Effect of offering dairy cows diets differing in phosphorus concentration over four successive lactations: 1. Food intake, milk production, tissue changes and blood metabolites

The loss of phosphates from dairy farms contributes to the eutrophication of waterways. Whilst reducing the phosphorus (P) content of dairy cow diets has the potential to help reduce phosphate losses, diets containing inadequate dietary P may have a negative effect on cow health and performance. To address this issue, 100 winter-calving Holstein-Friesian dairy cows were offered diets containing either 'high' or 'low' levels of dietary P. The experiment was conducted over a 4-year period, with 80 primiparous cows commencing the study in year 1, while a further 20 primiparous cows commenced the study in year 2. Rations offered during the winter comprised grass silage, maize silage (70 : 30 dry matter (DM) basis, approximately) and concentrates (10.0 to 12.0 kg/cow per day). During the summer periods in years 1 and 2, half of the cows grazed both day and night, while the remaining cows grazed by day, and were housed by night and offered grass silage. During years 3 and 4, all cows grazed both day and night during the summer period. Concentrate feed levels during the summer periods were 3.0 to 4.0 kg/cow per day. Different dietary P levels were achieved by offering concentrates containing either high or low P levels during the winter period (approximately 7.0 or 4.4 g P/kg DM respectively), and during the summer period (approximately 6.8 or 3.6 g P/kg DM, respectively). Total ration P levels averaged 4.9 and 3.6 g P/kg DM for the 'high' and 'low' P winter diets respectively, and 4.2 and 3.6 g P/kg DM for the 'high' and 'low' P summer diets respectively. A total of 95, 70, 50 and 22 cows completed each of lactations 1 to 4 respectively. Dietary P level had no significant effect on food intake, milk output or milk composition (P > 0.05). Plasma P concentrations were significantly lower with cows offered the 'low' P diet in each of lactations 1 to 4 (P < 0.05). In each of lactations 3 and 4, cows offered the 'low' P diet tended to have lower condition scores and live weights than those offered the 'high' P diet. The results of this experiment highlight that the P content of dairy cow diets can be substantially reduced with no detrimental effect on dairy cow performance.     

Spectroscopic properties and chromophore conformations of the photomorphogenic receptor: phytochrome.

Fluorescence lifetimes of 'large (mol. wt. 120,000) and 'small' (mol. wt. 60,000) phytochromes isolated from oat and rye seedlings grown in the dark have been measured at 199 K and 298 K. Phytochrome model compounds have also been studied by phase modulation fluorometrically at 77 K for comparison with lifetime data for phytochrome. It was found that the fluorescence lifetime of 'large' phytochrome was significantly shorter than that of 'small' phytochrome and its chromophore models. The phytochrome chromophore of Pr form has been analyzed by fluorescence polarization, CD, and molecular orbital methods. The fluorescence excitation polarization of 'small' phytochrome and the chromophore model in buffer/glycerol mixture (3 : 1, v/v) at 77 K is very hight (0.4) at the main absorption band and is negative (--0.1) and close to 0 in the near ultraviolet band, respectively. Analyses of the spectroscopic data suggest that the chromophore conformation of Pr and Pfr forms of phytochrome are essentially identical. The induced ellipticity of 'large' rye phytochrome in the blue and near ultraviolet regions was found to be significantly higher than that of 'small' phytochrome, indicating that the binding interaction between the phytochrome chromophore and apoprotein is much tighter in the former than in the latter. In addition, the excitation energy transfer does occur from Trp residue(s) to the chromophore in 'large' phytochrome but not in 'small' Pr. This illustrates one feature of the role played by the large molecular weight apoprotein in the binding site interactions and primary photoprocesses of Pr. Finally, a plausible model for the primary photoprocesses and the mechanism of phytochrome interactions triggered by the Pr leads to Pfr phototransformation have been proposed on the basis of the above results.     

