A General Measure of In Vitro Phototoxicity Derived from Pairs of Dose-Response Curves and its Use for Predicting the In Vivo Phototoxicity of Chemicals

1997 ◽  
Vol 25 (4) ◽  
pp. 445-462 ◽  
Author(s):  
Hermann-Georg Holzhütter
Keyword(s):  
Dose Response ◽  
Uv Light ◽  
Test System ◽  
Maximal Response ◽  
Response Curves ◽  
General Measure ◽  
Absolute Change ◽  
Response Data ◽  
Dose Response Curves ◽  
The Absolute

In pharmacology, it is common to evaluate the influence of external effectors (for example, temperature, pH, and presence of a second drug) on dose-response relations by the potency factor (PF50): [Formula: see text] where ED50 (± effector) denotes the 50% effective dose in the presence and in the absence of the effector, respectively. In this paper, the external effector is ultraviolet (UV) light, and PF50 is referred to as the photoirritancy factor (PIF). There are two parameters which limit the applicability and toxicological reliability of the PIF. Firstly, the physical properties (for example, water solubility) of the chemical tested and the constraints of the biological test system may make it difficult, or even impossible, to achieve sufficiently high doses to observe 50% of the maximal response. In such cases, no numeric value of the potency factor can be computed. Secondly, the potency factor does not take into account the absolute change in response induced by UV light, i.e. depending on the shape of the ±UV dose-response curves, the absolute change in response may be small although the PIF is large, and vice versa. This paper proposes a more general measure of phototoxicity, the mean photo effect (MPE), which can be assessed from pairs of dose-response curves, even if the 50% response level is not reached in one curve or in both. The MPE is a weighted average of PIFd values across different dose levels (d being common to both dose-response curves). The absolute response changes, ΔRd, i.e. the differences between the -UV curve and the +UV curve are used as weighting factors. The numerical computation of the MPE is based on theoretical curves obtained by fitting a mathematical model to the experimental dose-response data. Plotting PIFd and ΔRd versus the corresponding doses permits differences in the shapes of the two curves to be assessed, and possible alterations in the toxic mechanisms induced by UV light to be revealed. The variance of MPE is estimated by a bootstrap procedure. The use of the MPE is illustrated by its application to dose-response data obtained with a human keratinocyte assay of fibroblasts in the EU/COLIPA international validation project on photoirritancy.

Analysis and comparison of sigmoidal curves: application to dose-response data

1989 ◽  
Vol 257 (6) ◽  
pp. G982-G989 ◽  
Author(s):  
J. B. Meddings ◽  
R. B. Scott ◽  
G. H. Fick
Keyword(s):  
Nonlinear Regression ◽  
Dose Response ◽  
Numerical Solutions ◽  
Maximal Response ◽  
Response Curves ◽  
Experimental Conditions ◽  
Response Data ◽  
Accuracy And Precision ◽  
Analysis Technique ◽  
Dose Response Curves

A number of physiological or pharmacological studies generate sigmoidal dose-response curves. Ideally, data analysis should provide numerical solutions for curve parameters. In addition, for curves obtained under different experimental conditions, testing for significant differences should be easily performed. We have reviewed the literature over the past 3 years in six journals publishing papers in the field of gastrointestinal physiology and established the curve analysis technique used in each. Using simulated experimental data of known error structure, we have compared these techniques with nonlinear regression analysis. In terms of their ability to provide accurate estimates of ED50 and maximal response, none approached the accuracy and precision of nonlinear regression. This technique is as easily performed as the classic methods and additionally provides an opportunity for rigorous statistical analysis of data. We present a method of determining the significance of differences found in the ED50 and maximal response under different experimental conditions. The method is versatile and applicable to a variety of different physiological and pharmacological dose-response curves.

