The interdependence between electrogram, total electrode impedance and pacemaker input impedance necessary to obtain adequate functioning of demand pacemakers. Electrogram maximum derivatives (DMAX, SMAX) and electrogram amplitudes (AMAX, UMAX) (Figure 2), were studied in 71 cases during permanent pacemaker treatment. During the acute phase, (at first implantation), 29 patients were studied, and during the chronic phase, (at pulse generator replacement), 42 patients were studied. Of these patients, 27 (acute phase) and 36 (chronic phase) were studied for tissue impedance (RT) and interface impedance (Faraday resistance RF and Helmholtz capacity CH). DMAX and SMAX changed from 3.47 +/- 0.33 V/s (mean +/- SEM) to 2.46 +/- 0.23 V/s and 1.93 +/- 0.20 V/s to 1.32 +/- 0.12 V/s; p < 0.02; p < 0.01. AMAX and UMAX remained nearly unchanged from acute to chronic phase. A paired comparison in 13 patients showed almost identical results. Electrograms recorded in patients with bundle branch block showed no statistical difference in DMAX, SMAX, AMAX, and UMAX compared with electrograms recorded in patients with QRS-complexes of normal duration. No correlation was found between rise in myocardial threshold and fall in DMAX and SMAX from acute to chronic phase; p > 0.8, p > 0.5. Patients with coronary heart disease were found to have significantly higher AMAX than patients classified as having rhythm disturbances of primary cause; p < 0.01. Extremely low values of amplitudes and maximum derivatives were found in some patients with myocardial infarctions and cardiomyopathies. No difference existed in DMAX, SMAX, AMAX, and UMAX recorded from electrodes with a 8 mm2 area compared with a 12 mm2 area (p > 0.5). RT was statistically significantly higher on the smaller compared with the larger surface electrodes (p > 0.005). There was a slight but not statistically significant fall in RT from acute to chronic phase (p > 0.2). RF ranged from 2.0-94.6 kohms. There was no statistically significant differences between the 8 mm2 compared with the 12 mm2 electrodes (p > 0.2). CH varied between 0.7 and 37.0 microfarads, with significantly lower values for the smallest electrodes (p < 0.05). In patients with electrograms of borderline amplitudes and maximum derivatives for being sensed, the low CH found with the small tip electrodes, will gave a higher risk of demand failure if the input impedance in the sensing circuit of a demand pacemaker is too low.


Similar QRS morphology in sinus rhythm and ventricular tachycardia. A 74-year-old man with coronary artery disease and bifascicular block suffered recurrent tachycardia. Although the similarity of QRS morphology in tachycardia and sinus rhythm suggested a supraventricular origin, His bundle electrocardiography and programmed stimulation showed a ventricular origin. Digitalis toxicity was not an etiological factor and there was no evidence to suggest a His bundle tachycardia. The case illustrates that in occasional circumstances the onset of ventricular tachycardia may produce no change in QRS morphology thus making the diagnosis by conventional electrocardiographic criteria difficult.


Serum and urine beta-2-microglobulin and lysozyme concentrations in transplant rejection. We conclude that serum and urinary beta-2-microglobulin concentrations are useful in the diagnosis of acute and chronic renal transplant rejection. In acute transplant rejection, serum elevation in beta-2-microglobulins usually precede a rise in the serum creatinine. Increased urinary beta-2-microglobulin concentrations and elevated fractional excretion of beta-2-microglobulin occur in both acute and chronic transplant rejection. The finding of massive and sustained beta-2-microglobulinuria following acute rejection may herald recurrent clinical rejection episodes and eventual graft loss. Finally, serum and urine lysozyme levels appear to be less sensitive than the beta-2-microglobulins for diagnosing rejection and are often spuriously elevated in the presence of systemic or urinary tract infection.


Effect of 17 phenyl PGF2 alpha on nasal patency in man. The effect of three different doses (12.5 microgram, 25 microgram and 50 microgram) of 17 phenyl PGF2 alpha on nasal patency in fourteen patients with vasomotor rhinitis was studied on separate occasions. In all subjects the three doses of the prostaglandin analogue increased nasal patency for varying duration. With the 50 microgram single dose the duration of action ranged between three hours and more than seven hours. There were no side effects in most of the patients.


[Action of a dithiocarbamate (zinebe) on the liver biosynthesis of RNA and proteins in the rat (author's transl)]. Zineb (zinc ethylene bisdithiocarbamate) inhibits the transcriptional activity of an in vitro system functioning with either RNA polymerases of isolated rat liver nuclei or purified E. coli RNA polymerase. Zineb also inhibits the in vitro aminoacid incorporation mediated by a polysomal standard system, by interacting with some pH 5 enzyme components(s). When rats fed a balanced semi-synthetic diet containing zineb (5 200 mg/kg) for ten weeks, one may observe a decrease in the RNA and protein syntheses in liver. The consequences of the ingestion of small but repeated doses of zineb are discussed.


Immunodiffusion study on group-specific antigen of MC-29 chicken hepatoma. Rabbit serum against crude MC-29 hepatoma extract does not contain anti-avian gs antibodies detectable by gel diffusion methods. Rabbit serum containing antibodies against purified gs 1, gs 2, gs 3 and gs 4 antigens of avian myeloblastosis virus reveals gs 1 antigen in MC-29 hepatoma extract.


Comparison of a new microcrystalline dicoumarol preparation with warfarin under routine treatment conditions. 1 To determine whether two different oral anticoagulants show difference under routine clinical conditions, 71 patients were randomized to treatment with Apekumarol, a microcrystalline dicoumarol preparation, and 72 patients to treatment with warfarin. 2 During the inpatient phase of treatment both drug groups remained for about 94% of their treatment time within prothrombin value limits of 5-25% (Simplastin A). No statistically significant difference was found between the drugs. 3 While under outpatient care, both drug groups remained for about 80% of their treatment time within prothrombin value limits of 5-25%. No statistically significant difference was found between the drugs. 4 The intensity of control and number of prothrombin-determinations did not differ significantly between the groups. 5 Variations in the daily dose did not differ significantly between the groups. 6 The mean daily dose could not be correlated to mean body weight. 7 The mean daily dose decreased with age for the male patients taking warfarin, not for the female patients. There was no such decrease for either male or female patients taking Apekumarol. An additional 137 patients who at the time of the trial were under routine treatment with warfarin were also studied with regard to mean daily dose, age and sex. In this additional group the mean daily dose could be correlated with age in both males and females. 8 No difference between Apekumarol and warfarin could be demonstrated when tested under routine clinical conditions according to the design of the present study. Sensitivity for warfarin, but not for Apekumarol, seems to increase with age, this sensitivity has been demonstrated in both sexes.


A rat model (using a semipurified diet) of the fetal alcohol syndrome. A semipurified liquid diet was used to develop a chronic rat model of the fetal alcohol syndrome. Control female rats gained weight normally, reproduced normally, and gave birth to normal litters on this diet. Increased neonatal mortality, decreased neonatal weights, and altered sex ratios were observed in offspring of experimental alcoholic animals. Preliminary histological results include a mild delay in cell lamination patterns in the cerebral cortex at 4 days postnatal in alcoholic offspring as well as decreased formation of dendritic spines at 7 days postnatal.


