The specificity of the S1 and S2 subsites of elastase. Esters of tetrapeptides of the general formula ethoxycarbonyl-prolyl-alanyl-X-Y where either X or Y was an alanine residue were synthesised and their cleavage by elastase studied. It was found that variation of the alcohol moiety between methyl, cyclohexyl and nitrophenyl residues had no effect on the catalytic rate constant for cleavage of ethoxycarbonyl-prolyl-dialanyl-alanine esters demonstrating that acylation is much faster than deacylation for this system and also that non-productive binding is not kinetically significant. The effect of changing the amino acid residue in position X was small compared with that of change in position Y. The presence of valine and serine residues in position Y produced the highest specificity constant but the highest catalytic rate constant was found for a leucine residue in this position. The results are discussed in terms of the binding of the substrate to the enzyme.


The primary structure of the coat protein of alfalfa mosaic virus strain VRU. A hypothesis on the occurrence of two conformations in the assembly of the protein shell. The complete primary structure of the coat protein of strain VRU of alfalfa mosaic virus (AMV) is reported. The strain is morphologically different from all other AMV strains as it contains large amounts of unusually long virus particles. This is caused by structural differences in the coat protein chain. The amino acid sequence has mainly been established by the characterization of peptides obtained after cleavage with cyanogen bromide and digestion with trypsin, chymotrypsin, thermolysin or Staphylococcus aureus protease. The major sequencing technique used was the dansyl-Edman procedure. The VRU coat protein consists of 219 amino acid residues corresponding to a molecular weight of 24056. Compared to the coat protein of strain 425 [Van Beynum et al. (1977) Eur. J. Biochem. 72, 63-78], 15 amino acid substitutions were localized. Most of them have a conservative character and may be explained by single-point mutations. A correction is given for the AMV 425 coat protein: Asn-216 was shown to be Asp-216. The prediction of the secondary structure for the two viral coat proteins was not significantly influenced by the various amino acid substitutions except for the region containing residues 65-100. This led us to the hypothesis that the AMV coat protein may occur in two different conformations favouring its incorporation into either a pentagonal or hexagonal quasi-equivalent position in the lattice of the protein shell. The substitutions in the above-mentioned region of the VRU coat protein may have caused a strong preference for the hexagonal lattice conformation. The model is supported by preliminary sequence data of the same coat protein region in AMV 15/64, a strain morphologically intermediate between 425 and VRU.


Pattern of protein phosphorylation in intact stimulated cells: thyrotropin and dog thyroid. Two-dimensional, high-resolution electrophoretic technique of O'Farrell has been adapted to the analysis of thyroid phosphorylated proteins. Proteins were extracted from dog thyroid slices which had been incubated in the presence of [32P]phosphate with thyrotropin or with different agents which enhance the intracellular accumulation of cyclic AMP. About 350 phosphorylated polypeptides have been separated. Thyrotropin stimulates the phosphorylation of at least eight of these polypeptides. An increase in the phosphorylation of the same polypeptides was observed was observed when dog thyroid slices were incubated with dibutyryl adenosine 3':5'-monophosphate, cholera toxin or prostaglandin E1 instead of thyrotropin. Our results confirm that most of dog thyroid protein phosphorylation is independent of cyclic AMP. They offer a first link between the action of cyclic AMP on protein kinase and the physiological effects of thyrotropin. They strongly substantiate the hypothesis that most thyrotropin effects are mediated by cyclic AMP.


1H nuclear-magnetic-resonance studies of the porcine-pancreatic secretory trypsin inhibitor at 270 MHz. The pancreatic secretory trypsin inhibitor from porcine pancreas has been investigated by high-resolution 1H nuclear magnetic resonance (NMR) at 270 MHz. The presence of a number of slowly exchanging labile protons indicates that the protein is highly globular. Of the two tyrosyl rings, one is free-rotating and solvent-exposed while the other one is hindered in its mobility and buried in the interior of the protein. A lineshape analysis of the temperature dependence of aromatic resonances gave the dynamic parameters for activation of ring mobility. The inhibitor exhibits at least three well-resolved high-field ring-current-shifted methyl resonances. Form II of the inhibitor, that lacks the first four residues, has been compared with the intact form I. No detectable differences were found between the spectra of I and II, which indicates that the presence of the N-terminal tetrapeptide does not appreciably affect the overall conformation of the protein.


Genes and mRNAs coding for zein polypeptides in Zea mays. Zein messenger RNAs from maize endosperm were purified by successive oligo(dT)-cellulose chromatography and sucrose gradient centrifugation. Polyacrylamide gel electrophoresis under denaturing conditions revealed the presence of two size classes of zein messenger RNAs of Mr 3.5 x 10(5) and 4.10 x 10(5). The mRNA was shown to synthesize the major zein polypeptides, to have a base composition characteristic of a poly(A)-containing RNA and to be transcribed by reverse transcriptase into complementary DNA. The r0t1/2 of the hybridization curve of cDNA hybridized to an excess of mRNA was shown to be 7 x 10(-2) M . s indicating that about 15 non-cross-hybridizing sequences are present in the zein mRNA preparations. The kinetics of cDNA annealing with an excess of maize DNA from 2 n cells suggest a ten-times reiteration of each mRNA sequence. This result is confirmed from saturation experiments, where in cDNA excess to DNA, the number of zein genes per haploid maize genome was estimated as about 120 copies. Similar experiments carried out on DNA from normal and mutant endosperms (3n cells) indicate the absence of large amplifications or deletions of zein genes in the tissue devoted to zein synthesis.


Effects of amphiphiles on structure and activity of human erythrocyte membrane acetylcholinesterase. Detergents above their critical micellar concentration dissociate the aggregated forms of pure acetylcholinesterase from human erythrocyte membranes to a 6.5-S form, the protomer. This form is active only in presence of amphiphiles. 1. Uncharged (Triton X-100, Tweens, beta-D-octylglycoside), anionic (sodium dodecyl sulfate) and zwitterionic (lysophosphatidylcholine) detergents or bile salts (sodium cholate, deoxycholate) stabilize the 6.5-S enzyme at concentrations well below their critical micellar concentration. 2. Total erythrocyte lipids fully sustain catalytic activity of the 6.5-S form. 3. Protein-protein interactions stabilize the activity of the 6.5-S form of acetylcholinesterase. Above a critical acetylcholinesterase concentration (2.5 microgram/ml) enzyme activity no longer depends on the presence of an amphiphile as reaggregation to multiple molecular forms occurs. It is concluded that human erythrocyte membrane acetylcholinesterase is fully active only if the enzyme can undergo hydrophobic interactions with amphiphiles such as detergents, lipids or proteins.


The protein-chromophore bond in B phycoerythrin from Porphyridium cruentum. Radiosulfur labeling experiments. Red algae of the species Porphyridium cruentum were grown in a minimum sulfate medium containing 35SO42-. 35S-labeled phycoerythrin was extracted. B Phycoerythrin, b phycoerythrin and R phycocyanin could be separated from other proteins by using a carrier-free electrophoresis on columns. The final ratio A545/A280 of B phycoerythrin thus obtained was greater than or equal to 5. 35S-labeled B phycoerythrin was digested proteolytically with trypsin and pepsin. The resulting 35S-containing bilipeptides were separated by isoelectric focusing. Zones of enhanced chromophore concentration always showed an enhanced radioactivity. Peptide fractions with a low molar ratio sulfur/chromophore (1.1-1.8) were purified to remove sucrose and the carrier ampholyte. A modified, optimized Edman degradation followed. A butylacetate-soluble, red Edman product was obtained that contained most of the chromophore and the bulk of the radioactivity. This product was purified by two-dimensional thin-layer chromatography. The main spot of the chromatogram was subjected to acidic hydrolysis. The major part of the radioactivity in the hydrolysate cochromatographed with cysteine. That proves cysteine to be the binding amino acid in all cases investigated.


3H-actinomycin D binding to ultrathin section of plastic-embedded Locusta migratoria testicular tubules. Improvement of the technique and further characterization of the reaction. Semi-thin and ultrathin sections of locust testes have been incubated in 3H-actinomycin D solution and submitted to radioautography. The improved technical conditions described allow the reproducible obtainment of cell radioautographs with a moderate nuclear labelling and a very low nonspecific background which are usable for semi-quantitative results. Extraction with enzymes (DNase, RNase, pronase) or concentrated salt solution have been carried out before 3H-Actinomycin D treatment in order to characterize the reaction. The semi-quantitative results obtained at the light microscope level suggest that, in relation to the structural and chemical changes which occur in chromatin during spermiogenesis, some proteins may be easily hydrolysed in early spermatids. In ultrathin sections of spermatocytes the X chromosome is heavily "stained" with 3H-Actinomycin D, while 3H-uridine is not incorporated into the sex chromatin. These results are discussed in the light of current ideas on the constitution of active chromatin.


Plasmalemma invaginations, contraction and locomotion in normal and caffeine-treated protoplasmic drops of Physarum. The de novo formation of plasmalemma invaginations and vacuoles of light microscopic dimension in protoplasmic drops at different age-stages was studied quantitatively by applying morphometric methods and marker techniques. The study includes: i) the normal morphogenetic development to a drop age of 1 hour, ii) the influence of caffeine treatment, and iii) the effects of removal of this drug. In untreated drops, formation of invaginations and vacuoles is accomplished within 10 to 15 min. By application of 5 mM caffeine, the formation of plasmalemma invaginations is inhibited for 20 to 30 min. The onset of oscillating contraction activity is delayed, but not hindered by the drug. Drug removal 20 min after drop generation leads to an immediate initiation of plasmalemma infolding. Although caffeine does not hinder initiation of normal contraction activity, the locomotory ability of the drop is blocked if the drug is not removed from the drop and the substrate. Thus, caffeine uncouples motive force generation from locomotion in all plasmodial stages of Physarum. The cellular sites of drug action are discussed.


[Hopper-crystal-growth, basis of test morphogenesis in Patellina corrugata Williamson (Foraminifera, Protozoa) (author's transl)]. During the growth of Patellina corrugata's test platelike structures are formed showing typical characters of Hopper-crystals. These features indicate conditions of test development comparable to inorganic skeletal crystal growth. The whole test therefore can be regarded as a skeletal crystal (Hopper-crystal). Skeletal crystal growth is considered to be the basis of morphogenesis in the single-crystal test of Patellina.