FURTHER OBSERVATIONS ON THE OVARIAN ASCORBIC ACID DEPLETION TEST FOR LUTEINIZING HORMONE

1965 ◽  
Vol 32 (1) ◽  
pp. 1-7 ◽  
Author(s):  
E. T. BELL ◽  
J. A. LORAINE ◽  
S. MUKERJI ◽  
PACHARA VISUTAKUL
Keyword(s):  
Ascorbic Acid ◽  
Luteinizing Hormone ◽  
Dose Response ◽  
Intravenous Route ◽  
Response Curves ◽  
Standard Preparation ◽  
Routes Of Administration ◽  
Seasonal Factors ◽  
Dose Response Curves ◽  
The Absolute

SUMMARY A modification of the ovarian ascorbic acid depletion (OAAD) method for luteinizing hormone (LH) is described in which the standard and test materials are administered by the intraperitoneal rather than by the intravenous route. The potency of NIH-LH when administered i.p. and i.v. was the same and the slopes of the log dose-response curves for the two routes of administration were very similar. Studies on the effect of seasonal factors on the OAAD method showed that both the slopes of the log dose-response curves and the absolute levels of ascorbic acid varied from one assay to another. The necessity for using a standard preparation in all assays of LH activity by the OAAD method is emphasized.

Length-dependent sensitivity in vascular smooth muscle

1981 ◽  
Vol 241 (4) ◽  
pp. H557-H563 ◽  
Author(s):  
J. M. Price ◽  
D. L. Davis ◽  
E. B. Knauss
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Dose Response ◽  
Length Change ◽  
Maximal Response ◽  
Anterior Tibial Artery ◽  
Response Curves ◽  
Anterior Tibial ◽  
Dose Response Curves ◽  
Best Fit

Dose-response curves were obtained from dog anterior tibial artery rings at various lengths (L) to determine whether sensitivity to norepinephrine (NE) and potassium (K+) depends on arterial circumference. The dose for half maximal response (ED50) was determined by graphical estimation and by calculation from a best fit curve. For both NE and K+: 1) ED50 was lowest (most sensitive) at L for maximum active force (Lmax) and increased significantly as L decreased from Lmax; 2) ED50 at 1.0 and 1.15 Lmax was not significantly different; 3) ED50 of repeated dose-response curves at Lmax was not significantly different; and 4) when the direction of length change was reversed (from decreasing to increasing), the direction of change in ED50 was also reversed (from increasing to decreasing). Change in the dose for 10% maximal response was the same as ED50. The results did not depend on the method of determining ED50 or on whether responses were expressed as absolute values or as relative values. The results show that sensitivity of vascular smooth muscle depends on L and that the length-sensitivity relation is similar to the length-active tension relation. Similarity of results for NE and K+ indicate that length-dependent sensitivity does not depend on the method of stimulation.

Mechanical response of pulmonary artery under aerobic and hypoxic conditions

1977 ◽  
Vol 42 (3) ◽  
pp. 438-443
Author(s):  
J. F. Souhrada ◽  
D. W. Dickey
Keyword(s):  
Pulmonary Artery ◽  
Dose Response ◽  
Guinea Pigs ◽  
Main Pulmonary Artery ◽  
Maximal Response ◽  
Response Curves ◽  
Aerobic Conditions ◽  
Experimental Medium ◽  
Hypoxic Conditions ◽  
Dose Response Curves

The present study demonstrates the reactivity of isolated main pulmonary artery (MPA) from guinea pigs and rats to two vasoactive drugs, norepinephrine (NE) and histamine (H), in substrate-rich and substrate-free medium, under both aerobic (PO2 = 95 +/- 0.5 Torr) and hypoxic conditions (PO2 = 30 +/- 1 Torr). The sensitivity of MPA from guinea pigs to NE and H during aerobic conditions is not significantly affected by the absence of substrate in the experimental medium. Furthermore, it is demonstrated that in the substrate-rich experimental medium (5.5 mM glucose), the reactivity of MPA from guinea pigs to NE and H is not significantly affected by acute hypoxia as compared with the response of MPA during aerobic conditions. These experiments contrast with data obtained when substrate is absent from the experimental medium. The dose-response curves of MPA from guinea pigs to NE and H under this condition were significantly blunted during hypoxia. Following the completion of the dose-response curves during aerobic conditions, with both NE and H, spontaneous mechanical activities were seen in the guinea pig MPA. On the other hand, it was demonstrated that during aerobic and hypoxic conditions MPA's isolated from rats exhibit no physiological response to histamine even when administered in the dose required to produce the maximal response in MPA isolated from guinea pigs. The sensitivity of MPA from rats to NE during aerobic conditions is not significantly affected by the absence of substrate in the experimental medium. However, when the preparation was exposed to hypoxia, the presence of substrate failed to maintain the reactivity of MPA to norepinephrine. In addition, MPA isolated from rats demonstrated a smaller contractile response to NE than those from guinea pigs. Furthermore, no spontaneous mechanical activities were observed after norepinephrine or histamine administration. The present study, in addition to pointing out species differences, shows the important role of exogenous substrate in maintaining the reactivity of pulmonary vascular smooth muscle during hypoxia.