Gas chromatography and mass spectrometry of disaccharides from glycoproteins. Partial acid hydrolysis and methanolysis released disaccharides and disaccharide methylglycosides from the glycoproteins, ovomucoid and porcine gastric mucin in amounts of 0.5--7 microgram disaccharide per mg of glycoprotein. These disaccharides were fractionated by gas chromatography as the trimethylsilyl (Me3Si) derivatives. The composition of recovered disaccharides has been determined by hydrolysis and rechromatography of the Me3Si monosaccharides. The intersaccharide linkages of the disaccharides have been determined by electron impact mass spectrometry. This simple and rapid method can give structural information on small glycoprotein samples.


Isoelectric focusing of oligopeptides: detection by specific stains. By using ultrathin (350 micrometers) polyacrylamide gels, which at the end of the fractionation are pasted to filter paper and dried in an oven at 110 degrees C, and after isoelectric focusing it has been possible to detect oligopeptides in the di- to tetradecapeptide range, which could not be detected by protein staining techniques. This is achieved by developing a series of specific stains for the following amino acids: Arg, Tyr, His, Trp, Met and Cys. Except for Met and Cys, the detection limits appear to be in the order of 0.2--2 micrograms of free amino acid loaded in the gel. The Pauli reaction for His and Tyr and the Sakaguchi stain for Arg can be developed sequentially in the same gel, thus allowing the detection of four different amino acids since, under these conditions, also Trp reacts. Unfortunately, more general reactions, such as the permanganate, the 'Lowry' and the ninhydrin stains, cannot be utilized since the carrier ampholytes react very strongly with all these reagents.


The use of an oscillating-tube densitometer as a tool in enzyme kinetics. Determination of the influence of sodium ascorbate on invertase, dextransucrase and dextranase. The use of a commercial oscillating-tube densitometer with an accuracy of 4 . 10(-7) g/cm3 for the determination of enzyme-kinetics constants is tested. This method is applied to the investigation of the influence of vitamin C (sodium ascorbate) on the glycolytic enzymes invertase, dextransucrase and dextranase. Invertase is inhibited uncompetitively, dextransucrase non-competitively. There is no significant effect of the vitamin on dextranase. The comparison of the mechanisms of the three enzymes suggests that only those reaction steps are inhibited by vitamin C in which fructose is released from the enzyme.


Displacement chromatography of simple protein mixtures, using carboxymethyldextrans. Simple mixtures of proteins have been used as models to demonstrate that the fractionation of proteins by carboxymethyldextrans on an anion exchanger (Peterson, E.A. (1978) Anal. Biochem. 90, 767--784) is fundamentally a displacement process. Examples include the separation of two closely similar proteins, the A and B forms of beta-lactoglobulin. The use of carboxymethyldextran spacers was essential for good separation, since resolution based on protein--protein displacement was generally inadequate. The origins and effects of heterogeneity in the preparations used as spacers are discussed.


Silver staining of myelin by means of physical development. For staining myelin with silver a physical development technique has been devised that can render visible the thinnest fibers in various animal species, including fishes and reptiles, even in the early phase of myelination and may be applied to both frozen and embedded materials. Its principle is as follows: Myelin can form and bind colloidal silver particles in a 0.1% ammoniacal silver nitrate solution of pH 7.5. The production of metallic silver by other tissue elements is suppressed by the sections pretreated with a 2:1 mixture of pyridine and acetic anhydride for 30 min. The colloidal silver particles bound in the myelin are enlarged to microscopic dimensions by a special physical developer.


The natural course of central retinal vein occlusion. Thirty-five patients with central retinal vein occlusion had an extensive medical and laboratory evaluation to identify possible etiologic factors. We describe the natural course of the two forms of retinal venous occlusive disease, hemorrhagic retinopathy and venous stasis retinopathy, by observing a group of 25 untreated patients from one to eight years. Systemic vascular disease was most commonly associated with retinal venous occlusive disease. The visual prognosis is favorable in venous stasis retinopathy and extremely poor in hemorrhagic retinopathy. However, two of ten patients with venous stasis retinopathy developed hemorrhagic retinopathy with poor visual outcome.


Antigenic similarity between squamous cell carcinoma of horn (horn cancer) and normal bovine foetal tissues. Normal bovine foetal (liver and skin) and horn cancer tissue antigens were examined using double diffusion agar gel precipitation and immuno-electrophoretic tests to detect any cross reactivity among them. Rabbit horn cancer antisera absorbed with normal bovine liver, skin and horn core epithelium antigens, when tested with foetal skin and liver (4 to 6 months of gestation), revealed the presence of 2 foetal antigens in horn cancer. Immuno-chemically 2 of the horn cancer antigens were found to be identical to the bovine foetal antigens.


World veterinary manpower: comparative aspects. The number of veterinarians in relation to livestock units, human population, milk and meat production and gross national product for 93 countries is presented. The structure of veterinary manpower is discussed and information on veterinary involvement with companion animals and public health is given. For 23 countries representing all the continents, the number of livestock units, human population and gross national product per veterinary school are presented.


Some inherited lysosomal enzyme defects with special reference to the liver and prospects for their control. Lysosomes are subcellular organelles which contain hydrolytic enzymes. These enzymes catalyse the degradation of macromolecules which are: I. taken into the cell by endocytosis; II. derived from the cells own structural components by autophagy; III. unused secretory products in the case of exocrine or endocrine cells. Each of the lysosomal storage diseases is due to the mutation of a gene which directs the synthesis of a specific lysosomal enzyme. This alters the catalytic activity of the enzyme and specific, incompletely degraded, macromolecules accumulate. The lysosomal enzymes pass from cell to cell by a combination of exocytosis and endocytosis, and this makes enzyme replacement therapy potentially feasible. The on-going trial of fibroblast transplantation as a means of enzyme replacement for the treatment of Pfaundler-Hurler disease in the author's laboratory, is briefly described.


Hearing thresholds with outer and inner hair cell loss. Hearing impairment and related cochlear histopathologic changes were evaluated in experimental animals after treatment with aminoglycoside antibiotics or exposure to intense sound. In the course of treatment with kanamycin, neomycin, or dihydrostreptomycin, permanent hearing loss in monkeys and guinea pigs occurred first at the high frequencies and progressed toward the lows. Exposure to different octave bands of noise at 120 dB SPL in monkeys and chinchillas produced permanent hearing loss at frequencies related to the spectral characteristics of the octave band. In most instances loss of outer hair cells was substantially greater than that of inner hair cells. In fact, the pattern and location of missing outer hair cells on the basilar membrane were most often correlated with threshold shifts of 50 dB or less. Generally inner hair cell loss was observed when the threshold shift was greater than 50 dB. Our data support the place principle and the inference that the outer hair cells are essential for hearing from threshold to about 50 dB SL. The inner hair cells, if functioning normally, apparently take over above that level. Although there is little doubt that such a generalization will, in the long term, be found to have been greatly oversimplified, there is every reason to believe that a combination of behavioral and morphologic procedures, as used in this study, will play an important part in elucidating the differences in functional significance of the two types of hair cells.


