Functional organisation of the endomembrane network in the digestive gland of the Venus flytrap: revisiting an old story with a new microscopy toolbox.

Up-to-date imaging approaches were used to address the spatiotemporal organisation of the endomembrane system in secretory cells of Dionaea muscipula. Different 'slice and view' methodologies were performed on resin-embedded samples to finally achieve a 3D reconstruction of the cell architecture, using ultrastructural tomography, array tomography, serial block face-scanning electron microscopy (SBF-SEM), correlation, and volume rendering at the light microscopy level. Observations of cryo-fixed samples by high-pressure freezing revealed changes of the endomembrane system that occur after trap activation and prey digestion. They provide evidence for an original strategy that adapts the secretory machinery to a specific and unique case of stimulated exocytosis in plant cells. A first secretion peak is part of a rapid response to deliver digestive fluids to the cell surface, which delivers the needed stock of digestive materials 'on site'. The second peak of activity could then be associated with the reconstruction of the Golgi apparatus (GA), endoplasmic reticulum (ER) and vacuolar machinery, in order to prepare for a subsequent round of prey capture. Tubular continuum between ER and Golgi stacks observed on ZIO-impregnated tissues may correspond to an efficient transfer mechanism for lipids and/or proteins, especially for use in rapidly resetting the molecular GA machinery. The occurrence of one vacuolar continuum may permit continuous adjustment of cell homeostasy. The subcellular features of the secretory cells of Dionaea muscipula outline key innovations in the organisation of plant cell compartmentalisation that are used to cope with specific cell needs such as the full use of the GA as a protein factory, and the ability to create protein reservoirs in the periplasmic space. Shape-derived forces of the pleiomorphic vacuole may act as signals to accompany the sorting and entering flows of the cell.

Initiation of forward gait with lateral occurrence of emotional stimuli: general findings and relevance for pedestrians crossing roads.

With reference to theoretical models regarding links between emotions and actions, the present study examined whether the lateral occurrence of an emotional stimulus influences spatial and temporal parameters of gait initiation in 18 younger and 18 older healthy adults. In order to simulate road-crossing hazard for pedestrians, slides of approaching cars were used and they were presented in counterbalanced order with threatening slides from the International Affective Picture System (IAPS) and control slides of safe walking areas. Each slide was presented on the left side of the participant once the first step was initiated. The results evidenced medio-lateral shifts to the left for the first step (right foot) and to the right for the second step (left foot). These shifts were both modulated by the slide contents in such a way that the resulting distance between the screen and the foot (right or left) was larger with the IAPS and traffic slides than with the control slides. The slides did not affect the base of support, step length, step velocity and time of double support. Advancing age influenced the subjective impact of the slides and gait characteristics, but did not modulate medio-lateral shifts. The data extend evidence of fast, emotional modulation of stepping, with theoretical and applied consequences.

Enhanced precision of ankle torque measure with an open-unit dynamometer mounted with a 3D force-torque sensor.

PURPOSE: Many studies have focused on maximum torque exerted by ankle joint muscles during plantar flexion. While strength parameters are typically measured with isokinetic or isolated ankle dynamometers, these devices often present substantial limitations for the measurement of torque because they account for force in only 1 dimension (1D), and the device often constrains the body in a position that augments torque through counter movements. The purposes of this study were to determine the contribution of body position to ankle plantar-flexion torque and to assess the use of 1D and 3D torque sensors. METHODS: A custom designed 'Booted, Open-Unit, Three dimension, Transportable, Ergometer' (B.O.T.T.E.) was used to quantify plantar flexion in two conditions: (1) when the participant was restrained within the unit (locked-unit) and (2) when the participant's position was independent of the ankle dynamometer (open-unit). Ten young males performed maximal voluntary isometric plantar-flexion contractions using the B.O.T.T.E. in open and locked-unit mechanical configurations. RESULTS: The B.O.T.T.E. was reliable with ICC higher than 0.90, and CV lower than 7 %. The plantar-flexion maximal resultant torque was significantly higher in the locked-unit compared with open-unit configuration (P < 0.001; +61 to +157 %) due to the addition of forces from the body being constrained within the testing device. A 1D compared with 3D torque sensor significantly underestimated the proper capacity of plantar-flexion torque production (P < 0.001; -37 to -60 %). CONCLUSIONS: Assessment of plantar-flexion torque should be performed with an open-unit dynamometer mounted with a 3D sensor that is exclusive of accessory muscles but inclusive of all ankle joint movements.