Length-dependent sensitivity at lengths greater than Lmax in vascular smooth muscle

1983 ◽  
Vol 245 (3) ◽  
pp. H379-H384 ◽  
Author(s):  
J. M. Price ◽  
D. L. Davis ◽  
E. B. Knauss
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Dose Response ◽  
Muscle Length ◽  
Maximal Response ◽  
Anterior Tibial Artery ◽  
Active Force ◽  
Response Curves ◽  
Active Stress ◽  
Dose Response Curves

Previous work has shown that vascular smooth muscle sensitivity depends on muscle length (arterial circumference) at lengths equal to and less than that for maximum active force (Lmax). In the present study dose-response curves were obtained from dog anterior tibial artery rings at lengths equal to or longer than Lmax. The curves were compared with dose-response curves obtained at lengths less than Lmax. The agonist concentration for half maximal response (ED50) was determined by graphical estimation and by calculation from a best-fit curve. The results show that with norepinephrine (NE) stimulation 1) ED50 decreased significantly at each step when the rings were stretched from Lmax to 1.15 Lmax and then to 1.30 Lmax; 2) ED50 increased significantly when length was decreased from 1.15 to 1.00 Lmax; 3) ED50 decreased significantly at each step when the rings were stretched from 0.70 Lmax to Lmax and then to 1.30 Lmax; and 4) for NE concentration greater than the ED50 at Lmax, active stress was significantly higher at Lmax than at 0.70 Lmax or 1.30 Lmax. For an NE concentration less than the ED50 at Lmax, the active stress at 1.30 Lmax was higher than the active stress at Lmax and at 0.70 Lmax. The results show that sensitivity of vascular smooth muscle continually increases with stretch and does not have a maximum at the length for maximum active force.

STUDIES ON THE PITUITARY MELANOPHORE-EXPANDING HORMONE WITH REFERENCE TO ITS IDENTITY WITH ACTH

1954 ◽  
Vol 11 (1) ◽  
pp. 7-13 ◽  
Author(s):  
B. KETTERER ◽  
ELIZABETH REMILTON
Keyword(s):  
Linear Regression ◽  
Dose Response ◽  
Progressive Increase ◽  
Regression Curve ◽  
Response Curves ◽  
Experimental Conditions ◽  
Response Data ◽  
The Mean ◽  
Dose Response Curves ◽  
Linear Regression Curve

SUMMARY 1. The standard Xenopus method for the assay of pituitary melanophore-expanding hormone has been critically examined, and the results from various assay procedures are statistically analysed. 2. Log dose-response data are well fitted by a linear regression curve. Responses at 3 hr give a steeper curve than those at 1½ hr. 3. Results collected 6 months apart show that the mean and slope of dose-response curves remain constant when Xenopus are given regular dosage; there is, however, a progressive increase of variance with time shown by the colony under these experimental conditions. 4. Evidence is presented to show that Xenopus must be minimally disturbed during assay, and that assay doses must be given not less than 1 day apart.