The fibrinolysin system in otitis media with effusion. Middle ear effusions can cause the fibrinolytic dissolution of human fibrin clots. The data presented herein indicate that the highest level of fibrinolytic activity and general proteolytic activity is found in the serous effusions. The mucoid effusions show the lowest levels of fibrinolytic activity. The significance of the fibrinolytic activity in middle ear fluids is not fully understood. Plasmin, the active protease of the fibrinolysin system, is capable of activating a number of proteolytic systems, including the intrinsic coagulation system, the vasoactive peptide system, and the complement system. All the inflammatory mediator systems can promote increased vascular permeability, tissue edema, and leukocytic migration. The relation of these systems to inflammation in the middle ear may have a profound influence on the possible conversion of a simple middle ear effusion into chronic otitis media.


Immunological relationships among transaldolases in the genus Bifidobacterium. Antisera were prepared against electrophoretically homogeneous transaldolase (dihydroxyacetone transferase, E.C. 2.2.1.2.) of Bifidobacterium thermophilum (B. ruminale) RU326 (ATCC25866), B. cuniculi RA93 (ATCC27916) and B. 'minimum' (homology group) F392 (ATCC 27538). Crude extracts of eighty six strains previously assigned to twenty one species of the genus Bifidobacterium on the basis of deoxyribonouclelic acid (DNA) homology (DNA-DNA hybridization), were compared by double diffusion tests on Ouchterlony plates. Eight groups of identical antigenic specificity were recognized. By analysis of the spur formation, the groups of identical specificity were arranged in preliminary sequences of decreasing similarity to each of the three homologous transaldolases used as reference points. The relationships between immunological data and the genetic similarity among the species of the genus measured by means of DNA-DNA hybridization were discussed together with some relevant points of bifidal ecology.


Regulation of the immune response by macrophages. Regulation of the immune response by macrophages was studied with cellular resistance to Listeria monocytogenes as parameter. The use of agents which suppress macrophage activity during the induction-phase of immunity enabled the induction of protective immunity with killed listeria. Fractionation of the cell content of listeria yielded an RNA'se sensitive fraction which in a dose of 300 ng and in combination with the cationic surfactant dimethyl dioctadecyl ammonium bromide induced protective immunity against listeria.


Preliminary quantification of immunological relationships among the transaldolases of the genus Bifidobacterium. The immunological relatedness among the transaldolases (dihydroxyacetone transferase, E.C. 2.2.1.2) of twenty species of the genus Bifidobacterum has been tested by the microcomplement fixation method, using B. thermophilum (B. ruminale) RU326 (= ATCC 25866), B. cuniculi RA93 (= ATCC 27916) and B. 'minimum' (DNA homology group) F392 (= ATCC 27916) as references. Based on the serological relationships of the transaldolases, expressed either as indices of dissimilarity or as immunological distances, the twenty species of the genus Bifidobacterium were arranged into clusters. These clusters generally coincided with the immunological groups obtained previously by the immunodiffusion method (Sgorbati and Scardovi, 1979).


Hormonal control of gene expression: differential activation of rat bone marrow RNA polymerases by erythropoietin and testosterone. Hormones play a role in the regulation of gene expression by inducing changes in enzyme patterns in target cells mediated by the synthesis of specific RNA molecules. Erythropoiesis has been used as a system for studying the molecular mechanism of regulation of gene action by means of two hormones: erythropoietin and testosterone. Experiments designed to correlate the biochemical action of both hormones on rat marrow cells are herein reported. Both factors seems to act at different biochemical and citological levels. Erythropoietin triggers the erythropoietic process acting on the erythropoietin sensitive cells (ESC), in which the hormone induces the synthesis of a high molecular weight RNA, which is the precursor of a functional 9 S messenger RNA. Testosterone seems to act on polychromatophilic erythroblasts, in which the synthesis of ribosomal RNA or its precursor is stimulated. The steroid enhances the nuclear ribonuclease activity, which could represent a control mechanism for the processing (maturation) of high molecular weight RNAs. The incorporation of 3H-GTP and 3H-UTP into RNA by isolated rat bone marrow nuclei is stimulated by erythropoietin and testosterone. Using alpha-amanitine and different ionic strength conditions it was found that erythropoietin enhances preferentially RNA polymerase II activity while testosterone increases RNA polymerase I activity. It is postulated that erythropoietin and testosterone act synergically to create the biochemical machinery for hemoglobin synthesis, the macromolecule that characterizes the erythropoietic process.


Probing the program of gene expression utilized in early development. Somatic cell nuclei injected into amphibian oocytes enlarge, exchange protein with the surrounding cytoplasm, and actively synthesize RNA for weeks. The message activity of the RNAs synthesized within the oocytes can be detected as new proteins made a few days after the injection of somatic nucleic. The oocyte cytoplasm seems to reprogram the injected nuclei, allowing the expression only of those genes which are normally active in oocytes (De Robertis and Gurdon, 1977). Genes which are unexpressed in somatic cells can be activated by injecting Xenopus laevis somatic nuclei into oocytes of Pleurodeles waltlii (Urodela). The genes that were activated are normally expressed in Xenopus oocytes but not in somatic cells. Conversely, genes which are normally expressed in somatic cells but not in oocytes become inactive after injection into oocytes. We conclude that genes which become inactive during cell differentiation can be reactivated, in the absence of cell division, by normal components of oocyte cytoplasm. These components could turn out to be examples of the "determinants" of egg cytoplasm responsible for nuclear activity in early development.


Radioimmunoassay for the detection of HBeAg and anti-HBe. We describe a solid-phase radioimmunoassay, based on the "sandwich" principle, using anti-HBe1/anti-HBe2 containing IgG precoated to polystirene beads as the solid-phase, and 125I-anti-HBE as labelled antibody for the detection of HBeAg and a two steps competitive procedure for the detection of anti-HBe. Comparison of titre obtained by ID and RIA or reference sera containing HBeAg and anti-HBe revealed that RIA is about 200 fold more sensitive than ID in detecting both HBeAg and anti-HBe. Moreover an increase in the detection rate of HBeAg from 5.9% (ID) to 15% (RIA) and of anti-HBe from 47.8% (ID) to 74.2% (RIA) and consequently a reduction of the frequency of negative samples for both HBeAg and anti-HBe from 46.3% (ID) to 10.8% (RIA) was determined by testing sera from HBsAg asymptomatic carriers. With regard to specificity, the frequency of presumptive HBeAg positive samples not confirmed was of 1 out of 29 (3.4%). Technical improvements of sensitivity and specificity of the described RIA are under development.


Quantum chemical studies of the metabolism of polycyclic aromatic amines and the stabilities and electrophilicities of their arylnitrenium ions in relation to their mutagenic/carcinogenic potencies. Electronic parameters related to the cytochrome P450-catalyzed reactions of eight polycyclic aromatic amines have been calculated using all valence electron semiempirical molecular orbital methods. The reactions considered lead to the presumably carcinogenic arylnitrenium ions and to the competing hydroxylation and epoxidation products. The stabilities of the arylnitrenium ions relative to the N-hydroxylamines and their sulfate esters were also calculated, together with electrophilic reactivity parameters of the ions. The resulting parameters were used to predict major metabolites of the parent compounds and also to correlate with observed mutagenic activities of the four pairs of polycyclic aromatic amines studied. The major factor in determining mutagenic potencies of parent compounds appears to be the extent of N-hydroxylation and competing ring oxidations, as well as the electrophilic properties of the arylnitrenium ions.


