Radiation response of multicell spheroids--an in vitro tumour model.

Multicell spheroids provide an in vitro tumour model of intermediate complexity between tumours and standard cell cultures. Spheroids of several varieties of normal and tumour cells have been grown and Chinese hamster V79-171b cells have been studied in detail. Spheroids simulate conditions in those solid tumours which contain nodular areas with necrosis at a distance from blood vessels and show a decrease in growth fraction as they increase in size. They contain radiation resistant chronically hypoxic cells which develop spontaneously as the spheroids grow, and exist in a tumour-like environment, yet retain their growth potential. Several assays have been developed or adapted for use in spheroid experiments to study cell properties and responses to treatment. These include many of the techniques used in tumour biology to determine the net response of the tumour, such as volume changes, histology, and net cell survival as well as more direct assays involving separation of subpopulations of cells. Methods were also developed to study in situ repair and repopulation processes after ionizing radiation or other cytotoxic agents.


Separation and characterisation of tryptic fragments from the adenosine triphosphatase of sarcoplasmic reticulum.

The two halves of the ATPase, M, 115,000, from sarcoplasmic reticulum produ-ed by limited trypsin treatment have been purified in sodium dodecylsulphate. The fragment of Mr60,000 has been purified by electrophoresis on cellulose acetate slabs and that of Mr 55,000 by gel filtration. The two halves of the 60,000 Mr fragment (Mr33,000 and 24,000) produced by more extensive trypsin treatment have also been purified by gel filtration in sodium dodecylsulphate. The sum of the amino acid analyses of the constituent tryptic fragments is in good agreement with that for the whole ATPase. The amino acid compositions of the two halves of the ATPase were strikingly similar. N-terminal analysis shows that the ATPase and its constituent tryptic polypeptides all possess a single N-terminal alanine implying no further cleavage of the polypeptide by trypsin. Attempts to solubilize selectively the tryptic fragments from the membrane by a variety of denaturing and solubilising agents under a variety of conditions have proved unsuccessful, suggesting that the interaction between the tryptic polypeptides is stronger than between the lipid and the protein. The possibility that the interaction  between the tryptic polypeptides includes disulphide bonding has been eliminated.


Limited proteolysis of yeast phosphofructokinase by subtilisin. Alterations in enzyme activity, subunit composition, and hydrodynamic properties.

Yeast phosphofructokinase having a molecular weight of 750000--800000 (20 S) has been subjected to limited proteolysis by subtilisin and yeast proteases. Two steps of proteolytic degradation could be distinguished: in the first step, which is accompanied by an increase in molecular activity, the subunits alpha and beta (Mr 120000) are converted to alpha' and beta' (Mr approximately 900000), and in the second step, accompanied by a decrease in enzyme activity, alpha' is converted to alpha'' (Mr 80000) and two further fragments having Mr 45000 and 35000 become detectable. In the course of the conversion the sedimentation value of the undissociated enzyme drops from 20 S to about 17 S. The two substrates fructose 6-phosphate and ATP exhibit characteristic protective effects on enzyme activity and on subunit degradation. Whereas the first step is not strongly influenced by the substrates, fructose, 6-phosphate inhibits significantly the degradation of alpha' and beta', whereas ATP prevents only degradation of beta'. When in presence of ATP alpha' is degraded to alpha'', the quaternary structure of the 17-S enzyme is no longer stable and a dissociation of the molecule occurs to a 12-S form which is enzymically active and ATP-sensitive and in which the ratio of alpha'' to beta'' is one-to-one.


Immunochemical studies on tyrosinase induction in Neurospora.

An immunoassay for tyrosinase, using the modified bacteriophage technique, was developed: Tyrosinase of Neurospora was conjugated to bacteriophage T4 using glutaraldehyde as a cross-linking agent. The conjugated phage that survived the coupling process could be inactivated by antiserum raised in rabbits against pure tyrosinase, but not by normal serum. This inactivation was specifically inhibited by pure Neurospora tyrosinase, and the degree of inhibition was proportional to the concentration of tyrosinase within the range of 30-150 ng/ml. Crude mycelial extract possessing tyrosinase activity could similarly inhibit the inactivation of the conjugated phage by the antiserum. To evaluate the tyrosinase content of crude extracts their inhibitory capacity was compared to that of known amounts of pure tyrosinase, and the amounts thus calculated agreed with those predicted from an enzymatic assay. The tyrosinase-bacteriophage immunoassay was used for the quantitation of tyrosinase-antigen in crude extracts of Neurospora cultures that had been induced to form tyrosinase by the addition of ethionine. Enzymatic activity appeared after a lag of several hours, increased for 2-3days and then declined. Immunological assays of these cultures showed: (a) serologically reactive protein started to accumulate upon culture starvation and was evident during the lag period; (b) specific activity (units per mg antigen) was constant throughout induction; (c) at the phase of decrease in mycelial enzyme content, increasing amounts of serologically reactive protein were detected in the medium, indicating that some enzyme was eventually excreted. These results show that the lag is not a qualitatively distinct period, and support the previously forwarded notion that tyrosinase is synthesized de novo upon induction.


A new method of isolation and some properties of the heavy chain of human plasmin.

A method is described by which the heavy chain of human plasmin, obtained by partial reduction of urokinase-activated plasminogen with 2-mercaptoethanol, is adsorbed on lysine coupled to polyacrylamide. The heavy chain is recovered from the adsorbent by elution with 6-aminohexanoic acid (yield 60-65%). Sulfhydryl titrations of the heavy chain showed that the partial reduction involved primarily the cleavage of the sole interchain disulfide bridge of plasmin. Dodecylsulfate-polyacrylamide electrophoresis gave essentially a single band corresponding to a component of about 60000 molecular weight. The NH2-terminal amino acid was predominantly threonine. 6-Aminohexanoic acid at different concentrations caused significant variations of the sedimentation and diffusion constants of the heavy chain indicating inhibitor-induced conformational alterations of the protein. The present results suggest that in plasmin only the heavy chain is capable of interacting with 6-aminohexanoic acid, and it appears that it is primarily this chain which plays an important role in the inhibition of the enzyme by 6-aminohexanoic acid.


On the mechanism of protein-synthesis inhibition by abrin and ricin. Inhibition of the GTP-hydrolysis site on the 60-S ribosomal subunit.

The mechanism of protein synthesis inhibition by the toxic lectins, abrin and ricin, has been studied in crude and in purified cell-free systems from rabbit reticulocytes and Krebs II ascites cells. In crude systems abrin and ricin strongly inhibited protein synthesis from added aminoacyl-tRNA, demonstrating that the toxins act at some point after the charging of tRNA. Supernatant factors and polysomes washed free of elongation factors were treated separately with the toxins and then neutralizing amounts of anti-toxins were added. Recombination experiments between toxin-treated ribosomes and untreated supernatant factors and vice versa showed that the toxin-treated ribosomes had lost most of their ability to support polyphenylalanine synthesis, whereas treatment of the supernatant factors with the toxins did not inhibit polypeptide synthesis. Recombination experiments between toxin-treated isolated 40-S subunits and untreated 60-S subunits and vice versa showed that only when the 60-S subunits had been treated with the toxins was protein synthesis inhibited in the reconstituted system. The incorporation of [3H]puromycin into nascent peptide chains was unaffected by the toxins, indicating that the peptidyl transferase is not inhibited. Both the EF-1-catalyzed and the EF-2-catalyzed ability of the ribosomes to hydrolyze [gamma-32P]GTP was inhibited by abrin and ricin. An 8-S complex released from the 60-S subunit by EDTA treatment possessed both GTPase and ATPase activity, while the particle remaining after the EDTA treatment had lost most of its GTPase activity. Both enzyme activities of the 8-S complex were inhibited by abrin and ricin. The present data indicate that there is a common site on the 60-S subunits for EF-1- and EF-2- stimulated GTPase activity and they suggest that abrin and ricin inhibit protein synthesis by modifying this site.


Adenylate cyclase from rat-liver plasma membrane: inhibition by mersalyl and other mercurial derivatives.

The adenylate cyclase activity from a rat liver plasma membrane preparation was inhibited by low concentrations (1-10 muM) of the mercurial diuretic mersalyl. Complete inhibition was obtained with 0.1 mM mersalyl. Similar effects were observed whether the adenylate cyclase preparation was assayed in the presence of 10 muM GTP, 0.1 muM glucagon, 10 mM NaF or without any addition. The effect of mersalyl was not due to inhibition of the regenerating system present in the incubation medium, since the effect of mersalyl was preserved and even enhanced in its absence. The inhibition brought about by mersalyl was due to both a decrease of the maximal velocity of the reaction and of the affinity of the enzyme for the substrate. It was immediate, and irreversible spontaneously, but it was reversed by the simultaneous additions of 2-mercaptoethanol, in a dose-dependent fashion. Other -SH reagents were found to have an effect equal to, or lower than, that of mersalyl. Mersalyl had no effect upon Mg2+-ATPase, although it inhibited the (Na+-K+) activated ATPase. Since mersalyl is known to be a 'non-penetrant' reagent, it is postulated that a catalytically important, mercurial-sensitive, part of adenylate cyclase is at the surface of the plasma membrane. This view is supported by the following facts: (a) mersalyl acted with a similar dose-response curve upon an intact as well as a detergent-dispersed cyclase preparation while no effect was observed upon a solubilized Mg2+-ATPase preparation; (b) a covalent p-chloromercuribenzoate-Sephadex preparation (but not its supernatant) inhibited the cyclase from intact membranes. It is proposed that mercurial derivatives, by their relative specificity of action (no effect on Mg2+-ATPase), can serve as useful probes in the elucidation of the multicomponent structure of the cyclase system.


The effects of fusimotor stimulation during small amplitude stretching on the frequency-response of the primary ending of the mammalian muscle spindle.

Single fusimotor fibres to the de-efferented soleus of the anaesthetized cat were stimulated repetitively while recording the response of single primary afferents to small amplitude sinusoidal stretching at frequencies of 0-5--500 HZ. The response of the ending was determined by averaging its firing for many cycles to construct a 'cycle histogram'. When small enough, the stretching modulated the firing sinusoidally; as the frequency increased the requisite amplitude fell to a fraction of a micron. The amplitude of the modulation (in impulses/sec) divided by be amplitude of stretchin (in mm) gave the sensitivity of the ending for the particular frequency in question. 2. The passive frequency--response curve obtained in the absence of fusimotor stimulation agreed with those obtained before...


Effects of specific chemical modification of actin.