Application of Drug Receptor Theories to the Analysis of the Myotropic Effects of Bradykinin

1975 ◽  
Vol 53 (3) ◽  
pp. 345-353 ◽  
Author(s):  
J. Barabé ◽  
W. K. Park ◽  
D. Regoli
Keyword(s):  
Dose Response ◽  
Terminal Ileum ◽  
Maximal Response ◽  
Rat Stomach ◽  
Response Curves ◽  
Rat Uterus ◽  
Drug Concentrations ◽  
General Pharmacology ◽  
Dose Response Curves ◽  
Drug Receptor

Cat jejunum and terminal ileum, and rat stomach strip and rat uterus contract to bradykinin, while rat duodenum relaxes. Dose–response curves of classical hyperbolic shape are obtained in the first three preparations, but not in the others.The negative logs of the drug concentrations which give 50% of the maximal response. (pD2) values were, respectively, 7.68 and 7.77 in the cat jejunum and terminal ileum, 6.78 in the rat stomach strip and 8.64 in the rat uterus in estrus.Theoretical dose–response curves, constructed by using experimental pD2 values in the equation of Clark, (General pharmacology. Verlag Van J. Springer, Berlin, 1937), are superimposed to experimental curves, obtained in the cat jejunum and terminal ileum, but not in the rat stomach strip. This comparison was not made in the rat uterus and duodenum.The myotropic effect of bradykinin appears to be a direct one in the cat jejunum, the terminal ileum and the rat stomach strip, because it is not affected by anticholinergics, antiadrenergics, antihistaminics and indomethacin. pD2 values and the slope of the dose–response curves of the rat uterus were reduced by indomethacin.The results indicate that cat jejunum and terminal ileum are sensitive and specific for bradykinin and appear to be the most reliable preparations for studies on the structure–activity relationships of this peptide.

scholarly journals Cutaneous blood flow and sweat rate responses to exogenous administration of acetylcholine and methacholine

2007 ◽  
Vol 102 (5) ◽  
pp. 1856-1861 ◽  
Author(s):  
Kenichi Kimura ◽  
David A. Low ◽  
David M. Keller ◽  
Scott L. Davis ◽  
Craig G. Crandall
Keyword(s):  
Blood Flow ◽  
Dose Response ◽  
Sweat Rate ◽  
Molar Concentration ◽  
Maximal Response ◽  
Cutaneous Blood Flow ◽  
Response Curves ◽  
Forearm Skin ◽  
Dose Response Curves ◽  
Cutaneous Blood

The aim of this study was to evaluate cutaneous vasodilation and sweating responses to exogenous administration of acetylcholine (ACh) and methacholine (MCh), which have different sensitivities to endogenous cholinesterase. Four intradermal microdialysis probes were placed in dorsal forearm skin: two sites were perfused with ACh (1 × 10−7–1 M) and the other two with the same molar concentrations of MCh. Sweat rate (SR) and cutaneous blood flow were simultaneously assessed directly over each microdialysis membrane. Dose-response curves were constructed, and the effective concentration of the drug resulting in 50% of the maximal response (EC50) was identified. For SR and cutaneous vascular conductance (CVC), there were no significant differences in EC50 between sites receiving the same drug: −1.52 ± 0.18 and −1.19 ± 0.09 log-molar concentration of ACh at distal and proximal sites, respectively, and −2.35 ± 0.24 and −2.42 ± 0.23 log-molar concentration of MCh at distal and proximal sites, respectively, for SR ( P > 0.05) and −3.87 ± 0.32 and −3.97 ± 0.27 log-molar concentration of ACh at distal and proximal sites, respectively, and −4.78 ± 0.17 and −4.46 ± 0.16 log-molar concentration of MCh at distal and proximal sites, respectively, for CVC ( P > 0.05). However, the EC50 for CVC and SR was significantly lower at the MCh than at the ACh sites. A second procedure was performed to confirm that differences in responses between ACh and MCh could be attributed to different cholinesterase sensitivities. Similarly, four microdialysis membranes were placed in dorsal forearm skin: two sites were perfused with ACh and other two with MCh. However, one of each of the ACh and MCh sites was also perfused with 10 μM neostigmine (an acetylcholinesterase inhibitor). Neostigmine at the ACh site induced a leftward shift (i.e., lower EC50) of the SR and CVC dose-response curves compared with the site treated with ACh alone, resulting in no difference in the EC50 for SR and CVC between the ACh + neostigmine and the MCh site. These results suggest that elevations in SR and CVC occur earlier with MCh than with ACh treatment because of differences in cholinesterase susceptibility between these drugs.