Dynamic inorganic ion-protein interactions in structural organization of DNA of living cell nuclei. Staining polarization optical techniques showed differences in the structural organization of DNA of chromatin in interphase nuclei and in mitotic chromosomes. The DNA was non-birefringent in intact interphase cell nuclei, but birefringent in chromosomes and in isolated nuclei incubated in a physiological electrolyte solution. The birefringence of DNA appears to be related to an unfolding of DNA filaments induced by free cations and to the oriented binding of dye molecules to DNA phosphates. We propose that the actual concentration of free cations inside the living cell nuclei is regulated by a dynamic interaction between nuclear proteins and ions.


Histochemical characteristics of the anal sacs of the cat. The histochemistry of polysaccharides and proteins and the distribution of RNA were investigated in the glands and mature secretion of the anal sacs of the cat. Glycogen was extremely abundant in the epithelial lining of the lumen and efferent duct of the anal sac and also in ducts of sebaceous glands of the sacs constantly formed in the place of degenerating acini. Little glycogen was found in the unchanged sebaceous acini. The apocrine glands of the anal sacs were substantially richer in glycogen than the apocrine sweat glands of the skin and were in an active state. A high concentration of acid mucopolysaccharides was present in the mature secretion, and their source was the apocrine glands. The phases of the secretory cycle alternated in different cells and secretory portions of the apocrine glands highly asynchronously. Carboxylated acid mucopolysaccharides were more abundant than sulfated. Sialomucin, hyaluronic acid, and chondroitin sulfate could be identified in the secretion. Neutral and acid mucopolysaccharides were present in the mature secretion in the form of complexes with one another in approximately equal proportions; neutral polysaccharides consisted chiefly of glycogen. Besides polysaccharides, the apocrine glands produce large quantities of protein, in which the mature secretion was rich. This protein was predominantly basic, and the secretion contained very little acid protein. Maturing cells of the sebaceous acini, and also other specific sebaceous glands, were characterized by a much higher protein concentration than in ordinary sebaceous glands.


Pathogenesis of proliferative neovascular retinopathies and the role of vitrectomy. A hypothesis. Proliferative neovascular retinopathies may be caused by the release of a hypothetical vasoproliferative factor, but mechanical factors seem to self-perpetuate the disease as well. Contraction of proliferative tissue causes vitreous detachment and traction on the retina. Congestion of vascular fronds by traction further stimulates vascularization. Traction on vessels results in hemorrhages. Surgical removal of all intravitreal scaffolds by vitrectomy seems to eliminate the development of this vicious cycle. Results of vitreous surgery seem to indicate that the hypothesis of scaffold removal is valid.


The technique and indications for photocoagulation in diabetic retinopathy. II. The treatment of diabetic retinopathy. The beneficial effect of photocoagulation in diabetic retinopathy is now established (3-7, 10). Whereas the capability of providing photocoagulation is widespread, the indications and actual technqiues are poorly defined. In an earlier paper (1) we discussed the principles of photocoagulation. In this paper the detailed techniques required for the treatment of individual lesions and of different types of retinopathy will be discussed.


Role of nucleic acids and cytoskeleton in nuclear refringence phenomenon (nrp) of human peripheral blood lymphocytes. Nuclear refringence is a phenomenon visible by light microscopy that is correlated with lymphocyte maturation and activation. The present study, in employing isolated nuclei as well as intact lymphocytes, confirms the nuclear basis of the phenomenon. Enzymatic degradation of nucleic acids in isolated nuclei attenuates refringence, providing presumptive evidence for the involvement of chromatin in refringence. Cytoplasmic events involving RNA and protein synthesis and microtubular, but not microfilament, function are necessary for the maintenance of refringence. These studies provide the basis for evaluating refringence within the context of nuclear protein changes known to occur during lymphocyte activation and for determining its usefulness as a simple test of the dynamic state of the lymphocyte nucleus.


Ethanol, disulfiram, and pyrazole: effects on interferon production in mice. The effects of ethanol alone or combined with pyrazole or disulfiram were studied in reference to interferon production. Saline, ethanol (2 g/kg), pyrazole (2 mg/kg), or disulfiram (2 mg/kg) were injected, intraperitoneally, 3 hr after poly I:C, an interferon inducer. Drugs were administered either by a single injection, or twice daily for 7 days, and the mice were sacrificed 6 hr after poly I:C. Sera from mice were pooled, dialyzed, and assayed for interferon by the microplaque reduction method. Administration of ethanol alone reduced interferon production 76% and 66% after the single and the multiple injections, respectively. A similar inhibition of interferon occurred after either pyrazole or disulfiram, with a greater potency for pyrazole. Treatment with pyrazole prior to ethanol reduced interferon production further, but only after the 7-day schedule. All drugs tested affected cellular immunity as measured by interferon production. Moreover, reduced interferon production by these agents was associated with a loss of splenic lymphoid tissue, suggesting that ethanol, pyrazole, and disulfiram could affect susceptibility to viral infection.


Glucagonoma, chronic recurrent peptic ulcer disease, and enhanced amylase-creatinine clearance ratio. Report of a case with review of the literature. A 53-year-old white woman developed diabetes mellitus, migratory erythema, and anemia, clinical features suggesting the presence of a "glucagonoma." Ten years earlier, after laparotomy and pancreatic biopsy, she had been told that she had an inoperable pancreatic carcinoma. Review of that biopsy together with current hormonal assay now confirms the diagnosis of glucagonoma. The recurrent peptic ulcer in this patient despite high levels of glucagon, a gastric inhibitory agent, is noted but not explained. An enhanced amylase-creatinine clearance ratio supports the notion that glucagon increases the clearances of amylase.


Isoprinosine as an immunopotentiator.

Isoprinosine as an immunopotentiator. Isoprinosine is a compound developed for antiviral use. The effects of isoprinosine on mouse responses to sheep red blood cells were studied over a wide range of doses, from 0.5 microgram/kg to 5 g/kg, i.p. administered at the time of i.v. immunization or as pretreatment for 7 days before antigenic stimulus. Low doses, 50 microgram/kg to 50 mg/kg, significantly increased the numbers of IgM- or IgG-spleen antibody-forming cells. Large doses, such as the LD50 (5 g/kg) or pretreatments where unable to impair mouse immune responsiveness. Isoprinosine (< 500 mg/kg/day) orally administered at time of or one day after immunization stimulated immune responses. In vitro addition of isoprinosine to spleen lymphocytes augmented PHA- or Con A-induced proliferation over a concentration range from 10 to 150 microgram/ml, whereas isoprinosine had no effect in the absence of mitogens. These data, and the lack of immunodepressing effect, suggest that there is a need for further evaluation of isoprinosine as an immunopotentiator.


Differentiation and regulation of lymphocyte populations: evidence for immunopotentiator-induced T cell recruitment. Isoprinosine, the p-acetaminobenzoic acid salt of inosine dimethylaminoisopropanol (1:3 molar ratio) and sodium diethyldithiocarbamate are two immumopotentiators which share an ability to induce in vivo acquisition of a specific T-cell marker by undifferentiated precursor lymphoid cells of healthy nu/nu mice, without affecting the B-cell lineage. Serum from treated nu/nu mice tested in dual assays, contains a selective inducer of prothymocytes.


