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Food iron absorption in man. Applications of the two-pool extrinsic tag method to measure heme and nonheme iron absorption from the whole diet.

A new radioisotope method to measure iron absorption from the whole diet was used in this study. The method is based on the concept that food iron is absorbed from two pools, the heme iron pool and the nonheme iron pool, which can be especially labeled with two radioiron isotopes given as hemoglobin and as an iron salt. The purpose of this study was to test the accuracy of this two-pool extrinsic tag method. The meals served were composed as an average of 6 wk consumption in the present material of 32 young enlisted men. The mean and total heme and nonheme iron absorption in all the 32 young men was 1.01+/-0.11. This figure agrees well with the mean daily losses expected for this group of subjects (1.0 mg). The conclusion can therefore be made that there are no major systematic errors of the present method to measure the total iron absorption from a mixed diet. In one series a comparison was made of the absorption of heme and nonheme iron from the meals. A significant correlation between the absorption of the two kinds of iron was found. However, a much greater fraction of the heme iron was absorbed (37%) than of the nonheme iron (5%). The absorption both from breakfast and lunch was in two series found to give a good prediction of the total daily nonheme iron absorption. One series was designed to compare the effect of two levels of iron fortification. There was a significant increase in iron absorption when the level of iron fortification of the meals was increased.

Absorption↗

Intestinal absorption of several beta-lactam antibiotics. V. Effect of amino beta-lactam analogues and dipeptides on the absorption of amino beta-lactam antibiotics.

The absorption mechanism of amino beta-lactam antibiotics was investigated by using the whole small intestine of a rat. Mutual inhibition among amino beta-lactam analogues and the effects of dipeptides were studied. The influences of glycylglycine on the absorption of cephradine at the four different parts of intestine were also studied. Similarly to the case of cephalexin and cephradine, the absorption of amoxicillin was significantly inhibited by cyclacillin, cephradine, and cephalexin, but the absorption of ampicillin was not reduced by all tested antibiotics. In the experiments using dipeptides (6.0 mM), the absorption of cyclacillin was reduced significantly by glycylglycine, not by L-carnosine. And cephalexin absorption was influenced by L-carnosine (6.0, 10 mM), not by glycylglycine (6 mM). On the contrary, the absorption of cephradine was not reduced at all by these dipeptides. And from the experiment using the four different parts of intestine, it was shown that the transport interaction of glycylglycine with cephradine was observed in only one segment (the upper part of jejunum). These result suggest that the carrier-mediated transport system correlated to dipeptides participates only to a small degree in the common absorption mechanisms of these amino beta-lactam antibiotics.

Amoxicillin↗

Immunological control of drug absorption from the gastrointestinal tract: effect of local anaphylaxis on the intestinal absorption of low molecular weight drugs in the rat.

Intestinal absorption of various drugs was examined by means of in situ recirculation technique during local anaphylaxis. The antibody was determined by passive cutaneous anaphylaxis technique in rats immunized once or three times. The optimal condition of local anaphylaxis was determined by the leakage of Evans Blue. The most significant increase in leaks of the dye was observed by the intraluminal challenge with 400 mg of ovalbumin for 10 min in ovalbumin-immunized rats, and this condition was chosen as the optimal condition of local anaphylaxis. Under this condition, intestinal absorption of caffeine, phenylbutazone, and bromthymol blue (BTB) significantly decreased in ovalbumin-immunized rats compared with the control, whereas no significant effect was noted in the intestinal absorption of salicylic acid, quinine, pralidoxime iodide (2-PAM), tetracycline, and phenol red. In normal rats, no significant decrease was obtained in the intestinal absorption of caffeine, phenylbutazone, and BTB. On the other hand, the decreased absorption of BTB was not found in ovalbumin-immunized rats by the intraluminal challenge with bovine gamma-globulin. Furthermore, there was no significant change in the decreased absorption of BTB between rats immunized once and three times. The most effective condition for decreased BTB absorption was observed by the intraluminal challenge with 200 mg of ovalbumin for 10 min in ovalbumin-immunized rats, which almost correlated with the data of Evans Blue leakage. From these observations, it appears that the mucosal immune responses affect the intestinal absorption of low molecular weight drugs.

Anaphylaxis↗

Skin absorption of solvent mixtures--effect of vehicles on skin absorption of toluene.