Carbon tetrachloride induced proliferation of tight junctions in the rat liver as revealed by freeze-fracturing. Application of carbon tetrachloride produced a progressive proliferation of tight junctions in the rat liver. This system proved to be rapid and highly reproducable and affords the opportunity for tracing the fate of tight junctions in freeze-fracture replicas, facilitating investigations on their formation and function. Beginning on day one carbon tetrachloride treatments resulted in the progressive loosening and fragmentation of the junctional meshwork. After three to four days the membrane outside the zonulae occludentes was extensively filled with proliferated discrete junctional elements often forming complex configurations. From the fifth day on the zonulae occludentes were restricted again predominantly around the bile canaliculus margins. But the junctional meshwork of the zonulae occludentes remained loosened in comparison to those in the control rats. It could be shown that tight junction proliferation on the lateral surface of the plasmalemma occurred both through de novo formation from discrete centers of growth by addition of intramembranous particles and through reorganization of preexistent junctional strands of the fragmented zonulae occludentes bodies. Whereas the large gap junctions close associated with the zonulae occludentes remained more or less unaffected during the experiments, small gap junctions increased in number after five days and were located at the margin or in the tight junction domain. It is assumed that the degeneration of the tight junctions served as a pool for intramembranous particles which form the gap junctions. The results of these observations are discussed in relation to those obtained in other systems.


Comparison of the ultrastructure of adrenaline and noradrenaline storage granules of bovine adrenal medulla. The ultrastructure of the membranes of noradrenaline (NA) and adrenaline (A) granules of the bovine adrenal medulla (Terland, O., T. Flatmark, and H. Kryvi, Biochim, Biophys. Acta 553, 460--468 (1979)) was analyzed by transmission, negative staining and freeze-etch electron microscopy. The two types of storage granules can be distinguished mainly by two morphological criteria: (a) The NA-granules have a more electron dense matrix core than the A-granules, (b) the NA-granules revealed less asymmetry in the distribution of intramembrane particles (nPF:nEF = 4,5:1) than the A-granules (nPF:nEF = 9:1). Thus, the trilaminar structure, negative staining pattern and size distribution of the intramembrane particles of the two fracture faces on freeze-etch electron microscopy were very similar for the two types of granules. Freeze-etching revealed a wide range of the particle size distribution for both fracture faces in both types of granules, with an average diameter of 12.6 +/- 2.7 nm (A-granules) and 10.2 +/- 2.8 nm (NA-granules) for the E-fracture faces and 11.4 +/- 2.7 nm (A-granules) and 9.8 +/- 2.4 nm (NA-granules) for the P-fracture faces. Some of the particles on the P-fracture face (outer surface of the membrane) revealed a subunit structure, most clearly seen in the specimens of NA-granules. Morhpometric analyses of sectioned bovine adrenal medulla revealed that the chromaffin granules on an average account for approx. 13.5% of the cytoplasmic volume in the total population of chromaffin cells.


Nucleolar fibrillar centers in mouse spermatid nucleoli. The nucleoli of young spermatids of mice are described. They exhibit a very special shape resembling a "padlock" in which three different areas can be distinguished: (a) a compact zone corresponding to the fibrillar component, (b) the granular component and (c) a fibrillar center of low density. Fibrillar and granular components usually appear segregated. This nucleolus has been reconstructed based on serial sectioning. When the silver impregnation technique is employed, both fibrillar and granular components show a positive reaction, although the fibrillar center is free of granules. The morphology of the fibrillar center seems to be similar to that reported in other cells. The possibility that these fibrillar centers correspond to the nucleolar organizer is discussed.


Lamellar bodies at the cell periphery of human muscle fibers. The cytochemical and ultrastructural features of lamellar bodies in human skeletal muscle fibers were studied using tannic acid-glutaraldehyde, ruthenium red-glutaraldehyde fixation methods, conventional electron microscopy and the freeze fracture technique. The lamellar bodies consisted of concentric lamellae with a regular spacing of 6.5 +/- 0.2 nm. These structures were found preferentially at the cell periphery closely associated with the plasma membrane, near the nuclear poles and in the space between muscle fiber and satellite cell. The cytochemical and ultrastructural features of the lamellar bodies suggest they are largely composed of phospholipid. It is possible that these structures are involved in muscle membrane maintenance.


Ultrastructure and evolution of a sperm: phylogenetic implications of altered motile machinery in Ophryotrocha puerilis spermatozoon. Modifications to the mitochondria and flagellum of the Ophryotrocha spermatozoon render it immotile. The sperm may represent an evolutionary unstable intermediate between flagellate and aflagellate sperm types.


A morphometric analysis of microtubules in relation to the inhibition of lysosome movement caused by colchicine. Colchicine was administered intraperitoneally to rats in doses which are known to inhibit the basal migration of lysosomes in uterine epithelial cells. The fractional volume of microtubules in the cells was then measured by morphometry. Colchicine at 0.10 mg/kg reduced the microtubule content of the cells from 0.22% down to 0.15%, and 1.0 mg/kg reduced microtubule content to 0.03%. Microtubules were essentially absent from the cells after colchicine doses of 3.0 and 10.0 mg/kg. The microtubule content of uterine epithelial cells thus decreased in the colchicine dose range from about 0.10 to 1.0 mg/kg, the same dose range in which an inhibition of lysosome migration has been observed. These results support the suggestion that microtubules are necessary for the basal migration of lysosomes in uterine epithelial cells. In addition, colchicine at 1.0 mg/kg caused a redistribution of the Golgi complex and a class of electron-transparent, 130 to 450 nm vesicles. These organelles were restricted to the apical halves of the cells in untreated rats, but they were dispersed throughout the cells after drug treatment. The change in the position of the organelles may be caused by a loss of cytoskeletal function of the microtubules.


Radio-isotope bone scanning in suspected osteomyelitis in children. The usefulness of radio-isotope bone scanning in suspected osteomyelitis has been widely acclaimed. Fourteen children had rectilinear bone scans performed three hours after injection of Tc99m methylene diphosphonate. A diagnostic accuracy of 56% was achieved, which is lower than in other series. The reasons for this are discussed and the value of bone scanning in the evaluation of osteomyelitis is questioned.


113mIn-tripolyphosphate: a new radiopharmaceutical for bone scanning. A compound was synthetized, which by X-Ray diffraction studies proved to be hexahydrated sodium tripolyphosphate. By means of in vitro procedures, it was demonstrated that this compound was able to complex trivalent cations, and that 113mIn could be completely incorporated to it. In vivo studies in rats, and scans performed with this new radiopharmaceutical in humans showed its highly selective accumulation in metastasic bone tumors.


A single tracer method for background subtraction in 197HgCl2 renal uptake measurements. The 197HgCl2 renal uptake was measured in 70 patients in order to study the degree of the background activity and to determine the best correction method. The deduction of background activity was made: a) From a concentric perirenal region of interest with no additional correction. b) Using a double tracer technique with colloid 99mTc, subtracting consecutively the liver and spleen activity and the residual background. Comparison of the results revealed: a) The correction of background activity significantly altered the uptake estimations, and thus must include both the liver and spleen activity, and the residual background. b) The choice of a median region of interest for the residual background evaluation led to a systematic error. c) The assessment of background activity, based upon a perirenal region of interest provided a sufficiently precise and reliable correction of the total background, while avoiding the inconveniences associated with the double tracer method.


Calculation of net renal uptake curves without use of a blood pool agent. The usual method of generating net Hippuran renal uptake curves involves the administration of a blood pool agent. Two techniques are presented which permit the determination of net renal uptake curves from data intrinsic to the Hippuran study. This reduced the complexity of the procedure, as well as reducing its cost and decreasing the radiation exposure. The new procedures were tested in 9 patients and showed excellent correlation with the blood pool agent technique.


The application of radioactive labeled leucocytes for the proof of inflammations. The methods we have tested are reliable for the labeling of granulocytes and monocytes as well as with colloidal substances and with soluble 99mTc-phytate. Supravital colorations and electronmicroscopic pictures do not show any damage of the cells caused by the labeling procedures. In the clinical application after the reinjection of the labeled granulocytes and monocytes, inflammations in the abdomen and in the bone could be proved and localized exactly. A final evaluation of the described methods and its differentiation from other diagnostic possibilities will only be possible after further investigations.


Comparison of two different biliary agents in healthy subjects and in patients with liver disease. The kinetics of the iminodiacetic acids, dimethyl and diethyl IDA were studied in patients without and with liver disease. Diethyl IDA seems to be superior to dimethyl IDA in its diagnostic usefulness. It shows lower urinary excretion and a faster liver uptake, leading to more pronounced differences in liver peak time and in liver elimination half time and to faster visualisation of the gall bladder in patients with liver disease. The enterohepatic reabsorption is negligible as shown by enteral administration of the radiopharmaceutical and monitoring blood activity. Therefore it is not necessary to correct time activity curves of the liver for the IDA secreted into the gastrointestinal system. The most valuable diagnostic information with this substances may be gained in investigations on cholecystectomized patients with cholestasis.


The stability and distribution of Tc-HIDA in vivo and in vitro. Tc-HIDA is a promising new agent for imaging the biliary system. In this study radiochromatography on paper or Sephadex G25 gel has been used to measure the proportions of TcHIDA, 'hydrolysed Tc' and pertechnetate in solutions from a commercial kit (CIS TCK 15) and in body fluids from patients and rats receiving this radiopharmaceutical. The tissue distribution in male and female rats has shown the radiopharmaceutical to be rapidly removed from the blood by the liver and then excreted via the intestines. There appears to be a sex difference in the uptake in the kidneys and in the urinary excretion in both rats and patients.


Quality control of colloid and particulate 99mTc-labeled radiopharmaceuticals. A procedure for the radiochemical purity control of colloid and particulate 99mTc-labeled radiopharmaceuticals is described. The proposed technique is based on the use of two chromatograms, using in both, 15% H3PO4 as solvent and impregnated glass fiber media (Gelman ITLC type SG) as stationary phase. A pretreatment of the radiopharmaceutical with 6 N NaOH is involved prior to one chromatographic run. The procedure is fast and the different species (free pertechnetate, 99mTc-Sn-colloid and labeled 99mTc) in a 99mTc-labeled radiopharmaceutical can be determined accurately and with reliability.