The dependence of the incorporation of methamphetamine into rat hair on dose,frequency of administration and hair pigmentation

In this paper, the incorporation of methamphetamine (MA) into rat hair was studied. The main purpose of this study was to investigate whether MA can be detected or positive hair results can be obtained in hair of rats administered a single dose of MA. The relationship between dose and frequency of administration and the concentrations of MA and its metabolite, amphetamine (AP), in rat hair were evaluated and the MA and AP concentrations in white and pigmented hair were compared. MA was administered to rats as follows: low dose (0.5 mg/kg/day), medium dose (2 mg/kg/day) and high dose (10 mg/kg/day). The frequency of administration was one time per day for 1, 2, 3, 4, 5, 15 and 30 days. Hair and urine samples were collected from rats and analyzed by gas chromatography/mass spectrometry (GC/MS). MA could be identified in pigmented rat hair when MA was administered for 4 or more days at low daily dose and on day 1 following administration of medium and high daily doses. Positive results for MA were obtained from pigmented rat hair when MA was administered for 30 days at low daily dose, for 4 or more days at medium daily dose, or for 2 or more days at high daily dose. The concentrations of MA and AP found in rat hair were proportional to the dose and frequency of administration. The concentrations of MA and AP in pigmented rat hair were 2–10 times higher than those in white rat hair. The results of this study on the incorporation of MA into rat hair can serve as a model to better understand the incorporation of MA into human hair even though there are differences between animal models and human hair.     

Analysis of the negative inotropic effect of acetylcholine on frog atrial fibres

Voltage-clamp experiments have been performed on frog atrial preparations in order to study the mechanism of the inotropic effect of acetylcholine (ACh) at various concentrations. The amplitude of the slow inward current (Is) is reduced even at low ACh concentrations; such low concentrations have little or no effect on potassium permeability. Dose-effect relationships for Is inhibition (Is/Is max) by ACh show a half amplitude dose (K0.5 around 8 X 10(-8) M ACh. The reduction of Is is attributed largely to a decrease of the maximal conductance of the slow channel (gs). Steady-state activation and inactivation parameters are not affected by ACh. Experiments in a Na-free solution (Na replaced by Li ions) or in a Ca-free solution (with EGTA) indicate that the "slow sodium current" is more sensitive to ACh than the "slow Ca current", although these two currents both seem to flow through the slow channel. The decrease of the phasic component of contraction observed in the presence of ACh is very well correlated with the decrease of Is (K0.5 = 8 X 10(-8) M ACh), while the increase of the tonic tension may be related to the outward potassium current induced by high concentrations of ACh. The significant difference between the half amplitude dose (K0.5) observed in the dose effect curves with ACh for Is inhibition (K0.5 = 8 X 10(-8) M) and for ACh-induced extra-current (K0.5 - 10(-6) M) may indicate the presence of two muscarinic receptors.     

Kinetics of indole acetic acid-induced growth in hypocotyl sections of Vigna mdiata

The kinetics of auxin-induced elongation of segments from Vigna radiata (L.) Wilczek hypocotyls have been investigated using auxanometer measurements. Doseresponse curves were established for several well-defined parameters of the growth response. The experimental data revealed that different kinetic parameters were affected differently by increasing auxin concentrations. The dose-response curves are slightly sigmoid for fresh weight, nearly bell-shaped for total elongation or maximal elongation rate, and nearly linear for maximal growth acceleration. The effects of auxin concentration on regulation of growth orientation are discussed.
A biphasic response is observed mainly with segments taken from the middle of the hypocotyl ('C') which exhibit maximal growth rates. Segments from other levels, with lesser growth potentials, exhibit a very weak response. The two successive phases may then require different maturation states. Kinetics of the acceleration curves are very similar all along the hypocotyl and remain very homogeneous with increasing IAA concentrations.     

