a patient with Barth syndrome.

Barth syndrome is a rare X-linked disease in which mild hypocholesterolemia is observed in some patients. We investigated cholesterol biosynthesis in lymphoblasts from a normal and age-matched Barth syndrome patient. Control and Barth syndrome (DeltaTAZ1) lymphoblasts were incubated in the presence or absence of serum to induce cholesterol synthesis and hydroxymethylglutaryl-coenzyme A reductase activity and expression, and cholesterol biosynthesis from radioactive precursors was determined. Cholesterol biosynthesis from [2-14C]pyruvate was stimulated 2-fold in control cells, but was unchanged in DeltaTAZ1 lymphoblasts, and from [1-14C]acetate was stimulated 77% in control but only 26% in DeltaTAZ1 lymphoblasts upon serum removal, indicating a lower ability of DeltaTAZ1 cells to upregulate cholesterol biosynthesis. The reason was an inability to increase hydroxymethylglutaryl-coenzyme A reductase activity, which was already near maximum in DeltaTAZ1 lymphoblasts, in response to serum removal, compared with control cells. The reduced ability to increase hydroxymethylglutaryl-coenzyme A reductase enzyme activity in DeltaTAZ1 lymphoblasts was due to a decrease in hydroxymethylglutaryl-coenzyme A reductase messenger RNA. Although total cholesterol levels are similar under standard culture conditions, DeltaTAZ1 lymphoblasts have a diminished capacity to respond to increased demand for cholesterol biosynthesis because of an already elevated level of synthesis under standard culture conditions.

Effects of acetic acid and its assimilation in fed-batch cultures of recombinant Escherichia coli containing human-like collagen cDNA.

The primary processing problem in recombinant Escherichia coli fermentation is the production of acetic acid, which can inhibit both cell growth and recombinant protein production. The ability of E. coli to assimilate acetate permits it to solve this problem in a rather creative manner. In this study, the effects of acetic acid assimilation through a glucose starvation period at different cell growth phases were investigated in fed-batch cultures of recombinant E. coli. Experimental results showed that the human-like collagen (HLC) production could be improved by introducing glucose starvation at the end of batch culture and pre-induction phase, while the glucose starvation at the induction phase resulted in a poor HLC productivity. The acetic acid assimilation was observed during all the glucose starvation periods. In addition, a systematic study for evaluating the effects of acetic acid was carried out by adding acetate into culture media at different cell growth phases and then employing a glucose starvation after several hours. It was found that obvious acetate inhibition on cell growth occurred in the batch culture phases while its inhibitory effect on HLC expression occurred only in the post-induction phase. The longer the elevated acetic acid concentration maintained, the stronger the inhibitory effects were. These results are of significance for optimizing and scaling-up fermentation processes.

Changes in cell turgor pressure related to uptake of solutes by Microcystis sp. strain 8401.

Uptake of several naturally occurring organic solutes by the unicellular cyanobacterium Microcystis sp. caused changes in cell turgor pressure (p(t)), which was determined by measuring the mean critical pressure (p(c)) of gas vesicles in the cells. Cells had an initial p(t) of 0.34 MPa, which decreased to 0.08 MPa in 0.15 M sucrose. In solutions of polyols, p(t) gradually recovered as the solutes penetrated the cytoplasmic membranes. From measurements of the exponential rate of turgor increase, cell volume and surface area, the permeability coefficient of the cytoplasmic membrane to each solute was calculated. Permeabilities to amino acids, ammonium ions and sodium acetate indicated little passive movement of these substances across the cell surface from solutions at high concentrations. We looked for evidence of ion trapping of acetic acid: at low pH there was a rapid rise in turgor pressure indicating a rapid uptake of this weak acid. After 20 min the turgor was lost, apparently due to loss of integrity of the cell membranes. For cells in natural habitats, studies of the permeability of cells to solutes is relevant to the problem of retaining substances that are accumulated by active uptake from solutions of low concentrations in natural waters.

13C-acetic acid is more sensitive than 13C-octanoic acid for evaluating gastric emptying of liquid enteral nutrient formula by breath test in conscious rats.