Radioimmunoassay of lactate dehydrogenase, H forms. Antisera to H4-lactate dehydrogenase (LDH) were elicited in rabbits, against both human (h) and porcine (p) isoenzymes. 125I-labelled H4-LDH was prepared by electrolytic iodination. A simple and fast procedure (1-h incubation for clinical assays) was set up by using polyethylene glycol for the bound-free separation. The results obtained in the antiserum characterization indicated that the heterologous homotetramer, M4 was completely discriminated in the porcine system, while a weak cross-reaction with human antisera resulted. In both cases, for the hybrid forms, a cross-reactivity level related to the stoichiometric contents of the H-subunit in the tetramers was observed. The H4-LDH from other species was found to be much more effectively disinguished in the porcine than in the human system. The assay for human LDH was further validated in terms of analytical suitability and clinical response. For healthy subjects the mean concentration was 0.46+/-19 micrograms/ml (mean+/-SD). Patients with acute myocardial infarction had levels ranging from 1.2 to 5.9 micrograms/ml.


Results of regular study on radionuclidic purity of 99mTc obtained from 99Mo--99mTc generators. A total of 39 99mTc eluates obtained from 9 99Mo--99mTc generators delivered by The Radiochemical Centre Amersham during one year was studied with regard to their radionuclide purity. Using a Ge(Li) spectrometer the contaminants 60Co, 103Ru, 131I, 134Cs, 140Ba, 140La and 188Re were found in 99mTc-eluates with average levels ranging from 2.9 X 10(-3) to 2.8 X 10(-1) per cent of 99mTc activity. The additional total body absorbed dose caused by these contaminants, as calculated from their average content in 99mTc eluates, was less than 1% of the dose due to 99mTc pertechnetate.


Radiation exposure due to 99mTc and 131I manipulated in syringes. Dose rates around and at the surface of disposable polypropylene syringes of 1 and 2 ml volume filled with 99mTc and 131I solutions were determined by thermoluminescence dosimetry. Data for the efficacy of 0.5 and 1 mm lead shielding of 99mTc and for the dose contribution due to beta- -radiation from 131I are given.


Camera aperture to optimize data collection in nuclear medicine. Collection of data with a large field of view camera can cause problems when a small organ like the heart is to be imaged, especially when high activity is used. A simple, inexpensive mask is described that solves most of these problems.


Relationship between blood flow and radiostrontium uptake in the healing bone fracture. The healing of a diaphyseal tibia fracture was followed in the rat. Callus formation, blood flow and Sr-85 uptake were assessed by a ratio comparison of the fractured to the contralateral side. No correlation was found between blood flow and Sr-85 deposition in the callus and the adjacent bone of the same extremity. It is concluded that the deposition of a radiopharmaceutical in bone is primarily related to the presence of calcifiable organic bone matrix and only secondarily to the bone formation rate and to variations in the local blood flow.


Tumoral calcinosis: accumulation of bone-seeking tracers in the calcium deposits. Rapid and preferential uptake of 47 calcium 87m Strontium by the multinodular calcareous masses was seen in a patient with tumoral calcinosis. Using 87m Strontium, scintigraphic imaging of the tumors was achieved, while the skeleton was barely visible. More intense accumulation of 87m Sr. in one tumor suggested a higher rate of calcium depostion. This was in agreement with the observation that this tumor showed less tendency to decrease in size during calcium and phosphorus deprivation therapy. Quantitative measurements of the uptake of bone seeking tracers by the tumors may be of value in estimating calcium turnover in the calcified masses.


The short-term fixation of radiopharmaceuticals in bone. The relationship between local blood flow and bone clearance of 18 F, 32 P-P (orthophosphate), 32 P-PPi (pyrophosphate), 45 Ca, 85 Sr, 99mTc- and 113 Sn-EHDP was evaluated in the non-adult rat. During one single blood passage nearly 100% of 18 F was extracted in the femoral diaphysis, the extraction efficiency for the other radiopharmaceuticals was about 40%. Blood flow and bone clearance was higher and the extraction efficiency lower in the distal femoral epiphysis including the growth plate. The short-term bone uptake of 32 P-P was lowered considerably by the simultaneous application of inactive orthophosphate, whereas the bone uptake of 32 P-PPi was not suppressed. It is suggested that the rapid deposition of radiopharmaceuticals in bone is not caused by an ionic exchange process on crystal surfaces of already existing mineral but is due to an ionic exchange during the precipitation of calcium phosphate in the calcifying bone matrix. Only 18 F is exchanged with hydroxyl groups of already precipitated hydroxyapatite, whereas the calcium and phosphate ions in the precipitated hydroxyapatite are not further accessible to an exchange process.


A rabbit model to study the biokinetics of radiopharmaceuticals in bone. A technique is described using the rabbit as the 'animal model' to measure the rate of extraction of the bone-seeking agent, technetium labelled ethane-1-hydroxy-1, 1-diphosphonate (99Tcm-EHDP) following constant-rate infusion into the femoral nutrient artery. The extraction is shown to be approximately linear for a period of 60 min. Washout of the tracer from bone was followed for a further 60 min period and graphical analysis shows that each curve could be resolved into two distinct exponential functions. The 2-h residue of 99Tcm-EHDP in bone has been examined and is found to be dependent upon its route of administration. A high residue (43.7 +/- 5.1%) results when the infusate is delivered directly into the nutrient artery and the single muscular branch is tied off. Retrograde infusion via the femoral artery results in a lower residue (7.8 +/- 2.3%). The close correlation of these results in the rabbit with our previous findings in the dog suggested that the rabbit model described can be used to study the kinetics of other radiopharmaceuticals during their passage through bone.


Absorbed dose estimation of some 99mTc-hepatobiliary agents. The absorbed dose estimates of several organs after administration in humans of six new hepatobiliary agents (five 99mTc-Pyridoxal amino acid complexes and the 99mTc-HIDA) are presented in comparison with 131I-Rose-Bengal. The results indicated that the radiation doses absorbed by the total body and gonads as well as by the critical organ (upper part of the large intestine) are significantly lower than those of Rose Bengal. Therefore, the new 99mTc-agents can be safely applied to humans.


Synthesis of five new 99mTc-HIDA isomers and comparison with 99mTc-HIDA. Five new HIDA-compounds were synthesized, labelled with 99mTc and compared with 99mTc-(2,6-dimethyl)HIDA viz. (N-2,6-dimethylphenylcarbamoylmethyl)iminodiacetic acid by means of TLC and biodistribution studies on rabbits. The computerized method used by Britton and Brown during renal studies was successfully adapted to study the biodistribution of these agents.


Adrenal scintigraphy with 131I-19-iodochlesterol in the diagnosis of Cushing's syndrome associated with adrenal tumor. Seven patients with Cushing's syndrome secondary to adrenocortical tumors were studied using 131I-19-iodocholesterol. The diagnosis of all cases were verified histologically. In three cases with adenoma the uptake of the tracer was in the tumor only, while the two patients with adrenocortical carcinoma failed to show adrenal accumulation of the labelled compound. In two patients there was a hyperplasia-like scintigraphic pattern, while the stimulation and suppression biochemical tests suggested adrenal tumor. One of these cases was verified as a mixed form (adenoma plus hyperplasia), and the tumor bearing gland was significantly larger on the scan which helped the preoperative localization. In the other case, verified as bilateral multiple adrenocortical adenomas, the autonomus function of both adrenals was proved by dexamethasone suppression scanning. It seens reasonable to use the latter as an adunctive diagnostic procedure in patients where there is a discrepancy between the standart scintiscan and the biochemical indexes of adrenal hyperfunction.


Dose dependent uptake of 125I-toluidine blue of 75Se-selenomethionine in the parathyroids of rabbits. Different dosages of 125I-toluidine blue and 75Se-selenomethionine were injected intravenously into rabbits, and the concentration of radioactivity was measured in the parathyroids and surrounding tissues. The response of i.v. injected glucagon was also determined. It was found that 0.16 mg 125I-toluidine blue gave a concentration ratio of parathyroid/thyroid and parathyroid/muscle which was significantly higher than both lower and higher doses at 1 minute after i.v. injection of the labelled dye. Glucagon had little or no effect on the uptake of 125I-toluidine blue and 75Se-selenomethionine.


Transverse CAT of the myocardium with 201 T1. A new computerized axial tomographic (CAT) scanner (J&P Tomoscanner) which enables the transverse section viewing of any organ labelled by one of the monophotonic gamma-ray emitting racers commonly used in Nuclear Medicine, has been recently commercialized. Its abilities in visualising the 201 T1 labelled myocardium are evaluated with normal hearts and patients with documented infarctions. Positive results have been obtained, allowing an interesting approach to the estimation of the anatomical extent of necrosis.


RIA system programming by means of kinetic parameters. Insulin-125I antibody reaction was optimized by physical-chemical parameters. After the activation energies Ea and Ed for association and dissociation, respectively were calculated from the experimental data, the theoretical values of the reaction rate constants ka and kd were determined as well as equilibrium constants K. By means of the empirical formulae, the approximate incubation time for the RIA kit and maximal percent of insulin-125I binding to antibody (%B) in relation to temperature were computed. The proposed method may be applied to the new antigen-binder systems preparation (new antibodies, shortening of the incubation time, temperature changes, influence of different ions and kind of buffer).


Calculation of residence time distributions of intravascular radioactive tracers in fields of external registration. Because of the physiological significance of the mean velocity of blood flow, indicator dispersion models are of special interest and possess practical relevance, if biological and extraneous variables can be altered. The variables being considered are flow characteristics of the streaming blood, tracer distribution at the entrance into the flow system, and the area in which impulses are collected to form the time-activity curve. Using a simplified version of the general convective diffusion equation (diffusion model) in which the diffusion constant D includes all propagation and mixing of the tracer, a simple numerical method can be applied. The method is used to determine influences of injection and changed regions of interest on the time-activity curve and the following parameters: appearance times, peak times, mean circulation times, and the times of the first inflection points. For this purpose, the range of D was determined in 14 patients by applying experimental data to the model. The calculations on the variables show, that the advantage of this method is its applicability to any experimental case by simply adapting the input data to the recordings.


Sensitometry of films for nuclear medicine. A method is described for the determination of characteristic curves for films used in nuclear medicine imaging. Such characteristic curves are presented for a number of commercially available film types and their significance is discussed.