The skin absorption rates of toluene in various solvent mixtures were investigated in mice. The skin absorption rate of toluene in toluene/methanol mixtures exhibited a parabolic relationship to the mixed ratio. The maximum rate was obtained at a mixed ratio of 50% (V/V). The skin absorption rate of toluene at this point (50%) was about 4.7 times higher than that of pure toluene. The permeability coefficient (Kp) of toluene increased as the mixed ratio of methanol increased. Methanol enhanced skin absorption of toluene. The skin absorption rate of toluene in a toluene/benzene mixture was inversely proportional to the concentration of benzene. The Kp of toluene is kept constant through the mixed ratios of benzene, and benzene does not have an enhancing effect on the skin absorption of toluene contained in the toluene/benzene mixture. We examined the effects of various vehicles on the skin absorption rates of toluene in mixtures containing 50% (v/v) of toluene. Methanol was a good penetration enhancer for toluene, and its effect is similar to the effect of well-known skin penetration enhancers like DMSO, N,N-Dimethylacetamide, and N,N-Dimethylformamide. Therefore, it is necessary to take special precautions against the skin absorption of toluene when handing thinners that contain methanol.

Animals↗

Iron absorption in early pregnancy - a study of the absorption of non-haeme iron and ferrous iron in early pregnancy.

The absorption of non-haeme iron in the food and from a 3 mg reference dose of ferrous iron in solution was measured in 17 healthy women before the 12th week of gestation and in 13 of the women two months after legal abortion. The absorption of ferrous iron in solution was calculated from determinations of the 55Fe activity in blood samples. The food iron absorption was measured from 59Fe-labelled test meals using a highly sensitive whole-body counter. In addition bone-marrow smears and other haematological parameters were studied. The median value of the absorption from the non-haeme food iron was 2.5% and from the ferrous iron salt 10.0 % in early pregnancy in women with storage iron. Two months after abortion the absorption increased to 12.6 and 42.6 % respectively. In early pregnancy the absorption of iron was higher in women without stainable iron in bone-marrow smears. The present results confirm previous observations that the absorption of food iron in early pregnancy is lower than the basal daily requirements. The low absorption is only partly explained by the reduced requirements of iron in early pregnancy. Some other factors related to pregnancy seem also to be involved.

Abortion, Induced↗

Utility and limitation of calciuric response to oral calcium load as a measure of intestinal calcium absorption: comparison with isotopic fractional calcium absorption.

The intestinal absorption of calcium (Ca), indirectly measured from the calciuric response to oral Ca load (1g), was compared to the more directly obtained isotopic fractional absorption, alpha (from the fecal recovery of orally administered 47Ca). In 17 normal subjects and 30 patients with absorptive hypercalciuria (AH), there was a significant (P less than 0.001) correlation of alpha with the Ca load responses, (r = 0.81). However, this correlation was not observed in patients with renal hypercalciuria (RH), and those with AH receiving thiazide or orthophosphate. In RH, 38 per cent of patients had elevated Ca load responses, despite normal values for alpha. The point correlating the calciuric response and alpha in these patients was below the 95 per cent confidence limit of the line correlating alpha and the load response. Thus, Ca load response often overestimated intestinal Ca absorption, because of the high basal (fasting) urinary Ca. Thiazide therapy in RH improved the correlation between the two tests of Ca absorption. However, thiazide therapy in AH produced normal Ca load responses despite persistently high alpha in 60 per cent of patients. Similarly, 50 per cent of patients with AH receiving orthophosphate had normal Ca load response, although alpha remained elevated. Thus, Ca load response underestimated Ca absorption when patients with AH took thiazide or orthophosphate, probably because these drugs augment renal tubular reabsorption of Ca. These data support the Ca load test as a valid indirect measure of intestinal Ca absorption in normal subjects and patients with AH, in whom fasting urinary Ca is not elevated. In conditions of renal Ca, leak or with various drugs known to alter renal Ca handling, there seen to be large deviations of Ca load response from alpha. Care should be exercised before reaching conclusions regarding the intestinal Ca absorption in these situations.

Administration, Oral↗

STUDIES ON INHIBITION OF INTESTINAL ABSORPTION OF RADIOACTIVE STRONTIUM. I. PREVENTION OF ABSORPTION FROM LIGATED INTESTINAL SEGMENTS.