Macromolecular binding in assessing drug and chemical-induced tissue lesions. The general concepts that underlie the use of macromolecular binding in studies of chemical-induced tissue lesions are reviewed. Parameters for studying the formation of chemically reactive metabolites are discussed and the factors that alter the formation and covalent binding of reactive metabolites are selectively emphasized. Some of the experimental work that led to these concepts is presented.


[The pancreatico-respiratory syndrome. Problems of intensive therapy and illustration of 5 clinical cases]. A short account of the mechanisms responsible for pleuropulmonary affections in the course of pancreatitis is followed by the presentation of personal cases observed over the previous four years and reference is made to the relatively high frequency of pleuropneumopathy. Lastly, mention is made of the treatment of pancreatitis. Recent criteria lay down that this should be conservative and medico-intensive in the acute stage. Surgery should be left for cases of peritonitic abdomen (exploratory laparotomy) and chronic pancreatitis.


Metabolism of halogenated methanes and macromolecular binding. Dihalomethanes are metabolized to carbon monoxide both in vivo and in vitro. The reaction is catalyzed by a hepatic microsomal cytochrome P-450 dependent mixed function oxidase system. Bioorganic mechanism studies suggest an initial oxygen insertion reaction followed by rearrangement to a formyl halide intermediate which in turn decomposes to yield carbon monoxide. In vitro studies show that 14C-dichloromethane becomes covalently bound to both microsomal protein and lipid. The similar characteristics of metabolism to carbon monoxide and covalent binding suggests that a common intermediate, perhaps the formyl halide, may be involved. Dihalomethanes are also metabolized to formaldehyde, formic acid, and inorganic halide. A glutathione transferase, located in hepatic cytosol fractions, appears to be involved. Reaction mechanism studies suggest that a S-hydroxymethyl glutathione intermediate may yield formaldehyde or be diverted via formaldehyde dehydrogenase/S-formyl glutathione hydrolase to yield formic acid. Haloforms are also metabolized in vitro to carbon monoxide by a hepatic microsomal cytochrome P-450 dependent mixed function oxidase system. This reaction is a markedly stimulated by sulfhydryl compounds.


Late complications of retrolental fibroplasia. A modified classification of cicatricial RLF has been presented. Particular emphasis is placed on myopia, one of the hallmarks of grade i cicatricial RLF. Whether or not this is axial myopia is still open to question, but since the more myopic eye was usually longer by ultrasonic measurement than the less myopic eye, actual enlargement of the eye must play some role. Other features noted in grade I RLF were retinal pigmentation, vitreous membranes, and equatorial retinal folds. In grade II the most significant finding was dragging of the retina. It was noted that in all cases of dragging of the retina there were also peripheral retinal changes. Even more important, however, was the fact that peripheral retinal changes are often present when there are no changes in the posterior pole. Neovascularization, elevated retinal vessels, lattice degeneration, and retinal breaks were other features of grade II retinopathy. Significant was the fact that the incidence of lattice in patients with cicatricial RLF (15%) was more than twice that of the normal population. Grade III RLF was characterized by falciform retinal folds, and grade IV, by retinal detachment. Thirty-nine retinal detachments were rhegmatogenous and usually had temporal retinal breaks. Retinal detachments of a tractional exudative variety (14 eyes) tended to occur earlier in life than the rhegmatogenous variety. In infants, they were preceded by increased tortuosity of the vessels in the posterior pole and, sometimes, accumulation of exudation, both ominous prognostic signs.


Observations of vitamin E in experimental oxygen-induced retinopathy. Retrolental fibroplasia (RLF) is a proliferative retinopathy observed in premature infants. Its relationship to oxygen has led to both clinical and laboratory research into the potential benefits of the antioxidant tocopherol in this disorder. Although the clinical trials to date have only suggested a beneficial result, studies in the kitten model demonstrate two significant actions of tocopherol in protecting against this retinopathy, only one of which is predicted by theory. These data support the need for prospective controlled trials in human premature infants.


Clinical evaluation of the EMIT procainamide and N-acetylprocainamide assay. Procainamide and its major metabolite, N-acetylprocainamide, were measured by the homogeneous enzyme immunoassay technique (EMIT). The reagents for the EMIT assays were supplied as a separate matched set for each assay. There is no cross-reactivity by procainamide in the assay for N-acetylprocainamide or by N-acetylprocainamide in the assay for procainamide. Within-day precision determined by replicate analysis of samples in the therapeutic range gave a coefficient of variation of less than 5% for each assay. The day-to-day coefficient of variation was less than 6% for each assay. Quantitative results obtained by the enzyme immunoassay on serum samples from patients receiving procainamide were compared with the results obtained by a high pressure liquid chromatography procedure. For the procainamide assay the correlation coefficient (r) was 0.983; for the N-acetylprocainamide assay the correlation coefficient was 0.981. There was no false positives or false negatives. The immunoassay requires 50 microliters of serum and the enzyme activity is measured in a spectrophotometer. An individual determination requires only 1 min to perform; therefore, the procedure can be used for either emergency or routine analysis.


Structural characteristics of antigen-specific suppressor factors: definition of 'constant' region and 'variable' region determinants. Anti-suppressor factor antisera, prepared either in rabbits (R alpha SF) or in syngeneic (CBA) mice (M alpha SF) by repeated immunisation with antigen purified CBA antigen specific suppressor factor reactive to KLH was shown to abolish the suppression caused by suppressor factors (SF) in vitro. R alpha SF inhibited the function of all SF regardless of their strain of origin or their antigen specificity suggesting that it recognized 'constant region'-like determinants in SF molecules. It did not have any effect on antigen-specific helper factors. Syngeneic M alpha SF only abolished the function of suppressor (or helper) factors which were KLH-specific, and only provided they were derived from the appropriate strains of mice; thus resembling the effects of anti-idiotype antibody. No linkage to MHC could be demonstrated but there was some evidence of possible association with allotype. A schematic structure of the SF molecule is suggessted on the basis of these findings with antisera to SF.


Computed tomography in the evaluation of the suspected carcinomatous prostate. Twenty-six patients with physical findings suspicious for prostatic cancer were examined by contrast-enhanced computed tomography (CT) of the prostate region prior to prostatic biopsy of resection. Twelve had benign hypertrophy and/or prostatitis and fourteen had adenocarcinoma. Prostatic contour, density, seminal vesicle "angle," extraprostatic soft tissue "mass," and the pelvic fat planes were evaluated. A nodular prostatic contour was found only in patients with adenocarcinoma of the prostate, indicating a role for CT in the diagnosis of this disease. Two patients with benign prostatic disease had extraprostatic soft tissue "masses" identical to those seen in six patients with adenocarcinoma of the prostate, suggesting limited usefulness of CT in staging patients with known tumor.


Cerebellotrigeminal and focal dermal dysplasia: a newly recognized neurocutaneous syndrome. A female child with brachycephaly, hypertelorism, convergent strabismus, interstitial keratitis, analgesia on both sides of the face, absent corneal reflexes, and focal congenital alopecia of a zone of the occipital and posterior parietal scalp is presented. The patient also had generalized hypotonia in early life, and at age 4 years 9 months, she was found to be moderately mentally subnormal and to have severe cerebellar deficit consisting of gait and truncal ataxia. There was no clinical evidence of other cranial nerves being affected. It is postulated that the patient has a cerebellotrigeminal and focal dermal dysplasia due to a development arrest of the ectoderm, which gives rise to the alar plate of the rhombencephalon, the overlying epidermis, the motor nucleus of V, and the trigeminal placodes.