Breath test using 13C-labeled compound has been used as a convenient method to evaluate gastric emptying. 13C-Labeled acetic acid or octanoic acid has been used in clinic. However, there is few report comparing two compounds. This study aimed to compare 13C-acetic acid and 13C-octanoic acid in newly-constituted breath test for monitoring gastric emptying in conscious rats. After fasting, rats were orally administered Racol (liquid enteral nutrient formula) containing 13C-labeled compound (same molar of 13C-acetic acid or 13C-octanoic acid) and housed in a chamber. The expired air in the chamber was collected in a breath-sampling bag using a tube and aspiration pump. The level of 13CO2 in the expired air was measured using an infrared spectrometer at appropriate intervals for 120 min. Expired 13CO2 air from 13C-acetic acid changed at significantly higher levels than that from 13C-octanoic acid. Cmax and AUC120 min values of expired 13CO2 from 13C-acetic acid were significantly higher than those from 13C-octanoic acid, but Tmax was not different between them. These results show that 13C-acetic acid is more sensitive for monitoring gastric emptying than 13C-octanoic acid in liquid test meal although both acids clearly monitored gastric emptying.

Demineralisation of permeable hydroxyapatite with alternating water and acidic buffer: scanning microradiographic study of effect of switching period.

Permeable hydroxyapatite (HAP) blocks were exposed for equal times alternately to pH 4.0 buffer and water for 237 h. Rates of HAP loss with time (determined from changes in X-ray attenuation) were measured as a function of switching period tau (the time for a complete cycle) from 0.5 to 6 h and with a continuous buffer flow. The mean rate of HAP loss decreased markedly as tau increased, and for large tau was about half the rate for continuous buffer flow. We propose that demineralising conditions through the depth of the HAP are influenced by the extent of retention of buffer within its pores which will depend on tau. A mathematical model with parameters R(0) and Deltat was developed, where R(0) is the rate of demineralisation for continuous flow, and Deltat a time added to each tau/2 buffer exposure to account for its retention in the HAP pores. Experimental data fitted the model with Deltat approximately 8 to approximately 10 min and with R(0) close to the rate observed for continuous buffer flow. The model predicts that the rate decreases and approaches R(0)/2 as tau --> infinity, as was found experimentally to be the case. This type of study could potentially give information about subsurface porosity and transport processes during acidic dissolution of permeable solids, for example in dental caries and dental erosion.

In situ detoxification and continuous cultivation of dilute-acid hydrolyzate to ethanol by encapsulated S. cerevisiae.

Dilute-acid lignocellulosic hydrolyzate was successfully fermented to ethanol by encapsulated Saccharomyces cerevisiae at dilution rates up to 0.5h(-1). The hydrolyzate was so toxic that freely suspended yeast cells could ferment it continuously just up to dilution rate 0.1h(-1), where the cells lost 75% of their viability measured by colony forming unit (CFU). However, encapsulation increased their capacity for in situ detoxification of the hydrolyzate and protected the cells against the inhibitors present in the hydrolyzate. While the cells were encapsulated, they could successfully ferment the hydrolyzate at tested dilution rates 0.1-0.5h(-1), and keep more than 75% cell viability in the worst conditions. They produced ethanol with yield 0.44+/-0.01 g/g and specific productivity 0.14-0.17 g/(gh) at all dilution rates. Glycerol was the main by-product of the cultivations, which yielded 0.039-0.052 g/g. HMF present in the hydrolyzate was converted 48-71% by the encapsulated yeast, while furfural was totally converted at dilution rates 0.1 and 0.2h(-1) and partly at the higher rates. Continuous cultivation of encapsulated yeast was also investigated on glucose in synthetic medium up to dilution rate 1.0 h(-1). At this highest rate, ethanol and glycerol were also the major products with yields 0.43 and 0.076 g/g, respectively. The experiments lasted for 18-21 days, and no damage in the capsules was detected.

Fecal acetate is inversely related to acetate absorption from the human rectum and distal colon.

In humans, colonic bacteria ferment unabsorbed carbohydrates, producing the SCFA acetic, propionic and n-butyric acids. To test for interactions among the SCFA that may affect their absorption, healthy subjects (n = 10) were given 300-mL rectal infusions containing acetate (60 mmol/L), propionate (20 mmol/L) and butyrate (20 mmol/L), alone or in combinations of two or three. The solutions were retained for 30 min, and then subjects voided a sample for SCFA measurement. To examine the relationship between absorption and fecal SCFA concentrations, a fecal sample was collected at the end of the study. The mean percentage of butyrate absorption (30.2 +/- 4.6%) exceeded that of acetate (24.1 +/- 3.7%) (P < 0.05). Absorption tended to be less (P = 0.12) when a SCFA was infused alone (26.7 +/- 4.0%) than when all three were infused (32.0 +/- 5.7%). Bicarbonate concentration was higher after butyrate-containing infusions than after saline. The fecal molar acetate percentage was inversely correlated with the percentage of acetate absorption from the infusion of three SCFA (r = -0.834, P < 0.005). We conclude that there was no combination effect on SCFA absorption, and the chain-length effect suggests passive diffusion as a likely mechanism of absorption. Furthermore, fecal acetate may reflect absorption, rather than production of colonic acetate.

