Endogenous pyrogen formation by human blood monocytes stimulated by polyriboinosinic acid:Polyribocytidylic acid

The pyrogenic response to supernatants from human blood monocytes stimulated with polyriboinosinic acid:polyribocytidylic acid (poly I:C) was characteristic of a response to endogenous pyrogen in that it was brief and monophasic, and was destroyed by heating the supernatants at 70°C for 30 min. Pyrogen production was unimpaired when the incubations were carried out in the presence of cycloheximide (50 g/ml; an inhibitor of protein synthesis) or indomethacin (50 g/ml; an inhibitor of prostaglandin synthesis). Also, neither interferon, interleukins, tumor necrosis factor nor prostaglandin E₂ were detectable in the supernatants from the poly I:C-stimulated human monocytes.     

Unusual antimicrobial compounds fromAeollanthus buchnerianus

Using bioassay guided isolation, three novel 12 carbon polyoxygenated fatty acids and a novel abietane diterpene have been isolated from the the chloroform extract of aerial parts ofAeollanthus buchnerianus (Lamiaceae). Rigorous spectroscopic methods were used for compound identification. (Z,Z)-8-acetoxy-5-hydroxydodeca-2,6-dienoic acid and (Z,Z)-5,8-dihydroxydodeca-2,6-dienoic acid inhibited the spore germination ofCladosporium cucumerinum (both with Minimum Inhibitory Dose (MID) values of 1 g) andAspergillus niger (MID 5 and 25 g respectively). Further, they also reduced the hyphal growth ofPythium ultimum. (Z)-5-hydroxy-6,7,8-triacetoxydodeca-2-dienoic acid exhibited short term inhibition of the growth ofCladosporium cucumerinum. The novel abietane diterpenoid, (rel)-14-acetoxyabiet-7-en-18-oic acid inhibited the growth of the gram positive bacteriaBacillus subtilis, staphylococcus aureus andStreptomyces scabies (MIC values 80, 20 and 20 g ml^–1 respectively).     

Central nicotinic receptor blockade inhibits emotionally conditioned pressor responses in rats

A conditioned stimulus previously paired with electric footshock produced an increase in blood pressure in conscious, freely moving rats. The conditioned pressor response was reproducible. Intracerebroventricular injection of the nicotinic receptor antagonists hexamethonium (1–10 g) or pentolinium (10 g) but not the muscarinic receptor antagonist methylatropine (3 g) produced an inhibition of the conditioned pressor response, whereas intraarterial injection of hexamethonium (10 g) did not affect the response. Intraventricular injection of the cholinesterase inhibitor physostigmine (3–10 g) produced an enhancement of the conditioned pressor response. These results are consistent with the possibility that central nicotinic receptors play a role in the expression of the emotionally conditioned pressor response in rats.     

Antidiuretic effects of oxytocin in the Brattleboro rat

Summary The antidiuretic activity of oxytocin (OT) was measured in Brattleboro rats with congenital diabetes insipidus. A dose dependent antidiuretic response was found in animals receiving chronic infusions of 0.1 g/h, 1.0 g/h, and 5 g/h of OT. OT infused at the rate of 5 g/h over a 7-day period completely reversed the symptoms of diabetes insipidus. The results support the concept that OT serves as a weak agonist of vasopressin at the level of the kidney and at pharmacological levels exhibits antidiuretic activity.     

Repercussions de l'hyperthyroïdie sur le contenu total en ADN du cervelet de rat âgé de 6 et 35 jours. Effets comparés de la LT3 et de la DLT4

Summary The total cerebellar proteins RNA and DNA contents from DLT4- and LT3-treated rats was studied at 6 and 35 days of age. The effect of injections of 5 g/j of DLT4 is comparable to that of 25 g of LT3 at birth, followed by 0.5 g every 2 days. On the other hand, injection of 0.5 g of LT3 every 2 days does not induce any significant modification of the total DNA contents in the cerebellum.     

Deciduogenic effects of mediators of the polyphosphatidylinositol pathway in pseudopregnant mice

Intraluminal injections (15 l) of either concanavalin A (125 g) or ionophore A 23187 (0.01 mol) induced a decidual cell reaction (DCR) in the uterus of day 4.5 pseudopregnant mice. However, when these agents were administered in different combinations with each other or with CaCl₂ (15 mol) and phorbol-12-myristate-13-acetate (1.6 nmol), interacting effects occurred to either enhance or inhibit each of the others' independent deciduogenic capacities. The results suggest that the polyphosphatidylinositol pathway and Ca²⁺ are involved in the induction of the DCR in mice with complex interactions occurring between the active components of the pathway to modulate the outcome of the transformation process.     

