A Landau-de Gennes theory for hard colloidal rods: Defects and tactoids.

We construct a phenomenological Landau-de Gennes theory for hard colloidal rods by performing an order parameter expansion of the chemical-potential dependent grand potential. By fitting the coefficients to known results of Onsager theory, we are not only able to describe the isotropic-nematic phase transition as function of density, including the well-known density jump, but also the isotropic-nematic planar interface. The resulting theory is applied in calculations of the isotropic core size in a radial hedgehog defect, the density dependence of linear defects of hard rods in square confinement, and the formation of a nematic droplet in an isotropic background.

Implications of protein polymorphism on protein phase behaviour.

The phase behaviour of small globular proteins is often modeled by approximating them as spherical particles with fixed internal structure. However, changes in the local environment of a protein can lead to changes in its conformation rendering this approximation invalid. We present a simple two-state model in which protein conformation is not conserved and where the high-energy, non-native state is stabilised by pair-wise attractive interactions. The resulting phase behaviour is remarkably complex, non-universal and exhibits re-entrance. The model calculations show a demarcation between a regime where conformational transitioning is largely enslaved by phase separation and one where this is not the case. In the latter regime, which is characterised by a large free energy difference between the native and the non-native state, we deduce that the kinetics of the phase transition strongly depend on the average conformation of the proteins prior to their condensation. For condensation to occur in this regime within a dispersion of native proteins, nucleation of a cluster of proteins in the non-native state is required. We argue that our theory supports the distinction between common phase separation and the nucleated assembly of non-native supramolecular aggregates in protein dispersions.

Self-crowding induced phase separation in protein dispersions.

The coupling between protein conformation, molecular volume, and solution phase behaviour is studied theoretically for a two-state, coarse-grained protein model in which protein molecules can reversibly switch between a native and a non-native excited state. In the model, native and non-native conformers are represented by perfect spheres with different hard-core diameters. We presume the larger, non-native species to attract each other through some unspecified potential. We find that at low concentrations the native state is stabilised energetically and that at high concentrations the native state is again stabilised but this time by self-crowding, i.e., a lack of free volume. These two regimes are separated by two first-order transitions from a region where the non-native conformational state is prevalent, stabilised by attractive interactions between the proteins. The calculated phase diagram is very sensitive to even quite small differences in particle volumes and has unusual features, including the loss of a critical point if the size difference is sufficiently large.

Tactoids of plate-like particles: size, shape, and director field.

We studied, by means of polarized light microscopy, the shape and director field of nematic tactoids as a function of their size in dispersions of colloidal gibbsite platelets in polar and apolar solvents. Because of the homeotropic anchoring of the platelets to the interface, we found large tactoids to be spherical with a radial director field, whereas small tactoids turn out to have an oblate shape and a homogeneous director field, in accordance with theoretical predictions. The transition from a radial to a homogeneous director field seems to proceed via two different routes depending in our case on the solvent. In one route, the what presumably is a hedgehog point defect in the center of the tactoid transforms into a ring defect with a radius that presumably goes to infinity with decreasing drop size. In the other route, the hedgehog defect is displaced from the center to the edge of the tactoid, where it becomes virtual again going to infinity with decreasing drop size. Furthermore, quantitative analysis of the tactoid properties provides us with useful information on the ratio of the splay elastic constant and the anchoring strength and the ratio of the anchoring strength and the surface tension.

Osmotic compression of droplets of hard rods: a computer simulation study.

By means of computer simulations, we study how droplets of hard, rodlike particles optimize their shape and internal structure under the influence of the osmotic compression caused by the presence of spherical particles that act as depletion agents. At sufficiently high osmotic pressures, the rods that make up the drops spontaneously align to turn them into uniaxial nematic liquid-crystalline droplets. The nematic droplets or "tactoids" that are formed this way are not spherical but elongated, resulting from the competition between the anisotropic surface tension and the elastic deformation of the director field. In agreement with recent theoretical predictions, we find that sufficiently small tactoids have a uniform director field, while large ones are characterized by a bipolar director field. From the shape and director-field transformation of the droplets, we are able to estimate the surface anchoring strength and an average of the elastic constants of the hard-rod nematic.