Kinetic adaptations of the intact limb in transfemoral amputees using a microprocessor prosthetic knee.

BACKGROUND: In recent decades, high-tech prostheses, including microprocessor-controlled knee (MPK), have been developed to improve the functional abilities of lower limb amputees and to reduce gait asymmetry for the prevention of early joint degradation of the intact limb. The aim of this study was to determine the differences in joint moment and power of the intact limb of transfemoral amputees (TFAs) with an MPK compared to healthy individuals in 2 walking speed conditions. METHODS: Twenty-one TFAs with MPK and matched 21 healthy individuals performed a walking task at spontaneous and rapid self-selected speeds. Spatiotemporal gait parameters and intact limb kinetic data were recorded. RESULTS: The hip and knee moments in the frontal plane during rapid walking were not significantly higher than spontaneous walking in TFA group (respectively p = 0.08 and p = 0.48) and were lower than the control group. In the sagittal plane, the hip extensor moment in TFA was higher than the control group in the landing phase (p < 0.001 in both speed conditions). SIGNIFICANCE: The kinetics of the intact limb of active TFAs with an MPK showed a significant reduction at the knee internal abductor moment compared to the control, potentially limiting risk factors for knee osteoarthritis. However, in the sagittal plane, higher hip extensor moments could favor low-back pain appearance. Gait analysis of lower limb amputees should thus be performed to highlight these kinetic adaptations and then help to propose the most relevant rehabilitation and prevention exercises to limit the appearance of early musculoskeletal degeneration.

Transfemoral amputees adapt their gait during cross-slope walking with specific upper-lower limb coordination.

BACKGROUND: Unilateral lower limb amputees have asymmetrical gaits, particularly on irregular surfaces and slopes. It is unclear how coordination between arms and legs can adapt during cross-slope walking. RESEARCH QUESTION: How do transfemoral amputees (TFAs) adapt their upper-lower limb coordination on cross-slope surfaces? METHODS: Twenty TFA and 20 healthy adults (Ctrl) performed a three-dimensional gait analysis in 2 walking conditions: level ground and cross-slope with prosthesis uphill. Sagittal joint angles and velocities of hips and shoulders were calculated. Continuous relative phases (CRP) were computed between the shoulder and the hip of the opposite side. The closer to 0 the CRP is, the more coordinated the joints are. Curve analysis were conducted using SPM. RESULTS: The mean CRP between the downhill shoulder and the uphill hip was higher in TFA compared to Ctrl (p = 0.02), with a walking conditions effect (p = 0.005). TFA showed significant differences about the end of the stance phase (p = 0.01) between level ground and cross-slope, while Ctrl showed a significant difference (p = 0.008) between these walking conditions at the end of the swing phase. In CRP between the uphill shoulder and the downhill hip, SnPM analysis showed intergroup differences during the stance phase (p < 0.05), but not in the comparison between walking conditions in TFA and Ctrl groups. SIGNIFICANCE: TFA showed an asymmetrical coordination in level ground walking compared to Ctrl. Walking on cross-slope led to upper-lower limb coordination adaptations: this condition impacted the CRP between downhill shoulder and uphill hip in both groups. The management of the prosthetic limb, positioned uphill, induced a reorganization of the coordination with the upper limb of the amputated side. Identifying upper-lower limb coordination adaptations on cross-slope surfaces will help to achieve rehabilitation goals for effective walking in urban environments.

Walking abilities improvements are associated with pelvis and trunk kinematic adaptations in transfemoral amputees after rehabilitation.

BACKGROUND: Rehabilitation can be proposed to transfemoral amputees to improve functional abilities and limit the risk of early degeneration of the musculoskeletal system partly due to altered kinematic behavior. The main aim of this study was to assess the impact of functional rehabilitation on clinical walking tests, gait symmetry and pelvis and trunk kinematics in transfemoral amputees during overground walking. METHODS: Eleven transfemoral amputees followed a functional rehabilitation program with objectives aimed at improving walking abilities and gait symmetry. Clinical functional tests, symmetry between prosthetic and intact sides and trunk and pelvis motions were recorded before and after rehabilitation. FINDINGS: Clinical walking tests were improved after rehabilitation (p < 0.05), and step width was reduced (p = 0.04). Regarding symmetry between the single stances on the prosthesis and intact sides, only a significant decrease in trunk frontal inclination asymmetry was noted after rehabilitation (p = 0.01). Pelvic frontal obliquity was significantly increased during prosthetic (p = 0.02) and intact single stances (p = 0.005). INTERPRETATION: Our study showed a positive effect of rehabilitation on transfemoral amputees functional abilities. These improvements were associated with higher pelvic mobility in frontal plane and a more symmetrical redistribution of the frontal trunk sway around the vertical axis during gait. These results suggest the importance of a postural reeducation program for transfemoral amputees aimed at improving pelvic dynamic control while repositioning the trunk by postural corrections during gait.