Effects of resiniferatoxin desensitization of capsaicin-sensitive afferents on detrusor over-activity induced by intravesical capsaicin, acetic acid or ATP in conscious rats.

Recent studies have provided evidence for a major role of urothelially released ATP acting on a subpopulation of pelvic afferent nerves in mechano-afferent transduction in the bladder. We investigated whether desensitization of capsaicin-sensitive nerve fibres by systemic resiniferatoxin (RTX)-pretreatment can counteract the detrusor over-activity induced by intravesical capsaicin, acetic acid or ATP. Cystometric investigations were performed on awake female Sprague-Dawley rats before and 24 h after injection of RTX (0.3 mg/kg s.c.) or vehicle. The effects of intravesically instilled ATP (0.1 or 1.0 mM), capsaicin (30 microM) or acetic acid (pH 4.0) were compared with those of intravesical saline. RTX, but not its vehicle, significantly increased threshold pressure, voiding interval, micturition volume and bladder capacity. In the vehicle-pretreated rats, intravesical instillation of capsaicin or acetic acid significantly decreased voiding interval, micturition volume, and bladder capacity. However, in the RTX-pretreated rats, neither capsaicin nor acetic acid affected any parameter. On the other hand, intravesical ATP (0.1 mM) significantly decreased voiding interval and micturition volume in both groups of animals. At 1.0 mM, ATP also increased basal pressure and decreased the pressure threshold for micturition in both groups. The present results support the view that increased extracellular ATP has a role in mechano-afferent transduction in the rat bladder and that ATP-induced facilitation of the micturition reflex is mediated, at least partly, by nerves other than capsaicin-sensitive afferent nerves.

TCA cycle kinetics in the rat heart by analysis of (13)C isotopomers using indirect (1)H.

This study was designed to test the hypothesis that indirect (1)H[(13)C] detection of tricarboxylic acid (TCA) cycle intermediates using heteronuclear multiple quantum correlation-total correlation spectroscopy (HMQC-TOCSY) nuclear magnetic resonance (NMR) spectroscopy provides additional (13)C isotopomer information that better describes the kinetic exchanges that occur between intracellular compartments than direct (13)C NMR detection. NMR data were collected on extracts of rat hearts perfused at various times with combinations of [2-(13)C]acetate, propionate, the transaminase inhibitor aminooxyacetate, and (13)C multiplet areas derived from spectra of tissue glutamate were fit to a standard kinetic model of the TCA cycle. Although the two NMR methods detect different populations of (13)C isotopomers, similar values were found for TCA cycle and exchange fluxes by analyzing the two data sets. Perfusion of hearts with unlabeled propionate in addition to [2-(13)C]acetate resulted in an increase in the pool size of all four-carbon TCA cycle intermediates. This allowed the addition of isotopomer data from aspartate and malate in addition to the more abundant glutamate. This study illustrates that metabolic inhibitors can provide new insights into metabolic transport processes in intact tissues.

Metabolic fate of [14C]-ethanol into endothelial cell phospholipids including platelet-activating factor, sphingomyelin and phosphatidylethanol.

The metabolic fate of ethanol into the phospholipid pool of calf pulmonary artery endothelial cells was studied. [14C]-ethanol was incorporated into various endothelial cell phospholipids including phosphatidylethanol (PEth), which may represent a substantial fraction in microdomains of membrane phospholipids. The incorporation into phospholipids was reduced in the presence of pyrazole and cyanamide, inhibitors of ethanol metabolism. Wortmannin, the phosphatidylinositol 3-kinase inhibitor, increased [14C]-PEth formation. [3H]-acetate was also incorporated into endothelial cell phospholipids but in a different pattern. Distribution of [3H]-acetate and [14C]-ethanol into the fatty acyl moiety versus the glycerophosphoryl backbone of the phospholipids was also different. Stimulation of the endothelial cells with ATP increased [3H]-acetate incorporation into platelet-activating factor (PAF) and ethanol decreased it. Ethanol exposure increased ATP-stimulated [3H]-acetate incorporation into sphingomyelin. However, ATP had no effect on the incorporation of [14C]-ethanol into any phospholipids. The results suggest that the two precursors contribute to a separate acetate pool and that the sphingomyelin cycle may be sensitized in ethanol-treated cells. Thus, metabolic conversions of ethanol into lipids and the effect of ethanol on specific lipid mediators, e.g PAF, PEth and sphingomyelin, may be critical determinants in the altered responses of the endothelium in alcoholism.

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants.