Dynamic Surface Enrichment in Drying Thin-Film Binary Polymer Solutions.

Solution-cast, thin-film polymer composites find a wide range of applications, such as in the photoactive layer of organic solar cells. The performance of this layer crucially relies on its phase-separated morphology. Efficient charge-carrier extraction requires each of the components to preferentially wet one of the two electrodes. It is often presumed that the experimentally observed surface enrichment required for this is caused by specific interactions of the active ingredients with each surface. By applying a generalized diffusion model, we find the dynamics to also play an important role in determining which component accumulates at which surface. We show that for sufficiently fast evaporation the component with the smallest cooperative diffusivity accumulates at the free interface. Counterintuitively, depending on the interactions between the various components, this may be the smaller solute. Our comprehensive numerical and analytical study provides a tool to predict and control phase-separated morphologies in thin-film polymer composites.

Structuring of polymer solutions upon solvent evaporation.

The morphology of solution-cast, phase-separated polymers becomes finer with increasing solvent evaporation rate. We address this observation theoretically for a model polymer where demixing is induced by steady solvent evaporation. In contrast to what is the case for a classical, thermal quench involving immiscible blends, the spinodal instability initially develops slowly and the associated length scale is not time invariant but decreases with time as t(-1/2). After a time lag, phase separation accelerates. Time lag and characteristic length exhibit power-law behavior as a function of the evaporation rate with exponents of -2/3 and -1/6. Interestingly, at later stages the spinodal structure disappears completely while a second length scale develops. The associated structure coarsens but does not follow the usual Lifshitz-Slyozov-Wagner kinetics.

Diurnal prolactin surges and sexual differentiation of the rat hypothalamus.

Neonatal exposure to testicular androgens interferes with the ability in adulthood to release an ovulatory amount of LH, with the display of female sexual behavior, and also with control mechanisms of the activity of the corpora lutea. It is likely that effects on luteal activity involve, in part, effects on the control of PRL secretion. Therefore, serum concentrations of PRL were studied in ovarian graft-bearing male rats, castrated as neonates (NC-males) or as adults (AC-males), during a luteal phase induced and temporarily sustained by an ectopic pituitary graft. Before the removal of the pituitary graft, serum PRL and progesterone levels were high at all times when measured during the day in both AC-males and NC-males. Removal of the pituitary graft, 6-7 days after ovulation led to immediate cessation of luteal activity in AC-males but not in NC-males. In the latter animals, luteal activity was maintained for at least 6 days after pituitary graft removal by intermittently high levels of PRL, i.e. around 0400 and 1900 h. However, in AC-males, PRL levels were generally low after pituitary graft removal. It is concluded that the absence of neonatal exposure to testicular secretion contributes to the ability of the adult rat to release PRL in a diurnal surge-like manner. The inability to release PRL in this way may explain the failure of AC-males to maintain luteal activity.

Effects of the progesterone antagonist RU486 on ovarian activity in the rat.

Adult female rats were treated for 2 or 4 weeks with the progesterone antagonist RU486 to study its effect on the regulation of ovarian function. In rats with 5-day ovarian cycles, the vaginal cyclicity disappeared. Uninterrupted vaginal cornification emerged within 4 days after the start of treatment and cornification persisted for the whole period of treatment. It took more than 2 weeks after cessation of 2-4 weeks of treatment before 5-day vaginal cycles reappeared. Ovarian weights increased rapidly resulting from the accumulation of large numbers of corpora lutea. In addition, the ovaries developed occasional follicular cysts which could reach an extremely large size (2 mm or more). Analysis of serial histological sections of ovaries, combined with plasma concentrations of estradiol-17 beta and progesterone, indicated cyclic ovulation and corpus luteum formation together with persistence of functional activity of already existing and newly formed corpora lutea. RU486 seems to have the unique property of dissociating cessation of luteal activity and ovulation in rats. After treatment with RU486, pituitary enlargement and mammary gland alveolar development were observed. It is hypothesized that these effects result from unopposed estrogen action on PRL secretion. The effects of RU486 are reversible: 4 to 5 weeks after the end of treatment ovarian activity seems normal (as evidenced by reduction of ovarian weights and 5-day vaginal cycles) except for the presence of occasional large follicular cysts which may require longer periods for their regression.