Disruption of the secretion and action of 17,20ß-dihydroxy-4-pregnen-3-one in response to a rise in temperature in the Arctic charr, Salvelinus alpinus. Consequences on oocyte maturation and ovulation.

Plasma levels of 17,20ß-dihydroxy-4-pregnen-3-one (17,20ßP), and the timing of ovulation were investigated in female Arctic charr (Salvelinus alpinus) reared at 5°C and at 10°C during the pre-spawning period. The effects of switching from 5 to 10°C, and from 10 to 5°C were also investigated. 17,20ßP plasma levels were higher at 5°C than at 10°C. A switch from 10 to 5°C stimulated 17,20ßP secretion, whereas a switch from 5 to 10°C had the opposite effect. Ovulation occurred spontaneously in the females kept at 5°C, and in those switched from 10 to 5°C. In contrast, ovulation was inhibited in females reared at 10°C, and in those switched from 5 to 10°C. Oocyte maturation at 5°C and at 10°C in the presence of LH or of 17,20ßP was also investigated in vitro using donor females reared at 5 or 10°C. Both LH and 17,20ßP stimulated oocyte maturation more effectively in oocytes incubated at 5°C than at 10°C. At both incubation temperatures, the rearing temperature of the donor females had a significant impact on their responsiveness to LH stimulation, but had no effect on their responsiveness to 17,20ßP stimulation. In addition to the inhibition of LH secretion, which had already been reported, the results reported here show that in Arctic charr raising the temperature above the physiological range reduces both follicular responsiveness to LH stimulation and the sensitivity of oocytes to 17,20ßP stimulation.

Genotoxic pressure of vineyard pesticides in fish: field and mesocosm surveys.

The present study deals with the genotoxicity assessment of vineyard pesticides in fish exposed in the field or in mesocosm conditions. Primary DNA damage was quantified as strand breaks using the single cell gel electrophoresis assay (Comet assay) applied to fish erythrocytes. In a first experiment, a significant genotoxic effect was observed following an upstream-downstream gradient in early life stages of brown trout (Salmo trutta fario) exposed in the Morcille River contaminated by a mixture of vineyard pesticides during three consecutive years. The pronounced response in terms of DNA damage reported in the present study could argue for a high sensitivity of fish early life stage and/or a high level of exposure to genotoxic compounds in the Morcille River. This stresses the interest in using trout larvae incubated in sediment bed to assess genotoxic compounds in the field. In a second experiment, adult European topminnow (Phoxinus phoxinus) were exposed in water running through artificial channels to a mixture of diuron and azoxystrobin, two of the main pesticides detected in the Morcille watershed. As compared with the unexposed channel, a 3-5-fold increase in the DNA damage was observed in fish exposed to chronic environmental pesticide concentrations (1-2 microg L(-1) for diuron and 0.5-1 microg L(-1) for axoxystrobin). A single 6h pulse of pesticide (14 microg L(-1) of diuron and 7 microg L(-1) of azoxystrobin) was applied to simulate transiently elevated chemical concentrations in the river following storm conditions. It did not increase genotoxicity. After a 1-month recovery period, DNA damage in exposed fish erythrocytes recovered to unexposed level, suggesting possible involvement of both repair mechanisms and cellular turnover in this transient response. This work highlights that vineyard treatment by pesticides and in particular diuron and azoxystrobin can represent a genotoxic threat to fish from contaminated watershed rivers.

Immunoreactive collagenase and bone resorption.