A method is reported which permits selective suppression of absorption of radioactive strontium from ingested food material, permitting the calcium to be available to the body. Studies were carried out in vivo by injection of Sr(89) and Ca(45) in the presence of inert carrier into ligated intestinal segments in rats, and the amount of absorption was measured by standard monitoring techniques. The pattern of absorption of both ions is very similar but the rate of absorption is different. It was found that the polyelectrolyte, sodium alginate, obtained from brown algae (Phaeophyceae), injected simultaneously with radiostrontium effectively reduces the absortion of Sr(89) from all segments of the intestine by as much as 50-80% of the control values. No significant reduction in absorption of Ca(45) was observed in equivalent concentrations. The reduction in blood levels of Sr(89) and in bone uptake corresponded to the absorption pattern. The difference in the effect on strontium and calcium absorption may be due to differences in the binding capacity of sodium alginate from the two metal ions under the conditions present in vivo.

Absorption↗

Experimental studies on absorption, distribution and excretion of a new antibiotic, fosfomycin. II. Absorption of oral preparations of fosfomycin calcium salt in dogs (author's transl).

To establish the best usage and dosage of fosfomycin granule and capsules which had been prepared based on our fundamental experiences as described in the first report, absorption of calcium salt contained in both preparations was evaluated using dogs as test animals. (1) Granule containing the calcium salt equivalent to 200 mg of fosfomycin free acid per g showed almost the same absorption as the bulk (fosfomycin calcium), having no disadvantage due to processing. (2) Capsules containing the calcium salt equivalent to 250 mg and 500 mg of the free acid per capsule showed slightly more retarded absorption than the bulk, probably due to some inevitable factors such as disintegration rate of capsules and dispersion rate of the calcium salt. But, once dispersed, the calcium salt in capsules was well absorbed as well as the bulk material. (3) Gastrointestinal absorption of granule and the capsule contents was almost the same. (4) Simultaneous administration of capsules and water improved the absorption efficiency. Though administration after feeding caused somewhat retarded absorption of the drug, the serum levels were rather well sustained with a slight drop but sufficiency of absorption, suggesting better clinical advantages than in the fasted animals. (5) Fosfomycin calcium salt in both preparations was well absorbed in the test animals through gastrointestinal tract as well as the bulk calcium salt, without any possible disadvantage caused by processing. In addition, the absorption efficiency was improved by giving with water or meal to the animals.

Administration, Oral↗

Drug absorption VII: influence of mesenteric blood flow on intestinal drug absorption in dogs.

Intestinal absorption of sulfaethidole and haloperidol was determined using an in situ canine intestinal preparation. Intestinal absorption of sulfaethidole was determined at three or four mesenteric blood flow rates in each dog, ranging from unaltered flow (100%) to no flow (0%). A relatively small change in absorption rate occurred when the splanchnic blood flow rate was decreased about 35%. Further reductions in mesenteric blood flow resulted in progressive impairment of sulfaethidole absorption. The simultaneous measurement of sulfaethidole intestinal disappearance and appearance in blood indicates that sulfaethidole disappearance is equivalent to absorption. Haloperidol absorption also decreased with decreased intestinal perfusion but differed from sulfaethidole in that membrane storage of haloperidol appeared to take place during its absorption.

Animals↗

Effect of absorption promoters on subcutaneous absorption of human epidermal growth factor in rats.

Subcutaneous administration of human epidermal growth factor (hEGF) to rats gave a significantly smaller value of area under the curve (AUC) of concentration in plasma of immunoreactive hEGF versus time than intravenous administration, probably because the slow entry rate into the blood circulation and consequently the enzymic degradation of hEGF at the injection site. In the present study, absorption promoters such as sodium caprate, N-acylamino acids, disodium ethylenediamine-tetraacetate (EDTA), and sodium glycocholate were used because they were expected to inhibit the enzymic degradation of hEGF at the injection site and to facilitate the entry of hEGF into the blood circulation. Coadministration of an absorption promoter with hEGF significantly increased the entry rate and AUC value of immunoreactive hEGF compared with the case without the absorption promoter. The enzymic degradation of hEGF in the supernatant of the rat subcutaneous tissue homogenates and in the buffer solution containing leucine aminopeptidase or protease was markedly inhibited by the presence of the absorption promoters except EDTA. On the other hand, only EDTA increased the initial entry rate of FITC-dextran (M(r), 4000), which is not metabolized at the injection site, although all absorption promoters including EDTA markedly increased the extravasation of Evans blue. Thus, the increased subcutaneous bioavailability of hEGF in the presence of absorption promoters (except EDTA) was mainly attributed to the inhibitory effect of absorption promoters against the enzymic degradation of hEGF at the subcutaneous tissues.