Morphological and functional aspects of the glomerular basement membrane. Addition of Alcian Blue to the fixative reveals structural elements bearing negatively charged sites within the glomerular capillary wall of the rat kidney. According to previous research (review by Brenner et al., 1977), these charges represent one of the most important functional components of the glomerular filtration barrier. Alcian Blue also demonstrates negatively charged microfibrils (probably glycoproteins) of the mesangial matrix. Preliminary observations of glomeruli of lower vertebrates suggest that there is always a close correlation between the presence of microfibrils and mesangial cells.


[Preferential synthesis of fetal hemoglobin in in vitro cultures of erythroid precursors from peripheral blood of healthy persons and those with hemoglobinopathies]. The synthesis of fetal hemoglobin was investigated in in vitro cultures of erythroid precursors isolated from peripheral blood of normal individuals, newborns and of subjects with different hemoglobinopathies. Synthesis of hemoglobin was assessed by 35s-methionine labeling of cultures and measurement of the radioactivity incorporated into the hemoglobins A2, A, F and S isolated by column chromatography on DE 52 cellulose. The erythroid precursors from most of the studied individuals cultured in in vitro system responded with synthesized an average of 15% of Hb F while cultures from newborns produced an average of 60% of Hb F in comparison of 73% of Hb F in peripheral blood of the same newborns. Erythroid precusors from subjects heterozygotes for beta-thalassemia, heterozygotes for HPFH, and homozygotes for Hb S produced an average of 20%, 43% and 30% of Hb F, respectively, in comparison of 7%, 14% and 9% of Hb F, respectively, present in the RBC of the same individuals. These data support the previously published results (6-11) that erythroid bursts in culture reactivate the structural genes for the gamma chain synthesis.


[Use of a new radioenzymatic test in urticaria and drug allergy]. The Authors report preliminary data obtained with the histamine release test, evaluated with a radioenzymatic technique, in patients with a symptomatology highly indicative for penicillin allergy (21 cases) and in patients with urticarial forms (20 cases) or atopic dermatitis (5 cases). This technique, suitably modified, turned out very sensitive and able to detect histamine concentrations in the picogram range. The use of this test in type I immunoreactions, after Ig removal and fixation (IgE, IgG4) from basophils of atopic and normal subjects, pointed out its remarkable advantages (sensitivity, specificity, reliability) in comparison with other techniques. The results of our case-list, even if preliminary seem to be indicative for an extension of the trial to a wider number of patients both with allergies of pharmacological type and with urticarial forms or atopic dermatitis.


Different sensitivity of hemagglutinating and hemolytic activities of Sendai virus to non-antibody inhibitors. Research was carried out in order to verify if some components of human serum, which are capable of inhibiting the hemagglutination by Sendai virus, are active even towards the hemolysis by the same virus. It was ascertained that both orosomucoid and low-density lipoproteins obtained by floatation had no action on the hemolysis by Sendai virus while lipoproteins precipitated with dextrane sulphate showed a certain inhibiting effect. This action however is not due to lipoproteins but is probably related to some substances used in the method of precipitation.


Regulation of the mitochondrial anabolism at low concentrations of thyroid hormones and by some of their structural analogues. The Authors demonstrate that the in vitro stimulation of mitochondrial RNA synthesis produced by thyroid hormones takes place also at physiological levels, equal to those held in the liver cells of experimental animals. Two groups of male rats have been used: normal control animals (N) and animals surgically thyroidectomized on the 25th day of life (T). The animals were fed and kept in standard conditions and killed on the 85th day of life. The purification of mitochondrial samples and the determination of the mitochondrial RNA synthesis were carried out as previously described. The results suggest that the in vitro stimulation of mitocondrial RNA synthesis is already significant at the concentration of lnM. The trends are qualitatively comparable for either N or T animals. The structural analogues TRIAC (3,5,3'-triiodothyroacetic acid) and TRIPROP (3,5,3'-triiodothyropropionica acid) exhibit a clearly stimulatory effect on samples of N animals, while on samples of T animals is significant only for the first analogue. Similar trends are also observed on ADP/O ratio.


[Cell coat modifications in a series of gliomas as a function of malignancy]. We have compared the cell coat of benign gliomas and of malignant gliomas, by Ruthenium Red staining in vitro and by Thiéry's cytochemical reaction in situ. The thickness of the cell coat is not significantly increased by malignancy on the free surfaces of glioma cells. But in malignant gliomas only the cell coat disappears on the intercellular contact zones. These modifications are related to cell fusion characteristics of malignant transformation.


Studies on the clastogenic effects of biologic stains and dyes. We have surveyed the clastogenic potential of 12 different groups of stains and dyes totalling 48 compounds. We observed that 18 compounds induced significant increase in chromosome damage. Most of them were also found to be mutagenic, carcinogenic, or toxic in other reported studies. However, no significant studies were reported on six of them. It is concluded that these agents are potentially harzardous and should be studied further in detail.


Flow cytometry: general principles and applications to selected studies in tumor biology. The cell populations derived from normal tissues and solid tumors comprised many different cell types. Within each cell type there is a distribution of cells in different phases of the cell cycle and/or metabolic states (ie, differing rates of protein, RNA, and other macromolecular syntheses). Flow cytometry and companion instrumentation now promise to aid in rapid quantitative analyses of heterogeneous cell populations, thus finding broad applicability in many areas of cancer research and treatment. Since it is projected that this analytical technique will greatly expend our knowledge in tumor biology, it seems appropriate to review the basis principles of the methodology and to demonstrate recent applications in several areas of current research. After reviewing basis principles, a detailed description of one specific flow cytometer, the PHYWE-ICP-22, with its computer interface as developed in this laboratory is described. Subsequently, applications of this methodology to analyses of tumor cell kinetics, assays of blastogenesis, and studies of human colon cancer are presented as specific, current applications of flow cytometry. It is anticipated that this overview of flow cytometry along with some current applications will provide a background understanding for the inevitable rapid future developments in this area of research.


[Immunological tolerance: induction and abrogation]. The induction of a state of immune tolerance is a complex event which can take place at different stages of the immune response since it depends both upon the chemical structure and the experimental manipulation of the antigen. This paper deals mainly with the relationship between antigen structure and the kinds of tolerance induced.


[Endotoxic contamination of biological products (ribosomal vaccines, viral vaccines and interferon)]. The A.A. have examined by the Limulus assay the possible endotoxin contamination in some biological products (ribosomal vaccines, viral vaccines, interferon). While the preparations of influenza vaccines and a partially purified fraction of ribosomal vaccine from Pseudomonas aeruginosa exhibit a gelation of lysate with high levels of endotoxin, rubeola vaccines, interferon and a purified fraction of ribosomal vaccine, presented a negligible amount of endotoxin. The results are discussed with the aim to examine the possible role of contaminating endotoxin in the mediation of some adverse effects and of the unsuspected extrinsic adjuvant activities developed in clinical and experimental use of these preparations.