G-actin has been nitrated with tetranitromethane in conditions that lead to the modification of one tyrosine residue. The reactive residue was found by earlier workers to be Tyr-69. The nitrated actin is conformationally similar to native G-actin, as judged by sedimentation velocity and circular dichroism analysis. A small proportion only is in the form of covalently linked dimers and trimers. The nitrated G-actin will polymerise to form filaments, indistinguishable in the electron microscope from those of native F-actin, but the polymerisation process is slower. Reduction of the nitrophenol group to the corresponding aminophenol leaves the properties of the protein in respect of polymerisation unchanged. When a dansyl group is introduced at the same point, however, the ability of the actin to polymerise is lost. The nitrated actin and its reduced counterpart will also bind heavy meromyosin, and the characteristic arrowhead formation of the bound molecules along the filaments can be seen in the electron microscope. Neither of the modified F-actins, however, significantly activates or inhibits the myosin ATPase activity. The fluorescence of nitrated actin is strongly quenched through the presence of the nitrophenol chromophore. In soluble complexes with heavy meromyosin the fluorescence is indistinguishable from the sum of the separate contributions of the two protein components. There is thus no measurable excitation transfer between any tryptophan residues on the myosin heads, such as that inferred to be present in the ATPase site, and the nitrotyrosine in position 69 of the actin sequence. Implications of this observation are considered in relation to the different interaction sites in myosin and in actin. The activation of heavy meromyosin ATPase by copolymers containing actin and nitroactin in different proportions has been measured, and is not proportional to the fraction of native actin. The results are consistent with the view that the function of actomyosin depends on the interaction of the myosin heads with more than one actin subunit.


Recycling of glucose by rat hepatocytes.

1. The metabolism of glucose labeled uniformly with 14C, and in positions 2, 3 and 5 with tritium by hepatocytes from fed and fasted rats were studied. Cells were incubated with glucose as sole substrate, or with glucose and a variety of glucose precursors, and uptake or production of glucose, and the utilization of the isotopes was determined. 2. There was no uptake of glucose at concentration of up to 15 mM, and net glucose synthesis in the presence of precursors. 14C was however recovered in CO2, lactate and amino acids, and tritium in water. Considerable incorporation into glycogen from 14C and 3H-labeled glucose occurred at high (above 20 mM) glucose concentrations. 3. The yield in water always exceeded that in 14C-labeled products. The yield in 3HOH from [2-3H] glucose exceeded that from [5-3H] glucose, and the latter was greater than from [3-3H] glucose. 4. Utilization of labeled glucose does not follow Michaelis-Menten kinetics. The fractional rate of uptake of 14C and tritium-labeled glucose increases with glucose concentration with a maximum at about 15 mM and then declines. 5. The effect of numerous gluconeogenic substrates on the isotope utilization and the 3H/14C ratio in glycogen was studied. The uptake of 14C was always depressed. Addition of lactate and dihydroxyacetone has little effect on the detritiation of [2-3H] glucose, but it is depressed by other substrates. The detritiation of [3-3H]-and[5-3H]glucose is depressed in gluconeogenesis, that from [3-3H]glucose usually more than from [5-3H]glucose. In the presence of lactate detritiation of [3-3H]glucose is about half that from [5-3H]glucose. 6. Equations to calculate the phosphorylation of glucose and fructose 6-phosphate in the presence of futile cycling between glucose and glucose 6-phosphate and fructose 6-phosphate and fructose 1,6-bisphosphate were derived. 7. The estimate of glucose phosphorylation requires determination of the specific activity of glucose 6-phosphate from [2-3H]glucose. It appears that futile cycling between glucose and glucose 6-phosphate is extensive in cells with a high glycogen content, but is low in cells from starved rats and nearly absent in those from diabetic animals. 8. The estimation of the phosphorylation of fructose 6-phosphate in the presence of cycling requires knowledge of the specific activities of fructose 6-phosphate and fructose 1,6-bisphosphate from [3-3H]glucose. At present there are no adqquate data to calculate phosphorylation and recycling of fructose 6-phosphate, but under some conditions the rate may be quite high.


Self-association of human erythrocyte phosphofructokinase. Kinetic behaviour in dependence on enzyme concentration and mode of association.

The kinetic behaviour of human erythrocyte phosphofructokinase has been analyzed over a relative wide range of enzyme concentration (0.01 -- 1.7 mug/ml). The kinetic cooperativity which becomes apparent when the enzymic reaction rate is plotted versus the fructose 6-phosphate concentration decreases with increasing enzyme concentration. Simultaneously, a decrease of the half-saturation concentration for fructose 6-phosphate [S]0.5 is observed. Maximum velocity passes through a maximum at increasing enzyme concentrations. Sets of curves representing specific enzymic activity of phosphofructokinase versus enzyme concentration obtained at various fixed concentrations of fructose 6-phosphate and ATP are analyzed. The shapes of these curves are interpreted in terms of an association model of human erythrocyte phosphofructokinase, in which an inactive dimer (Mr 190000) and active multimers of the dimeric form are involved. The conclusion is drawn that the sigmoidal shape of the plots of the enzymic reaction rate versus fructose 6-phosphate concentration is partially caused by a displacement of the equilibrium between different states of association of phosphofructokinase to multimers by this substrate. On the other hand, the inhibition of the enzyme by high concentrations of ATP may be partially caused by a shift of this equilibrium to the state of the inactive dimer.


Active Ca transport of sacroplasmic reticulum during experimental uremia. Changes in kinetics and lipid composition.

The Ca-transport system of sarcoplasmic vesicles of rabbits is altered by experimental uremia. 1. The influx rate constant of the experimental membranes decrease with a resulting decrease of the calcium influx rate. 2. The experimental membranes transport a smaller amount of Ca2+ per mol of ATP split than the controls, i.e. their transport ratio is discussed. 3. The calcium permeability of the experimental membranes increases with a resulting decreased concentrating ability. 4. The phosphatide content but not the cholesterol content of the experimental membranes decreases with a consequent increase of the cholesterol/phosphatide ratio. 5. The fatty acid pattern of total phosphatides of the experimental membranes changes. A relative decrease of palmitic acid and oleic acid occurs and a relative increase of stearic, arachidonic and higher unsaturated fatty acids. 6. The altered lipid composition of the membranes does not change the temperature dependence of the kinetics.


Postnatal decline of (Ca2+ + Mg2+)-activated membrane ATPase in cattle red cells.

Acitivity of membrane bound (Ca2+ + Mg2+)-stimulated ATPase, associated with Ca2+ outward transport, in calf red cells is high at birth and declines with a rate constant of 0.041 d-1 after the 3rd week. The decline parallels the disappearance of fetal hemoglobin.


The solubility properties of granulopoiesis inhibiting factor.

The solubility of granulopoiesis inhibiting factor (GIF) in various aqueous and organic solvents was investigated. GIF is soluble in water, 10% acetic acid, and 10% pyridine. It is not extractable by 1-butanol at low and high pH. A high solubility was found in polar organic solvents (ethanol-acetone 9:1, and chloroform-methanol 1:1), whereas GIF seems to be insoluble in pure chloroform.


Effect of anti-Ia sera on mixed lymphocyte reaction in mice.

Anti-I-region associated (Ia) antigen sera specificially supress the stimulation reaction in unidirectional mouse MLR. No effect on the response was observed with these antisera. In addition, anti-H-2 sera did not affect stimulation, however, in some cases diminished the response. The data indicate that Ia antigens and LAD might be identical. Stimulation in mixed lymphocyte culture reaction (MLR) or graft vs host reaction has been shown to be governed by genes located in the I-region (immune response region) of the H-2 complex in mice (Bach et al. 1972 and Klein and Park 1973) and it appears that the H-2K and H-2D regions play only a minor role in that reaction. Recently, antigens have been described (Hauptfeld et. al. 1973, Götze et al. 1973, David et al. 1973) which are controlled by the I-region, so-called I-region associated equals Ia antigens. If these antigens are related to determinants important for the MLR, then one might predict inhibition of the MLR. Abbasi et al (1974) reported that poly-specific anti-H-2 alloantisera inhibited the H-2 complex controlled stimulation in MLR. Using antisera produced in I-region congenic strains, we have been able to show complete and specific inhibition of the stimulation in MLR, without affection of the response, thus indicating that Ia antigens and lymphocyte activating determinants might be identical.


Antimicrobial effectiveness of Locacorten-Vioform cream in Secondary infections of common dermatoses.

In a double-blind study involving 430 patients, Locacorten (0.02%)-Vioform (3%) cream was found to be highly effective in the treatment of dermatological conditions complicated by secondary bacterial involvement. Themicrobiological conversion and clinical improvement in patients treated with Locacorten-Vioform combinations were markedly greater than those achieved in the groups medicated with (1) Vioform 3%, (2) Locacorten 0.02%, and (3) placebo cream. Staphylococcus aureus was the most prevalent organism isolated from the skin lesions. All strains were sensitive to Vioform, as compared to 62% found to be resistant to other antimicrobials.


Occupational cobalt and chromium dermatitis in an offset printing factory.

A case history of three offset printers and the atomic absorption analysis of their contact materials is described. Though the factory direction and personnel felt strongly about cooperating in order to combat the disease, it became apparent that the prescribed measures had not been effectively carried out. The chemical analysis of the materials proved to be a necessary supplement to the patch testing procedure.


Increased hemoglobin AIc in diabetic mice.

The minor hemoglobins AIa, AIb, and AIc were studied in mice with either genetic or chemically induced diabetes. Hemoglobin AIc was elevated approximately twofold in all the phenotypically diabetic mice studied (C57BL/KsJ-db/db, C57BL/KsJ-ob/ob, C57BL/6J-db/db, and alloxan- and streptozotocin-treated mice). Elevation of the hemoglobin AIc in C57BL/6J-db/db mice was of short duration, reflecting the transitory diabetes characteristic of these mice. The degree of increase of hemoglobin AIc levels was unrelated to severity of hyperglycemia, duration of diabetes, age of mouse, or body weight. It is not known what factor(s) dictates the steady-state concentration of hemoglobin AIc.


Fetal pancreas transplantation for reversal of streptozotocin-induced diabetes in rats.

Streptozotocin-induced diabetes in rats was completely reversed by transplantation of syngeneic fetal pancreases placed beneath the kidney capsule. To accomplish complete reversal of diabetes, four or more pancreases were necessary; three resulted in partial reversal, and two produced a slight but significant effect in some recipients. Removal of the transplants resulted in the prompt return of diabetes. The islets of Langerhans in the transplants functioned homeostatically; this was indicated by regular normal blood glucose values, in addition to normal findings in blood IRI response and glucose disappearance rate after glucose injection. Disappearance of exocrine elements, with only ducts and fibrous tissue remaining, resulted in a pure endocrine organ. The advantages of this technie, such as ease of accessibility for placement, observation, and removal, are of great importance for possible application to humans.