Exostoses induced by 224Ra (ThX) in children. Exostoses are benign cartilaginous tumors of bone. They can occur naturally or be induced by radiation therapy during the time of skeletal growth. We have observed exostoses in 28 of 218 children given repeated injections of radioactive bone-seeking 224Ra. The younger the age at irradiation, the higher the incidence of exostoses. Boys are more susceptible than girls. To our knowledge, none of these radiation-induced exostoses have become malignant, although 36 of these children have developed bone sarcomas elsewhere in the skeleton.


Dubin-Johnson syndrome in a neonate.

Dubin-Johnson syndrome in a neonate. We described the clinical and biochemical findings in a 32 day-old boy with the Dubin-Johnson syndrome. Only two other patients diagnosed as having the Dubin-Johnson syndrome during neonatal period have been reported in the literature. The ratio of urinary coproporphyrin isomer I of our patient was 97% and that of his parents were carrier level, confirming that increased urinary excretion of coproporphyrin isomer I is of diagnostic value in neonates with the Dubin-Johnson syndrome.


Anxiety and depression in young and middle aged hypertensive and normotensive subjects. This study examined the effects of essential hypertension on measures of anxiety and depression for two age groups of hypertensive (free from hypertension-related pathology and/or cardiovascular disease) and normotensive subjects. Hypertensive subjects had significantly higher State Anxiety scores and Zung Depression scores than did normotensive subjects. These differences between the blood pressure groups were due largely to the scores of the younger hypertensive subjects. The results of the present study are consistent with previous results from our laboratory that have found that younger hypertensives differed (relative to controls) from middle aged hypertensives on measures, such as, symptoms reported on the Cornell Medical Index and WAIS Performance scores. The results of the present study were discussed within the context of age associated differences in response to hypertension and factors that might account for these differences.


Comprehension and memory of prose materials by young and old adults. Twenty-seven young (X- = 27.3 years) and twenty-seven old (X- = 67.7 years) female volunteers participated in a study of the effect of vocabulary level upon memory and comprehension of prose materials. Comprehension was measured by presenting five multiple choice questions simultaneously with half of the prose passages. Memory was measured with the rest of the materials by presenting questions only after a passage was completed and removed from sight. The results indicated that age-related differences varied inversely with vocabulary levels but did not interact with type of task. No significant differences were found between age groups at the highest vocabulary level. The parallel findings for the two tasks suggest that inadequate comprehension represents one major factor underlying age-related differences in the retention of meaningful materials. A second memory task, not dependent upon accuracy of comprehension, indicated age-related differences at all vocabulary levels. This suggests that old adults may also hve deficits in the ability to maintain in store and/or retrieve meaningful materials.


Future time perspective in later adulthood: review and research directions. The degree of experimental and quantitative detail which can be imposed on older adults' temporal experiences remains a significant area for investigation. This paper surveys material related to this general area of study, taking primarily a psychological focus. Review of this literature suggests that while a basic future time orientation is not significantly affected in later life, older adults are at-risk of developing a less extended and less full personal future. However, research has been almost exclusively cross-sectional,, and our empirical knowledge concerning the functional impact of such changes, should they occur, is minimal. In addition, although several correlates of older adults' future time perspective have been reported, normative directions of causal relationship have not been determined. Consequently, while much of our data are varied, interesting, and insightful, they are at present difficult to organize or apply. Research should be pursued on several fronts, including prospective designs, the use of time perspective as a predictor and outcome variable, model development, and close association with temporal concepts in social gerontology.


Q-technique methodology in the study of healthy aged men: Part I. The paper describes the application of the Q-technique to the study of healthy aged men. The research is an NIMH project, and the Q-technique was performed on the initial study done in 1957 (47 Ss across 550 variables). The paper focuses on clusters of interdisciplinary relationships which contained biological and cognitive influences. Twenty-eight out of the 64 clusters fell within this category. This first paper concentrates on methodological precedures and issues, and also presents 6 clusters of output derived from the first two Q-factors. The remaining 22 clusters will be described in subsequent papers.


Measurement of self-concept and self-esteem in older people: state of the art. Self-concept and self-esteem instruments used in gerontological research were reviewed. Instruments were found to vary substantially in their conceptualization of self-concept and/or self-esteem as well as in the amount of attention given to assessing their psychometric properties. The assumptions made about the relationship of age to self-concept and/or self-esteem also varied. More careful attention to selection, construction, interpretation, and continued validation of instruments is recommended. Several instruments are particularly recommended for further use in gerontological research.


Formal education and intellectual functioning in the immigrant Cuban elderly. Formal educational experience during childhood and adolescence is an important modifier of intellectual functioning in North American elderly populations. The culture-independence of this relationship was examined with 31 immigrant Cuban elderly men and women who received their education in Cuba. The amount of formal education was significantly related to performance on measures of classification, combinatorial analysis (males only), hypothesis testing, and matrices. An index of fluid intelligence (matrices) also was significantly related to performance on the classification and hypothesis testing tasks. All participants conserved both liquid and mass, thus showing no regression on these two concerete operational tasks.


Effects of word frequency on adult age differences in word memory span. The present study tested the hypothesis that the age deficits in memory span commonly found in elderly adults are attributable, at least in part, to elaborative rehearsal processes. The test required an analysis of age differences between young and elderly adult subjects in word span for list types varying in word frequency. Significant age deficits were found for all list types, with the magnitude of the deficit increasing as the frequency of intrastring items increased. The pattern of this interaction indicated the involvement of a decreased proficiency in elaborative rehearsal with increasing age as a contributor to the age deficit in word span.


Minority women's attitudes about aging.

Minority women's attitudes about aging. Many factors combine to make middle age a critical period with respect to views about aging. To examine the effects of culture, background, and familial characteristics on attitudes toward old age, 304 middle-aged women from Black, Cuban, American Indian, Chicano, and white cultures were studied. A stepwise multiple regression analysis examined the contribution of each variable in the prediction of attitudes toward old age. Attitude toward death, attitude toward family, and church affiliation were found to be important predictors of attitude toward old age. Thus, clinicians dealing with middle-aged women having negative attitudes or fears about aging and old age in general may need to examine family dynamics as well as views about death and dying for possible problems. Where religion is already an integral part of a client's life, perhaps clinicians could use this orientation for added support.


Information feedback, reaction time and error rates in young and old subjects. Reaction times and error rates in a letter classification task were examined in two groups of subjects, one with a mean age of 20.7 years, the other with a mean age of 63.6 years, under conditions where specific feedback regarding the speed and accuracy of performance on each trial either was or was not provided. Feedback produced a speed-accuracy trade-off for younger subjects. This finding offers no support for the position that feedback results in a general improvement in performance. Older subjects did not show a speed-accuracy trade-off. Rather, feedback resulted in trends toward both longer reaction times and higher error rates. These results are interpreted as showing that feedback has a phasic alerting effect in the younger subjects, but not in the older subjects.


Age, sex, and hemisphrere asymmetry differences induced by a concurrent memory processing task. Subjects in three age groups matched simultaneously presented pairs of visual words or geometric shape stimuli as belonging in a "same" or "different" category. No age effects were observed unless subjects were also required to repeat the sequence of the last three "same-different" responses immediately following each trial. Cerebral asymmetry effects were expected for the word and shape dimensions, but were not observed. Asymmetry effects for the same-different dimensions were noted and allowed inference regarding serial and parallel processing effects. When required to repeat the sequence of past responses following each trial, older males showed an increase in time to make the same judgments. Older females showed increased right vs left visual field processing times when required to repeat the sequence of same-different responses following each trial.


Intellectual and personality differences between community-living and institutionalized older adult females. The purpose of the present study was to investigate simultaneously differences between normal institutionalized older adults and community-living older adults with respect to intelligence/cognitive test performance and personality. Participants were 25 community-living females (M age = 72.9 yrs, SD = 6.34) and 25 institutionalized females (M age = 80.0 yrs, SD = 6.46). Intellectual/cognitive ability was assessed by the WAIS, Stanford-Binet Intelligence Scale (Form L-M), Ravens Coloured Progressive Matrices; personality was assessed by the Hand Test, a projective technique. Several multivariate analyses (discriminant analysis) were conducted. Results suggested that even when controlling for age and level of education, institutionalization appears to be associated with intellectual/cognitive as well as personality deficits. The findings were discussed in terms of the potential implications for the professional working with institutionalized older adults.


Dimensional preferences in middle and old age. Dimensional preferences in 40 middle-aged (M = 41.62 years) and 40 elderly (M = 72.22 years) females were assessed using a dimensional choice task. Significant age differences in reaction times of choice but not in number of dimensional choices were obtained. There was a perfect rank-order correspondence between the two age groups in dimensional choices with form being the most preferred and color the least preferred perceptual dimension.


Stimulus persistence and age.

Stimulus persistence and age. Stimulus persistence theory states that the effects of stimulation are longer lasting in the nervous system of the old person than the younger one. This concept and its relation to perceptual functioning and age was examined via two perceptual aftereffect tasks, the Spiral and the Waterfall. Twenty-four elderly adults (mean age = 65.5) and twenty-four young adults (mean age = 24.8) were each exposed to both stimulus displays, at four exposure durations: 45, 60, 90, and 120 seconds. Following the offset of each display, the duration of any apparent motion aftereffect was recorded. Although on the basis of stimulus persistence theory it was expected that the elderly would exhibit longer mean aftereffects than the young, no support for this hypothesis was noted. Possible explanations of these results and their relation to previous findings, in terms of fatigue effects and sample characteristics, are discussed.


EEG alpha slowing, refractory period, and reaction time in aging. The present study was undertaken to determine if a model accounting for reaction time (RT) on the basis of the EEG alpha rhythm and refractory period (RP) could predict RT in young and old adult subjects. Ten male university undergraduates and ten community residing males over  the age of 65 had resting EEG recorded at right and left occipital sites. Mean alpha frequency was 1.2 Hz slower in old subjects. RT and RP were assessed in a warned  RT paradigm. Sixty-four trials of each of a 50, 100, 250, and 500 msec inter-stimulus interval (ISI) were presented in random order. The model was useful in predicting the magnitude of the age difference in RT and the difference in RT between the 50 and 100 msec ISI. However, old subjects were not disproportionately slowed at the 100 msec ISI as the model would predict and all RTs were underestimated. Thus, while a model using EEG alpha period and RP to predict age differences in RT has some weaknesses, it is useful enough to warrant additional examination.