Tumor markers for ovarian cancer.

Tumor markers for ovarian cancer. The literature on tumor distinctive markers in ovarian cancer has been reviewed. Various immunological and biochemical approaches have been attempted for the diagnosis and management of patients with ovarian cancer. The complex spectrum of antigens that can be detected in human ovarian cancer consists of several tumor-associated antigens, fetal or carcinoembryonic antigens, carcinoplacental markers, and normal tissue antigens. We have described and partially characterized two ovarian tumor-associated antigens designated as OCAA and OCAA-1, which seem to have potential for the immunodiagnosis of ovarian cancer. Several other investigators have carried out similar studies, but in general their serological characterization of these antigens has been limited. The well-defined embryonic proteins that have been examined in the ovarian cancer include carcinoembryonic antigen (CEA), alpha-fetoprotein (alpha-fp), beta-oncofetal antigen (BOFA), Regan and Nagao isoenzymes and human chorionic gonadotropin (HCG). The presence of pregnancy-zone protein (PZP) has also been reported in ovarian cancer. In addition, several normal tissue components include fibrin-fibrinogen degradation products (FDP), alpha 1-globulin, and urokinase have been found associated with ovarian cancer. Both humoral antibodies and cell-mediated immune responses against tumor-associated antigens can be measured in ovarian cancer patients. In addition, serum factors, which block cellular immune reactions, have been identified. However, progress in this area has been hampered by the complexity of the antigens associated with ovarian tumors and the lack of standardized, well-characterized sources of antigens or target cells. Enzymes, especially those involved in glycoprotein biosynthesis, (eg, glycoprotein:glycosyltransferases and glycosidase) have been explored as possible early biochemical indicators of ovarian neoplasia. A serum specific deficiency of alpha-L-fucosidase has been found in patients with ovarian cancers. Of all the glycoprotein:glycosyltransferases studied, galactosyltransferase has been found to be the best enzyme marker for ovarian adenocarcinoma. The determination of serum levels of this enzyme reflected the clinical status of the patient with respect of tumor progression as well as tumor burden. Recently, assay of a phosphodiesterase, which specifically hydrolyzes cytidine 5'-monophospho-N-acetylneuraminic acid, has been found promising in the detection and management of patients with ovarian cancer.


[General considerations on the interferon system]. In this review some interesting aspects of interferon and its present perspectives for a practical utilization in human viral infections and in oncological therapy are described. After an evaluation of general characteristics of IF, some particular problems are considered about this system as that of IF and poly I:C interaction with cellular membranes, genetic control of IF system, IF and its inducers influence on immune reactivity and human tumours cells IF inducing ability and sensitivity. Finally are described some data obtained in various Institutes with IF on human virosis and those obtained on human and animal oncology with IF and interferon inducers.


An endoscopic staining method for detection and operation of early gastric cancer. The long term survival rate of gastric cancer has been much improved, and the 5-year survival rate in our institute was 42% with a significant difference between 29% in the advanced stage and 94% in the early stage. This result means that gastric cancer must be detected and treated early as possible. However, the diagnosis of early gastric cancer may remain extremely difficult even for an excellent endoscopist. Therefore, in order to clearly recognize early gastric cancer, an endoscopic staining method with methylene blue has been studied. In this method, one capsule of 150 mg methylene blue is swallowed with a small amount of proteinase solution 3 hours before endoscopy. Gastroscopy is performed routinely after this preparation. This procedure was performed on 153 gastric cancers and 137 of them (89.5%) have been successfully dyed in dark blue. In several cases, with this method, the actual borderline between the normal mucosa and the malignant extent was clearly recognized and resection line was decided. Introduction of methylene blue into the stomach could also stain the intestinalized epithelium of the gastric mucosa. Differential diagnosis of the dyed intestinal metaplasia and the dyed carcinoma seems to be very easy, because both gastric lesions have the characteristic dyed patterns. Mechanism of this phenomenon has been considered to be due to an absorption of the dye in the intestinal metaplasia, and in the gastric cancer, many factors may be involved, among which are the infiltration or diffusion of the dye into the cancerous tissue, the absorption from the abnormal epithelium, and the staining of the necrotic tissue.


Computer-controlled double-beam scanning microspectrophotometry for rapid microscopic image reconstructions. A method for automated collection of various specific data from an entire microscopical preparation and their quantitative evaluation is described. Its application to the study of neuronal connections is discussed in some detail. Brain sections are scanned using a computer-controlled microscope for reflectance, fluorescences or absorbance signals. Two illuminating beams are used, one of them being amplitude modulated. By means of a synchronous detection the two signals are recorded simultaneously: for example, in an autoradiograph, the reflectance (measuring the density of the silver grains in emulsion) and the absorbance (allowing to localize the underlying counterstained cells). The data are stored in a computer. Various off-line processing schemes allow the reconstruction of the data with respect to the corresponding spatial coordinates. Thus pseudo-three-dimensional, analogue or digital, graphic displays may be obtained in which the patterns of neuronal connections can be recognized and interpreted. A method for the detection of weakly labelled nerve fibres based on digital filtering is presented. The whole processing for a frontal section of the mouse brain (7 X 10 mm area) takes less than 1 h. In addition to the evaluation of microscopically labelled material (grains of autoradiographs, horseradish peroxidase, nucleic acids) the technique described has been successfully used for the study of naturally fluorescent intracellular components in living tissue cultures.


Guinea-pig keratinocytes and melanocytes in tissue culture: scanning, transmission and high voltage electron microscope observations. Keratinocytes and melanocytes cultured from guinea-pig epidermis were studied with scanning, transmission and high voltage electron microscopy to characterize the surface and internal morphology. Keratinocytes exhibited contact-inhibition and a range of surface structures consistent with cell-cycle dependent changes. Stereoscopic analysis of high voltage electron micrographs indicated regular oval nuclei with nucleoli at different depths, while thin sections revealed local channels in the nuclei. Secondary cultures differed from primary cultures in the disorder of the microfilaments, in the failure to form desmosomes, and in the failure of melanocytes to persist in culture. The beaded surface of melanocytes was indicative of underlying melanosomes that were seen in high voltage micrographs. Melanocytes were rounded with moderate ruffles or were dendritic with ruffles on the termini. These findings are discussed in relation to the observational techniques and in relation to modes of locomotion of and pigment transfer to epidermal cells.


[Chemotherapy for digestive carcinomas. Results of a phase II clinical trial (author's transl)]. A so-called "phase II" clinical trial was carried out in 89 patients with digestive carcinomas who were treated either by chemotherapy alone or by a combination of surgery and chemotherapy. Thirty-five patients with inoperable tumours received chemotherapy alone without any noticeable improvement. Thirty-two patients had tumours which were operable but with inoperable extension. Palliative chemotherapy, given after surgery to reduce the size of the tumour, resulted in a prolongation of survival. Twenty-two patients with operable tumours without inoperable extension benefited from routine adjuvant chemotherapy.


[Indications for normovolemic hemodilution]. So long as normovolaemic conditions are respected and the haematocrit is not reduced to less than 30 per cent, there are a large number of surgical and even medical indications for deliberate haemodilution. Amongst the several contraindications, coronary insufficiency must be considered as being formal.