Secretion of hydrochloric acid in Ghosh-Lai rat preparation following stimulation with histamine and pentagastrin and inhibition by gastrone and atropine.

In this study of 48 successfully operated rats, we evaluated the applicability of the Ghosh-Lai stomach preparation to the quantitation of the HCl output following stimulation with histamine and pentagastrin and inhibition with gastrone and atropine. The usefulness of this rat preparation was well demonstrated and good duplication of results was obtained with various doses of histamine and pentagastrin. In this rat preparation, the HCl output following an optimal pentagastrin dose of 6 mug/kg/30 min i.v. was almost twice higher than that after an optimal histamine dose - for this rat preparation - of 0.2 mg/kg/30 min i.v. The inhibitory effect of gastrone B on histamine-stimulated HCl output has been confirmed in this rat preparation and shown to extend also on the pentagastrin-stimulated HCl secretion. The comparison of the atropine and gastrone inhibition of the pentagastrin-stimulated HCl output in the Ghosh-Lai preparation indicates that the inhibitory effect of 40 mug gastrone i.v. is similar in rat to that of 0.4 mg atropine sulfate i.v. under these conditions.


Lymphocyte function in patients with Hodgkin's disease.

Lymphocyte functions in vitro were tested in patients with Hodgkin's disease. The PHA-induced lymphocyte stimulation and the responding capacity in the one-way mixed lymphocyte culture reaction was found to be severely depressed. In contrast, the PHA-induced production of a human mononuclear leukocyte chemotactic factor was not significantly impaired.


Adaptive significance of vegetative incompatibility in Neurospora crassa.

Certain features reminiscent of sexuality occur in the vegetative life cycle of some filamentous fungi such as Neurospora crassa. Hyphal fusions can occur between genetically different individuals, thereby endowing the new composite mycelium, a heterokaryon, with some of the advantages of heterozygosity usually associated with diploid organisms. In N. crassa, however, there are a number of incompatibility loci which prevent formation of heterokaryons unless the alleles at the incompatibility loci are identical in the two mycelia. The selection pressures that maintain incompatibility polymorphisms are not known. We suggest here that they are maintained because they prevent a kind of exploitation of heterokaryons by nuclei that are nonadaptive in homokaryons but that enjoy a proliferative advantage over other nuclei in heterokaryons. A mathematical model that abstracts the major features of the vegetative life cycle of Neurosopra crassa has been developed, and the action of selection in this model and various extensions of it is such as to maintain polymorphisms of vegetative incompatibility factors.


[Demonstration of a post-traumatic, transdiaphragmatic liver prolapse by combined hepatic and lung scans (author's transl)].

A case of right-sided traumatic diaphragmatic rupture with transdiaphragmatic liver prolapse is reported. The situation could be clarified by combined hepatic and lung scans. The advantages of isotope examinations, particularly in patients with multiple trauma, are stressed.


[Angiography, scintigraphy and ultra sound in the diagnosis of splenic or hepatic disease in Hodgkin's disease (author's transl)].

The combined use of selective angiography, scintigraphy and ultra-sound is not reliable enough for the certain exclusion of visceral involvement in Hodgkin's disease. Since this is of importance in the planning of treatment, it confirms the recommendation made by Wagenknechts, Glatsteins and others that a diagnostic splenectomy be carried out in order to determine the presence of visceral involvement.


[Hypotonic oesophagography using propanthelin bromide (Pro-Banthine) (author's transl)].

Hypotonic oesophagography was performed in 149 patients after the intravenous injection of propanthelin bromide (Pro-Banthine). An injection of 0.25-0.50 mug/kg body weight led to hypotonia of the oesophagus in 96% of cases. During hypotonia, it was possible to demonstrate oesophageal varices in 24 out of 27 patients, and small hiatus hernias in 10 out of 23 patients. Conventional examination demonstrated oesophageal varices in only 13 of the 27 cases and hiatus hernias in four out of the 23 cases. Hypotonic oesophagography thus leads to an improvement in the accuracy in the diagnosis of oesophageal varices and small hiatus hernias. Following injections of Pro-Banthine, transient side effects were observed consisting of tachycardia, difficulties in accomodation, reduction in salivary excretion and difficulties in micturition. Pro-Banthine is contra-indicated in the presence of glaucoma, severe cardiac and circulatory diseases and urinary obstruction.


[Intravenous cholegraphy with Chologram (a double blind comparison with Bilivistan) (author's transl)].

A double blind comparison between Ioglycamid (Bilivistan) and the new I-V cholegraphic medium Iotroxamid (proposed trade name Chologram) is reported. Chologram produces better pictures and it is better tolerated. Pharmaco-kinetic data are published for the first time.


[Thyroid phlebography and selective venous sampling for parathormone estimations in primary hyperparathyroidism (author's transl)].

Selective venous sampling for parathormone estimations has become an important method in the diagnosis of primary hyperparathyroidism and, together with thyroid phlebography, is an accurate means for localising hormone active parathyroid tissue. Thirty-three patients were examined by this technique and in 25 of these an exploration was carried out subsequently. Twenty-one had primary hyperparathyroidism. In one case hormone analysis lead to a false positive finding. In 18 of these 21 patients, the hormone-active parathyroid tissue was correctly localised. Phlebography showed evidence of an adenoma in five patients, but is of most use for localising the source of the hormone.


[Radiological findings following post-traumatic bleeding into porencephalic cysts (author's transl)].

Six operatively treated patients are described who had sustained post-traumatic bleeds into a porencephalic cyst. The plain films of the skull showed asymmetries of the sphenoidal wings, bulging of the middle temporal fossae, circumscribed protrusions in the temporal region and unilateral enlargement of the paranasal sinuses. These changes are pathognomonic of a cystic malformation. Angiography shows an almost vertical origin of the middle group of vessels and a paucity of vessels in the area of the cyst. An accurate radiological diagnosis can be made pre-operatively, subdural haematomas and hygromas, arachnoid cysts and tumours entering into the differential diagnosis. Clinical evidence of raised intracranial pressure is an indication for performing burr holes; a porencephalic malformation does not in itself require surgical treatment.


[Radiculo-tomography for lumbosacral myelography (author's transl)].

The use of tomography can increase the value of lumbosacral myelography in certain circumstances. Post mortem studies have resulted in standardisation of the examination, which will combine optimal diagnostic information with the least amount of effort and with an acceptable amount of stress on the patient. 110 tomographic examinations were performed amongst 680 lumbosacral myelograms (Dimer X). On the basis of this experience, the value of this procedure is illustrated from various points of view and its indications are discussed.


[The effect of haemodynamic factors on tomograms during angio-tomography (author's transl)].

Haemodynamic factors may influence the appearance of the tomograms during the performance of angio-tomography. Model experiments have shown that the thickness of section able to demonstrate vessels may depend on the relationship between vessel opacification and the timing of the cut. The clinical relevance of this is illustrated by tomographic studies of the renal and cerebral circulations.


[Sterno-costo-clavicular hyperostosis (author's transl)].

A case is described in which bilateral subclavian vein occlusion was caused by fusiform expansion of the clavicle with hyperostosis, ankylosis of the sterno-clavicular joint, widening and thickening of the sternum and ossification of the costal cartilage of the first ribs. Histologically this proved to be due to an hyperostotic spongiosclerosis.


[Mesomelic dwarfism (author's transl)].

Mesomelic dwarfism of the Langer type is described. This is characterised by shortening and bending of the middle portions of the extremities. This is compared with three cases of "mesomelic" changes in the forearm. The classification of this condition is discussed.


[False aneurysms and bone deterioration (author's transl)].

Two comparable cases of false aneurysm were studied. The different reaction of the adjacent bone is compared with experimental studies of constant and oscillating pressure effects to the bone.


[The intraosseus ganglion (author's transl)].

The intraosseous ganglion is a not very well-known cystic benign bone lesion, which has to be included in the differential diagnosis of benign bone tumors. The ganglion's pathogenesis is still uncertain. Radiologically it presents all signs of a benign epiphyseal cystic tumor and can be easily differentiated from malignant changes. The characteristic X-ray signs, together with the typical location, and especially when situated in the distal medial epiphysis of the tibia, will allow a preoperative diagnosis.


[Comparative investigations concerning osteotropic radiopharmaceuticals. III. Scanning with 18F and 99mTc-malignant diseases (author's transl)].

Total body scans under identical conditions and using 18F and 99mTc-EHDP were carried out in patients with known, or subsequently proven, metastases. There was no statistical evidence for any difference in the number of scintigraphically demonstrable lesions. Patients with internal prostheses who developed infections of the prosthesis show no significant difference in the uptake of either nucleid. Soft tissue lesions showed a significantly higher uptake of 99mTc-EHDP than of 18F. 18F can be replaced for nearly all problems by 99mTc-EHDP without loss of diagnostic information.


[New basic considerations and theories for improving angio-cine-densitometry. IV. The effect of discontinuous, pulsatile flow on angio-cine-densitometric measurements (author's transl)].

Various expressions of the speed of flow of blood in arteries are discussed, such as instantaneous speed, integral speed, pulse wave speed (phase speed) and velocity of a moving front. The flow profile of the abdominal aorta was obtained by cine-densitometry, bearing in mind the above considerations, and the densitometric curves have been analysed. Differences of the flow velocities in different persons have been interpreted as a result of these theoretical considerations. The effects of the point of measurement, the distance over which the measurement was carried out and the pulse wave length on velocity were examined. In order to obtain a median blood stream velocity by angio-cine-densitometry, repeated injections of small volumes of contrast medium (random sampling method) are recommended.


Cellular survival and isoeffect formulae: a study in applied radiobiology.

The Ellis formula for the sterilization of tumors is compared with isoeffect curves for sterilizing doses as a function of the fractionation number N. Using cell parameters from human kidney cells irradiated in cell cultures we get a good agreement over a wide range of N values. The exponents obtained from the calculations are 0,26-0,30 and 0,36 for 200 keV X-rays, Co-60 rays and high energy electrons respectively. A reduction of tumor cells to 10(-8) seems sufficient for tumor sterilization. For normal tissue Ellis has proposed another formula (for X-rays) where a time factor T0,11 concerns homeostatic repair and proliferation of normal alls. The Ellis formula can be simplified when we assume that all treatment schemes be given with the same weekly dose-rate, i.e. have the same "hardness". Cell survivals according to this formula have been calculated and tolerance doses for other radiations giving the same effect computed. Only for electrons do the tolerance doses exceed the sterilizing doses. The Ellis formula does not represent isoeffects and can only be regarded as an approximation. Tolerance doses can be calculated giving the same cell survivals for all N-values, i.e. isoeffects, and a comparison with sterilizing doses shows a good agreement with clinical experience. However, the influence of the overall-dose rate still is an unsolved problem which can only be solved experimentally.