1. Active mouse bone collagenase is excluded from its inhibitory antibody by preincubation of that antibody with various forms of inactive enzyme, e.g. 'procollagenase', some collagenase-inhibitor complexes or partially denatured or degraded collagenase. This property allows the detection of several enzymatically inactive forms of collagenase. 2. The accumulation of immunoreactive collagenase in the culture fluid of mouse bones occurred only in the presence of heparin and was not correlated with bone resorption induced by parathyroid hormone. These experiments provide further (see Lenaers-Claeys, G. and Vaes, G., Biochim. Biophys. Acta (1979) 584, 375-388), more conclusive evidence that the critical role in the resorption of the organic matrix of these explants may be due to another enzyme system than collagenase.

Partially reversible osteopenia after surgery for primary hyperparathyroidism.

Bone mineral content of the radius in 30 patients with primary hyperparathyroidism was determined by single-photon absorptiometry before and one year after the removal of a parathyroid adenoma. When compared with normal age- and sex-matched subjects, preoperative bone mineral content was decreased by more than 1 SD in 24 patients 3 cm from the styloid apophysis (distal site) and in 23 patients 8 cm from the styloid apophysis (proximal site). The decrease at the distal site, where there is more trabecular bone, was significantly greater than at the proximal site (74% +/- 13% of the normal mean vs 79% +/- 13%). Bone mineral content values were not different in the female and male populations. One year after surgery, the bone mineral content was significantly increased but was still lower than normal at both sites (81% and 84% of the normal mean, respectively). In some patients, seen up to three years after surgery, the bone mineral content remained lower than normal. We conclude that primary hyperparathyroidism causes an osteopenia that affects both cortical and trabecular bone and that is only partially reversible, even in the long term.

Long-term irreversibility of bone loss after surgery for primary hyperparathyroidism.

We reported previously that radial bone mineral content was decreased in patients with primary hyperparathyroidism and remained subnormal 1 year after surgery. In this study, we reviewed the results of sequential measurements of the radial bone mineral content, performed up to 107 months after removal of the parathyroid adenoma in 71 patients suffering from primary hyperparathyroidism. Bone mineral content increased during the first year after surgery. During the period 1 to 8 months after removal of the adenoma, the mean monthly increment was 0.009 +/- 0.0022 g/cm for the radial epiphysis and 0.0084 +/- 0.0023 g/cm for the shaft. However, in 39 patients seen at the end of 1 year after surgery, the bone mineral content of the epiphysis remained more than 1 SD below the normal mean in 61% (24) of the patients and more than 2 SDs in 36% (14) of the patients. For the shaft, those percentages were 59% (23) and 26% (10), respectively. Thereafter, the monthly increment rate of bone mineral content decreased rapidly with time, so that only minor further increase could be expected. Data show that patients with a low bone mineral content when diagnosed with primary hyperparathyroidism will conserve life long an irreversible loss of bone as compared with a matched control population.

Oleate Abrogates Palmitate-Induced Lipotoxicity and Proinflammatory Response in Human Bone Marrow-Derived Mesenchymal Stem Cells and Osteoblastic Cells.

Osteoporosis is a metabolic bone disease associated with unequilibrated bone remodeling resulting from decreased bone formation and/or increased bone resorption, leading to progressive bone loss. In osteoporotic patients, low bone mass is associated with an increase of bone marrow fat resulting from accumulation of adipocytes within the bone marrow. Marrow adipocytes are active secretory cells, releasing cytokines, adipokines and free fatty acids (FA) that influence the bone marrow microenvironment and alter the biology of neighboring cells. Therefore, we examined the effect of palmitate (Palm) and oleate (Ole), 2 highly prevalent FA in human organism and diet, on the function and survival of human mesenchymal stem cells (MSC) and MSC-derived osteoblastic cells. The saturated FA Palm exerted a cytotoxic action via initiation of endoplasmic reticulum stress and activation of the nuclear factor κB (NF-κB) and ERK pathways. In addition, Palm induced a proinflammatory response, as determined by the up-regulation of Toll-like receptor 4 expression as well as the increase of IL-6 and IL-8 expression and secretion. Moreover, we showed that MSC-derived osteoblastic cells were more sensitive to lipotoxicity than undifferentiated MSC. The monounsaturated FA Ole fully neutralized Palm-induced lipotoxicity by impairing activation of the pathways triggered by the saturated FA. Moreover, Ole promoted Palm detoxification by fostering its esterification into triglycerides and storage in lipid droplets. Altogether, our data showed that physiological concentrations of Palm and Ole differently modulated cell death and function in bone cells. We therefore propose that FA could influence skeletal health.