Suckling stimulus, lactation, and suppression of ovulation in the rat.

Lactation pseudopregnancy in rats suckling a 5-pup litter lasted 22.0 +/- 0.4 days (mean +/- SEM; n = 11). By day 13 of lactation (day 1 of lactation = day of parturition), the continuation of lactation pseudopregnancy was dependent on the suckling stimulus, as litter removal on day 13 resulted consistently in ovulation on day 16. Measurements of various hormones before and after litter removal revealed high concentrations of progesterone and PRL during lactation and a rapid drop of both hormone concentrations after litter removal. Lactation pseudopregnancy in rats suckling a 10-pup litter lasted 26.1 +/- 0.9 days (n = 16). After litter removal on day 13 of lactation, the lactation pseudopregnancy continued for a further 7- to 11-day period, as evidenced by daily vaginal smears which remained mucified during that period. Measurements of hormone concentrations revealed continuously high concentrations of PRL before litter removal and a pattern of PRL secretion characterized by at least two diurnal peaks during the first days after litter removal. Progesterone concentrations decreased by 50% after litter removal, but the levels then remained constant and well above those found after the removal of 5-pup litters. It is argued that the different response to litter removal on day 13 of lactation between rats suckling 5 or 10 pups is due to the initiation of PRL peaks in rats with 10-pup litters: these PRL peaks are able to maintain luteal function for some period. It is further argued that the initiation of PRL peaks in rats with 10-pup litters is due to the high blood concentrations of progesterone at the time of litter removal compared to those of rats with a 5-pup litter.

Effect of thyroid status and paraventricular area lesions on the release of thyrotropin-releasing hormone and catecholamines into hypophysial portal blood.

TRH is a potent stimulator of pituitary TSH release, but its function in the physiological regulation of thyroid activity is still controversial. The purpose of the present study was to investigate TRH and catecholamine secretion into hypophysial portal blood of hypothyroid and hyperthyroid rats, and in rats bearing paraventricular area lesions. Male rats were made hypothyroid with methimazole (0.05% in drinking water) or hyperthyroid by daily injections with T4 (10 micrograms/100 g BW). Untreated male rats served as euthyroid controls. On day 8 of treatment they were anesthetized to collect peripheral and hypophysial stalk blood. In euthyroid, hypothyroid and hyperthyroid rats plasma T3 was 1.21 +/- 0.04, 0.60 +/- 0.04, and 7.54 +/- 0.33 nmol/liter, plasma T4 50 +/- 3, 16 +/- 2, and 609 +/- 74 nmol/liter, and plasma TSH 1.58 +/- 0.29, 8.79 +/- 1.30, and 0.44 +/- 0.03 ng RP-2/ml, respectively. Compared with controls, hyperthyroidism reduced hypothalamic TRH release (0.8 +/- 0.1 vs. 1.5 +/- 0.2 ng/h) but was without effect on catecholamine release. Hypothyroidism did not alter TRH release, but the release of dopamine increased 2-fold and that of noradrenaline decreased by 20%. Hypothalamic TRH content was not affected by the thyroid status, but dopamine content in the hypothalamus decreased by 25% in hypothyroid rats. Twelve days after placement of bilateral electrolytic lesions in the paraventricular area plasma thyroid hormones and TSH levels were lower than in control rats (T3: 0.82 +/- 0.05 vs. 1.49 +/- 0.07 nmol/liter; T4: 32 +/- 4 vs. 66 +/- 3 nmol/liter; TSH: 1.08 +/- 0.17 vs. 3.31 +/- 0.82 ng/ml). TRH release in stalk blood in rats with lesions was 15% of that of controls, whereas dopamine and adrenaline release had increased by 50% and 40%, respectively. These results suggest that part of the feedback action of thyroid hormones is exerted at the level of the hypothalamus. Furthermore, TRH seems an important drive for normal TSH secretion by the anterior pituitary gland, and thyroid hormones seem to affect the hypothalamic release of catecholamines.

In vivo hypothalamic release of thyrotropin-releasing hormone after electrical stimulation of the paraventricular area: comparison between push-pull perfusion technique and collection of hypophysial portal blood.