Subunit structure of 6-phosphofructokinase from brewers' yeast.

An analysis of 6-phosphofructokinase from brewers' yeast in the presence of sodium dodecylsulfate reveals the occurrence of four components with the following molecular weights: alpha = 140000, beta = 130000, and alpha' = 92000, beta' = 87000. It was found that the alpha- and beta-components can be converted to the alpha' and beta' components by treatment of the native preparation with hyaluronidase. A comparison of the molecular weight obtained by ultracentrifugation and gel filtration with the results obtained by dodecylsulfate electrophoresis after treatment with hyaluronidase reveals that the alpha' and beta' components are the smallest molecular structures obtained upon dissociation of the native enzyme. The mechanism of action of hyaluronidase suggests a desensitization of the alpha and beta components of the enzyme towards dodecylsulfate. Thus, in the absence of hyaluronidase treatment; only an apparent molecular weight for the alpha and beta component is obtained. The analysis indicates that the native enzyme might be composed of four different subunits with an alpha, beta, alpha' and beta' configuration. It is not excluded that the native enzyme consists only of alpha- and beta-chains.


Ca2+-ATPase activity and lipid composition of sarcoplasmic reticulum of the gastrocnemius muscle of denervated frog.

Abnormalities have been noticed in the phospholipid and cholesterol composition of the atrophied gastrocnemius muscle of frog denervated for 1 month. Cholesterol : phospholipid molar ratios in the muscle increased on denervation. Sphingomyelin and cardiolipin fractions increased in contrast to phosphatidyl choline, phosphatidyl serine and phosphatidyl ethanolamine in the sarcoplasmic reticulum (SR) of denervated muscle. Na-azide sensitive Ca2+ ATPase activity of the mitochondria did not alter whereas that of SR decreased on denervation. Phospholipase C digestion impaired the organelle Ca2+-ATPase activity. The above abnormalities in enzyme activities have been correlated to the changes in the lipid composition of the denervated muscle. On the basis of these changes it is discussed that the primary change in the muscle due to denervation is the change in the permeability of the membrane.


Assessment of cost related characteristics and conditions of long-term care patients.

The present federal policy that requires a cost differential to be imposed between intermediate and skilled care residents/patients is based on a number of unproven assumptions that need to be tested. Are there groupings of cost related characteristics and conditions that allow appropriate distinctions between intermediate and skilled nursing care? If so, is the difference in the cost of care greater than the cost of collecting the data? Do the intangible costs exceed the benefits, especially if forced transfer of residents/patients is involved? If groupings do exist, will facilities reimbursed for high-cost patients be encouraged to maintain them in a state of dependence? Further, can the decisions be made reliably, even in communities where a spectrum of alternative resources is not available? Are there political reasons why objective decisions would not be acceptable to state administrators trying to balance their budgets? At the least, it appears that further research is needed in all of these areas. Before the results are in, perhaps a careful, judicious interpretation of the law is needed in order to positively affect the quality of life available to the persons consuming nursing home services.


Inactivator of the first component of human complement (C1INA). Enhancement of C1INA activity against C1s by acidic mucopolysaccharides.

Acidic mucopolysaccharides enhanced C1INA activity against C1s. The presence of heparin in the reaction mixture of C1INA and C1s markedly enhanced the inactivation of C1s by potentiating C1INA activity. Treatment of serum with C1s resulted in the inactivation of C4 hemolytic activity, but this inactivation of C4 by C1s was prevented by the presence of heparin presumably due to the enhancement of C1INA activity normally present in serum. The other acidic mucopolysaccharides, chondroitin sulfates A, B and C, were also effective on potentiating C1INA activity against C1s, but they were less active than heparin. The enhancement of C1INA activity by heparin was not observed in the reaction between C1INA and plasmin.


American and Soviet medical manpower: growth and evolution, 1910-1970.

Between 1910 and 1970 the number of physicians in the United States increased 2.5 times, in Soviet Russia almost 25 times. The number of physicians per constant unit of population remained fairly stable in the United States, rising slightly in the last few years. In the U.S.S.R. that number increased 16 to 18 times, and now stands about 50 per cent higher than in the United States. About 10 per cent of American physicians are women; in the U.S.S.R. it is about 70 per cent. Neither society has resolved the problem of deploying physicians to the rural areas. American physicians are more specialized than their Soviet colleagues. The article concludes with general remarks about the two health systems, pointing out resemblances and divergences. The hypothesis of a possible "convergence" is entertained.


Experimental immune complex glomerulopathy: immunomorphological aspects and correlations with human disease.

Thirteen rabbits received parenteral injections of human serum albumin (HSA) in regular doses and intervals during 6-32 seeks. The antigen load elicited immune responses of variable degree, giving a good correlation with the extent and quality of immune complex (IMC) deposition, as shown by immunological methods, electron and fluorescence microscopy and by histoautoradiography. Morphology of immune deposits (IMD) depended on the composition of the IMC and produced a continuous scale of glomerular involvement, ranging from minimal and focal to massive and diffuse deposition. Probable rudimentary traces of IMD were apparent in the ultrastructure even in animals with negative results of both immunological and immunfluorescence examinations. The relationships are discussed between experimental IMC-induced glomerulopathy on the one hand, and some problems of development and bioptic evaluation of human glomerulonephritis.


Biosynthesis and metabolism of androgenic hormones in cancer of the prostate.

The conversion of dehydroepiandrosterone sulfate into testosterone, delta4-androstenedione, and dihydrotestosterone has been measured in minced prostatic adenocarcinoma tissue from three untreated patients, two patients treated with diethylstilbestrol, and one treated with cyproterone acetate. The results obtained show that in both treated and untreated patients the dehydroepiandrosterone sulfate is actively transformed into dehydroepiandrosterone. The subsequent conversion into testosterone and dihydrotestosterone is less evident and inconstant in untreated patients and is inhibited in patients treated with diethylstilbestrol or cyproterone acetate. No formation of delta4-androstenedione was demonstrated in either treated or untreated patients.


Natural remission of nephrotic syndrome in a dog with immune-complex glomerular disease.

A nephrotic syndrome caused by immune-complex glomerular disease was diagnosed in a 4-year-old male Great Dane. The syndrome was characterized by proteinuria, hypoproteinemia, hypoalbuminemia, hypercholesterolemia, and subcutaneous edema. Renal biopsy revealed segmental membranous glomerular disease. The edema underwent complete remission 18 days after admission. Two months after admission, there was no clinical or laboratory evidence of glomerular disease. Periodic reevaluation of the dog during the next 2 years revealed recurrence of proteinuria, but no other clinical or laboratory abnormalities. Serial renal biopsies revealed persistence, but no appreciable increase, in the severity of the segmental membranous glomerular disease. The natural course of the nephrotic syndrome and immune-complex glomerular disease has been associated with unpredictable variability. It was concluded that the widespread use of corticosteroid or immunosuppressant therapy in dogs with immune complex glomerular disease should be withheld until the natural course of the disease has been evaluated in a significant number of patients and until the results of well-controlled clinical studies confirm or deny their therapeutic value.


Spectinomycin modification. I 7-EPI-9-deoxy-4(R)-dihydrospectinomycin.

7-EPI-9-deoxy-4(R)-dihydrospectinomycin (10) had been prepared and its structure firmly established by complete analysis of its pmr spectrum. This analog of spectinomycin is devoid of antibiotic activity.


Spectinomycin modification. II. 7-EPI-sectinomycin.

7-Epi-spectinomycin (9) and 7-epi-4(R)-dihydrospectinomycin (10) have been prepared and their structure firmly established by proton magnetic resonance. Both of these spectinomycin analogs are devoid of antibiotic activity.


Sucrose-dependent spectinomycin-resistant mutants of Escherichia coli.

Spectinomycin-resistant (Spcr) mutants of Escherichia coli were isolated from nutrient agar plates containing 20% sucrose and 100 mug of spectinomycin per ml. About one-third of the Spcr mutants thus obtained were sucrose dependent (Sucd) and were classified into two types: I, those unable to grow on sucrose-free medium in the presence of spectinomycin; and II, those unable to grow on sucrose-free medium irrespective of the presence of spectinomycin. Most of these mutants were hypersensitive to antibiotics, dyes, and detergents and were abnormal in cell morphology, suggesting changes in cell envelopes. Reversion experiments indicated that the sucrose-dependent spectinomycin resistance and hypersensitivity to various chemicals were not independently induced properties. The Sucd-Spcr mutations of type I mutants were transducible by phage P1 and were mapped at the strA-aroE region.


Synthesis of polyadenylic acid-containing ribonucleic acid during the germination of Neurospora crassa conidia.

Ribonucleic acid (RNA) synthesized during the first 1 h of conidial germination (15 to 20, 25 to 30, and 55 to 60 min) has been characterized by sucrose-sodium dodecyl sulfate gradient centrifugation, binding to polyuridylic acid filters, and oligo(dT)-cellulose chromatography. At all labeling periods examined, polyadenylic acid-containing RNA is synthesized, processed, and incorporated into polysomes. Approximately 40% of the labeled RNA sedimenting between 5 and 17S binds to polyuridylic acid filters. RNA which binds to oligo(dT)-cellulose displays a heterogeneous distribution in sucrose-sodium dodecyl sulfate gradients with a major, broad peak at 10-16S. In addition, some polyadenylic acid-containing RNA sediments beyond the 25S marker. Approximately 3% of the [3H]adenosine in pulse-labeled polysomal RNA is in polyadenylic acid segments resistant to pancreatic and T1 ribonucleases.


Energy coupling in the active transport of amino acids by bacteriohodopsin-containing cells of Halobacterium holobium.

Growth of Halobacterium halobium under illumination with limiting aeration induces bacteriorhodopsin formation and renders the cells capable of photophosphorylation. Cells depleted of endogenous reserves by a starvation treatment were used to investigate the means by which energy is coupled to the active transport of [14C]proline, -leucine, and -histidine. Proline was readily accumulated by irradiated cells under anaerobiosis even when the photophosphorylation was abolished by the adenosine triphosphatase inhibitor N,N'-dicyclohexylcarbodimiide (DCCD). The uptake of proline in the dark was limited except when the cells were allowed to accumulate adenosine 5'-triphosphate (ATP) by prior light exposure or by the oxidation of glycerol. DCCD inhibited this dark uptake. These findings essentially support Mitchell's chemiosmotic theory of active transport. The driving force is apparently the proton-motive force developed when protons are extruded from irradiated bacteriorhodopsin or by the dydrolysis of ATP by membrane adenosine triphosphatase. Carbonylcyanide m-chlorophenylhydrazone (CCCP), a proton permeant known to abolish membrane potential, was a strong inhibitor of proline uptake. Leucine transport was also apparently driven by proton-motive force, although its kinetic properties differed from the proline system. Histidine transport is apparently not a chemiosmotic system. Dark- or light-exposed cells show comparable initial rats of histidine uptake, and these processes were only partially inhibited by DCCD or CCCP. The histidine system apparently does not utilize ATP per se since comparable rates of uptake were exhibited by cells of differing intracellular ATP levels. Irradiated cells did effect a greater total accumulation of histidine than dark-exposed cells. These findings suggest that ATP is needed for sustained transport.