Arrhenius curves of hydrogen transfers: tunnel effects, isotope effects and effects of pre-equilibria

In this paper, the Arrhenius curves of selected hydrogen-transfer reactions for which kinetic data are available in a large temperature range are reviewed. The curves are discussed in terms of the one-dimensional Bell-Limbach tunnelling model. The main parameters of this model are the barrier heights of the isotopic reactions, barrier width of the H-reaction, tunnelling masses, pre-exponential factor and minimum energy for tunnelling to occur. The model allows one to compare different reactions in a simple way and prepare the kinetic data for more-dimensional treatments. The first type of reactions is concerned with reactions where the geometries of the reacting molecules are well established and the kinetic data of the isotopic reactions are available in a large temperature range. Here, it is possible to study the relation between kinetic isotope effects (KIEs) and chemical structure. Examples are the tautomerism of porphyrin, the porphyrin anion and related compounds exhibiting intramolecular hydrogen bonds of medium strength. We observe pre-exponential factors of the order of kT/h congruent with 10(13) s-1 corresponding to vanishing activation entropies in terms of transition state theory. This result is important for the second type of reactions discussed in this paper, referring mostly to liquid solutions. Here, the reacting molecular configurations may be involved in equilibria with non- or less-reactive forms. Several cases are discussed, where the less-reactive forms dominate at low or at high temperature, leading to unusual Arrhenius curves. These cases include examples from small molecule solution chemistry like the base-catalysed intramolecular H-transfer in diaryltriazene, 2-(2'-hydroxyphenyl)-benzoxazole, 2-hydroxy-phenoxyl radicals, as well as in the case of an enzymatic system, thermophilic alcohol dehydrogenase. In the latter case, temperature-dependent KIEs are interpreted in terms of a transition between two regimes with different temperature-independent KIEs.     

Hydrogenosomal succinate thiokinase in Tritrichomonas foetus and Trichomonas vaginalis

Succinate thiokinase displays a diversity of nucleotide specificity and molecular size throughout Nature. Eukaryotes and Gram-positive bacteria possess distinct 'small' (dimeric) thiokinase enzymes which are specific for adenine (ADP) or guanine (GDP) nucleotides, whereas Gram-negative bacteria contain a single 'large' (tetrameric) enzyme which utilizes both nucleotides. Succinate thiokinase activities, both ADP- and GDP-dependent, were shown to be hydrogenosomal in Tritrichomonas foetus and Trichomonas vaginalis. Surprisingly, the 'small' enzyme was found in T. foetus whereas T. vaginalis contained a 'large' enzyme.     

Antidepressant action of St. John's Wort, Hypericum perforatum: a test of the circadian hypotheses.

Extracts of the plant St. John's Wort, Hypericum perforatum, are effective for treatment of mild depression. It has been hypothesised that H. perforatum may be acting on the circadian timing system either directly or via a photosensitising action to produce changes in mood. Two experiments were conducted to test these hypotheses. Under constant dark (Experiment 1) or low constant light (Experiment 2) rats were permitted to free-run. Rats were then treated with a 'high' (616mg/kg/day; n = 8 per experiment) or 'low' (308 mg/kg/day; n = 8 per experiment) dose of H. perforatum or a control solution (n = 8 per experiment) in drinking water, and circadian locomotor rhythms examined for alterations of period. A minor shortening of mean period (2.4 min) was observed on cessation of H. perforatum treatment in the low-dose group in Experiment 2, and was considered to be a measurement artifact and of no clinical value. Otherwise, no significant differences in free-running period between treatment groups were observed in either study, indicating that H. perforatum has no direct or photosensitising effect on the mammalian circadian system. These results suggest that the antidepressant action of H. perforatum is not mediated by a circadian mechanism.     