Effect of ergot alkaloids on serum prolactin in non-psychotic organic brain syndrome of the elderly. A compound of ergot alkaloids (Hydergine-Sandoz) or placebo was given to sixty elderly nursing home patients with non-psychotic organic brain syndrome. Subsequent to 12 weeks of treatment, the mean serum prolactin of the drug was significantly lower than that of the placebo group. No correlation was found between changes in prolactin and changes in behavior using Sandoz Assessment of Clinical Status Rating Form-Geriatric [SCAG]. It may be that a higher dose of Hydergine with an accompanying greater drop in prolactin would be required to observe this effect.


Relationship of age and hypertension to neuropsychological test performance. Young adult (X- = 29) and middle aged (X- =50) hypertensive and normotensive subjects were compared with respect to seven neuropsychological test scores derived from tests on the Halstead-Reitan battery. Age main effects, with inferior performance for the middle aged subjects, were observed for the localization and time portions of the Tactile Performance Test (TPT) and for the Trail Making A test. The multivariate age effect was significant for the composite of seven scores. A multivariate blood pressure main effect was obtained and main effect blood pressure was significant for the category test; hypertensives made more errors than normotensives. A blood pressure by age interaction was observed for finger tapping scores and the TPT-Memory scores with larger differences between hypertensives and normotensives for the younger than for the middle aged group. Results were discussed in terms of previous studies of age and hypertension with the WAIS, the Primary Mental Abilities Test and serial reaction time measures. The poor prediction of hypertensive status from individual neuropsychological test scores was emphasized and readers were cautioned not to conclude that essential hypertensives, as a group, can be characterized as brain damaged.


Age dependent changes in the sperm population and fertility in the male rat. The entire reproductive tract was devoid of spermatozoa during the first 42 days of life. The first appearance of spermatozoa was detected in the caput epididymidis of 45-day old rats and in the cauda epididymidis of 52-day or older rats. The number of spermatozoa in the reproductive tract increased with age. The sperm population reached its maximum in the caput epididymidis by day-72 and in the caud epididymidis by day-100. These high levels were maintained beyond 450 days of age. Fifty-two day old male rats failed to impregnate the female. Although male rats of 62-500 days were able to sire litters, the peak reproductive period was found to be between days 100-270, during which the number of young per litter ranged between 11-16. Towards advanced age the litter size dropped to 8. The lower fertility in the males of advanced age might be due to a lower proportion of fertile spermatozoa, a change in the physiology of epididymis or an altered sperm transport in the reproductive tract.


A questionnaire assessment of age differences in retention of recent and remote events. Age-related differences in memory for recently and remotely dated events have been evaluated through questionnaire assessment. Results from previous studies employing the technique differ in the analysis of retrieval of recent and remote events. In attempting to reconcile extant results from the literature, the following study employed a group of healthy males ranging in age from 26 to 69. The results showed: 1. The elderly exhibited no impairment of the ability to remember recent colloquial events. 2. The elderly's surprisingly accurate recall of remote events was not a product of selective and frequent rehearsal.


Age differences in dark-interval threshold across the life-span. The threshold for the dark interval between two flashes was used to examine differences in stimulus persistence in 72 subjects aged 20 to 79 years. The data were then combined with results from two previous studies involving children and adolescents; thus, a cross sectional, adult, life-span view of differences in the dark-interval threshold was presented. Results showed that during childhood and adolescence there is a decline in stimulus persistence as a result of decreased receptor sensitivity. In adulthood, however, the dark-interval threshold remains fairly stable until the 70s where it increases significantly. Two possible explanations for this increase in stimulus persistence in old age are discussed.


A brief note on a preliminary study of life satisfaction for male university faculty of differing chronological ages. Male faculty of a large southeastern university were surveyed in order to determine the relationship between life satisfaction and chronological age for a relatively homogeneous population. It was found that satisfaction increased linearly with age for this group. However, the relationship was so slight it was suggested that differences in chronological age did not account for a meaningful amount of the variance in life satisfaction. This is consistent with results obtained for larger segments of the population. Suggestions for further research are provided.


Effects of labetalol and propranolol on blood pressure at rest and during isometric and dynamic exercise. The influence of intravenous labetalol and propranolol on the blood pressure response to isometric and dynamic exercise was examined in a double blind study in eight, young, normotensive volunteers. Effects were recorded after propranolol 7.5, 15 and 30 mg i. v., and after labetalol 30, 60 and 120 mg i. v. In control experiments saline was administered. Mean blood pressure rose with successive handgrip tests following saline and propranolol, but not after labetalol, and the difference was significant. The total dose of each drug produced the same reduction in heart rate during sub-maximal bicycle exercise. The exercise-induced systolic blood pressure response did not differ between the drugs.


Effect of oral labetalol on plasma catecholamines, renin and aldosterone in patients with severe arterial hypertension. Arterial blood pressure and plasma catecholamines, renin activity and aldosterone concentration in 12 patients with severe essential hypertension were studied before and after combined alpha- and beta-adrenergic receptor blockage induced by oral labetalol treatment for 2 months. Furosemide in a fixed dose was employed as a basic antihypertensive agent throughout the study. Blood pressure was adequately controlled in only 6 patients. Mean body weight increased by 1.8 kg and there was a rise in body weight which was inversely correlated with the fall in standing mean blood pressure. The mean plasma noradrenaline concentration decreased from 0.30 to 0.20 ng/ml, whereas plasma adrenaline did not change significantly. Plasma renin activity and aldosterone concentration varied greatly, but the mean values did not change significantly. Change in body weight was correlated inversely with changes in plasma noradrenaline and renin. The results suggest that labetalol, through its combined alpha- and beta-adrenergic receptor blocking action, induces a rise in body weight, probably due to sodium and fluid retention, which partly counterbalances the antihypertensive effect of labetalol, and partly modifies both renin and sympathetic nervous activity.


Food-induced reduction in bioavailability of atenolol. The influence of food intake on the bioavailability of the beta-adrenoceptor blocker atenolol was assessed by measurement of its single-dose kinetics in ten healthy volunteers, who took 100 mg both in the fasting state and together with a standardized breakfast. Food intake significantly shortened the time to reach peak concentration (2.7 h vs 1.5 h), but caused a significant reduction in AUC values, the mean decrease being 20%. The elimination half-life was unaffected. Atenolol, which is relatively hydrophilic, is incompletely absorbed in the fasting state, and escapes first-pass metabolism. The present findings indicate that food intake causes further impairment of its absorption, even though the absorption rate may initially be enhanced. This contrasts with previous observations on the more lipophilic beta-adrenoceptor blockers propranolol and metoprolol.


Multicompartment pharmacokinetics of netilmicin. The pharmacokinetics of a single dose of netilmicin (NM) was studied in 6 healthy volunteers. Elimination of the drug was followed in serum and urine for 24 h and 72 h, respectively. NM concentrations were measured with a modified radioenzymatic assay. A three compartment open model was employed to calculate the pharmacokinetic parameters. Following the rapid initial distribution, biphasic elimination with half lives of 1.99 (t 1/2 beta) and 36.89 h (t 1/2 gamma) was demonstrated. Measurable amounts of NM were excreted in the urine for up to 72 h. The volume of distribution at steady-state (Vdss) of 0.68 l/kg was 3 to 4 times larger than previously reported for this antibiotic. NM plasma clearance was 91 ml/min and the renal clearance was 67 ml/min. The data indicate that on repetitive dosing the amount of drug in the body would be considerably underestimated if the prolonged terminal elimination phase were not taken into account. During prolonged treatment, accumulation of NM in renal and other tissues is likely to occur, as has been described for other aminoglycosides. The possible consequences of this pharmacokinetic behaviour are discussed.


The binding of gitoxin to human plasma proteins. The binding of gitoxin, digitoxin and digoxin to human plasma proteins was measured by ultracentrifugation and equilibrium dialysis. At concentrations in the range of therapeutic plasma levels, protein binding amounted, respectively, to 85, 92 and 20%, the last two values being consistent with data reported in the literature. The affinity of purified human albumin was not significantly different for the three cardiac glycosides tested. No other protein than albumin was found to bind gitoxin in human plasma.


Bioavailability of oral testosterone in males. Twenty-six male volunteers received a single oral dose of testosterone as free crystals or as the undecanoate ester. The latter was given either in crystalline form or in arachis oil. All preparations were tested three times in the same individual, whilst fasting on 2 days and on one day together with a breakfast rich in fat. Serum testosterone concentration was measured at intervals for up to 6--24 h after the dose. A significant and reproducible rise in serum testosterone level was found after ingestion of free testosterone. Testosterone esterified with undecylenic acid was only effective when administered in arachis oil. The meal increased the bioavailability of the ester, but had hardly any effect on that of the free hormone. It is concluded that bioavailability of oral testosterone can be improved by pharmaceutical means to an extent sufficient to produce adequate blood levels in substitution therapy.


Method of estimating the disposition of drugs in the Danish population by sampling reimbursed prescriptions. A method by which for a small cost it is possible to obtain a quantitative expression for the utilization, disposition and cost of drugs in a Danish county of 500,000 inhabitants. The procedure makes it possible to follow reimbursed preparations, i. e., drugs considered essential for the treatment of diseases. The sale, in D kr., of reimbursed preparations to individuals comprises about 80% of the sale of preparations requiring prescription. And in turn the sale of the latter amounts to 80% of total drug sales to single subjects. The method is based on systematic utilization of all the information available on each prescription; concepts of 'Entry Parameters', 'Calculated Units' and 'Derived Units' was discussed; the 'units' concern frequency of prescription and age- and sex specific dose-frequencies (weight units or defined daily doses -- DDD's). The possibility of creating a base for calculation of the frequency of certain diseases and of side-effects of drugs is mentioned, and examples of the application of the method to certain categories of patients and to certain drugs are given. From limitations of the method it is concluded that it should be considered as a supplement to other complete methods for the study of drug consumption.