Nature of the energy requirement for the irreversible adsorption of bacteriophages T1 and phi80 to Escherichia coli.

The nature of the energy requirement for irreversible adsorption of phages T1 and phi80 was studied by using various specific energy inhibitors and mutants lacking either the Ca2+, Mg2+-adenosine triphosphatase or the ability to produce cytochromes in the absence of added 5-aminolaevulinic acid. It was found that irreversible adsorption could be energized both through the electron transport chain and from adenosine 5'-triphosphate via the Ca2+, Mg2+-adenosine triphosphatase, indicating the involvement of the energized membrane state. These results and the discovery that phages T1 and phi80 adsorb reversibly to the isolated tonA gene product are discussed in terms of the possible involvement of functions expressed by the tonB gene region in irreversible adsorption and the relationship to iron transport.


Mutant of Escherichia coli defective in response to colicin K and in active transport.

A mutant of Escherichia coli has been isolated that grows poorly on succinate and exhibits a markedly reduced sensitivity to colicin K. This mutant is also deficient in the respiration-linked transport of proline and thiomethyl-beta-D-galactoside but appears normal for the adenosine triphosphate-dependent transport of glutamine and arginine. A temperature-conditional revertant of the mutant grows on succinate and is sensitive to colicin K at 27 C, but fails to grow on succinate and is insensitive to colicin K at 42 C. Proline transport in the temperature-conditional revertant is reduced at 42 C when either glucose or succinate is used as energy source. Glutamine transport, on the other hand, is normal at 42 C with glucose as energy source, but is reduced with succinate, although not to the same extent as is proline transport. The lack of growth on succinate and the deficiencies in transport at 42 C are not due to a temperature-dependent lesion in either the electron transport chain or in Ca2+, Mg2+-activated adenosine triphosphatase activity. Membrane vesicles prepared from the temperature-conditional revertant are impaired in proline transport at both 27 and 42 C. These findings suggest the existence in the cytoplasmic membrane of E. coli cells of a component, presumably protein, that is required for colicin K action and that functions in respiration-linked and, to a lesser degree, in adenosine triphosphate-dependent active transport systems. This protein may serve as the primary target of colicin K action.


Coordinated alterations in ribosomes and cytoplasmic membrane in sucrose-dependent, spectinomycin-resistant mutants of Escherichia coli.

Alterations in cytoplasmic membrane and ribosomes from sucrose-dependent spectinomycin-resistant (Sucd-Spcr) mutants of Escherichia coli, mutants that are resistant to spectinomycin in the presence of 20% sucrose but sensitive in the absence of sucrose, were studied. The protein composition of cytoplasmic membrane was analyzed by gel electrophoresis on polyacrylamide gel containing 8 M urea and 0.5% sodium dodecyl sulfate, which assured the reproducible separation of 28 protein bands. A major protein band, I-19, was missing in all cytoplasmic membrane preparations from 10 Sucd-Spcr mutants. Besides protein I-19, proteins I-13 and I-24 were missing in some mutants. On the other hand, the protein composition of cytoplasmic membrane from a sucrose-independent spectinomycin-resistant mutant was indistinguishable from that from the wild-type strain. The polypeptide synthetic activity of ribosomes from Sucd-Spcr mutants was resistant to spectinomycin. Studies on a revertant obtained from one of these mutants without any selection for sensitivity to spectinomycin revealed that a single mutation was responsible for both the ribosomal alteration, i.e., spectinomycin resistance, and the lack of protein I-19 in the cytoplasmic membrane. Studies on a transductant obtained with a Sucd-SPcr mutant as the donor also confirmed the single-mutation concept. It was concluded that in Sucd-SPcr mutants an alteration in the ribosomes caused the deletion of protein I-19 from cytoplasmic membrane.


Inactivation of carotenoid-producing and albino strains of Neurospora crassa by visible light, blacklight, and ultraviolet radiation.

Suspensions of Neurospora crassa conidia were inactivated by blacklight (BL) radiation (300 to 425 nm) in the absence of exogenous photosensitizing compounds. Carotenoid-containing wild-type conidia were less sensitive to BL radiation than albino conidia, showing a dose enhancement factor (DEF) of 1.2 for dose levels resulting in less than 10% survival. The same strains were about equally sensitive to shortwave ultraviolet (UV) inactivation. The kinetics of BL inactivation are similar to those of photodynamic inactivation by visible light in the presence of a photosensitizing dye (methylene blue). Only limited inactivation by visible light in the absence of exogenous photosensitizers was observed. BL and UV inactivations are probably caused by different mechanisms since wild-type conidia are only slightly more resistant to BL radiation (DEF = 1.2 at 1.0% survival) than are conidia from a UV-sensitive strain (upr-1, uvs-3). The BL-induced lethal lesions are probably no cyclobutyl pyrimidine dimers since BL-inactivated Haemophilus influenzae transforming deoxyribonucleic acid is not photoreactivated by N. crassa wild-type enzyme extracts, whereas UV-inactivated transforming deoxyribonucleic acid is photoreactivable with this treatment.


Immunoferritin determination of the distribution of (Na+ + K+) ATPase over the plasma membranes of renal convoluted tubules. I. Distal segment.

The distribution of (Na+ + K+) ATPase over the plasma membranes of distal convoluted tubules from canine kidney has been determined. This enzyme is responsible for the coupled active transport of Na+ and K+ across animal cell membranes. Ultrathin frozen sections were cut from fragments of renal cortex and specifically stained with antibodies, which recognize antigenic sites on the enzyme, and ferritin-conjugated goat antirabbit gamma-globulins. It is demonstrated that (Na+ + K+) ATPase is distributed uniformly and at high concentration over the plasma membranes which form the intercellular spaces of this epithelium. The enzyme is located on the luminal surface of the tubules as well but at a much lower concentration. These results, in combination with those of previous determinations of the cation fluxes across this epithelium, can be used to formulate a complete description of the cation movements through this tissue.


Immunoferritin determination of the distribution of (Na+ + K+) ATPase over the plasma membranes of renal convoluted tubules. II. Proximal segment.

The distribution of (Na+ + K+) ATPase over the plasma membranes of the proximal convoluted tubule from canine renal cortex has been determined. Ultrathin frozen sections of this tissue were stained with rabbit antibodies to this enzyme and ferritin-conjugated goat antirabbit gamma-globulin. It is demonstrated that high concentrations of this enzyme uniformly line the intercellular spaces of this epithelium. The consequences of this observation are discussed in terms of the low resistant tight junctions of these tubules and the isotonic fluid transport which they support. Furthermore, antibodies to (Na+ + K+) ATPase recognize an antigen on the luminal surfaces of the tubules within the brush border. It is proposed that the enzyme is present in this region of the plasma membrane as well, although at much lower concentration. To further substantiate this conclusion, a brush border fraction has been purified from rabbit kidney and been shown to contain significant (Na+ + K+) ATPase. These results contradict earlier conclusions about the location of (Na+ + K+) ATPase in this tissue.


Pyrolysis gas chromatography as an aid to the identification of Penicillium species.

Using pyrolysis gas chromatography it was possible to identify each of a series of eleven Penicillium species and the related species Aspergillus niger CMI 31821 and Neurospora crassa CMI 75723, based on relative peak heights and retention times of the most reproducible peaks in each pyrogram.


Method for the measurement of plasma dehydroepiandrosterone by gas-liquid chromatography with electron capture detection.

A method is described for the measurement of plasma dehydroepiandrosterone, as the iodomethyldimethylsilyl ether derivative, by gas-liquid chromatography and electron capture detection using the relatively new and highly stable stationary phase Dexsil 300. Preliminary purification of the plasma extract was required and alumina column chromatography was utilised, both before and after derivatization of the steroid extract. Specificity, precision, sensitivity and accuracy were all satisfactory. The method was used to study the relationship between age and the level of plasma dehydroepiandrosterone in a group of normal women. A significant negative correlation was observed.


In vitro steroid metabolic studies in testicular 17 beta-reduction deficiency.

In order to document testicular 17beta-reduction deficiency (17RD) and to search for additional metabolic aberrations possibly associated with this disorder, the metabolism of 14C-labeled pregnenolone (delta5P), 17-HYDROXYPROGESTERONE (17OHP), dehydroepiandrosterone (DHEA), androstenedione (A), testosterone (T) and estrone (E1) was studied in testicular minces from a 46-year-old male pseudohermaphrodite (MPH) with highly elevated testicular A and minimal T secretion but normal extragonadal conversion of A to T. Testicular minces from a 20-year-old MPH with apparently normal testicular T biosynthesis served as a control. The results of this investigation show that the 17RD testes metabolized delta5P along delta5- and delta4- pathways but, in contrast to the control, converted more 17OHP, metabolizing it predominantly to A rather than T, failed to reduce DHEA to androst-5-ene-3beta,17beta-diol, metabolized DHEA very efficiently to A and produced little T, and converted only minimal quantities of A and E1 to their 17beta-reduced counterparts. 17beta-Reduction increased slightly but was far from being restored to control levels upon addition of NADH plus NADPH. However, oxidation of T to A by 17RD testicular minces, with and without additional NAD plus NADP, was comparable to that by the control. These results document 17RD for A, DHEA and E1 and suggest that the lack of elevated 17OHP and DHEA secretion by the 17RD testes was due to increased 17, 20-lyase and perhaps elevated 3beta-hydroxysteroid dehydrogenase and/or isomerase activity. The observation that 17beta-reduction was only slightly increased upon addition of NADH plus NADPH, but that oxidation of T to A was normal, is consistent with the assumption that more than one 17beta-hydroxysteroid dehydrogenase may be involved in testicular 17beta-reduction and/or 17-oxidation, and that the 17RD testes studied either lacked the enzyme which acts predominantly as 17beta-reductase or that the affinity of this 17beta-reductase for reduced cofactor(s) and/or substrates was abnormal.


The Raji cell  radioimmune assay for detecting immune complexes in human sera.