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 microns-thick oriented PP) into the food stimulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.

Determinants of the acetate recovery factor: implications for estimation of [13C]substrate oxidation.

When using (13)C or (14)C tracers to study substrate metabolism, an acetate correction factor should be applied to correct for loss of label in the exchange pathways of the tricarboxylic acid cycle. We have shown recently that the [(13)C]acetate recovery factor has a high inter-individual variability and should therefore be determined in every subject. In the present study we examined the factors that might explain some of the variability between subjects in acetate recovery factor. Data were pooled from four different studies with identical protocols, in which the acetate recovery factor was measured, prior to an intervention, to correct plasma fatty acid oxidation rates. Acetate recovery was measured after 2 h of [1, 2-(13)C]acetate infusion at rest followed by 1 h of cycling exercise at 40-50% of maximal oxygen uptake. Inter-individual variance in acetate recovery was 12.0% at rest and 16.1% during exercise. Stepwise regression revealed that, at rest, 37.1% of the acetate recovery could be accounted for by basal metabolic rate adjusted for fat-free mass, percentage body fat and respiratory quotient (RQ). During exercise, 69.1% of the variance in acetate recovery could be accounted for by energy expenditure adjusted for fat-free mass, % body fat and RQ. In conclusion, we show that the acetate recovery factor has a high inter-individual variability, both at rest and during exercise, which can partly be accounted for by metabolic rate, RQ and % body fat. These data indicate that the acetate recovery factor needs to be determined in every subject, under similar conditions as used for the tracer-derived determination of substrate oxidation. Failure to do this might result in large under- or over-estimation of plasma substrate oxidation, and hence to artificial differences between groups.

Utilization and transport of acetic acid in Dekkera anomala and their implications on the survival of the yeast in acidic environments.

The yeast Dekkera anomala IGC 5153 exhibited a restricted ability to use weak acids as the only carbon and energy sources. Of the monocarboxylic, dicarboxylic, and tricarboxylic acids tested, only acetic acid was used in such a way. The cells were able to grow at acetic acid concentrations of 0.1 to 3% (vol/vol) over a pH range of 3.5 to 5.5, and the specific growth rates decreased exponentially with the increase of the undissociated acetic acid concentration in the culture medium. Transport assays carried out in cells that exhibited higher specific growth rates showed the presence of an acetate-proton symport associated with a simple diffusion component of the undissociated acetic acid, the weight of the latter increasing with the undissociated acid concentration in the culture media. The acetate carrier was shared by propionic, formic, and sorbic acids and was inducible and repressed by glucose and concentrations of undissociated acetic acid in the culture medium above 0.3% (vol/vol). In undissociated acetic acid repression conditions, the lowest values for the yeast specific growth rates were obtained, and the simple diffusion of the undissociated acid was the only mechanism involved in the acetic acid uptake by the cells. The results will be discussed in terms of the high tolerance of D. anomala to the acidic stress conditions present in wine.

A biologically based risk assessment for vinyl acetate-induced cancer and noncancer inhalation toxicity.

The 1990 Clean Air Act Amendments require that health risk from exposure to vinyl acetate be assessed. Vinyl acetate is a nasal carcinogen in rats, but not mice, and induces olfactory degeneration in both species. A biologically based approach to extrapolating risks of inhalation exposure from rats to humans was developed, which incorporates critical determinants of interspecies dosimetry. A physiologically based pharmacokinetic (PBPK) model describing uptake and metabolism of vinyl acetate in rat nose was validated against nasal deposition data collected at three airflow rates. The model was also validated against observations of metabolically derived acetaldehyde. Modifying the rat nose model to reflect human anatomy created a PBPK model of the human nose. Metabolic constants from both rats and humans specific for vinyl acetate and acetaldehyde metabolism enabled predictions of various olfactory tissue dosimeters related to the mode of action. Model predictions of these dosimeters in rats corresponded well with observations of vinyl acetate toxicity. Intracellular pH (pHi) of olfactory epithelial cells was predicted to drop significantly at airborne exposure concentrations above the NOAEL of 50 ppm. Benchmark dose methods were used to estimate the ED10 and LED10 for olfactory degeneration, the precursor lesion thought to drive cellular proliferation and eventually tumor development at excess cellular acetaldehyde levels. A concentration x time adjustment was applied to the benchmark dose values. Human-equivalent concentrations were calculated by using the human PBPK model to predict concentrations that yield similar cellular levels of acetic acid, acetaldehyde, and pHi. After the application of appropriate uncertainty factors, an ambient air value of 0.4 to 1.0 ppm was derived. The biologically based approach supports a workplace standard of 10 ppm.