Unilateral electrical stimulation for 15 min of the paraventricular area of anesthetized rats induced a 2- to 3- fold increase in plasma TSH levels and caused an increased release of TRH into hypophysial stalk blood from 217 +/- 25 to 530 +/- 90 pg/15 min (n = 6). This experimental model was then used to determine the in vivo hypothalamic release of TRH by push-pull perfusion of either the mediobasal hypothalamus (MBH) or anterior pituitary (AP). Before stimulation, TRH release per 15 min was 4.2 +/- 0.7 pg from the MBH (n = 18) and 3.5 +/- 0.3 pg from the AP (n = 13). Unilateral electrical stimulation of the paraventricular area led to higher plasma TSH levels in 27 of 31 rats, and levels during stimulation increased from 0.89 +/- 0.04 to 1.86 +/- 0.10 ng/ml (n = 31). No significant increase in TRH in the perfusates was observed when push-pull perfusion was done in the MBH contralateral to the site of stimulation (n = 6). However, TRH release increased 2- to 3-fold during the perfusion of the MBH ipsilateral to the site of stimulation (15.4 +/- 4.3 pg/15 min; n = 13). In conclusion, push-pull perfusion of the MBH or AP can be used to estimate hypothalamic TRH release. However, the output of TRH by push-pull perfusion is low and varies considerably between individual rats. Thus, the practical value of push-pull perfusion for measurement of in vivo TRH release seems limited.

Hypothyroidism may account for reduced prolactin secretion in lactating rats bearing paraventricular area lesions.

Lesions in the paraventricular area (PVA) of lactating rats have been found to inhibit PRL release. We have examined whether this reduced PRL release is due to hypothyroidism resulting from destruction of the PVA. Rats were made hypothyroid by thyroidectomy on day 15 of pregnancy or by methimazole treatment from the day of parturition. Electrolytic lesions were placed bilaterally in the PVA on day 15 of pregnancy. The following variables were studied: weight gain of the pups, nursing behavior, thyroid status, and release of PRL. The treatments did not affect the time the mothers spent with the pups but reduced the daily weight gain of the pups. Rats with PVA lesions had reduced PRL and TSH levels during lactation compared with controls. Suckling-induced PRL release after 6 h of separation of mothers and pups was less in PVA-lesioned rats than in controls, but T4-treatment did overcome this blunted response in rats with lesions. Levels of T3 and T4 in PVA-lesioned rats were lower than those in controls. In rats made hypothyroid by thyroidectomy or treatment with methimazole, PRL levels were lower and TSH levels higher than those in euthyroid mothers on days 8, 15, and 22 of lactation. Suckling after 6 h of separation of pups and mothers raised PRL levels both in control and methimazole-treated rats, but in the latter animals the response was blunted. It is suggested that the reduced PRL release in lactating rats with PVA lesions could be due to hypothyroidism resulting from these lesions.

Control of prolactin release induced by suckling.

In the present study, the role of dopamine and TRH in suckling-induced PRL release was investigated. Bupropion, a dopamine reuptake blocker, increased hypophysial stalk dopamine levels and inhibited suckling-induced PRL release. A short period of suckling, thought to induce a transient decrease in hypothalamic dopamine release, led to higher PRL levels following an iv injection of TRH than in rats which had not nursed their young for a short period after 4- to 6-h separation. These results, in combination with previous data, suggest that a decrease in hypothalamic dopamine release is important for suckling-induced PRL release. Increased PRL release may be in part due to an augmented hypothalamic release of TRH. Since serotonergic mechanisms seem involved in TRH release, lactating rats were treated with drugs acting on serotonergic pathways. Parachlorophenylalanine and pizotifen did not alter suckling-induced PRL release. Methysergide, a serotonin receptor blocker, prevented this PRL release when administered ip but not when injected into the lateral brain ventricle. Since methysergide is converted peripherally into metabolite(s) with dopamine agonistic activity, its effect on suckling-induced PRL release may be due to this action, rather than to its action on serotonin receptors. Thus, these data do not indicate that serotonergic mechanisms are important for suckling-induced PRL release. Passive immunization against TRH inhibited suckling-induced PRL release, indicating that TRH is a hypophysiotropic mediator of this PRL release.