A sensitivie and simple procedure for the detection and quantitation of soluble complement (C)- fixing immune complexes in sera of patients with various disease states has been developed by utilizing C receptors on Raji cells. These cells lack membrane-bound immunoglobulin but have receptors for IgG Fc, C3b, C3d, and possibly with other C proteins. Uptake experiments showed that both aggregated human gamma globulin (AHG) and 7S IgG bound to receptors for IgG Fc; however, AHG reacted with C bound to cells only via receptors for C and this binding was much more efficient than via IgG Fc receptors. AHG was used as an in vitro model of human immune complexes and its uptake by Raji cells was quantitated by 125I-radiolabeled antihuman IgG. The limit of sensitivity of this test was 6 mug  AHG/ml serum. The ability of Raji cells to detect AHG in serum depended on the amount of radioactive antibody used and the size of aggregates. The presence of an excess of C somewhat inhibited binding of AHG containing C to Raji cells. The efficient binding of AHG by receptors for C on Raji cells was used for the detection and quantitation of immune complexes in human sera. Raji cells were incubated with sera to be tested and then reacted with excess radiolabeled antihuman IgG; the amount of radioactivity bound to the washed cells was determined and referred to a standard curve of radioactive antibody uptake by cells previously incubated with increasing amounts of AHG in serum. Thereby immune complexes were detected and quantitated in serum hepatitis, systemic lupus erythematosus, vasculitis, subacute sclerosing panencephalitis, dengue hemorrhagic fever, and malignancies.


Light and immunofluorescent study of the Arthus reaction in the rabbit lung.

A localized Arthus reaction was produced in the lung of sensitized rabbits by delivery of antigen into a lower lobe bronchus using a method of selective bronchial catheterization under fluoroscopy. The rabbits were sensitized with bovine immunoglobulin G (B-IgG) in incomplete Freund's adjuvant (IFA) to produce precipitating antibody without classic delayed hypersensitivity. Pulmonary histopathology was studied at intervals following antigen challenge, using light and immunofluorescent microscopy. Gross lesions peripheral to the lower lobe bronchus receiving antigen were found within 12 hr. Subsequent necrosis resulted in a dense scar by 6 wk. Microscopically, early lesions were typified by localized bronchitis, bronchiolitis, alveolitis, and vasculitis with exuberant exudates containing predominantly polymorphonuclear leukocytes. Extensive focal necrosis was present by 72 hr. Immunofluorescent studies revealed the presence of B-IgG, rabbit IgG, and complement (C3) in and around bronchi, bronchioles, alveoli, and vessels. No granulomatous lesions were found, and proliferation of alveolar lining cells was not detected in these studies. Thus, the lung can participate in an acute Arthus reaction following local antigen challenge in systemically sensitized animals. The pathology more closely resembles a necrotizing bacterial pneumonia than an interstitial or hypersensitivity pneumonitis under the conditions of this experimental system. Implications for human disease are speculative.


In vitro generation of suppressor cell activity: suppression of in vitro induction if cell-mediated cytotoxicity.

It was observed that when normal mouse spleen cells were cultured alone in vitro (precultured) for 3 to 7 days, these cells lost the ability to generate cell-mediated cytotoxicity (CML) during subsequent in vitro sensitization with allogeneic spleen cells, trinitrophenyl (TNP)-modified syngeneic spleen cells, or syngeneic tumor cells. These precultured cells, which were themselves unable to generate CML, were also shown in mixing experiments to suppress, actively, the generation of CML by freshly explanted spleen cells. Suppression occurred at the sensitization phase of CML, and not at the effector level; supernatants from suppressive precultured cells were not suppressive. Suppression was totally abrogated by the treatment of spleen cells with a T cell-specific rabbit anti-mouse brain serum and complement (RalphaMB+C) either before or after preculturing, suggesting that a T cell eas essential both to the generation of suppressor activity and to its expression. Suppressor activity was entirely absent in precultured nylon wool column-nonadherent spleen cells, a T cell-enriched population containing most of the RalphaMB+C-sensitive cells in the spleen. Precultured nylon column-adherent cells (T cell-depleted) did have suppressive activity, and a mixture of nylon-adherent and nylon-non-adherent cells was a suppressive after preculture as the precultured unseparated spleen. Moreover, the ability of nylon-adherent spleen cells to generate suppressive activity during preculturing was abrogated by treatment with RalphaMB+C. Thus, the "spontaneous" generation of CML-suppressive activity was dependent upon a limited subpopulation of splenic T cells isolated in the nylon column-adherent fraction. The relationship of these data to a previously described synergy between subpopulations of normal spleen in the generation of CML is discussed, and the findings related to other suppressor systems described in the literature.


Stimulation of lymphoid cells from normal and immune mice by syngeneic BALB/c plasma cell tumors.

Lymphoid cells from normal and immunized BALB/c mice could be stimulated in vitro by syngeneic PCT contrasted with an absence of response to a number of other tumors. Maximal responses of normal cells to PCT were found to occur 5 days after the initiation of the cultures at an optimal responding:stimulation cell ratio of 1:2. MLTI activity of normal cells could not be blocked or enhanced by PCT myeloma protein products indicating that MLTI reactivity was directed against non-idiotypec cell surface determinants. Lymphoid cells from immunized mice demonstrated increased MLTI responses to cells of the immunizing tumor but not to other PCT, indicating that the post-immunization MLTI responses were primarily to individual rather than shared tumor cell surface antigens. Activity of both normal and immunized spleen cells was found to involve thymus-derived lymphocytes. The persistence of residual MLTI activity after treatment with anti-theta serum and complement, however, implicated participation of non-theta antigen-bearing cells in MLTI reactivity. From these data, we conclude that lymphoid cells from un-immunized mice are capable of T cell-dependent reactivity to syngeneic PCT-associated antigens and that elevations in these reactivities after immunization may reflect specific cellular immune responses.


The effects of cell density, incubation temperature, syngeneic serum, and syngeneic red blood cells on mouse lymphocyte responses in vitro.

Various parameters affecting the quantitative assessment of mouse mixed lymphocyte response (MLR) in microplates were investigated. In agreement with the findings of others cell density was found to be particularly critical, maximal proliferative efficiency occurring at 3 X 10(6) viable cells/cm2. Slight reduction in the degree of viable cell--cell interaction caused by either reduction in cell density or insertion of irradiated cells into the cultures greatly reduced the efficiency of the proliferation. An incubation temperature of 33-34 degrees C was found to have distinct advantages over 37 degrees C, there being improved cell survival and reproducibility at this lower temperature. Transformation of mouse lymphocytes was strongly inhibited by low concentrations of normal syngeneic serum, the degree of inhibition depending on the concentration of foetal calf serum (FCS) in culture and on the method of preparation of serum. Inhibition of mouse cell responses was also caused by syngeneic red blood cells (RBC), in contrast to the pronounced enhancement of guinea pig lymphocyte responses to phytohaemagglutinin (PHA) in the presence of syngeneic RBC.


Single and multiple drug therapy in autologous immune complex nephritis in rats.

Autologous immune complex (AIC) nephritis is a form of chronic renal disease with remarkable similarities to idiopathic membranous nephropathy occurring in man. AIC nephritis was induced in 160 gram Lewis rats with a single footpad injection of tubular brush-border antigen (FxIA) in complete Freund's adjuvant. When killed at 8 weeks, 85 per cent of the rats demonstrated typical diffuse glomerular deposits of immunoglobulin G and B1C (C1/3 component of complement) by immunofluorescent microscopy, and subepithelial electron-dense deposits by electron microscopy. Both immune complex disease and significant proteinuria occurred in two-thirds of these animals. An attempt to modify the natural course of established AIC nephritis using large doses of potent glucocorticoids (methyl-prednisolone), anti-inflammatory agents (acetylsalicylic acid, indomethacin, and cyproheptadine), and immunosuppressive drugs (cyclophosphamide, azathioprine) was begun 4 weeks after initial immunization and continued for 4 more weeks. None of the single drug nor multiple drug protocols employed was of demonstrable benefit in ameliorating the immune events operating in AIC nephritis. Cyclophosphamide and indomethacin, when used singly, were associated with significant mortality in the animals studied. All combined drug protocols involving glucocorticoids and antimetabolites were associated with unacceptable mortality as well. Of interest, immune complexes could not be demonstrated in the vascular choroid plexus of any rat with AIC nephritis. This failure to modify the course of established renal disease (AIC) in an experimental animal with generally available pharmacologic agents, is similar to the usual results of such treatment in chronic renal disease (idiopathic membranous nephropathy) in man. It is possible that new and more potent anti-inflammatory agents employed singly or in various combinations, will permit more successful manipulation of the host's immunologic system to prevent or modify immune injury of the renal glomerulus.


Electrocardiogram during experimental coronary thrombolysis.

The electrocardiogram (ECG) was analyzed before and after enzymatic lysis of an intracoronary thrombus in six closed-chest dogs, and in a control group of five dogs without a thrombus. Thrombolysin (18,000 to 50,000 Merck units in 250 ml of 5% dextrose in water) was infused directly into the anterior descending coronary artery in both groups under the same conditions. Complete recanalization of the obstructed artery was achieved in all cases after development of an occluding thrombus and signs of myocardial infarction. In the experimental group, statistically significant differences were found in the mean HR and the R-R,QRS, Q-T and Q-Tc intervals. Changer in the mean values for these parameters were also observed in the control groups without coronary thrombosis but were not significant. Sinus rhythm was maintained in both groups. Histologic findings of the affected myocardium following Thrombolysin infusion was compatible with those seen in acute myocardial infarction. No evidence of necrosis, hemorrhage or inflammation was observed in the control hearts. Infusing Thrombolysin directly into the anterior descending artery had no adverse effect on the myocardium.


Hyphal wall growth in Neurospora crassa and Geotrichum candidum.

Growth of the walls of hyphae of Neurospora crassa and Geotrichum candidum was studied using longitudinal and serial transverse sectioning methods. Rigidification of the hyphal wall below the extension zone did not appear to involve the gross formation of a secondary wall since the transition from extensible to non-extensible wall was not associated with an increase in thickness. However, behind the extension zone the walls leading hyphae of N. crassa increased in thickness until eventually they attained a thickness which was up to five times that of the tip wall. A hypothesis of hyphal wall growth is proposed.


Morphology and growth kinetics of hyphae of differentiated and undifferentiated mycelia of Neurospora crassa.

A comparison was made of the morphology and growth kinetics of hyphae of differentiated and undifferentiated mycelia of Neurospora crassa. Undifferentiated mycelia were formed during exponential growth on solid media or submerged culture. Hyphae at the margin of differentiated mycelia (colonies) differed from undifferentiated mycelia in diameter, extension rate, extension zone length, and intercalary and apical compartment length. The mean hyphal extension rate (E) of an undifferentiated mycelium was a function of the length of the mycelium's hyphal growth unit (G) and the organism's specific growth rate (alpha). Thus, E=Galpha.