Storage of irradiated human blood; a source of error in quantitative chromosome analysis

Summary Human whole blood was irradiated with 2.5 Gy of 220 kVp X-rays and stored before culture with 9.7 M BrdU and 19.4 or 38.7 M BrdU for 0, 24, 48 and 72 h. The frequency of dicentrics and ring chromosomes was determined in cells staining as first division (M1) metaphases with the fluorescence plus Giemsa technique. Storage had no influence on the observed aberration yields in 44 h cultures containing 9.7 M BrdU. In 66 h cultures at 19.4 M BrdU the observed yields after 2 and 3 days' storage were significantly lower as compared to cultures from fresh blood. No storage effect was revealed in 66 h cultures containing 38.7 M BrdU. In cases where cytogenetic radiation dosimetry has to be carried out using blood samples which have been in transit for 2–3 days, the findings are of relevance for a correct determination of the chromosome damage in M1 cells.     

Giant pigment granules in dermal melanocytes of rat scrotal skin

Summary Giant melanosomes, ellipsoidal (1.5 m×1.3 m) or spherica (1.1 m–1.4 m in diameter) exist in the scrotal skin of the black pelted Long Evans rat. They are longer and wider than normal stage IV melanosomes (0.7 ×0.4 m) found in theses dermal melanocytes.The authors acknowledge with gratitude, the technical assistance of Evelyn Ann Flynn. This work was supported by grant AM 20669-16 from the National Institute of Arthritis, Metabolism, and Digestive Diseases, USPHS.     

Adrenocortical metabolism and plasma corticosterone in soman intoxicated rabbits

Summary The organophosphate neurotoxin soman produced impairments in adrenocortical RNA and protein metabolism. Fasciculate and reticular cell RNA and protein contents were supporessed with sublethal to acutely lethal dosages (20, 30 and 40 g/kg, s.c.) during the acute excitatory phase of intoxication and at 6–8 h post injection. All three dosages produced ca 90% inactivation of plasma cholinesterase. A transient elevation of plasma corticosterone occurred with 20 g/kg soman whereas there was a protracted increase with 30 g/kg. Corticosterone was not significantly elevated with 40 g/kg, but death occurred at 13±4 min. Thus, the magnitude and/or nature of soman-induced metabolic impairments does not appear to prevent adrenal activation.Supported by US Army Medical Research and Development Command Contract DAMD 17-81-C-1202.     

The length-tension diagram of single muscle fibres of the crayfish

Zusammenfassung An isolierten Muskelfasern vonAstacus fluviatilis wurde mittels Kaliumkontraktur die Spannung-Sarkomerlängenrelation untersucht. Maximale Faserspannung wird bei einer Sarkomerlänge von 10,5 entwickelt. Werden Fasern zu einer Sarkomerlänge von 16,5 gedehnt, so ist die Spannung nicht mehr messbar. Diese Sarkomerlänge ist etwas grösser als diejenige (14.45), die aus der Länge der dicken Filamente (3.95) und der optimalen Länge hervorgeht.     

Failure of somatostatin to influence experimental tumor cell growth in vivo and in vitro

Summary The influence of somatostatin on tumor cell growth was studied in vivo in mice (sarcoma 180 ascites tumor and Lewis lung tumor) and in vitro on nontransformed and polyoma-transformed cell lines. 4 or 20 g/100 g of cyclic somatostatin and 4 g/100 g of linear protamin Zn-bound somatostatin were injected s.c. twice daily in the in vivo study. Cyclic somatostatin (1, 4 or 10 g/ml) was added twice daily to the cell cultures. Somatostatin administration influenced neither the survival of animals nor the growth rate of cultured cell lines. *** DIRECT SUPPORT *** A2025117 00011     

Effect of dipyridamole on glomerular mesangial cell ecto-5′-nucleotidase expression

Although dipyridamole has been extensively studied as an anti-aggregating agent, its mechanism of action has not been elucidated. Cultured mesangial cells were treated with dipyridamole 1–100 M from 6–72 h. Ecto-5-nucleotidase activity approximately doubled (from 115±11 to 226±14 nmol/min/mg) after treatment with 100 M dipyridamole for 72 h. This effect was concentration- and time-dependent. Cycloheximide, an inhibitor of protein synthesis, did not alter basal 5-nucleotidase activity. However, it prevented stimulation by 5 M dipyridamole. Adenosine availability at the receptor sites was increased by dipyridamole and S-(p-nitrobenzyl)-6-thioinosine (NBTI), which inhibit adenosine uptake into the cell. Addition of dipyridamole or NBTI to the adenosine-treated mesagial cells produced an additive increase in ecto-5-nucleotidase activity. Dipyridamole, through its effect on extracellular adenosine and ecto-5-nucleotidase, may have an influence upon regulation of the glomerular microcirculation.     

A sensitive protein assay method using micro-titer plates

Summary A commercially available protein-assay, based on the reaction between bicinchoninic acid (4,4-dicarboxy-2,2-biquinoline) and copper was adapted for use in microtiter plates. The assay is sensitive between 10 m/ml and several mg/ml, and it uses only 10 l of sample material. The results obtained with the method were in accordance with other protein determination methods. The assay was shown to be reproducible, reliable and insensitive to non-ionic detergents.Acknowledgments. The authors are grateful to Miss Barbara Moser who skillfully performed a number of these experiments. This research was supported by the Swiss National Science Foundation, Grant No 3.300–0.82, to U.B.     