Cytotoxicity of human mononuclear cells against chicken and human red blood cells, induced by treatment of the effector cells with phospholipase C. Human mononuclear cells from peripheral blood which were treated with phospholipase C (PLC), became cytotoxic against human or chicken red blood cells. PLC-induced cellular cytotoxicity against human red blood cells was further analyzed and compared to anti-D-mediated, antibody-dependent cellular cytotoxicity (ADCC), using the same target cells. ADCC, but not cytotoxicity of PLC-treated effector cells, was inhibited by free IgG. In addition, iodoacetate strongly enhanced PLC-induced cytotoxicity, but blocked ADCC completely. Addition of fetal calf serum or human AB serum impaired PLC-induced cytotoxicity. A similar inhibition was found by adding lecithin liposomes suggesting that the inhibitory effect of sera was also due to their phospholipid content. The data show that cytotoxicity of PLC-treated effector cells can be clearly distinguished from cellular cytotoxicity, occurring spontaneously or induced by target cell antibodies. We favor the notion that cytotoxicity of PLC-treated effector cells against human erythrocytes is due to the action of PLC, adsorbed to the effector cells.


Regulation of tumor growth and antibody clone expression by antigen and anti-idiotype antibody ligands with specificity for receptor-binding sites. Dextran ligands, modified to increase epitope reactivity with receptors, were more effective in suppressing BALB/c mouse plasmacytomas MOPC 104 E and J-558, which bind alpha (1 leads to 3) dextran and have an idiotype (Id) in the common, than autoantibody (Ab) against the Id unique to each of the proteins secreted by the two tumors (the (IdI). BALC/c immunized with 104 E myeloma protein and expressing an antibody response to the 104 E IdI exhibited a specific, anti-104 E IdI transplantation resistance to lethal grafts of 104 E, but not J-558, tumors notwithstanding the shared common Id and similar ability to bind alpha(1 leads to 3) dextran. This autoantibody did not prevent modulation of the 104 E tumor to variant forms or the growth of the variants. On challenge with alpha (1 leads to 3) dextran, the immunized mice expressing the anti-104 C IdI responses failed to express the 104 D IdI-like antibody clone present in the normal, anti-alpha (1 leads to 3) dextran antibody repertoire. Passive, iso-anti-104 E IdI antibody had a transitory suppressive effect on the normal, 104 E IdI-like antibody clone but failed to circumvent 104 E tumor growth. It is apparent that the greater effectiveness of ligands strongly reactive in a nonphysiological manner with the tumor receptors lies in the stabilization of the tumor load without inducing variant escape or a disturbance of the immune network, and that receptor expression and malignancy state are not necessarily co-extensive functions.


Effect of isoproterenol on the isolated pregnant rat myometrium. In pregnant rat myometrium, isoproterenol (10(-8) M) inhibited spontaneous contractions without causing hyperpolarization. Isoproterenol (10(-6) M) relaxed the depolarized muscle without affecting the membrane potential. The presence of 80 mM Na+ did not affect the degree of high-K+ depolarization. It was also without any influence on the effects of isoproterenol on the depolarized uterus. The results are consistent with the concept that hyperpolarization is not a prerequisite for beta-adrenoceptor induced relaxation of uterine smooth muscle.


Glycine stimulates the spontaneous release of newly synthesized 3H-dopamine in rat striatal slices. Glycine (5 X 10(-5) M) stimulated the spontaneous release of 3H-DA continuously synthesized from 3H-tyrosine in striatal slices. This effect was calcium-dependent and significantly reduced in the presence of strychnine (10(-5) M). The glycine-evoked release of 3H-DA was no longer seen in the presence of tetrodotoxin (5 X 10(-7) M).


Antagonism by d,1-propranolol of imipramine effects in mice. Three agents with known or suspected antidepressant activity, imipramine, salbutamol and dexamphetamine, were active in animal tests predictive of an antidepressant effect in man: antagonism of the hypothermia induced by reserpine, by oxotremorine or by a high dose of apomorphine, and the potentiation of the yohimbine-induced toxicity. These effects were antagonized by d,1-propranolol, suggesting that the stimulation of beta-adrenergic receptors could be a common mechanism underlying their effects. These results agree with the noradrenergic hypothesis of the pathophysiology of affective disorders.


The effect of verapamil on the gastrocnemius and soleus muscles of the cat in vivo. The effects of verapamil on the force of contraction of soleus and gastrocnemius muscles of the cat were examined in vivo. The rapid intra-arterial administration of verapamil (100-800 micrograms/kg) produced a dose-dependent increase in the contractile force of soleus and gastrocnemius muscles stimulated supramaximally via their nerves. Verapamil also potentiated twitch tension in d-tubocurarine-treated, directly stimulated soleus preparations. This agent had no effect, however, on the contractile force of 14- to 16-day chronically denervated, directly stimulated soleus muscle. Verapamil (400 microgram/kg, i.a.) also produced a significant increase in muscle action potential duration while having no effect on amplitudes, rise times or resting membrane potential. It is concluded that verapamil acts directly on innervated skeletal muscle to increase muscle force and it is suggested that the probable site of action is at the muscle membrane.


Effects of clonidine and chlorpromazine on a sympathetic-cholinergic reflex. Intravenous administration of clonidine and chlorpromazine resulted in a dose-dependent inhibition of the amplitude of reflexly evoked electrodermal responses in intact and spinal cats. Yohimbine pretreatment (0.5 mg/kg, i.v.) antagonized the effects of clonidine but not chlorpromazine in both preparations. These findings confirm and expand previous observations that both clonidine and chlorpromazine inhibit the amplitude of centrally evoked responses in this sympathetic-cholinergic system. In addition, both drugs appeared to have a spinal site of action. The antagonism of the effects of clonidine by yohimbine suggests that the mechanism of the action of clonidine may be a result of activation of central inhibitory alpha-adrenergic receptors. The failure of yohimbine to antagonize the effects of chlorpromazine suggests that clonidine and chlorpromazine may depress these sympathetic reflexes by different mechanisms.


Antimyoclonic action of clonazepam: the role of serotonin. Clonazepam (5-(2-chlorophenyl)-1,3-dihydro-7-nitro 2H-1,4 benzodiazepin-2-one) (2 mg/kg) reduced a p,p'-DDT-induced myoclonus in mice by 50%. This antimyoclonic action of clonazepam was counteracted by the serotonin (5-HT) receptor blockers methysergide, metergoline and cinnanserin and potentiated by the 5-HT uptake inhibitors fluoxetine and chlorimipramine. Clonazepam (4 mg/kg) reduced plasma tryptophan by 27%, but had no effect on brain tryptopham, 5-HT, 5-hydroxyindoleacetic acid, 5-HT synthesis and 3H-5-HT receptor binding. Clonazepam (10(-5) M) inhibited brain synaptosomal 3H-5-HT uptake by 23% and increased 3H-5-HT release by 24%. However, 2-8 mg/kg of clonazepam administered intraperitoneally had no effect on 5-HT uptake or release. gamma-Aminobutyric acid (GABA) agonists (muscimol, acetylenic GABA, amino-oxyacetic acid) and the GABA antagonists bicuculline and isoniazid had no effect on p,p'-DDT-induced myoclonus. Furthermore, bicuculline did not counteract the antimyoclonic effect of clonazepam. We suggest that the antimyoclonic action of clonazepam is mediated by enhancement of serotonergic rather than GABAergic neurotransmission.


Species variation in dopamine receptor binding. Binding of 3H-spiroperidol, 3H-apomorphine and 3H-ADTN (2-amino-6,7-dihydroxytetrahydronaphthalene) associated with dopamine receptors has been evaluated in corpus striatal membranes of calf, rat and human brains. Substantial species differences are apparent for numberous agonists and antagonists in competing for receptor binding. In general, dopamine receptor antagonists are more potent in rat and agonists more potent in calf. In competing for 3H-spiroperidol binding sulpiride, molindone and metaclopramide show the most pronounced species differences, being 3--10 times more potent in rat and human than in calf. In all three species agonists compete for 3H-spiroperidol binding with Hill coefficients less than one while antagonists inhibit 3H-spiroperidol binding with Hill coefficients of about 1.0. Conversely, 3H-apomorphine and 3H-ADTN binding in all three species is inhibited by antagonists with Hill coefficients less than 1.0 while agonists display Hill coefficients of about 1.0. In general agonists are more potent in competing for binding of 3H-apomorphine and 3H-ADTN than 3H-spiroperidol. However, a small component of dopamine, apomorphine and ADTN inhibition of 3H-spiroperidol binding displays very high affinity (IC50 about 1 nM). In human amygdala 3H-spiroperidol appears to label serotonin receptors predominantly.


Antagonism of cholecystokinin-like peptides by opioid peptides, morphine or tetrodotoxin. Morphine, beta-endorphin, Met-enkephalin, and Leu-enkephalin antagonized intestinal actions of cholecystokinin octapeptide (CCK-8), caerulein, and pentagastrin in a manner partly suggesting physiologically competitive antagonism. Further, these acidic peptides (CCK-8, caerulein, pentagastrin) were much more sensitive to the actions of opioids than was angiotensin. Tetrodotoxin also caused changes in the concentration-effect curves, but these were different from the shifts due to the opioids and differentiated between CCK-8, caerulein, and pentagastrin. Naloxone did not modify the response to CCK-8 and caerulein, but completely abolished the antagonistic influence of the opioids. The potencies of morphine and the opioid peptides as antagonists of CCK-8, were of nearly the same order of magnitude. This and the presence in gut and brain of both CCK-like and opioid peptides suggests the hypothesis that these two groups of peptides interact on both myenteric and central nervous system receptors, and thus are directly involved in the regulation of both intestinal motility and satiety.


Animal maturity influences prostaglandin effects on gallbladder fluid transport and smooth muscle. Prostaglandin (PG) A, E and F compounds applied serosally inhibited net fluid absorption and increased intraluminal pressure in guinea-pig isolated gallbladder. The inhibition was dose-dependent for a given animal body weight but sensitivity varied inversely with animal body weight; relative potencies were PGE2 greater than F2alpha greater than A1. In heavier animals the inhibition was preceded by an apparent increase in fluid absorption, due to fluid extrusion following muscle contraction. Net fluid secretion was observed at higher concentrations of PGE2 or F2alpha in lighter animals. Mucosally applied PGs less potently inhibited absorption, with relative potencies of PGE2 greater than A1 greater than F2beta greater than F2alpha. The spasmogenic effect was estimated by measuring intraluminal pressure (PGE2 greater than F2alpha greater than A1).


