Characterization and immunocytochemical demonstration of glucocorticoid receptor using antisera specific to transformed receptor

Cytosolic and nuclear forms of the glucocorticoid receptor were characterized using immunochemical techniques. Antibodies were raised in rabbits to an M_r 58,000 fragment of the transformed (DNA-binding) glucocorticoid receptor purified from rat liver cytosol by DNA-cellulose chromatography and polyacrylamide gel electrophoresis. Antibodies reacted with the transformed receptor form in a radioimmunoassay for glucocorticoid receptor. Western blot analysis of antibody reactivity revealed a single M_r 185,000 receptor form in rat liver cytosol but a smaller M_r 85,000 form in nucleosol, indicating the M_r 85,000 form is the transformed receptor. Furthermore, western blot analysis indicates that the M_r 185,000 receptor undergoes proteolysis during receptor purification and in vitro transformation processes by generating immunochemically similar proteins of smaller molecular weights. An identical M_r 185,000 glucocorticoid receptor was detected in cytosols of four rat tissues; liver, brain, adrenal medulla, and thymus. The glucocorticoid receptor was localized to the cytoplasm and nucleus of rat adrenal medulla cells by immunohistochemistry, demonstrating the existence in vivo of the transformed receptor and translocation of the receptor from cytoplasm to nucleus.     

Serum testosterone, agonistic behavior, and dominance in inbred strains of mice

Social situations in which male mice establish dominant/subordinate relationships were utilized in an attempt to correlate circulating testosterone (T) titer with agonistic behavior. Two long-term (several months) and two short-term (3- and 5-day) situations in which dominance was verified by severity of body scarring or individual aggression scores indicated no consistent correlation of dominance with serum T levels.     

Familial Down's syndrome.

A family is reported in which the same mother conceived two children with trisomy 21. The pregnancy with the second affected child was interrupted after diagnostic amniocentesis. Maternal chromosome analysis was normal. This family and those previously reported suggest that there is an increased recurrence risk of trisomy 21 after the birth of an affected individual, possibly caused by a genetic tendency for non-disjunction. After the birth of a child with Down's syndrome, amniocentesis and chromosome analysis of cultured amniotic fluid cells is indicated in each further pregnancy, irrespective of maternal age.     

Biomass production in mass cultures of marine phytoplankton at varying temperatures

Natural populations of marine phytoplankton obtained from a large outdoor pond were grown on waste water-sea water mixtures in laboratory continuous cultures in the temperature range 5–33 °C. Virtually all of the influent inorganic nitrogen (14.0 mg l^?1) was assimilated at every temperature tested. There was, however, a distinct change in dominant species with temperature: below 19.8 °C Phaeodactylum tricornutum was dominant, at 27 °C Nilzschia sp. was the main species, and as the temperature increased above 27 °C a blue-green alga, Oscillatoria sp., became increasingly dominant. There is some indication that the excellent growth of P. tricornutum below 10 °C was related to a dramatic increase in the nutrient content per cell as the temperature decreased. Thus at low temperatures reduced division rates are compensated for by increased nutrient uptake rates. It follows that there is a transfer of phytoplankton protein from numerous small cells at intermediate temperatures to large cells that are reduced in numbers at lower temperatures but which represent the same total organic matter. The effect of this phenomenon on annual food chain efficiencies in both controlled and natural marine ecosystems is unknown.     

Effects of gonadal steroid hormones on hibernation in the Turkish hamster (Mesocricetus brandti)

Summary The Turkish hamster (Mesocricetus brandti) is a photoperiodic species in which exposure to a short day photoperiod induces gonadal atrophy. When hibernating male Turkish hamsters were administered testosterone via subcutaneous silastic capsules hibernation was abruptly terminated. The threshold serum testosterone level required to prevent hibernation was intermediate between the levels observed in males with fully-regressed and fully-functional testes. Dihydrotestosterone was as effective as testosterone in blocking hibernation. Estradiol had a partial inhibitory effect. Progesterone and corticosterone implants had no effect on hibernation. Similar results were obtained in female hamsters. The total period of hibernation for untreated males under laboratory conditions was usually 5–6 months. In most animals testicular growth began to occur toward the end of that period. As gonadal growth continued bouts of torpor decreased in frequency and duration, and hibernation was terminated when the testes approached reproductive size. Castrated males continued to hibernate much longer than intact animals. These observations suggest that in nature decreasing day length in the fall initiates gonadal regression which is followed by cold-induced hibernation. In the spring spontaneous recrudescence of the testes may lead to termination of the dormancy period. This mechanism may enable the species to prepare for the reproductive season with maximum efficiency.Supported by NIH Research Grant HD-10478 and by a grant from the Connecticut Research FoundationThe data in this paper were presented in part at the Annual Meeting of the American Society of Zoologists, Toronto, December 26–30, 1977     

Alterations of the carrier-mediated transport of an anionic solute,methotrexate, by charged liposomes in Ehrlich ascites tumor cells