Effects of dihydrotestosterone and oestradiol on sexual differentiation in male rats.

Adult male rats which had been castrated at birth and treated with the non-aromatizable androgen dihydrotestosterone propionate (DHTP) showed incomplete copulatory behaviour. When tested with oestrous female rats during treatment with testosterone propionate (TP) they readily mounted these females and showed frequent penile intromissions but rarely ejaculated. In a long series of observations the proportion of ejaculating rats in tests of 30 min did not exceed 50%. Neonatally castrated rats treated with DHTP during infancy thus seemed to be capable of ejaculation in adulthood during treatment with TP, but the threshold for the occurrence of the ejaculatory reflex seemed to be higher than in normal male rats. By replacing treatment in adulthood with TP by a combined treatment with DHTP and oestradiol benzoate (OB), the frequency of ejaculation was not increased. It was concluded that the incomplete copulatory behaviour was not due to reduced efficiency of aromatization of androgen within the brain of these rats. The addition of OB to DHTP during the neonatal period of treatment enhanced the frequency of ejaculation in adulthood. The combined treatment of 0.1 mg DHTP on days 1, 3 and 5 with 0.01 mg OB on day 1 made adult copulatory behaviour during treatment with TP indistinguishable from that of rats castrated on day 10 or rats castrated at birth and treated with TP during infancy. It was concluded that the masculine organization of systems and structures involved in the display of male copulatory behaviour occurs under the influence of both non-aromatizable androgen and oestrogen, oestrogen being most likely the substance required to 'organize' the central nervous aspects of the regulation of this behaviour. The absence neonatally of non-aromatizable androgen and/or oestrogen results in specific deficiencies in adult copulatory behaviour as compared with the behaviour of normal male rats.

Delay of ovulation in rats with sodium pentobarbitone: apparent differences between rats with 4- or 5-day reproductive cycles.

In rats with 4- or 5-day reproductive cycles various responses to the blockade of ovulation with sodium pentobarbitone at pro-oestrus (or at pro-oestrus and on the next day) were compared. The blood concentrations of oestradiol decreased rapidly during the 24 h period after injection of sodium pentobarbitone at pro-oestrus in rats with 5-day cycles. In those with 4-day cycles this response took almost a day to develop. Injection of sodium pentobarbitone at pro-oestrus and on the next day interfered with ovulation of the present crop of large follicles in rats with 5-day cycles. In 4-day cyclic rats this procedure delayed ovulation of these follicles by 48 h. Receptive behaviour was absent on the day after the second injection of sodium pentobarbitone in rats with 5-day cycles; some receptivity was, however, induced by the injection of gonadotrophin. The latter injection resulted in the release of a low number of eggs; fertilization of these eggs, however, did not occur. In 4-day cyclic rats receptive behaviour was recorded on the day after the second injection of sodium pentobarbitone: fertilization of delayed ovulated eggs took place normally but pregnancy was seen only rarely. The results indicated clear differences in responses to blockade of ovulation with sodium pentobarbitone between rats with 4- or 5-day cycles. The differences most probably result from a more advanced 'age' of preovulatory follicles at pro-oestrus of 5-day cycles compared with those of 4-day cycles. Experimental delay of ovulation reveals ageing changes and the probable onset of atresia at an earlier time after blockade in 5-day cyclic than in 4-day cyclic rats.

Plasma oestradiol and delayed ovulation after administration of sodium pentobarbitone to pro-oestrous 5 day cyclic rats.

Injection of sodium pentobarbitone (Nembutal) into rats between 12.00 and 13.00 h on the day of pro-oestrus was fully effective in blocking the expected ovulation. In 75% of the rats, ovulation of the present generation of large follicles occurred 24 h later (delayed ovulation). Injection of Nembutal between 12.00 and 13.00 h on the day of pro-oestrus and at the same time on the subsequent day was fully effective in blocking the ovulation twice: in only two rats out of 13 did ovulation of the present generation of follicles still occur. When unilateral ovariectomy was performed immediately after injection of a single dose of Nembutal into pro-oestrous rats, delayed ovulation was significantly inhibited. However, after inhibition of ovulation by either two injections of Nembutal or one injection and unilateral ovariectomy, delayed ovulation could be induced by treatment with a small dose of oestradiol benzoate during the Nembutal-induced anaesthesia. It thus seemed that delayed ovulation failed because of disruption of oestrogen production after administration of Nembutal. The concentration of oestradiol-17beta in the plasma of Nembutal-treated pro-oestrous rats decreased rapidly during the 24 h after treatment. It is concluded that this decrease in the concentration of oestradiol is due to the inherent ageing of preovulatory follicles manifesting itself when exposure to the ovulatory surge of LH is inhibited.