Zur Umwandlung von β-Pyridylcarbinol in Nicotinsäure im tierischen Organismus

Summary By way of the perfusion technique, it is shown that the liver of the rat readily converts-pyridyl-carbinol to nicotinic acid. The turnover attains a rate of 0.5M/g/min.     

Uptake of D-glucose anomers by rat retina

Summary Uptake ofd-glucose anomers by isolated rat retina was studied. After 3 min incubation at 37°C in the presence of or anomer (750 g/ml), a significantly greater uptake (1.32 mg/g wet tissue) of -anomer was observed compared with that of -d-glucose (1.11 mg/g wet tissue). This result and other data suggest that the carrier ford-glucose transport in the retina prefers the -anomer stereospecifically.Acknowledgment. The authors are grateful to Mr S. Suzuki for his technical assistance.     

Different effects of D-glucose anomers for respiration of bacterial germinated spores

Summary Effects of - or -D-glucose on the respiration of germinated spores (only germinated spores not including swollen spores and elongated spores) ofBacillus subtilis andB. megaterium were studied. In our conditions, net amount of oxygen consumed by 10¹⁰ germinated spores ofB. subtilis per min after addition of - or -D-glucose was 1.6 g or 6.6 g (/=4.13), while that byB. megaterium was 4.5 g or 6.8 g (/=1.51), respectively. However, the net amounts of oxygen consumed by 10¹⁰ vegetative cells per min after addition of - or -D-glucose were identical, forB. subtilis in both cases 443.0 g and forB. megaterium in both cases 604.4 g.     

Artificial spawning effected in the fresh water teleost,Cyprinus carpio by clomiphene citrate

Summary Triweekly i.m. injections of clomiphene citrate (group 1, 25 g/0.5 ml and group II, 50 g/1.0 ml) were administered for a period of 3 months during the preparatory period to female fresh water teleosts exhibiting ovarian recrudescence, while a control group received 0.5 ml of physiological saline throughout the period of experimentation i.e., from February through April. 50 g clomiphene citrate treatment brought about a steady increase in ovarian size, and oocytes began to enlarge and mature and finally ovulation took place in April. This is 4 months ahead of their normal occurrence.The authors wish to express their gratitude to Richardson Merrel Co. for supplying the sample of clomiphene citrate, and Dr K.N. Udupa, Institute of Medical Sciences, Banaras Hindu University, Varanasi for facilities.     

Lack of effect of cortisone,thyroxine and insulin on the developmental pattern of mouse intestinal glucose-6-phosphatase activity

Summary Daily administration for 3 days of cortisone (25 g/g b.wt), thyroxine (1 g/g b.wt) or insulin (12.5 mU/g b.wt) to 8-day-old suckling mice does not induce a premature decrease of the phosphohydrolase activity of intestinal glucose-6-phosphatase.Supported by grant MA-5969 from the Medical Research Council of Canada (D.M.). C.M. is a recipient of a studentship from MRC of Canada. D.M. is a chercheur-boursier du Conseil de la recherche en santé du Québec.     

Disruption of mitochondrial function as the basis of the trypanocidal effect of trifluoperazine onTrypanosoma cruzi

Summary The tricyclic anti-calmodulin drug trifluoperazine (TFP) inhibited growth and motility of epimastigotes ofTrypanosoma cruzi, at concentrations lower than 100 M, and motility and infectivity of the bloodstream trypomastigote form at 200 M. Electron microscopy of TFP-treated epimastigotes showed that the major effect was at the mitochondrial level, with gross swelling and disorganization. The oligomycin-sensitive, mitochondrial ATPase was completely inhibited by 20 M TFP, and the same drug concentration caused a 60% decrease in intracellular ATP content. The results suggest that the trypanocidal effect of TFP may be related more to mitochondrial damage than to the well-known anticalmodulin effect of the drug.     

The use of 5-fluorocytosine and ketoconazole in the culture of the erythrocytic stages ofPlasmodium falciparum and some tumor cell lines

Summary In vitro culture systems are often contaminated by bacteria and fungi. It is therefore often necessary to supplement culture media with agents such as penicillin/streptomycin, gentamycin or amphotericin B. The latter cannot be used in the in vitro culture of erythrocytic stages ofP. falciparum, and thus anti-fungal agents have not been regularly used in this system. We describe the prophylactic use of 5-fluorocytosine (5-FC) and ketoconazole (KTZ) in tissue cultures at concentrations up to 300 and 10 g/ml respectively which have no effect on the growth ofP. falciparum (FCR-3 strain). A melanoma cell line (C32) and a line of uterine carcinoma (C41) were also unaffected by similar concentrations of 5-FC and KTZ. When dissolved in complete culture medium (RPMI 1640) with 10% human plasma, the minimum inhibitory concentration of 5-FC for a susceptible strain ofCandida remained below 2 g/ml. These experiments suggest that 5-FC (at 50 g/ml) alone or in combination with KTZ (at 1 g/ml) is a useful addition to the armamentarium of antimicrobials available to the tissue culture biologist for a variety of cell culture systems.