Analytical theory for the fluence,planar fluence,energy fluence,planar energy fluence and absorbed dose of primary particles and their fragments in broad therapeutic light ion beams

The purpose of the present work is to develop analytical expressions for the depth variation of the fluence, planar fluence, the energy fluence, planar energy fluence, the mean energy and absorbed dose of primary ions and their associated fragments in tissue-like media with ranges of clinical interest. The analytical expressions of the primary ions and associated fragments take into account nuclear interactions, energy losses, range straggling and multiple scattering. The analytical models of the radiation field quantities were compared with the results of the modified Monte Carlo (MC) code SHIELD-HIT₊. The results show that the shape of the depth absorbed dose distribution of the primary particles is characterized by an increasingly steep exponential fluence decrease with depth as the charge and atomic weight increase. This is accompanied by a compensating increased energy loss towards the Bragg peak as the charge of the ion increases. These largely compensating mechanisms are the main reason that the depth absorbed dose curve of all light ions is surprisingly similar. In addition, a rather uniform dose in the plateau region is obtained since the increasing fragment production almost precisely compensates the loss of primaries. The dominating light fragments such as protons and alpha particles are characterized by longer ranges than the primaries and their depth dose curves to some extent coincide well with the depth fluence curves due to a rather slow variation of mean stopping powers. In contrast, the heavier fragments are characterized by the build up of a slowing down spectrum similar to that of the primaries but with initially slightly shorter or longer ranges depending on their mass to atomic number ratio. The presented analytical theory for the light ion penetration in matter agree quite well with the MC and experimental data and may be very useful for fast analytical calculations of quantities like mean energy, fluence, energy fluence, absorbed dose, and LET.     

The immunogenic and serological properties of the O-specific polysaccharide (L-hapten) in Shigella]

O-specific polysaccharide (L-hapten) was isolated earlier (Zh. mikrobiol. epidemiol. immunobiol., 1989, No. 11, pp. 8-11). In this paper L-hapten was shown to be unable, even at high concentrations (up to 2,000 micrograms/ml), to sensitize sheep red blood cells for passive hemagglutination by O-antibodies. At the same time classical LPS and heat-activated LPS were active at concentrations ot 32 and 8 micrograms/ml respectively. The O-antibody-neutralizing activity of L-hapten was lower than that of LPS 10(3)-10(4) times in the passive hemagglutination test and 25-50 times in competitive ELISA. The immunogenicity of isolated L-hapten was very weak: primary response in mice to the i.v. injection of 1-10 micrograms of L-hapten was similar to the effect produced by 10(-3)-10(-4) micrograms of LPS. No protective activity of L-hapten was noted in mice when the challenge dose of virulent shigellae was 16 LD50 or more, and only a weak protective effect was observed with a low challenge dose (8 LD50). The molecular basis of low serological and biological activity of L-hapten is discussed. The most probable explanation of the results obtained in this study is that L-hapten contains some nonspecific carbohydrates, inserted in or complexed with the O-side chain. Despite its low immunogenicity, L-hapten can be an important component of effective bacterial vaccines provided it is included into a suitable delivery system as is the case with Shigella ribosomal vaccine.     

The effect of misonidazole as a hypoxic radiosensitizer at low dose

Using an automated low dose survival assay, the radiosensitizing effectiveness of misonidazole at low radiation dose (0-6 Gy) was measured in cultured mammalian cells. Also measured was its effectiveness at high doses of radiation (0-35 Gy) using the conventional survival assay. In both cases, several concentrations of the drug from 0 to 5 mM were used. The data at low doses were analyzed by a two-parameter mathematical equation with linear and quadratic dose terms, S = e-alpha D-beta D2, which proved to be a good fit to the experimental data at all misonidazole concentrations. It is shown that whereas the coefficient of the quadratic dose term, beta, increases significantly with increasing misonidazole concentration, the drug does not significantly affect the coefficient of the linear term, alpha. The enhancement ratio (ER) of misonidazole is shown to be decreased at lower doses. The clinical implications of this result are discussed.     