Biogenesis of mitochondrial membranes in Neurospora crassa during cellular differentiation: changes in oxidative phosphorylation and synthesis of mitochondrial phospholipids.

Changes in the capacity of mitochondria to carry out oxidative phosphorylation and in the rate of synthesis and incorporation of phospholipids into mitochondria were measured during the germination of conidiospores of Neurospora crassa. The competence of isolated mitochondria to carry out coupled respiration was very low during the first 3 h growth, but it increased rapidly, reaching maximal levels at 5 to 6 h growth. Changes in mitochondrial function were the same in cells grown in 2% sucrose- or 15% glucose-supplemented medium. The rate of synthesis of mitochondrial phospholipids was very low during the first 2 h growth and increased to maximal levels between 3 and 5 h. The rate of synthesis of mitochondrial phospholipids was approximately three times higher in cells grown in 15% glucose than in those grown in 2% sucrose. The maximal rate of synthesis of mitochondrial phospholipids occurred during spore germination and preceded attainment of full competence for oxidative phosphorylation. The lipid-rich condition of the mitochondrial resulting from the high rate of synthesis of phospholipids in glucose-grown cells is postulated to be related to the whorled inclusions observed in thin sections of Neurospora cells.


Biogenesis of mitochondrial membranes in Neurospora crassa during cellular differentiation: ultrastructural changes accompanying differentiation.

The ultrastructural characteristics of Neurospora cells during dedifferentiation and redifferentiation of conidiospores into vegetative cells have been determined. This germination process occurs between 2 and 5 h after inoculation; by 3-5 h, approximately 50% of the cells have germinated. The cells enter the exponential phase of dry-weight gain between 4 and 5 h after inoculation. Several unusual structures are observed in Neurospora cells during germination. Whorled structures are frequently seen in the cytoplasm during germination, and occasionally at other times. They appear to be derived from the cytoplasmic membrane. Whorled structures of different appearance were observed in the mitochondria between 2 and 4 h after inoculation. Their number was related to the level of metabolizable carbohydrate, and was higher in 15% glucose-than in 2% sucrose-supplemented medium, and very low in medium containing 15% mannitol, or 2% sucrose+13% 2-deoxyglucose, or no added carbohydrate. The mitochondrial inclusions were osmiophilic and could be removed by treatment with 90% aqueous acetone in the cold, indicating that they were composed at least in part of lipid. The strong dependence of the number of mitochondrial inclusions on time and on carbohydrate supplementation, suggests that there is a physiological basis for these structures and that they reflect changes occurring in the mitochondria at times significant to cellular differentiation.


Biogenesis of mitochondrial membranes Neurospora crassa during cellular differentiation: ultrastructural changes accompanying differentiation.

The ultrastructural characteristics of Neurospora cells during dedifferentiation and redifferentiation of conidiospores into vegetative cells have been determined. This germination process occurs between 2 and 5 h after inoculation; by 3-5 h, approximately 50% of the cells have germinated. The cells enter the exponential phase of dry-weight gain between 4 and 5 after inoculation. Several unusual structures are observed in Neurospora cells during germination. Whorled structures are frequently seen in the cytoplasm during germination, and occasionally at other times. They appear to be derived from the cytoplasmic membrane. Whorled structures of different appearance were observed in the mitochondria between 2 and 4 h after inoculation. Their number was related to the level of metabolizable carbohydrate, and was higher in 15% glucose-than in 2% sucrose-supplemented medium, and very low in medium containing 15% mannitol, or 2% sucrose +13% 2-deoxyglucose, or no added carbohydrate. The mitochondrial inclusions were osmiophilic and could be removed by treatment with 90% aqueous acetone in the cold, indicating that they were composed at least in part of lipid. The strong dependence of the number of mitochondrial inclusions on time and on carbohydrate supplementation, suggests that there is a physiological basis for these structures and that they reflect changes occurring in the mitochondria at times significant to cellular differentiation.


The effect of canaline on some events in vaccinia virus replication.

Canaline, a pyridoxal phosphate antagonist, is shown to inhibit two distinct events in the replication of vaccinia virus in HeLa cells. Initial events proceed in the presence of the inhibitor leading to the formation of DNA-containing, cytoplasmic inclusions. However, further DNA synthesis is required for the subsequent production of infectious progeny following the reversal of canaline inhibition by pyridoxal phosphate. Inhibition of a separate, maturation event is shown by the delayed addition of canaline resulting in the failure to coat virus-specific DNA synthesized previously in the absence of the inhibitor. Thus, the replication of vaccinia virus is sensitive to inhibition by canaline at an early and a late stage in the replication cycle. Reversal is accomplished alternatively by the addition of 'non-essential' amino acids suggesting that the effects of canaline result from inhibition of specific protein functions.


Ragged-red fibers. A biochemical and morphological study.

Histochemical, ultrastructural and biochemical studies were performed on muscle biopsy specimens from a 30-year-old man with proximal limb weakness. Modified Gomori trichome stains of anterior tibial muscle revealed accumulations of red-staining material in the subsarcolemmal and intermyofibrillar regions of virtually every fiber (ragged-red fibers); these accumulations were rich in oxidative enzymes. Electron microscopy of this muscle showed that the red-staining areas consisted of large collections of abnormal-appearing mitochondria. Mitochondria isolated from the quadriceps muscle showed lack of respiratory control with alpha-glycerophosphate as substrate. However, the lack of respiratory control with alpha-glycerophosphate must be interpreted with caution since the quadriceps muscle was severely degenerated, and biochemical alterations of mitochondria may occur secondary to muscle degeneration itself. Nevertheless, this is the second reported case of lack of respiratory control with alpha-glycerophosphate in a patient with ragged-red fibers. Further interpretation of this defect and its significance must await more studies to determine whether this muscle mitochondrial abnormality is a common finding in the disorders in which ragged-red fibers are encountered.


A new approach to the surgical treatment of lumbar spondylosis.

The authors discuss routine removal of inferior articular processes and their facets during laminectomy, with wide visualization of the lumbar nerve roots, which has proven to be most successful in relief of the symptoms of lumbar spondylosis. No instance of postoperative vertebral instability has been found. In a consecutive series of 70 patients averaging 58.3 years of age, 91% of patients adequately followed did well. Of six patients who did poorly, three had apparent explanations and, presumably, such errors are avoidable. Despite the generally advanced age of this series (12 patients aged over 70 years) there was no mortality or undue morbidity. Patients previously operated on for disc herniation, as a group, were somewhat younger and the spondylosis tended to be more localized (to the area of previous surgery) than in patients operated on for the first time.


Cinematographic analysis of contractile events produced in intrafusal muscle fibres by stimulation of static and dynamic fusimotor axons.

1. Muscle spindles with an intact blood supply and uninterrupted connexions with ventral and dorsal spinal roots (Bessou & Pagés, 1967, 1972) have been prepared in cat's tenuissimus muscles with the aim of cinephotographically recording intrafusal movements induced by the stimulation of single static or dynamic gamma axons; the time cours of these movements and the morphological kind of activated intrafusal muscle fibres have been established. 2. Displacements of spindle guiding marks in the equatorial region elicited by stimulating single static gamma axons are 4-20 times greater in amplitude than the ones elicited by stimulating dynamic gamma axons at the same frequency. 3. The dynamic gamma axons induced a contraction only in nuclear bag fibres which, in addition, never received any static gamma innervation. The static gamma axons evoked contractions either in nuclear bag fibres alone, or in nuclear chain fibres alone, or in both types of intrafusal fibres. Two thirds of static gamma axons supplied nuclear bag fibres. For various reasons, one half only of static gamma axons innervating nuclear bag fibres could be shown to simultaneously innervate nuclear chain fibres. Consequently, about one third of static gamma axons supplied both nuclear bag fibres and nuclear chain fibres, but it is highly probable that this latter figure is an underestimate. One third of static gamma axons produced contraction in nuclear chain fibres only. In this work, the distribution of fusimotor axons has been established in only one muscle spindle of the cluster of muscle spindles that each fusimotor axon is generally innervating. 4. Generally speaking, a static gamma axon elicits contraction of several intrafusal fibres whereas a dynamic gamma axon innervates only one intrafusal fibre and frequently only one pole of the fibre. 5. One third of static gamma axons evoked contractions in nuclear chain fibres that seemed to involve the whole pole. The other static gamma axons and all dynamic gamma axons produced, in the intrafusal fibres that they supplied, one or several foci of localized contractions. 6. The nuclear chain fibres contract and relax faster than nuclear bag fibres. The contractions of nuclear bag fibres supplied by static gamma axons are stronger and faster than those of nuclear bag fibres innervated by dynamic gamma axons. Nearly all nuclear bag fibres innervated by static gamma axons, like the nuclear chain fibres, show transient contractions at each pulse of a stimulation at low frequency (2-20/sec). 7. The results are discussed taking into account the available anatomical and physiological data on the muscle spindle. Their consequences with regard to intrafusal working are briefly considered.


Load compensation in human masseter muscles.

The reaction of masseter and digastric muscles to changes in the load applied during a movement was studied in human volunteers. 2. Rapid stretch of the jaw-closing muscles evoked a monosynaptic jaw-jerk reflex response in the masseteric electromyogram (e.m.g.) after a delay of 6-8 msec. 3. The averaged integrated e.m.g. activity of the masseter was increased 5-10 msec after the rapid application of a load during voluntary closure of the jaw. It was not necessary to stretch the muscle to obtain this apparently monosynaptic response. 4. A compensatory increase in the velocity of movement followed 6-10 msec after the e.m.g. peak. 5. Unloading the masseter muscle during contraction was followed by a fall in e.m.g. activity, beginning after a latency of 6-5--1 msec, with later phases of depression beginning 27--36 and 60--70 msec after unloading. 6. The spindle-poor digastric muscles were not monosynaptically activated during loading, but responded with a latency of 24--34 msec. 7. It is concluded that monosynaptic e.m.g. changes are a prominent feature of the load compensation system in human jaw-closing muscles but not in jaw-opening muscles.


The use of IQ as a measure of problem solving ability with mongoloid and nonmongoloid retarded children.

A major purpose of this study was to determine the relationship of MA and IQ to learning a task, among retardates, involving conceptualization. Such tasks are considered by Jensen, in his theory on mental abilities, to test Level II abilities as opposed to Level I abilities which involve rote learning and primary memory. The theoretical basis for this part of the experiment are the opposing hypotheses of Weir and Zigler concerning the use of IQ or MA as the best measure of learning rate. Specifically, a dimension-abstracted oddity task was presented to 40 mongoloids and 40 nonmongoloids. The results supported Weir's hypothesis, as IQ was found to determine the rate of learning the task. A second major finding was that the nonmongoloids learned the task faster than the mongoloid Ss.


