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Biomedical subjects

R K Andjus

Publications and source records attributed to R K Andjus.

13 recordsLinked to original sources

Effects of xanthine derivatives on electroretinographic responsiveness.

In view of the use of synthetic propentofylline (PPF) as a protective agent in brain ischemia, its possible side effects on vision capacities have been explored by electroretinography in comparative experiments with theophylline. We used eyecup preparations of small-spotted dogfish sharks and of European eels, particularly suitable for long-lasting experiments. The drug exerted profound but reversible modifications of ERG records: (1) a dose-dependent increase of the amplitude and duration of the chemically isolated late receptor potential (LRP), (2) a partial unmasking of LRP, (3) a strong potentiation of the LRP-unmasking effect of low temperature, (4) a potentiation of light adaptation effects, and (5) a strong potentiation of the post-illumination hyperexcitability. The effects were explicable as due to a strong phosphodiesterase (PDE) inhibiting, cyclic guanosine monophosphate (cGMP) promoting, action of the drug. The effects were considerably stronger, or even of opposite sign, in comparison to those of the chemically related theophylline. PPF did not seriously affect the ERG c-wave originating in the pigment epithelium. The results suggested that the effects of PPF on vision may not seriously hamper the therapeutic use of the drug. They indicated, on the other hand, that PPF was a retinoactive drug of potential usefulness in the exploration of the complex biochemical events underlying visual transduction.

Adaptation, Ocular↗

In vitro and in vivo thermal activation of steroid-receptor complexes from rats and ground squirrels (Spermophilus citellus).

Using 3H-labelled triamcinolone acetonide (3HTA, synthetic steroid hormone), it was shown that the in vitro time course kinetics of thermal activation of 3HTA-receptor complexes exhibited the same temperature dependence in liver cytosols prepared from hibernating ground squirrels (Spermophilus citellus) as in cytosols from the rat. When 3HTA was injected in vivo to animals hibernating with a body temperature of 3 degrees C, the activation and nuclear uptake of the in vivo formed steroid-receptor complexes proceeded at a slow rate, comparable to the one predicted by in vitro studies. In the hibernator, the results are not indicative of adaptive modifications at the level of thermal activation, but prove that steroid action does proceed at a temperature incompatible with hypothermic survival in the nonhibernator.

Acclimatization↗

Biophysical models of protein denaturation. I. An improvement of the model of two states.

The model of two states (native and denatured), frequently used for the description of protein denaturation, has been complemented by relations defining, on theoretical grounds, the temperature dependence of the relevant thermodynamic functions. In essence, this was achieved by assuming that the temperature dependence of Gibbs free energies of the native protein and of denaturation can be approximated, within the interval (0 degree C, 100 degrees C), by second-order partial sums of Taylor series. The improved model operates with four parameters: the temperature of denaturation, the heat capacities of the native and denatured protein at the temperature of denaturation, and the entropy of denaturation at that temperature. A theoretical treatment is included of the temperature dependence of total heat capacity, the variable recorded in the form of continuous thermograms by means of differential scanning calorimetry. Our model correctly reproduces experimental thermograms of proteins and provides for the biophysical interpretation of a number of their geometric components. Fitting procedures were complemented by a newly devised method for estimating starting values of model parameters from calorimetric data. The phenomenon of cold denaturation was also reproduced quantitatively by our model, which supplies explicit proof of the exothermal nature of this phenomenon. Finally, the relationship between temperature profiles of thermodynamic functions describing denaturation has been defined by sequences of profile magnitudes at points where the profiles intersect the temperature axis and/or cross each other. Model-derived sequences of profile magnitudes, representative of cross-point temperatures and of intervals in between, together constitute a general characteristics of denaturation, uninfluenced by differences in thermodynamic stability between protein species.

Mathematics↗

Biophysical models of protein denaturation. II. Effects of denaturants and of pH.

In order to broaden the scope and increase the utility of differential scanning calorimetry, a theoretical model of calorimetric thermograms is presently proposed which facilitates their biophysical interpretation and accounts explicitly for their modifications induced by denaturing agents and/or pH. The model rests mainly on statistical-physical considerations, the denaturation-linked increase of the number of binding sites for denaturants (including H+) serving as the conceptual basis for thermogram modelling. Denaturants were envisioned as contributing indirectly to thermal denaturation by forming complexes preferentially with unfolded protein molecules, shifting thus the equilibrium towards the denatured phase. After postulating the probability of complex formation, mean numbers of the relevant molecular species were computed by ensemble averaging. Finally, an eight-parameter expression has been derived defining protein heat capacity as a function of both temperature and denaturant concentration (or pH), each of the eight parameters having a distinct biophysical meaning. The model has been tested by applying it to the prediction of the pH-dependence of thermograms. Four proteins have been considered (lysozyme, myoglobin, apomyoglobin, and ribonuclease A), each represented by a series of three to four published thermograms recorded under different pH conditions. Model equations, fitted simultaneously to all thermograms in a pH series, reproduced correctly experimental tracings. Parameter values obtained as best-fit requirements (particularly those representing the number of binding sites unmasked by denaturation and the free energy of ion binding) were in close agreement with empirical, mainly potentiometric, data from literature. The empirically established pH-independence of the total enthalpy of denaturation, the phenomenon of cold denaturation, the pH-dependence of the Gibbs free energy of denaturation, of the melting temperature and of the temperature of cold denaturation, were all correctly predicted by the model. Combined effects of multiple denaturants, including the effects of pH in the presence of denaturants other than protons, are also predictable by the model.

Calorimetry↗

Developmental pattern of the testicular androgen response to gonadotrophin stimulation in vitro and its modification by chronic hypoprolactinaemia.