The effect of dose and time on the induction of genetic alterations insaccharomyces cerevisiae by aminoacridines in the presence and absence of visible light irradiation in comparison with the dose-effect-curves of mutagens with other type of action

Summary Aminoacridines induce frameshift mutations and are photodynamically active, depending on whether visible light is absent or present. Therefore, a test system which allows to compare quantitatively the genetic effects of aminoacridines irradiated or unirradiated by visible light ought to be susceptible to the different DNA alterations which can be induced by these substances. For this reason in most experiments mitotic gene conversion and only in some selected experiments reverse mutation was chosen as the indicator of genetic activity. In contrast to mutation systems mitotic gene conversion has never shown a response specific to only some types of mutagens. The three aminoacridine derivatives used-acridine orange (AO), proflavine (PF), and acridine yellow (AY)—were successful in the induction of convertants at two different loci. No locus-specificity could be observed. The time-dependent induction of convertants proceeds quickly but soon reaches—especially after treatment without light—a saturation point. The dose/effect-curve after treatment in the dark has a slope increasing with increasing concentration. Irradiation with visible light results in a dose/effect-curve consisting of three parts. At first the increase of convertants is nearly linear extending one (AY) to three (AO) orders of magnitude. After that a saturation effect begins at the point at which an effectiveness of the acridines in the dark is apparent. At high concentrations an induction of convertants can again be observed which is nearly the same as that after treatment in the dark. To determine whether the dose/effect-curves obtained for gene conversion refer to similar curves for gene mutations after treatment with AO at the same locus not only gene conversions but also reverse mutations were scored for. AO-treatment in the dark is ineffective in inducing reverse mutations. Irradiation with visible light results in a dose/effect-curve beeing parallel only in its first part to the dose/effect-curve obtained for gene conversion, while in its second part a mutation frequency decline can be observed. Comparing the dose/effect-curves of AO resulting from the induction of gene conversion and gene mutation, and taking into account that no mutants can be induced by AO-treatment in the dark, the increase in convertants at high acridine-concentrations can be explained as an addition of light-dependent and light-independent effects. That means, in mutation systems at low concentrations of aminoacridines irradiation with visible light should cause transitions, transversions and microlesions, at intermediate concentrations frameshift lesions should begin to appear, and at very high concentrations nearly exclusively frameshift lesions should occur. The dose/effect-curves of aminoacridines compared with those of other mutagens are very complex. The dose/effect-curves of the mutagens of other type of action tested are linear in a double logarithmic scale, and parallel for induced gene conversion and induced gene mutation. These results indicate that the gene conversion ability of a given compound depends on its mutagenic property. That means, many mutagens may exert specific genetic effects not directly but mainly in indirect ways by leading to DNA damage, a situation for repair synthesis resulting as well in mutations as recombinations.     

Dormancy breaking and frost resistance induction in apple embryos as related to changes in reserve and polar lipids

Detailed dose-response curves were obtained for cytokinin-induced retention of chlorophyll in detached leaves of barley, Hordeum vulgare L. cv. Møyar, over a wide range, 10⁻¹⁰– 2.5 × 10⁻⁴ M , of external concentrations of benzyladeninc or kinetin. The over-all shape of the curves was log-linear and the response was subsensitive. Benzyladenine was somewhat more effective than kinetin. In the curves there were pronounced local minima and maxima both at low, intermediate (10⁻⁷– 10⁻⁵ M ) and high concentrations. At low and high concentrations these deviations from the best smooth curves were reproducible, statistically significant and occurred at the same concentrations for benzyladenine and kinetin. In the intermediate range the deviations from the log-linear line were marked and statistically highly significant in some but not all experiments. The cause of the complexities in the dose response curve is not known, but multiphasic uptake of the cytokinins may be involved.     

Detection of hormesis effect in longevity: simulation approach for heterogeneous population.