Social and play behaviors of institutionalized mongoloid and nonmongoloid retarded children.

A total of 24 institutionalized retarded children, 12 mongoloid (Down's syndrome) and 12 nonmongoloid, were observed in dyadic interaction with peers in a free-play situation, and a number of specific peer-social and non-social behaviors were recorded as they occurred. Differences between mongoloid and nonmongoloid Ss were most apparent on several social behavior categories which support the stereotypic conception of mongoloids as more sociable, gregarious, etc. Differences were most apparent for the mongoloid boys. The possible influence of tranquilizer drugs and cottage placements on the observed differences was discussed.


Follicular growth: the basic event in the mouse and human ovary.

Follicular growth is described as a continuum. It goes on at all times, at all ages, uninterrupted by pregnancy or other periods of non-ovulation. A distinction is made between the continuum at the beginning of follicular growth and events concerning the cyclicity at the end of follicular growth, i.e. ovulation. Follicles grow sequentially. Also large follicles continue to grow until they become atretic or ovulate. No evidence for a pool of large follicles held in reserve could be found. Examination of the effect of PMSG on the growth of large follicles showed that this hormone provented the degeneration of large follicles, thus allowing more follicles to grow further. As in the mouse, follicular growth occurs during human infancy and is the normal event during childhood. Ovaries without signs of follicular growth are uncommin in the child and are apparently connected with certain systemic diseases.


The fine structure of the glycocalyx of equine spermatozoa: a high-resolution cytochemical study.

Equine spermatozoa were obtained from ejaculates of young stallions. The seminal plasma was removed and the sperm pellets washed three times with 0-15 M-NaCl solution before final centrifugation at 4500 g for 15 min. The pellets were fixed in a mixture of 2-5% glutaraldehyde in 0-1 M-cacodylate buffer, pH 7-4, with 0-5% Alcian blue and post-fixed in 1% osmium tetroxide with 1% lanthanum nitrate; other samples were treated with ruthenium red. All samples were dehydrated in ascending concentrations of ethanol, embedded in araldite and thin sections examined in an electron microscope. Electron dense deposits of lanthanum were present on the surface plasmalemma of the head, mid-piece and tail of 70% of mature spermatozoa, and similar deposits were seen in ruthenium red-treated samples. No glycocalyx was observed in untreated spermatozoa.


An autoregulatory role for prostaglandins in placental hemodynamics: their possible influence on blood pressure in pregnancy.

This review asserts that prostaglandins of the E family play a vital regulatory role in uteroplacental blood flow. This role appears to be similar to activity within other organs, especially the kidney, in a process referred to as autoregulation of blood flow. This mechanism may serve as a system for ehmodynamic homeostasis in pregnancy. Moreover, hypertension in pregnancy (toxemia) may be due to a specific or generalized abnormality in this system.


Mutagenic evaluation of antischistosomal drugs and their derivatives in Neurospora crassa.

The mutagenic activities of lucanthone, hycanthone, niridazole, and the indazole analogs of lucanthone (IA-3 and IA-5) or hycanthone (IA-4 and IA-6) were studied by assaying for the induction of specific locus mutations in the ad-3 region of N. crassa. The results show that lucanthone, hycanthone, and their indazole analogs (IA-3 through IA-6) are all mutagenic in N. crassa when conidia are treated with any of these compounds. On a per mole basis, hycanthone is the least toxic and mutagenic, whereas IA-3 is the most toxic and mutagenic compound among the six closely related agents. In general, compounds with a methyl group at the C-4 position are more mutagenic than compounds with a methanol group; 6-chloroindazole analogs are more mutagenic and more toxic than nonchlorinated analogs. Niridazole is not mutagenic when conidial suspensions are treated. However, the mutation frequency increased more than 50-fold when niridazole was added to the medium used to grow vegetative cultures. Thus, it appears that the mutagenic activity of this latter compound requires metabolic activation.


Chloroquine-resistant malaria in Burma.

Two field trials to detect chloroquine-resistant malaria were conducted according to WHO recommendations in a malaria free area near Rangoon. Peripheral blood smears were examined for asexual forms of P. falciparum on day one through to day seven, on day 14, 21, and 28 after a standard dose of 1500 mg. of chloroquine base. Haskins test to detect chloroquine in urine was done on all cases and plasma chloroquine levels were measured in some. Out of 105 patients tested RI resistance was detected in 66, RII in 19 and RIII in three. Subsequent trials with other anti-malarial drugs indicated that the chloroquine-resistant P. falciparum were also resistant to one day therapy with pyrimethamine 50 mg. or sulphamethoxypyridazine 1 G given singly; and resistant to one day therapy with combinations of pyrimethamine 50 mg. plus sulphamethoxy pyridazine 1 G, pyrimethamine 13 mg. plus dapsone 100 mg., and trimethoprim 320 mg. plus sulphamethoxazole 1600 mg. All those tested were sensitive to quinine sulphate, 0-6 G given three times a day for 10 days, and were also sensitive to one day therapy with combinations of trimethoprim 500 mg. plus sulphalene 1 G, and pyrimethamine 50 mg. plus sulphamethoxine 1 G. Pyrimethamine 12-5 mg. plus dapsone 100 mg. in weekly doses was shown to be an effective chemoprophylaxis. Quinine was tested on 38 subjects while other drug schedule were tested on six to eight subjects.


Renal transplantation in prune belly syndrome.

The first reported case of renal transplantation in a patient with prune belly syndrome is presented. Transplanatation should be considered for patients with this syndrome in whom renal failure develops despite appropriate urologic reconstruction. A thorough preoperative evaluation ascertaining the status of the lower urinary tract is especially important in these patients.


Chlorpropamide-induced granulomas. A probable hypersensitivity reaction in liver and bone marrow.

Anicteric hepatitis, associated with fever and exfoliative dermatitis, developed in a diabetic patient two weeks after intake of a long-acting sulfonylurea, chlorpropamide (Diabinese). Granulomas showing heavy infiltration with eosinophils were found in the liver and bone marrow. These were interpreted as manifestations of an allergic reaction. The clinical signs, abnormal laboratory findings, and hepatic lesions subsided spontaneously on withdrawal of the drug. Bone marrow changes, however, persisted seven months after cessation of the drug. To our knowledge, this is the first report of a patient with liver and bone marrow inflammation characterized by granulomas with eosinophilic infiltration following intake of chlorpropamide.


Rates of cutaneous reactions to drugs. A report from the Boston Collaborative Drug Surveillance Program.

Rates of allergic skin reactions to commonly used drugs were estimated from data obtained on 22,227 consecutively monitored medical inpatients. A total of 57 drugs were implicated with skin reactions. Five or more reactions were attributed to each of 22 drugs (or drug groups). Many commonly used drugs did not appear to cause any allergic reactions. The study provides the practicing physician with drug-specific quantitative data that can be used to evaluate the causes of drug-induced rash, itching, or hives.


Endoscopic complications. Results of the 1974 American Society for Gastrointestinal Endoscopy Survey.

Esophagogastroduodenoscopy (211,410 examinations) had a complication rate of 1.3/1,000 cases. Duodenoscopy with cannulation was performed 3,884 times and had a complication rate of 21.6/1,000 examinations. Diagnostic coloscopy (25,298 examinations) had a complication rate of 3.4/1,000. Polypectomies during coloscopy (6,124 cases) had a complication rate of 23.3/1,000 cases. Esophageal dilations (13,139 cases) had a complication rate of 4.25 with mercury bougies, and in 9,431 cases metal olives produced a complication rate of 6.1/1,000 treatments. Dilation for achalasia in 1,224 patients produced a complication rate of 18.4/1,000 procedures. Peritoneoscopy (4,404 examinations) produced a complication rate of 5.4/1,000 patients. The value of these diagnostic and therapeutic procedures is now well established but must be weighed against a potential risk of complications.


Effects upon spindle discharges of electrical stimulation of static fusimotor fibers with concomitant application of muscle vibration.

Discharges of single afferent fibers from the primary endings of the soleus muscle spindles were recorded from thin dorsal root filaments in cats anesthetized with urethane and chloralose. The distal cut end of the ventral root was split into fine filaments to obtain functionally single fusimotor fibers. The fusimotor fibers obtained in this study were of the static type. The soleus muscle was sinusoidally stretched at 70 Hz with various amplitude concurrently with 100 Hz electric stimulation of fusimotor fiber. The spindle afferent discharges were analysed by compiling inter-spike interval histograms and cross-correlograms between the afferent spikes and the stimulus pulses applied to the fusimotor fiber. The same analysis was also made between the afferent spikes and peak extensions of muscle yielded by vibratory stimulation. One-third of the fusimotor fibers were capable of driving the spindle afferents. The driving of fusimotor stimulation was replaced by driving by muscle dibration of more than 10 mum amplitude applied concurrently with fusimotor stimulation. The remaining two-thirds of the fusimotor fibers could not drive the spindle afferents. In this case, the driving by muscle vibration was obtained when vibration of more than 5 mum amplitude was applied concurrently with fusimotor stimulation. It was suggested that fusimotor fibers which produced driving of the spindle afferents would terminate on nuclear chain fibers and those not producing driving on nuclear bag fibers, or the latter would terminate relatively distant from the primary ending as compared with the former.


[The ultrastructive of spotty corneal dystrophy (Groenouw II) (author's transl)].

Examination of the ultrastructure of cases of spotty corneal dystrophy (Groenouw II) showed polymorphous results in various layers of the cornea. Inclusions of acidic mucopolysaccharides occur in membrane surrounded storage granules and also in the free state. The results from sections of the central and peripheral regions are extraordinarily variable. The present investigations showed for the first time, that numerous vesicular inclusions can occur also in the boundary region between the stroma and Descemet's membrane.


Calcium and magnesium ATPases of the spectrin fraction of human erythrocytes.

Using a rapid method of preparation, spectrin has been isolated from human erythrocytes and its ATPase activity investigated. The ATPase activity with calcium has two distinct components, one with optimal activity when calcium and ATP are of equal concentration (low-Ca-ATPase) and another which is activated above 1 mM CaCl2 and is maximal at 100 mM CaCl2. There is also a Mg-ATPase with maximal activity at 10 mM MgCl2. The high-Ca-ATPase of spectrin, but not the low-Ca-ATPase, is inhibited by magnesium, while the Mg-ATPase is inhibited by Ca in excess of ATP. None of these activities exhibits the calcium-stimulated magnesium-dependent activity characteristic of the red cell calcium pump.