Effect of acute hypercalcemia on thyrotropin (TSH) and triiodothyronine responses to TSH-releasing hormone in man.

In chronic hypercalcemia, basal TSH has been found to be low, with normal serum circulating concentrations of T3 and T4. This observation suggested a potentiation by hypercalcemia of the thyroid secretory response to TSH. The present study was undertaken to assess the possible influence of hypercalcemia on the T3 secretory response to TSH. Since T3 secretion was studied after stimulation of endogenous TSH by TRH, it was first necessary to find a protocol enabling us to study the effect of calcium on T3 release without affecting TSH secretion. Eighteen subjects underwent two TRH tests, with and without simultaneous calcium infusion, at 2-week interval and in a randomized order. In group A (five subjects) calcium infusion started 1 min after TRH, in group B (five subjects) 10 min after TRH, and in group C (eight subjects) 20 min after TRH. In groups A and B, TSH secretion was markedly blunted by hypercalcemia. In contrast, when calcium infusion was started 20 min after TRH (group C), the TSH secretion profile was no longer different from that in the control study. However, in this situation the increments of T3 and free T3 120 and 180 min after TRH were significantly higher when the subjects were rendered hypercalcemic than in the control study. These findings suggest that calcium might act at two different levels, to enhance the thyroid secretory response to TSH and decrease TSH secretion by acting directly on the pituitary gland. Both effects would produce the association of low serum TSH and normal levels of T3 and T4 observed in chronic hypercalcemia.

Differential expression of the 67-kD laminin receptor and 31-kD human laminin-binding protein in human ovarian carcinomas.

The expression of the 67-kD laminin receptor (67LR) and the 31-kD human laminin-binding protein (HLBP31), two proteins involved in cancer cell laminin interaction, was evaluated on 30 ovarian cancer specimens. Expression of the 67LR was increased (up to 2.5-fold, in 87% of the patients), while HLBP31 expression was downregulated in cancer cells compared with the normal tissue, as detected by northern blotting and immunohistochemistry. The immunohistochemical study demonstrated that the 67LR was significantly overexpressed (P < 0.05) in the group of patients whose cytoreductive surgery was suboptimal, and those with poor clinical outcome. No correlation was observed between HLBP31 expression and clinicopathological features. Increased expression of the 67LR appears to correlate with the invasive phenotype of ovarian cancer cells and suggests a role of the latter in ovarian cancer invasion.

Reliability of amidolytic assays for progressive antithrombin and heparin cofactor activities on capillary blood.

Amidolytic assay of antithrombin III on capillary blood can validly substitute for similar assays performed on venous blood, as an excellent correlation exists (r = 0.95). For the amidolytic assays of heparin cofactor activity, a much less satisfactory correlation is found (r = 0.81). Results are far more dispersed and a decrease is observed in late capillary samples. Using a low heparin concentration to assay heparin cofactor activity leads to surprisingly high activities for capillary blood. The same type of discrepancy is observed during the earliest stages of clotting of venous blood.

Outcome of treatment in alcoholic women.

One hundred and seventy-eight females (mean age 40.6 +/- 10.2 years) were retrospectively studied by questionnaires for a mean duration of 46 (17-75) months. Sixty-two percent were married or living maritally. One third were working. The mean alcohol intake was 157 +/- 76 g/day and 57.3% had alcohol dependence for less than 5 years. Twenty-seven patients (15%) were lost to follow-up; out of the 151 remaining patients, 7 (4%) refused to answer and 18 (12%) died. Suicide and alcoholism complications were a frequent cause of death. One hundred and twenty-six questionnaires were obtained. Twenty-eight women (22%) were abstinent. A good outcome determined by the state of alcoholization (abstinence or moderate consumption) and the improvement of quality of life, was found in 44% of patients. Absence of marital life and greater alcohol intake were related to a poor outcome, whereas enrollment in a fellowship of recovering alcoholics was more frequent in abstinent patients. The mortality rate was important in alcoholic females. A number of factors were related to the outcome.

Site-specific counterion binding: application of the standard Poisson-Boltzmann cell model to ionic polysaccharides of the plant cell wall.