Adjuvants, Pharmaceutic↗

Lateral hypothalamic responses to pre-absorptive and post-absorptive signals related to amino acid ingestion.

This paper reports an investigation of the comparative effects of pre-absorptive and post-absorptive stimuli associated with amino acid ingestion on cat and rat lateral hypothalamic neuronal activity. Pre-absorptive signals, conveyed by the vagus nerve and originating from the small intestine while the amino acids are about to be absorbed, have been previously identified electrophysiologically in our laboratory [3]. They were found to selectively modulate the neuronal activity of some lateral hypothalamic (LHA) cells [4]. The effects of both pre-absorptive and post-absorptive signals associated with amino acid ingestion were analyzed with regard to their degree of convergence at the lateral hypothalamic level. The pre-absorptive signals consisted of either chemospecific information arising from intestinal chemoreceptors and related to the chemical nature of the ingested nutrient, or mechanical information arising from intestinal mechanoreceptors and related to distension. The post-absorptive signal consisted of a microiontophoretic application of amino acids known to selectively affect the neuronal activity of LHA amino acid-sensitive cells [15]. The convergence on these cells of the intestinal afferent amino acid-sensitive units was postulated in this study.

Amino Acids↗

Age-dependent alterations of intestinal absorption. II. A clinical study using a modified D-xylose absorption test.

With advancing age functional and morphological changes take place within the intestinal tract. The age-dependency of intestinal absorption has been studied using a D-xylose absorption test. The absorption rate (constant k12), the time of appearance of the relative maximum of the D-xylose concentrations in the blood serum after oral administration (time tm), and the extent of the relative absorption in the small intestine as a function of time RA(t) were determined by graphic-mathematical evaluation of the blood level kinetics of D-xylose after oral and intravenous administration. Within the age range of 3 to 96 yr, it has been proven that a slowing down of intestinal absorption occurs with advancing age. By prolonging the period during which the food components are absorbed in the small intestine (shift of tm towards the absorption processes take place at a slower rate. Absorption disorders in old age are due to morbidity and not, sui generis, to involution.

Adolescent↗

Intestinal oxalate absorption is higher in idiopathic calcium oxalate stone formers than in healthy controls: measurements with the [(13)C2]oxalate absorption test.

PURPOSE: We assessed the importance of oxalate hyperabsorption for idiopathic calcium oxalate urolithiasis, oxalate absorption in healthy volunteers and recurrent calcium oxalate stone formers was compared. MATERIALS AND METHODS: The [(13)C2]oxalate absorption test, a standardized, radioactivity-free test, was performed. On 2 days 24-hour urine was collected and an identical standard diet containing 800 mg Ca daily was maintained. On the morning of day 2 a capsule containing 0.37 mmol sodium [(13)C2]oxalate was ingested. A total of 120 healthy volunteers (60 women and 60 men) and 120 patients (30 women and 90 men) with idiopathic CaOx urolithiasis (60% or greater CaOx) were tested. RESULTS: Mean intestinal oxalate absorption in the volunteers was 8.0 +/- 4.4%, and in the patients was 10.2 +/- 5.2% (p <0.001). There was no significant difference in mean absorption values between men and women within both groups. A high overlap between the absorption values of volunteers and patients was found. Only in the patient group did absorption values greater than 20% occur. Oxalate absorption correlated with oxalate excretion in the patients, r = 0.529 (p <0.01) and in the volunteers, r = 0.307 (p <0.01). CONCLUSIONS: In high oxalate absorbers dietary oxalate has a significant role in oxalate excretion and, therefore, increases the risk of calcium oxalate stone formation.

Adolescent↗

Absorption of angiotensin II antagonists in Ussing chambers, Caco-2, perfused jejunum loop and in vivo: importance of drug ionisation in the in vitro prediction of in vivo absorption.