Developing male rats were treated chronically with bromocriptine (BR, 3 mg/kg b.w. daily) to maintain severe hypoprolactinaemia throughout postnatal development. This treatment induced a precocious increase in Leydig cell numbers per testis and caused substantial, but age-dependent, modifications of the androgenic responsiveness of incubated hemi-testis preparations to stimulation with hCG. Most conspicuous were: (i) a decrease in sensitivity of the testis to hCG at the approach of adult age (due presumably to reduced responsiveness of the Leydig cells), and (ii) a precocious increase in the steroidogenic maximum of the testis at peripubertal age. This probably resulted from the precocious increase in Leydig cell numbers, which was able to mask the negative consequences of reduced androgenic capacity per Leydig cell. The precocious increase in number of Leydig cells induced by hypoprolactinaemia could have resulted from facilitation of the proliferative action of the high prepubertal LH levels on Leydig cell numbers. There were no clear-cut indications for an important effect of BR-induced changes in LH levels, or of a direct effect of BR on the testis.

Androgens↗

Rapid naloxone-induced alterations of androgen variables in the growing male rat.

Contrary to earlier views on the inability of naloxone to affect androgen variables by way of general circulation, systemically applied naloxone (2.5 mg/kg body weight, single i.p. or i.v. injection) has been shown to rapidly induce (within an hour) a significant fall (-35.7% on the average) of the concentration of serum androgen (testosterone and dihydrotestosterone; (T + DHT) in peripubertal rats (51-58 days old). Such a response to the opiate antagonist was absent, however, in low-androgen prepubertal animals (37-44 days old) and in those among peripubertal rats which still showed subcritical initial levels of androgen in circulation (less than 1.5 ng/ml; experiments with repeated blood sampling in catheterized animals). In peripubertal rats naloxone was also shown to induce a significant decrease (-36%) in basal in vitro androgen production by testes removed 15 or 30 min following the intraperitoneal administration of the opiate antagonist. Such an inhibitory effect on basal steroidogenesis has not been observed in control multiple-dose experiments in which incubated testes from naloxone-naive rats have been directly challenged with naloxone; on the contrary, enhancing direct effects were recorded, but only with the highest concentration of naloxone tested (10(-4) M). The possibility thus remains open that indirect inhibitory effects of injected naloxone may be operational in intact animals. Hypoprolactinemia, known to interfere in an age-dependent manner with the responsiveness of Leyding cells to luteinizing hormone (LH), may be of particular relevance.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Effects of bromocriptine-induced hypoprolactinaemia on the developmental pattern of androgen and LH levels in the male rat.

In immature male rats, receiving daily injections of bromocriptine (3 mg/kg bw), serum prolactin (Prl) remains low throughout development. In such hypoprolactinaemic males androgen variables are affected: a) the normally low pre-pubertal serum androgen (testosterone, dihydrotestosterone-T, DHT) is considerably increased, in correlation with a precocious development of the testicular Leydig cell population; b) the peri-pubertal rise of androgen is not prevented, but is is followed by significantly lower post-pubertal T, DHT levels, in correlation with moderately reduced Leydig cell counts and with strongly attenuated growth rates of sex accessory organs. Observed alterations of the post-natal androgen pattern cannot be related to LH changes since developmental LH values remained essentially unaltered. Results are in concordance with a marked age-dependent sensibility of androgen variables to a lack of Prl in the developing male.

Androgens↗

Effects of short-term and long-term hyperprolactinemia on the developmental pattern of androgen and LH levels in the immature male rat.

Developmental patterns of serum levels of androgens and of the luteinizing hormone (LH) were studied comparatively in: i) long-term hyperprolactinemic (LT) immature male rats bearing ectopic pituitary grafts since the age of 21 days and examined at weekly intervals thereafter; and ii) in short-term hyperprolactinemic (ST) males grafted at several postnatal intervals and sacrificed 7 days postoperatively. In both ST and LT rats serum LH was markedly reduced, but only during the early prepubertal period (30 to 37 days), characterized in normal rats by conspicuously high LH levels. During the next pubertal phase of development (51 to 58 days), normally characterized by a steep rise of serum androgens in the presence of relatively low LH, the androgen surge was significantly attenuated in LT, but not in ST animals. This suggests that elevated PRL, if maintained long enough prior to the pubertal age, may significantly attenuate or delay the intensified secretion of androgens characteristic of puberty, presumably by suppressing the intensive prepubertal LH secretion. In LT rats the testicular growth was moderately retarded, but the growth of the dorsal prostate was enhanced, suggesting a PRL-induced increase of responsiveness to androgen.

Aging↗

Electroretinographic evaluation of spectral sensitivity in yellow and silver eels (Anguilla anguilla).

Although differences in visual pigments between developmental stages of the European eel are well known, the expected differences in spectral sensitivity have not been demonstrated at the electrophysiological level. In fact, one past electroretinographic study led to the conclusion that in eels there is no change in scotopic sensitivity, with increasing sexual maturity. In the present experiments, electroretinograms (ERGs) were recorded from in situ eyecups of immobilized eels Anguilla anguilla (L.) caught in coastal running waters. It was shown that the ERG b-wave is as good an indicator of spectral sensitivity as the unmasked late receptor potential (LRP) which directly reflects the responsiveness of photoreceptors. Complete spectral-sensitivity curves, based on b-wave thresholds and on thresholds of LRP subsequently isolated by means of sodium iodate, have been obtained in the same eel. Using fitted amplitude-log intensity functions for threshold calculation, and two models for computer-assisted fitting of spectral-sensitivity curves, significant differences in lambda max were found between yellow and silver developmental stages of the eel, identified by ocular index measurements.

Anguilla↗