Effects of L-glutamic acid and kainic acid on central cardiovascular control. L-Glutamic acid and kainic acid injected into the cisterna magna of dogs, produced a dose-dependent increase in blood pressure and a decrease in heart rate. In contrast, intravenous injection of both compounds was ineffective. The hypertension was probably due to an increase in sympathetic tone as guanethidine prevented the rise in blood pressure induced by central administration of L-glutamic acid and kainic acid. Kainic acid was 1 000 fold more potent than L-glutamic acid.


[Effect of stimulation of splanchnic mechanoreceptors on the activity of purkinje cells (author's transl)]. Climbing and mossy fibre activity in Purkinje cells of cat cerebellum (vermis part of lobules V and VI) were recorded by means of extracellular microelectrodes. Responses were obtained by stimulation of different types of mechanoreceptors in the gastro-intestinal tract: muscular receptors connected with C fibres, peritoneal movement receptors connected with A gamma delta or B fibres, Pacinian corpuscules connected with A beta fivres. The characteristics of the splanchnic fibre projections on this cerebellar area were defined. Splanchno-somatic, splanchno-visceral, and splanchno-cortical convergences were demonstrated.


Anatomtic and physiologic development of the photoreceptor of the kitten. The postnatal development of the photoreceptor of the cat was studied using physiological and anatomical methods. The late receptor potential (LRP) was recorded in vitro and the threshold and maximum amplitude determined. The same specimens used in the electrophysiological studies were then prepared for microscopy, and rod cell outer and inner segment length and diameter, photoreceptor density, and inter-receptor distance were determined. A small LRP was first recorded at 9--10 days, but only at very high stimulus intensities. Thereafter, there was a rapid decrease in the threshold and an increase in the amplitude of the LRP. The threshold reached adult values at 17--18 days, while the amplitude of the LRP was adultlike at 23--26 days. Of the anatomical parameters examined, inter-receptor spacing and rod cell diameters seem to be most clearly associated, respectively, with the attainment of adult LRP threshold and amplitude. Outer segment length was adult-like at 35--43 days of age and thus postdated physiological maturity of the photoreceptor. These observations suggest that the surface area of the rod cell outer segment tips is more critical in the development of the adult LRP than is the number of discs in the outer segment. In addition, changes over time in the mean diameter and length of rod cell inner segments follows the pattern of ontogenetic changes in LRP amplitude. These findings imply a close relationship during ontogeny between the metabolic functions of the inner segment and phototransduction at the outer segment disc.


Visual resolution, contrast sensitivity, and the cortical magnification factor. This study shows that photopic contrast sensitivity and resolution can be predicted by means of simple functions derived by using the cortical magnification factor M as a scale factor of mapping from the visual field into the striate cortex. We measured the minimum contrast required for discriminating the direction of movement or orientation of sinusoidal gratings, or for detecting them in central and peripheral vision. No qualitative differences were found between central and peripheral vision, and almost all quantitative differences observed could be removed by means of a size compensation derived from M. The results indicated specifically that (1) visual patterns can be made equally visible if they are scaled so that their calculated cortical representations become equivalent; (2) contrast sensitivity follows the same power function of the cortical area stimulated by a grating at any eccentricity; (3) area and squared spatial frequency are reciprocally related as determinants of contrast sensitivity; and (4) acuity and resolution are directly proportional to M, and the minimum angle of resolution is directly proportional to M-1. The power law of spatial summation expressed in (2) and (3) suggests the existence of a central integrator that pools the activity of cortical neurons. This summation mechanism makes the number of potentially activated visual cells the most important determinant of visibility and contrast sensitivity. The functional homogeneity of image processing across the visual field observed here agrees with the assumed anatomical and physiological uniformity of the visual cortex.


An estimation and application of the human cortical magnification factor. Comparisons of the published data on the density D of receptive fields of retinal ganglion cells and on the cortical magnification factor M indicated that M2 is directly proportional to D in primates. Therefore, the human M can be estimated for the principal meridians of the visual field from the density-distribution of retinal ganglion cells and from the density of the centralmost cones. Using the previously published empirical data, we estimated the values of the human M and express the values in four simple equations that can be used for finding the value of M for any location of the visual field. The monocular values of M are not radially symmetric. These analytically expressed values of M make it possible to predict contrast sensitivity and resolution for any location of the visual field. We measured contrast sensitivity functions at 25 different locations and found that the functions could be made similar by scaling the retinal dimensions of test gratings by the inverse values of M. Visual acuity and resolution could be predicted accurately for all retinal locations by means of a single constant multiplier of the estimated M. The results indicate that the functional and structural properties of the visual system are very closely and similarly related across the whole retina. Visual acuity, e.g., bears the same optimal relation to the density of sampling executed by retinal ganglion cells at all locations of the visual fields.


Correlations among climbing fiber responses of nearby cerebellar Purkinje cells in the immature rat. Correlations between pairs of spontaneous climbing fiber responses (CFRs) recorded from couples of nearby Purkinje cells (PCs) were studied in immature rats by using cross-correlograms between CFR pairs, and compaired to those in adult animals. Correlations were found as early as day 3. Some days later, on PN days 7--9, these correlations were higher than in the adult. In most cases, this was apparently not due to the multiple innervation of PC by climbing fibers (CFs) which normally occurs during this immature stage since: 1) temporal relationships between the paired CFRs varied by more than 30 ms and 2) thresholds for pairs of graded CFRs and additional components of the responses evoked in the 2 PCs by juxtafastigial or olivary stimulation were different. Synchronizing mechanisms were therefore likely to be already located at the olivary level. However, in 3 couples of multiply innervated PCs whose spontaneous CF activities were highly correlated, stimulation experiments revealed a common innervation of the 2 cells by branches of the same CF. In multiply innervated cells, spontaneous responses mediated through distinct CFs were also synchronized, suggesting that these fibers originate from neighboring neurons of the inferior olive. Finally, in 7 to 9-day-old rats, correlations among CFR pairs were much more restricted in the longitudinal axis of the folium than in the transverse one. On the whole, the present study indicates that correlations among CFRs of nearby PCs exist as soon as CF-PC synapses are established and the latter are already organized in sagittal strips at early stages of development.


Apparent movement and appearance of periodic stripes during eye movements across a stroboscopically illuminated random dot pattern. When the eyes follow a small target moving across a stationary random dot pattern illuminated stroboscopically at 3--45 flashes . s-1, the structure of the random dot pattern is seen as moving in the direction of the eye pursuit movements (sigma-movement). At flash frequencies above 9.5 flashes . s-1, periodic stripes oriented perpendicularly to the direction of the pursuit eye movements appear and are also seen as moving in the direction of the eye pursuit movement. The period Ps of the apparent stripes depended on the flash frequency fs and the angular speed of the eyes Ve: Formula: see text. Apparent sigma-movement and apparent stripes were also seen when eye pursuit movements were initiated outside of the random dot pattern and they continued autonomously without a moving target across the pattern. The angular velocity of the sigma-pursuit movement across the random dot pattern depended on the flash frequency and the angular velocity of the initiating target. With the eyes stationary but a moving random dot pattern, the apparent stripes also appeared at flash frequencies fs greater than or equal to 9.5 flashes . s-1. Eq. (1) was valid when Ve was replaced by the target angular velocity Vs (k = 1).


Brain-pineal nervous connections in the rat: an ultrastructure study following habenular lesion. The neural connections of the pineal gland with the central nervous system (CNS) were studied in rats by stereotaxic lesioning followed by electron microscopic determination of nerve terminal degeneration. Electrolytic lesions were placed in the right medial habenular nucleus and after 3--14 days survival, the animals were prepared for electron microscopy. Control animals contained nerve terminals with 40--60 nm dense core vesicles located in the perivascular space of the pineal and in the intraparenchymal area. In the lesioned animals many electron-lucent degenerating nerve terminals could be observed intermingled with normal nerve terminals in the perivascular spaces. These degenerating terminals were often swollen and contained one side of the terminal. Both normal and degenerating nerve terminals were also observed in the parenchyma between the pinealocytes. The present findings indicate that nerve fibers from the habenular area actually terminate in the rat pineal gland.


An analysis of the variability of unit activity in the cat's visual cortex. The spontaneous firing of single units in the cat's visual cortex, and their responses to repeated presentations of a constant visual stimulus, have been analysed quantitatively. Although the responses of most cells showed some habituation to the stimulus, this was less extensive than the fluctuations in responsiveness which occurred from one set of presentations to the next, over the course of several minutes. Fluctuations occurred largely independently in both the spontaneous and the stimulus-evoked firing of the cell. This suggests that separate mechanisms determine spontaneous and evoked firing. Quantitative estimates of the reliability (the reciprocal of the variability) of responses were higher if the peak firing rate of the cell was measured, rather than the total number of spikes fired by the cell. Thus, peak firing rate is more likely than total spike count to be the signal of biological significance.


Interstitiospinal action on forelimb, hindlimb, and back motoneurons. Responses of motoneurons supplying muscles of the forelimbs, hindlimbs, and back to stimulation of the interstitial nucleus of Cajal (INC) were recorded intracellularly in cats under chloralose anesthesia. Stimulation of the ipsilateral and contralateral INC evoked predominantly excitatory postsynaptic potentials in these motoneurons. Response latencies and properties of responses to multiple shock stimuli indicated that the responses were evoked by a di- or polysynaptic pathway. Stimulation of the anterior MLF (P2), which should have activated the entire interstitiospinal tract, but few reticulospinal or vestibulospinal fibers, evoked only polysynaptic responses. These results indicate that the INC does not establish direct synaptic connections with limb and back motoneurons.


14C-deoxyglucose mapping of orientation subunits in the cats visual cortical areas. The orientation domain in the cortical visual areas of anesthetized cats has been investigated by employing the 14C-deoxyglucose technique (Sokoloff et al., 1977). Orientation subunits (OS) are seen in the first (V1), the second (V2) and the third visual area (V3) as well as in the visual areas of the suprasylvian sulcus. In the latter regions OS are less elaborated than in V1, V2, and V3. The OS are continuous through all cortical layers; in V1 however, only weak label is detected in layer 4C. In V1, V2, and V3 the width of the OS is about 0.4 mm and the average distance between two OS centers is 0.9 mm. The spatial pattern of the OS seems to be more regular in the visual field periphery than in regions representing the vertical meridian.


Deformed optic nerves in coisogenic albino and pigmented C57BL/6J--c2j mice. The incidence of deformed optic nerve (DON) in C57 BL/6J--c2J mice is similar to that of a pigmented substrain of Long-Evans rats and much less than that of an albino substrain of Sprague Dawley rats. C57BL/6J--c2J albinos have no more DON than do coisogenic black mice. This suggests that albinism is not a factor in DON and that observed DON in Sprague Dawley rats are strain or substrain characteristics.