The aims of this study were (i) to compare the absorption of three closely related inhibitors of angiotensin II, RU60018, RU60079 and HR720, in various in vitro and in vivo models, and (ii) to explain the differences in the results and to assess the importance of drug ionisation to predict absorption. Drug absorption was investigated in Ussing chambers, Caco-2 cell monolayers, perfused rat jejunum loops and in vivo after oral, intraduodenal or intravenous administration. In Ussing chambers, the analogues showed the same site-related absorption profile and a common mechanism involving the paracellular pathway. At pH 7.4 in Ussing chambers, perfused jejunum loop or Caco-2 transport studies, the three compounds exhibited low and comparable permeability values suggesting that a similar level of oral absorption may be expected for all three compounds. However, after oral or intraduodenal administration, only HR720 was significantly absorbed. The in vivo results can be explained by the ionic distribution profile which indicated that only HR720 possessed a significant amount of uncharged species at pH values close to that found in the upper part of intestinal tract. Hence, it is expected that in this part of the intestine, only HR720 absorption is favoured. This is supported by Caco-2 transport studies performed when the pH of the apical medium was lowered from 7.4 to 6.0, in which a dramatic increase in permeability was observed for HR720 compared to those of the other analogues. This study highlights the usefulness of different absorption models for drug screening and demonstrates that ionisation profiles must be carefully considered to avoid rejection of promising compounds.

Administration, Oral↗

Improved nasal absorption of drugs using poly-L-arginine: effects of concentration and molecular weight of poly-L-arginine on the nasal absorption of fluorescein isothiocyanate-dextran in rats.

The effects of the concentration and molecular weight of poly-L-arginine (poly-L-Arg) on the in vivo nasal absorption of fluorescein isothiocyanate-labeled dextran (MW, 4 kDa, FD-4) in rats were studied. When poly-L-Arg with a range of different molecular weights (MW, 8.9, 45.5 and 92.0 kDa) was applied intranasally at various concentrations, the bioavailability (F(0-9 h)) of FD-4 increased with the increasing concentration of poly-L-Arg. The enhanced absorption was also dependent on the molar concentration, in that the poly-L-Arg with a higher molecular weight increased F(0-9 h) at a lower molar concentration. In addition, for each applied concentration, the poly-L-Arg exhibited a molecular weight-dependence as far as the enhancement of FD-4 absorption was concerned. On the other hand, the maximum absorption rate (MAR) of FD-4, calculated by means of a deconvolution method, tended to reach a maximum plateau level at a lower applied concentration for the poly-L-Arg with the highest molecular weight, but this plateau level was almost the same for poly-L-Arg with molecular weights of 45.5 and 92.0 kDa. Moreover, the simulated absorption profiles of FD-4 indicate that the degree of enhancement (the level of MAR and the subsequent reduction in the absorption rate) was dependent on the molecular weight of poly-L-Arg, while the effect of poly-L-Arg was maintained for a longer period, depending on the applied concentration, although the MAR was relatively similar. These results indicate that the molecular weight of poly-L-Arg appears to affect both the enhancing efficiency (absorption rate) and the time-frame of this enhancing effect, whereas the concentrations of each poly-L-Arg system applied only have an effect on the time-frame. These effects may also be associated with the charge density of a poly-L-Arg molecule.

Absorption↗

Solute absorption from the airways of the isolated rat lung. V. Charge effects on the absorption of copolymers of N(2-hydroxyethyl)-DL-aspartamide with DL-aspartic acid or dimethylaminopropyl-DL-aspartamide.

PURPOSE: To determine the effects of ionized substituents upon the pulmonary absorption of 6-8 kDa synthetic, hydrophilic polypeptides. METHODS: Fluorophore-labeled poly (hydroxyethylaspartamide), F-PHEA (neutral at pH 7.4) and its copolymer derivatives poly (hydroxyethylaspartamide-co-dimethylaminopropylaspartamide), F-P(HEA-DMAPA) (positive at pH 7.4) and poly (hydroxyethylaspartamide-co-aspartic acid), F-P(HEA-AA) (negative at pH 7.4) were synthesized and administered in different concentrations to the airways of the isolated rat lung preparation. The time and molecular weight dependencies of polypeptide absorption into perfusate were determined at intervals by gel permeation chromatography. RESULTS: For all polypeptides, molecular weights in perfusate were about 1 kDa less than those which were administered, due to preferential absorption of smaller molecules. The absorption, up to 70% of the administered dose over 3 h, of the anionic F-P(HEA-AA), was significantly faster than that of the neutral F-PHEA or the polycationic F-P(HEA-DMAPA). The latter derivative produced greatest edema in the lung. Absorption showed both active [dose-dependent kinetics] and passive [diffusive] components for all three polymers. CONCLUSIONS: Pulmonary absorption of similarly sized macromolecular PHEA derivatives, either neutral, positively or negatively charged, occured via carrier-mediated and diffusive mechanisms. The highest rate of absorption was observed with the polyanionic derivative.