Manifestation of hormesis in longevity was modelled by modification of the mortality rate during and after the period of a stress factor action. In heterogeneous population this can lead to observation of unchanged mortality during action of the stress and decrease in mortality after stress period. Stochastic simulations were made to investigate the possibility of detecting the hormesis effect on the basis of the stress-control longitudinal data. The goal of the stochastic simulation was to investigate the role in the hormesis detection of control and stressed group size, of population heterogeneity variance value, of stress and hormesis attributable risks as well as the role of a prior information about the survival in the control group. It was demonstrated that if the attributable risks for stress and hormesis effects are approximately equal, then in both 'high' and 'low' heterogeneous populations the hormesis phenomenon is detected with probability higher than 75% even in relatively 'small' groups of 50 subjects. In case of 'weak' effect the hormesis phenomenon is not detected in a 'highly heterogeneous' population even in a group composed of 1000 subjects. In a 'low heterogeneous' population the hormesis phenomenon is detected with probability higher than 70% when the group size is not less than 200 subjects. Information about the survival in control group did not play a critical role in all experiments and exact survival curve may be replaced by the traditional Kaplan-Meier estimate.     

Changes in prolactin in peripheral plasma during lactation in the brushtail possum Trichosurus vulpecula

A heterologous double-antibody radioimmunoassay has been validated for prolactin in plasma and pituitary preparations of T. vulpecula. Serial dilutions of crude pituitary homogenates and plasmas from several marsupials and purified prolactin from the tammar, Macropus eugenii, showed parallel dose response curves. In both male and female possums plasma prolactin concentrations increased in response to a single intravenous injection of thyrotrophin releasing hormone. Plasma prolactin concentrations were measured in six lactating females (June-November) and in four non-lactating females (July-October). In the following year prolactin levels were also measured in 11 possums with young less than 50 days old and in 24 possums with young aged between 100 and 145 days. In early lactation prolactin concentrations were low (less than 8 ng/ml) but increased to high levels (greater than 30 ng/ml) by 120 days and remained high until about 160 days of lactation. Thereafter concentrations declined although the young continued to take milk from the mother for a further 30-50 days. The changes in plasma prolactin concentrations throughout lactation are very similar to those described for the tammar, and this unusual pattern appears to be common to marsupials. Non-lactating possums showed no consistent changes in plasma prolactin concentrations between July and October.     

Molecular size diversity of citrate synthases from Pseudomonas species

Two forms of citrate synthase (EC 4.1.3.7) have been found in several species of Pseudomonas, a 'large' form (Mr congruent to 250,000) which is generally inhibited by NADH and reactivated by AMP, and a 'small' form (Mr congruent to 100,000) which is insensitive to these nucleotide effectors. Other species of Pseudomonas were found to contain either the 'large' or the 'small' form. Gel filtration and ion-exchange with the technique of fast protein liquid chromatography were used to resolve the enzymes. Where both citrate synthases were present, there did not appear to be an equilibrium between the two forms. The results reveal a new and complex diversity of citrate synthase within the genus Pseudomonas.     

Multiple,concentration-dependent effects of sucrose,auxins and cytokinins in explant cultures of kale and tobacco

We describe complex multiple concentration dependencies for the response of isolated pith tissues to plant biologically active substances. Kale and tobacco stem pith explants were cultured on agar media containing combinations of sucrose, cytokinin [kinetin or benzyladenine (BA)] and auxin [indole-3-acetic acid (IAA) or naphthaleneacetic acid (NAA)]. Absorption of these components by explants and their effects on explant mass, contents of soluble proteins, starch and sugars, and activity of ADP-glucose pyrophosphorylase (AGPase) were studied in relation to their concentration. Up to ten pronounced statistically significant maxima (peaks or waves) were repeatedly detected in the dose–response curves over a concentration range of several logarithmic orders. Slight maxima were observed in the corresponding absorption curves. Pronounced maxima of sucrose absorption were induced by IAA and BA, and those of NAA absorption were induced by sucrose. Both types of multiple maxima (in dose–response and absorption curves) may be due to changes in concentration of intracellular solutes (sugars, auxins and cytokinins), thereby affecting metabolic processes that act as sinks for external solutes and elicit feedback appearance of maxima in absorption curves. Good correspondence between external concentrations at which maxima of different compared curves occur in addition to statistical significance of individual maxima and repeatability of experimental results supports the conclusion that the multiple maxima exhibited are genuine. We consider it possibile that the multiple maxima are associated with endopolyploidy or mixoploidy and/or epigenomic diversity of pith cells that show different sensitivities to biologically active solutes.     