Dioestrous progesterone and pro-oestrous luteinizing hormone in 4- and 5-day cycles of female rats.

Female rats with showed 5-day ovarian cycles spontaneously, but 4-day cycles for periods of variable length after induction of pseudopregnancy, were examined. While the time of onset of the ovulatory LH surge at pro-oestrus was the same in both 4- and 5-day cycles, dioestrous progesterone concentrations were dissimilar. The most prominent feature was a sharp increase during the night of the first day of dioestrus of the 4-day cycle. The results are interpreted as suggesting that dioestrous steroid concentrations rather than pro-oestrous LH concentrations are important in establishing cycle duration.

Regulation of luteal activity in adult female rats treated neonatally with testosterone propionate.

The present study was concerned with the control of luteal activity in female rats which had been treated neonatally with 1.25 mg testosterone propionate (TP). Treatment of such rats in adulthood with 15 i.u. human chorionic gonadotrophin induced ovulation followed by a period of luteal activity. The two daily surges of prolactin secretion, typical for a period of luteal activity in the normal female rat, were not observed in TP-treated females. Instead, higher basal levels of prolactin were observed in TP-treated females than in normal female rats. Furthermore, uterine traumatization at 5 days after ovulation did not result in the formation of decidual tissue. In intact TP-treated females luteal activity, induced and temporarily sustained by an ectopic pituitary transplant, persisted after removal of the pituitary graft. In contrast, in TP-treated females which had been ovariectomized on day 25 of age and had received an ovarian transplant before induction of the luteal phase, luteal activity ended within a week after removal of the ectopic pituitary gland. Females treated with TP which had been ovariectomized on day 25 of life had lower plasma levels of prolactin and higher levels of dopamine in hypophysial stalk plasma than intact TP-treated females when measured at 4 months of age. Treatment of ovariectomized rats with oestradiol-17 beta increased levels of prolactin in plasma and lowered levels of dopamine in hypophysial stalk plasma. It is concluded that the control of luteal activity in TP-treated females shows 'male' characteristics. However, the presence of the ovaries in such rats leads to decreased hypothalamic release of dopamine and increased plasma levels of prolactin, probably due to increased oestrogen levels. These increased levels of prolactin are sufficient to maintain luteal activity.

Reduction of 5-day cycle length of female rats by treatment with bromocriptine.

Rats with 5-day ovarian cycles were injected daily with 1 mg bromocriptine. This treatment resulted in a change of cycle length from 5 to 4 days and a rapid increase in ovarian weight. The increase in ovarian weight resulted from the accumulation of large numbers of corpora lutea. Normal numbers of corpora lutea were formed during each cycle but luteal bodies did not disappear subsequently. Luteolysis affected only minor foci of luteal tissue and the majority of luteal tissue remained histologically intact throughout the further period of study. The reduction of cycle length from 5 to 4 days occurred when bromocriptine was administered from the day of ovulation only. If treatment was commenced at a later time during the cycle it was not effective. Treatment with bromocriptine appeared to affect the concentrations of progesterone in the blood during dioestrus. During treatment the rats showed the pattern characteristic for 4-day cycles: typically, the high concentrations of progesterone on the day after metoestrus remained absent. These data suggest (1) that the latter part of the production of progesterone during dioestrus by 'non-functional corpora lutea' is dependent on prolactin and (2) that prolongation of high progesterone production after metoestrus plays an important role in changing the length of the cycle from 4 to 5 days. Treatment with bromocriptine did not significantly affect the rate of maturation of follicles destined for the next ovulation. It is possible that follicular maturation is not among the critical variables which determine whether normal ovulatory cycles will last for 4 or 5 days.