Absorption↗

Evidence for dissolution rate-limited absorption of COL-3, a matrix metalloproteinase inhibitor, leading to the irregular absorption profile in rats after oral administration.

PURPOSE: This study was undertaken to elucidate the underlying mechanism of the irregular absorption profiles of COL-3, a matrix metalloproteinase inhibitor, with a double- or plateau-peak concentration after a single oral dose administration of COL-3 suspension to rats. METHODS: The gastrointestinal absorption profiles of COL-3 in rats were assessed by comparing serum drug concentration curves after the following various modes of drug administration: oral and intraduodenal doses, oral doses of COL-3 in fine and coarse suspensions, intraduodenal dosing to the bile-duct intact and cannulated (BDC) rats, and oral doses with and without food. In addition, the biliary excretion of COL-3 in the BDC rats was examined. RESULTS: Neither variable gastric emptying nor enterohepatic recycling was the source of the irregular gastrointestinal absorption of COL-3 in rats. Reduction in particle size, presence of food and endogenous bile emerged as the determinants of the oral absorption of COL-3 by enhancing the dissolution of the solid drug in the gastrointestinal fluids. Flip-flop of the absorption and elimination rate constants was noted only for COL-3 after intraduodenal administration of the coarse suspension to the BDC rats with the bile flow diverged out of the body. CONCLUSIONS: Variability in dissolution rate-limited absorption was the main cause of the irregular absorption of COL-3 after oral administration of its solid dosage form.

Administration, Oral↗

Hydrothermal treatment and malting of barley improved zinc absorption but not calcium absorption in humans.

OBJECTIVE: To study whether hydrothermal treatment or malting of barley (cv. Blenheim) improves zinc and calcium absorption in humans. DESIGN: : Two groups of 10 and 12 healthy subjects, respectively, were in a period of 2 months in a fasting state, served two single meals each containing porridge or breakfast cereals prepared from processed or unprocessed (control) barley (60 g). The meals included 200 g of milk, extrinsically labelled with (65)Zn and (47)Ca. Whole-body retention of both minerals was measured. SETTING: The study was carried out at the Department of Radiation Physics, Sahlgrenska University Hospital, Göteborg. SUBJECTS: The subjects were recruited among students at the Göteborg University. None dropped out. INTERVENTIONS: The activities of (65)Zn and (47)Ca were measured by whole-body counting four to five times over a 4-week period after each meal. RESULTS: Zinc absorption from hydrothermally treated barley porridge, containing 28 mg P as inositol tri- to hexaphosphates (InsP(3)-InsP(6)), was significantly higher (P<0.001) than from control porridge containing 111 mg P as InsP(3)-InsP(6), 25.2+/-6.9 vs 11.0+/-2.5% (n=12). Calcium absorption did not differ (P>0.05), 21.1+/-6.8 vs 19.5+/-4.7% (n=12). Zinc absorption from breakfast cereals of malted barley with phytase activity and containing 70 mg P as InsP(3)-InsP(6,) was significantly higher (P<0.05) than from flakes of barley, containing 108 mg P as InsP(3)- InsP(6) and no phytase activity, 22.9+/-5.8 vs 14.8+/-4.6% (n=10). The calcium absorption was 21.3+/-6.5 vs 18.5+/-4.3% (n=10) and did not differ significantly (P>0.05). CONCLUSION: Improvements of zinc absorption in breakfast meals can be achieved by optimised hydrothermal treatment or malting of barley. Calcium absorption was not influenced in the meals in this study. SPONSORSHIP: Supported by Semper AB, Sweden, Oy Lahden Polttimo, Finland, the SL-Foundation, Sweden, Swedish National Board for Industrial and Technical Development (NUTEK), the Nordic Industrial Foundation, Swedish Council for Forestry and Agricultural Research (SJFR, project no 50.0306/97).

Adolescent↗