A numerical method is presented for analysing the effects of the competitive processes of dissociation and complexation that occur at the level of the functional groups of charged polysaccharides during ion exchange experiments carried out on plant cell walls. The interactions between the exchange sites and the mobile ions are handled by the standard Poisson-Boltzmann cell model. The dissociation and the complexation are described by mass-action laws. Uncomplexed counterions are involved in the formation of a cylindrical double layer. This latter type of interaction determines the dissociation degree of the polyion. A least-squares algorithm is used to evaluate the complexation constants of the counterions from experimental data. The need of introduction of the complexation constant of the divalent counterions has been clarified by a comparison between theoretical titration curves calculated with and without specific site binding. The fraction of complexed sites does not agree with that predicted from Oosawa's theory of the condensation. The accumulation coefficients, calculated from the optimised theoretical titration curves, clearly demonstrate the competition between the protonation and the complexation processes at the level of the pectic acids of the wall.

Serial evaluation of dilated cardiomyopathy with exercise thallium-201 tomography: correlation with the evolution of left ventricular parameters.

The purpose of this prospective study was to correlate (1) the initial findings of exercise thallium-201 tomography with the evolution of left ventricular parameters at long term follow-up in patients with dilated cardiomyopathy and (2) the changes of exercise thallium-201 tomography repeated 1 year later. We studied 19 men with dilated cardiomyopathy and normal coronary angiogram. Two patients died and three patients had heart transplantation during follow-up. The other 14 patients were assessed at baseline and 1-year follow-up. Thallium-201 tomograms were divided into 20 segments for each patient. Two groups were defined according to the evolution of left ventricular ejection fraction: group 1 (n = 7) had unchanged or decreased ejection fraction at follow-up (24 +/- 11% at baseline versus 22 +/- 11% at follow-up, ns) and group 2 (n = 7) had improved ejection fraction at follow-up (25 +/- 9% at baseline versus 49 +/- 8% at follow-up, P < 0.03). The number of total abnormal segments at stress were not statistically different at baseline between groups 1 and 2, and in group 1 between baseline and follow-up. Group 2 at follow-up had a reduced number of total abnormal segments (P < 0.03). The percentage of reversibility was similar in both groups at baseline and follow-up. On exercise thallium-201 tomography, neither the presence nor the reversibility of stress myocardial perfusion abnormalities can predict improvement of left ventricular ejection fraction in dilated cardiomyopathy. However, regression of dilated cardiomyopathy is accompanied by a reduction of stress myocardial perfusion abnormalities.

Use of transepithelial electrical resistance in the study of pentachlorophenol toxicity.

The toxicity of pentachlorophenol (PCP), a polluting substance believed to exert a narcotic effect, was assayed using the Caco-2 cell line as a model. In order to assess this toxicity as fully as possible, several viability tests, each examining different endpoints, have been used. Neutral red uptake was found to be more sensitive to PCP than MTT and Alamar Blue tests. Transepithelial electrical resistance (TEER) was shown to be the most sensitive to PCP at concentrations and exposure times where the Alamar Blue, LDH leakage and Blue Dextran passage did not evidence any effect. Blue Dextran passage and optical microscopy revealed cellular detachment at concentrations where LDH and Alamar Blue showed little or no cytotoxicity. Thus, PCP seems to affect the integrity of the intestinal barrier at levels where no cytotoxicity is seen. Our results support the notion that TEER can be used as a very sensitive method for evaluating membrane-perturbing toxicants.

Use of the caco-2 model in the screening of polluting substance toxicity.

The aim of this work was to investigate the oral toxicity of representative chemicals chosen from each class of the list of 132 substances present in industrial effluents after the EEC Directive 76-464. Owing to its characterization as a model of the intestinal epithelium, the CaCo-2 cell line model was chosen. Cytotoxicity was assayed using the tetrazolium blue (MTT) test. For most of the substances, a linear correlation was observed between the octanol/water partition coefficient (log Kw) and the median inhibition concentration (IC(50)). This relationship between lipophilicity and toxicity is the hallmark of a narcotic mechanism of action. However, diethylamine appeared more toxic than the correlation would predict. Other amines were then tested (tert-butylamine, n-butylamine and benzylamine). All of these did not fit into the baseline correlation. The IC(50) were corrected by taking into account only the non-ionized, lipid insoluble, concentration at pH7.3. The amines still did not fit into the correlation, reinforcing the idea of a non-narcotic mechanism. The toxicity of a large number of substances can thus be predicted from their physico-chemical properties only when the substances exert a direct and non-specific effect. The amines appeared more toxic than substances with the same partition coefficient, showing that knowledge of the only lipophilicity is too restrictive to predict toxicity.