Summary Interaction of positively charged liposomes with Ehrlich ascites tumor cells increases the bidirectional transmembrane fluxes of the anionic folic acid analog, methotrexate. Negative liposomes reduce methotrexate influx. Stimulation of methotrexate influx by positively charged liposomes is time and concentration dependent, requiring at least a 5-min incubation with 2.5mm phosphatidylcholine containing 20% stearylamine for maximum effect. Stimulation is not appreciably reversed by washing the cells. Similar increases are observed for influx and efflux so that there is no change in the steady-state methotrexate electrochemical-potential difference across the cell membrane. The increase in influx appears to be a stimulation of the carrier-mediated transport process for methotrexate since both control and stimulated influx are abolished by the competitive inhibitor, 5-formyltetrahydrofolate or the sulfhydryl group inhibitor,p-chloromercuriphenylsulfonic acid and the Q₁₀ of the system remains unchanged. Influx of 5-methyltetrahydrofolate, which shares the same transport carrier as methotrexate, is also stimulated. However, the transport of folic acid, which is structurally similar to methotrexate but does not utilize the carrier, is unaffected. The kinetic change induced by positively charged liposomes is an increase in theV _ma ⁱⁿ , while theK _t ⁱⁿ remains unchanged. Trans-stimulation of methotrexate influx by 5-formyltetrahydrofolate occurs to the same extent in the presence or absence of positively charged liposomes. The liposomes have no apparent effect on the intracellular water, the extracellular space, or the chloride distribution ratio. The data suggest that interaction of positively charged liposomes with Ehrlich ascites tumor cells accelerates the rate of transposition of the membrane carrier system for methotrexate, altering the kinetics of transport without a change in transport thermodynamics.     

Problems in estimating growth rates of marine phytoplankton from short-term14C assays

Growth rate estimates () of phytoplankton populations that were sampled from nitrogen-limited continuous cultures and then incubated for short durations in batch culture with added¹⁴C-HCO₃ ^– were significantly different than steady-state growth rates () for 3 of 5 marine phytoplankton species. Two diatoms,Thalassiosira weissflogii andChaetoceros simplex, displayed virtually identical growth rates (=) over a wide range of, whereas for a third diatom,Phaeodactylum tricornutum, was overestimated by an average of 40% compared to. In contrast, was underestimated by the¹⁴C technique for the two remaining species: up to 40% at a steady-state of 1.0 day^–1 for the chlorophyteDunaliella tertiolecta and up to 100% at of 1.4 day^–1 for the haptophytePavlova lutheri. For the latter two species the divergence between and appeared to increase with increasing steady-state. A simple model of labeled and total carbon flow between the aqueous phase and cellular biomass was constructed to demonstrate that respiration was negligible when=, but was significant when>. In the cases in which<, a rapid physiological alteration presumably took place once the steady state was disturbed and cells were placed in the incubation chambers, which perhaps was related to the nutritional state of the cultures at the time of sampling. Questions thus are raised regarding our ability to measure accurately primary productivity from shipboard experiments with confined samples of phytoplankton from nutrient-impoverished waters that probably are less hardy than the laboratory cultures used in these studies.     

Respiratory responses in reversible diaphragm paralysis

The effects of diaphragm paralysis on respiratory activity were assessed in 13 anesthetized, spontaneously breathing dogs studied in the supine position. Transient diaphragmatic paralysis was induced by bilateral phrenic nerve cooling. Respiratory activity was assessed from measurements of ventilation and from the moving time averages of electrical activity recorded from the intercostal muscles and the central end of the fifth cervical root of the phrenic nerve. The degree of diaphragm paralysis was evaluated from changes in transdiaphragmatic pressure and reflected in rib cage and abdominal displacements. Animals were studied both before and after vagotomy breathing O2, 3.5% CO2 in O2, or 7% CO2 in O2. In dogs with intact vagi, both peak and rate of rise of phrenic and inspiratory intercostal electrical activity increased progressively as transdiaphragmatic pressure fell. Tidal volume decreased and breathing frequency increased as a result of a shortening in expiratory time. Inspiratory time and ventilation were unchanged by diaphragm paralysis. These findings were the same whether O2 or CO2 in O2 was breathed. After vagotomy, no significant change in phrenic or inspiratory intercostal activity occurred with diaphragm paralysis in spite of increased arterial CO2 partial pressure. Ventilation and tidal volume decreased significantly, and respiratory timing was unchanged. These results suggest that mechanisms mediated by the vagus nerves account for the compensatory increase in respiratory electrical activity during transient diaphragm paralysis. That inspiratory time is unchanged by diaphragm paralysis whereas the rate or rise of phrenic nerve activity increases suggest that reflexes other than the Hering-Breuer reflex contribute to the increased respiratory response.     

Plasma exchange in the long-term management of Waldenstr?m's macroglobulinemia.

Two patients with Waldenström''s macroglobulinemia (WM), which had become resistant to cytotoxic drugs, were treated for features of the hyperviscosity syndrome by repeated plasma exchange with the continuous-flow blood-cell separator over periods of 36 and 28 months, respectively. After four initial weekly plasma exchanges the procedure was carried out every 4 to 6 weeks and both patients tolerated it well. Relative viscosity of the serum was maintained within the normal range in one patient, and both patients remained free of symptoms of the hyperviscosity syndrome. The results suggest that treatment of WM by long-term "maintenance" plasma exchange alone should be considered in any patient with complications due to chemotherapy or whose disease fails to respond to chemotherapy.     

Differential scanning calorimetry and enzymic activity of rat liver microsomes in the presence and absence of Δ1-tetrahydrocannabinol

The thermal transitions of rat liver microsomes and isolated lipids were investigated by using differential scanning calorimetry. Endothermic transitions at ≈?5°C and between ≈18° and 40°C were detected in the membranes and at ≈?10°C and between ≈10° and 20°C in the extracted lipids.Interaction with $\text{Δ}{}^1\text{-}\text{tetrahydrocannabinol}$ of microsomal membranes and of extracted lipids influences the thermotrophic behaviour as revealed by differential scanning calorimetry and eliminates the break in the Arrhenius plot of the enzymic activity of $\text{O-}\text{demethylase}$.