The diversity of marine sterols and the role of algal bio-masses; from facts to hypothesis. Modern analytical methods have revealed the great variety of marine sterols, which possess many different side chains and unsaturation patterns. Such biochemical transformations require well-defined mechanistic pathways, and there must be some 'Raison d'Etre' for a situation which has withstood evolution and adaptive changes. However, in this area, insufficient and sometimes not very substantial experimental work has made it difficult to see correlations, and hence to form a solid hypothesis. A review is now presented in which the molecules dissolved in sea water, or found in marine organisms are considered with particular reference to algal production, and to the bio-ecological significance of the main sterols.


Substance P afferents from the habenula innervate the dorsal raphe nucleus. The lateral habenula nuclei of the diencephalon innervate the median and dorsal raphe nuclei of the brainstem. Habenula lesions lead to decreased substance P levels in the dorsal but not median raphe within 24 hours. From this data, we propose a peptidergic innervation of the dorsal raphe nucleus by the habenula nuclei.


Concerning the anchoring of acetylcholinesterase in biomembranes. Highly purified acetylcholinesterases from various sources do not contain hydroxyproline and hydroxylysine, indicating the absence of a collagenlike 'tail'. The enzymes examined are therefore bound to the membrane in a different way compared with acetylcholinesterase isolated from electric organ.


Causes of artificially high blood glucose values in experiments with diabetic rats and mice. Unusually high blood sugar values may be caused by the slightest contamination of the blood obtained after puncture of the tail vein by diabetic urine, that has dried onto the tails of the animals. This can be avoided by collecting blood that is allowed to drip from the severed tip of the tail or by carefully cleaning the tail before puncturing the tail vein.


Purification of an L-fucose binding lectin from Ulex europeus by affinity column chromatography. An L-fucose binding lectin from Ulex europeus was purified by affinity column chromatography using an L-fucose-starch complex. The lectin thus purified had a mol. wt of 60,000, and consisted of 2 glycoprotein subunits with mol. wt 29,000 and 31,000, respectively.


Effect of the ionic strength on the kinetic properties of the mitochondrial L-malate dehydrogenase. Increase of the ionic strength inhibits the catalytic activity of the mitochondrial MDH, reduces substrate inhibition and decreases the affinity of the substrates for the enzyme.


Reduction in dietary vitamin E prevents onset of hypertension in developing spontaneously hypertensive rats. Reduction in dietary vitamin E intake in developing spontaneously hypertensive rats abolished the onset of hypertension which is normally evident by 3 months of age.


The importance of the terminal hemi-acetal group for the ionophoric properties of nigericin. Opening of the terminal hemi-acetal in dihydronigericin drastically reduces the ionophoric properties of nigericin and dehydroxymethylnigericin with 6 intact heterocycles. This is shown by 2 complementary methods, first with a liquid membrane electrode system, secondly by testing their ionophoric activities in rat liver mitochondria.


[Spontaneous and induced quadriradial figures on swinechromosomes in lymphocyte cultures (author's transl)]. In chromosome preparations from swine lymphocyte cultures, quadriradial figures and homolog associations were seen only of chromosome 10 and at a low frequency. The frequency of quadriradial figures was substantially increased by mitomycin C treatment, whereby 26.2% of the quadriradials were formed from the same chromosome pair with the number 10.


5-Methylangelicin: a new highly photosensitizing angular furocoumarin. 5-Methylangelicin, a new highly photosensitizing angular furocoumarin, was studied in 2 different biological systems, the T2 phage and Ehrlich ascites tumor cells; in comparison with angelicin, the parent compound, it was several times more active.


Regeneration induced in the forelimbs by treatment with vitamin A in the froglets of Rana breviceps. Young froglets Rana breviceps do not possess the power to regenerate forelimbs after amputation through metacarpals. However, 5 min immersion of limb stumps in an oily solution of 150,000 IU vitamin A palmitate immediately after amputation, and once on each of the 3 subsequent days, induced definite though hypomorphic regeneration in 100% cases.


Comparison of the in vitro effects of colchicine and its derivative colchiceine on chondrocyte morphology and function. Colchiceine is a colchicine-metabolite which has been reported to inhibit axonal transport although not binding to brain tubulin. In the present study, colchiceine was shown not to depolymerize cytoplasmic microtubules, nor to mimic other effects of colchicine on the ultrastructure of cultured chondrocytes. In addition, the synthesis of proteoglycans was inhibited by colchicine but slightly stimulated by colchiceine. These results support the idea that the disturbances in cultured chondrocytes caused by colchicine are specifically related to a loss of cytoplasmic microtubules.


Selective enrichment technique for isolation of methanol-utilizing yeasts. A simple, selective enrichment technique was developed for isolation of methanol-utilizing yeasts by supplementing gentamicin or tetracycline in the medium.


Cell size and cell division of the anterior pituitary: time course in the growing rat. Pituitary cells increase their numbers more than 3-fold during the 1st 10 days of life while maintaining the same cell size ratios. In the 25-day-old animal, the rate of cell division slows and there is a slight increase in the number of large cells. An increase in adult weight is attributed to hyperplasia and a shift to a population of larger cells.


Lymphocytes, but not cancer cells are able to penetrate into the rat embryo yolk sac wall. Normal rat lymphocytes and cells of 2 highly invasive tumors, the L5222 rat leukemia and the VW rabbit carcinoma, were inoculated in vitro on the mesothelial surface of the visceral wall of the rat embryo yolk sac. After 48 h, lymphocytes, without any damage being inflicted on the mesothelial cells, had penetrated deeply into the yolk sac wall, whereas both kinds of cancer cells had destroyed the mesothelial cells, but not advanced beyond the basal lamina.


Sarcomere shortening and tension development during 'isometric' tetanus of muscle. Laser diffraction and tension measurements from frog muscle during isometric tetanus reveal that the sarcomeres contract more rapidly than tension develops.


Electron probe X-ray microanalysis and cryoultramicrotomy of unstained myocardial sarcoplasmic reticulum, in situ and fragmented. Myocardial sarcoplasmic reticulum of cats in situ and fragmented sarcoplasmic reticulum (FSR) were analysed using X-ray microanalysis, cryoultramicrotomy and scanning transmission electron microscopy. 2 types of FSR vesicles can be distinguished morphologically and by their different elemental composition especially by different Ca loading. The Ca content of the sarcoplasmic reticulum can also be detected in situ.


Regularly firing neurones in the rat suprachiasmatic nucleus. The spontaneous discharge of some suprachiasmatic neurones in vivo and in vitro was found to exhibit a very constant interspike interval. In vivo these cells were comparatively rare and appeared to be mutually coupled. The findings are discussed in relation to coupled oscillator theories of circadian rhythm generation.


High shortening velocity of isolated single arterial muscle cells. Surprisingly high shortening velocities (less than 200 msec contraction-relaxation cycles) were found in isolated vascular muscle cells cultured from rat or chick aorta. All of the small fraction cells with such quick contractions had membrane excitation by short duration spikes, rather than the slower graded depolarization of the other cells which produced 20-fold slower contractions.


Hepatic blood flow in acute myocardial ischemia. Hepatic blood flow was monitored in cats during myocardial ischemia (MI). Increased plasma CPK activity, the S-T segment of the electrocardiogram, and hepatic flow was reduced by 5 h to 40% of control. The results suggest that MI can influence organs distant from the original ischemic episode.


Fine structural correlations between the muscle pathology of amyotrophic lateral sclerosis (ALS) patients and experimental Ca-Mg deficient rats. Nonspecific myofibrillar changes such as streaming of the Z-line, formation of rod-like structures, satellitosis, proliferation of sarcolemmal nuclei and papillary projection of the sarcolemma were recognized as a disorganization of the muscle itself. In addition, fine structural pathology in ALS specimens showed characteristically a pig-tail formation - 'Zopfformation' - which has been considered to have a neurogenic origin.


Acute stress reduces the sensitivity of the vasculature to sympathetic control. Vascular smooth muscle from rabbits subjected to acute severe stress exhibits decreased sensitivity to sympathetic regulation. Stimulation of the sympathetic innervation of isolated vascular segments resulted in a similar subsensitivity as did exposure to norepinephrine (NE) but not histamine. Periodic contraction of these segments caused an increase in their maximum ability to contract independent of the constrictor procedure used. These results suggest that the increase in sympathetically mediated NE release that occurs in stress and some other pathological conditions may result in a blunting of neural control and possibly resistance to certain therapeutic agents.


Selective destruction of intestinal nervous elements by local application of benzalkonium solution in the rat. Intestinal aganglionosis produced by serosal application of 0.1% benzalkonium solution to the colon of the rat was studied electronmicroscopically, and it was concluded that a higher susceptibility to the agent and a lower recovering ability of the nerve elements might be responsible for the phenomenon.


Radioimmunoassay of progesterone in uterine flushings of buffalo (Bubalus bubalis). The concentration of progesterone as determined by radioimmunoassay varied in accordance with the phase of ovarian activity.


In vitro esterification of plant sterols by the esterifying enzyme of the small intestine of rat. The in vitro esterification of plant sterols, beta-sitosterol, campesterol and stigmasterol, by the esterifying enzyme of the small intestine of rat was studied in the presence of saturated and unsaturated fatty acids. Campesterol esterification was highest, followed by sitosterol and stigmasterol irrespective of the type of fatty acid. Both campesterol and sitosterol esterification was greater with unsaturated fatty acids than with saturated fatty acids.


Isometric relaxation in rat myocardium: load dependence and influence of caffeine. When caffeine plus calcium is added to the perfusing medium, isometric relaxation of rat myocardium is no longer affected by length changes occurring during the twitch. The dependence of isometric relaxation on the initial muscle length is still present and more pronounced after caffeine addition.


Effect of hormones on adenyl cyclase activity of rabbit mammary gland in vitro. Ovarian hormones namely beta-estradiol and progesterone were observed to stimulate the activity of adenyl cyclase of the mammary gland from pregnant rabbit in vitro, unlike the lactating tissue where it was inhibited. On the other hand, non-ovarian hormones like hydrocortisone, prolactin and insulin did not have a similar effect on this enzyme.