"Infant mortality in Armenia 1992-2003": a comment.

The purpose of this comment is to counsel caution in some of the conclusions drawn in an otherwise fine article recently published in Economics and Human Biology on infant mortality in Armenia by Hakobyan, Mkrtchyan and Yepiskoposyan. These relate first, to the reliability of estimates and trends in infant mortality estimated from DHS data; second, to the interpretation of what the authors consider to be a 'low' infant mortality rate in former communist countries given their level of economic development; and third, to the role of the health care infrastructure in countries of the former Soviet Union in producing these 'low' infant mortality levels. This comment argues that trends in infant mortality in Armenia and other CIS countries, although probably declining, are perhaps less certain than the authors allow, that existing evidence does not suggest that they are uniformly low by global standards, or that the health care systems in CIS countries are uniformly effective in reducing infant deaths.     

Ionic selectivity,saturation, and block in gramicidin a channels

Summary A theory, recently developed by Sandblom, Eisenman and Neher (1977) for the conductance of single gramicidin A channels predicts three limiting behaviors of the relation between conductance and salt concentration. These are: (i) a saturating behavior resembling a simple adsorption isotherm at medium and high concentrations, (ii) a decrease in conductance at the highest obtainable concentrations and (iii) deviations from the isotherm at very low concentrations. Features i and ii have been described before. Experimental evidence for point iii is given here. The new feature points towards interactions among ions in the channel at ionic concentrations as low as 1–10mm.Particular emphasis is given to the behavior at very low salt concentrations and the experimental problems encountered in this situation. In addition, mutual blocking effects among monovalent ions in symmetrical salt mixtures are characterized and found to be in satisfactory agreement with theoretical expectations, based upon the single salt conductance data presented here and zero-current potentials in salt mixtures to be described in a subsequent paper.     

Fibrin assembly in human plasma and fibrinogen/albumin mixtures

Magnetic birefringence is used to monitor the kinetics of thrombin-catalyzed fibrin polymerization in model systems of increasing complexity (i.e., fibrinogen solutions, fibrinogen/albumin mixtures, and plasma anticoagulated with citrate) and in plasma containing free calcium which is the physiological condition. The introduction of albumin into fibrinogen solutions shortens the lag period and enhances fiber thickness. The polymerization progress curves are sigmoidal at zero or low albumin concentrations, but at physiological and higher concentrations, they become hyperbola-like from the end of the lag period. High albumin concentration has thus induced a change in the assembly kinetics. The progress curves from plasma in which the cascade is dormant are also hyperbola-like although they round off more quickly because of antithrombin activity. In plasma containing free calcium, thrombin is endogenously produced, and the progress curves are nearly linear; hence, the assembly kinetics are very different from those of the model systems. The curves are not influenced by calcium-dependent cross-linking involving factor XIIIa. The progress curves are also linear when polymerization is induced with Russell's viper venom, which by directly activating factor X circumvents earlier steps in the cascade. This implies that linear polymerization is caused by events posterior to factor X activation and are thus likely to be largely dependent on the functioning of the prothrombinase complex. Addition of thrombin to plasma containing free calcium reduces the lag period. At low exogenous thrombin levels, the polymerization rate is increased, and the progress curves remain linear. However, at higher levels, the curves become more complicated and, paradoxically, full polymerization takes longer.(ABSTRACT TRUNCATED AT 250 WORDS)