Effects of inhaled substance P on airway responsiveness to methacholine in asthmatic subjects in vivo

1994 ◽  
Vol 77 (3) ◽  
pp. 1325-1332 ◽  
Author(s):  
D. Cheung ◽  
H. van der Veen ◽  
J. den Hartigh ◽  
J. H. Dijkman ◽  
P. J. Sterk
Keyword(s):  
Substance P ◽  
Dose Response ◽  
Forced Expiratory Volume ◽  
Maximal Response ◽  
Response Curves ◽  
Airway Narrowing ◽  
Dose Response Curves ◽  
High Doses

We tested the hypothesis that the inhaled tachykinin substance P (SP) can induce hyperresponsiveness to methacholine in asthmatic subjects in vivo. Nine atopic nonsmoking asthmatic males with normal forced expiratory volume in 1 s (FEV1; > 80% predicted) and increased methacholine sensitivity [provocative concn causing 20% fall in FEV1 (PC20) < 8 mg/ml] participated in a two-period placebo-controlled crossover study. Dose-response curves to SP (0.25–8 mg/ml) and placebo were recorded on 2 randomized days at least 1 wk apart, and methacholine tests were done 24 h before and 2 and 24 h after these challenges. The responses were measured by FEV1 (%fall from baseline). The position of the methacholine dose-response curves was expressed by the PC20 FEV1 and by the maximal response by the plateau level (MFEV1). SP caused a dose-dependent fall in FEV1 (P < 0.001). There was a slight increase in the PC20 FEV1 at 2 and 24 h, which was not significantly different between placebo and SP. Similarly, there was a reduction in MFEV1 at 2 h after both pretreatments. However, at 24 h after SP inhalation, MFEV1 increased compared with placebo. These changes in MFEV1 were significantly different between SP and placebo by 5.2 +/- 2.2% fall (SE) (P < 0.05). We conclude that 1) a bronchoconstrictive dose of SP, compared with placebo, enhances maximal airway narrowing to methacholine in asthma 24 h after inhalation and 2) tolerance develops to high doses of inhaled methacholine. These findings are suggestive of a role of SP in causing excessive airway narrowing in asthma by inflammatory mechanisms.

Stimulation of oxyntic and histaminergic cells in gastric mucosa by gastrin C-terminal tetrapeptide

1986 ◽  
Vol 251 (4) ◽  
pp. G529-G537 ◽  
Author(s):  
M. C. Ruiz ◽  
F. Michelangeli
Keyword(s):  
Gastric Mucosa ◽  
Histamine Release ◽  
Dose Response ◽  
Extracellular Calcium ◽  
Maximal Response ◽  
Response Curves ◽  
Direct Stimulation ◽  
Oxyntic Cell ◽  
Dose Response Curves ◽  
Stimulation Of

Gastrin tetrapeptide (TG) stimulation of H+ secretion and histamine release and their relationship were characterized in isolated amphibian gastric mucosa. TG released histamine and stimulated H+ secretion. Dose-response curves to TG showed nonparallel increases in both parameters. In tachyphylaxis experiments, there was no correlation between histamine release and H+ secretion. Comparison of cimetidine inhibitory dose-response curves of maximal TG or histamine stimulation showed a much higher sensitivity for TG-stimulated H+ secretion. This suggests that the pericellular histamine concentration released by TG was lower than that needed for maximal response. In mucosae maximally stimulated by histamine, TG induced a further increase in H+ secretion. In the presence of cimetidine, there was potentiation between TG and dibutyryl cAMP that was dependent on the presence of extracellular calcium. The results are interpreted to indicate that TG (or gastrin) stimulates H+ secretion through different mechanisms: 1) direct stimulation of the oxyntic cell dependent on extracellular calcium, 2) potentiation between gastrin and histamine, and 3) release of histamine from paracrine cells. Direct stimulation of the oxyntic cell by TG is not sufficient to trigger H+ secretion. Potentiation between TG and histamine at the oxyntic cell may result from interaction between TG-induced increases in intracellular Ca2+ and histamine-induced increases in intracellular cAMP.

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