[Replacement solutions and hemodilution]. By definition, haemodilution involves the use of filling solutions, but in this technique the role of different solutions has yet to be clearly defined. Amongst the criteria of choice used at the present time, two would seem to be essential: good tolerance, limiting the risks of complication and side effects to an absolute minimum; and a sufficiently long intravascular half-life ensuring good control of blood volume. On this basis, amongst the crystalloids Ringer lactate offers definite advantages. It is almost always used in association with a colloidal solution, very often human albumin or a dextran.


Intra-arterial chemotherapy of head and neck tumours. The benefits and complications of regional chemotherapy in the treatment of head and neck tumours are discussed. Intra-arterial chemotherapy has been employed in 72 cases of preoperative, postoperative and palliative management. Cytostatic treatment consisted of combined Vincristine, bleomycin, methotrexate, and mitolactol administration by a Watkins-USCI or Sharp chronofusor. In cases of preoperative treatment the tumour regression was in the range of 50-80%. Tumours of the gingiva, parotid gland, maxilla and tonsil responded very well, tumours of the tongue less well to the treatment. The most dangerous complication is thrombosis of the common carotid artery; to avoid this complication the prothrombin index was reduced and kept at the 30-40% level.


Biochemical and energetic gradients in the mucosa of stomach and duodenum of patients with antral ulcer. Separation and measurement of adenine-adenosine, adenosine monophosphate (AMP), adenosine diphosphate (ADP), adenosine triphosphate (ATP), lipid phosphates, RNA and DNA, further, separation and measurement of Mg2+-dependent, total (Mg2+-dependent plus Na+--K+-dependent, Mg2+-Na+-K-dependent) and Na+-K+-dependent ATPases were carried out in the mucosa and muscles of the corpus, antrum and duodenum of 68 patients with antral ulcer. It was found that (1) the substrate levels of adenosine nucleotides, lipid phosphates and RNA were significantly higher in the corpus mucosa than those in the mucosa of the antrum and duodenum (calculated for 1.0 mg DNA content); (2) the amounts of adenosine nucleotides and the sum of ATP + ADP + AMP did not alter significantly in the muscles of the corpus, antrum and duodenum; (3) the levels of adenosine nucleotides, sum of ATP + ADP + AMP, lipid phosphates and RNA, further the activities of Mg2+-dependent , total (Mg2/-dependent plus Na+-K+-dependent) and Na+-K+-dependent ATPases were significantly higher in the corpus mucosa than those in the corpus muscles. It was concluded that (1) energetic and biochemical gradients are present between the substrate levels in the mucosa of the corpus, antrum and duodenum; (2) the neural and/or humoral regulatory mechanisms differ in the mucosa and muscles of the corpus, in order to their biochemistry.


Dextran anaphylactoid reaction in Sprague-Dawley CFY rats. From a closed colony of randomly-bred Sprague-Dawley CFY rats about 23% failed to respond to intravenous dextran with the characteristic generalized anaphylactoid reaction, but still exhibited an inflammatory response when dextran was given into the foot pad. Brother-sister mating of rats showing the most expressed generalized reaction (reactor rats) yielded good responder offsprings, while the non-reactors had descendants completely unresponsive to systemic dextran. Brother-sister mating of selected non-reactor rats led to a gradual decrease in dextran paw oedema in the subsequent generations, and after the third mating, a complete local non-reactivity developed. In these rats the intradermal injection of dextran failed to increase vascular permeability, while the inflammatory response evoked by histamine, 5-HT, bradykinin, and compound 48/80 remained unchanged as compared to that of the reactor animals. These result show that the anaphylactoid reaction in Sprague-Dawley CFY rats is under genetic control.


Some characteristics of the insulin-induced potentiation to anaphylactoid reaction in Sprague-Dawley rats. The insulin-induced sensitization to generalized and local anaphylactoid reaction evoked by dextran was studied in Sprague-Dawley CFY rats. The generalized reaction was shown to be potentiated by insulin given subcutaneously in a dose-related manner. The minimum effective dose was as low as 0.04 U/kg. When this dose was injected intravenously, a marked but short-lived potentiation was observed. The insulin response could be elicited throughout the whole year. The local oedema induced by subplantar injection of dextran was found to be much less sensitive to insulin. Potentiation was observed during the period from March to October, while in the intermediate months, no such effect could be seen. The seasonal refractory state to insulin was abolished by bilateral adrenalectomy, and daily pretreatment of the rats with insulin for several days. Actinomycin D prevented the restorative effect of insulin pretreatment. Sensitization by a single insulin dose to both systemic and local dextran was suppressed in rats older than 6 months, and the refractoriness was in part reversed by adrenalectomy.


[Surgery of hepatic hydatic cyst. Per-operative complications. Possibility of anaphylactic etiology (author's transl)]. Among fifty cases hepatic hydatid cysts, we have observed four serious complications during operation. These complications appeared at the time of the puncture of the cysts with a severe shock in three cases. Anaphylactic etiology can be suspected but not proved without specific tests. Because the severity of these accidents (one death), anesthesiologist must be aware of the possibility of such complications: it is important to recognize anaphylactic reactions during surgical procedure to institute immediately a treatment to restore both circulatory competency and cardiac out put. Vasopressors with both alpha and beta properties are most useful under the conditions of massive peripheral vasodilatation and bronchospasm which occur in anaphylaxis.


[Hemodilution in vitro and parameters of hemostasis]. High molecular weight (approximately 60,000) dextran having been selected in a protocol of per-operative hemodilution, a prior study was undertaken in vitro in order to measure the variations in parameters of hemostasis. Factor of primary hemostasis, of plasma coagulation and fibrinolysis were thus evaluated in control blood samples, not diluted and diluted to 1/5 th by dextran or by a buffer solution. Statistical analysis was carried out using the t-test for pairs of variables. The following emerge from the results: 1 - an absence of difference with regard to the fibrinolytic activity of the plasma, but marked fragility of the fibrin clot in the presence of dextran; 2 - variations for plasma coagulation factors due to dilution; 3 - acceleration of the phase of polymerisation of fibrin monomeres and a decrease in platelet stickiness and aggregation due to dextran.


[Changes in physicochemical parameters and coagulation factors in hemodilution with dextran of molecular weight 60,000]. In 18 subjects undergoing aorto-bifemoral by-pass, pre-operative isovalaemic hemodilution was carried out (using a dextran of molecular weight 60,000) at the same time as venous section of a mean value of 1087 ml reducing the hematocrit to 0.30. The following were studied before, immediately after and 24 hours after hemodilution: blood viscosity (at variable shearing rates), oncotic pressure, sedimentation rate, clotting factors, plasma electrolytes. These results showed that at the end of hemodilution, oncotic pressure was maintained and even increased despite the fall in blood protein levels, with also a decrease in blood viscosity, a tendency to dynamic hypocoagulability (thrombocytopenia, fall in fibrinogen and increase in cephalin kaolin time and Quinck time) without any increase in lytic activity. 24 hours later, these parameters tended to return to normal. Discussion on the basis of a study carried out in vitro, seeks to demonstrate which of these changes were related to the dextran molecule and which were related to the hemodilution.


[Prevention of thromboembolism by normovolemic hemodilution]. In general surgery, routine thromboprophylaxis is strongly recommended in order to reduce the risk of thrombo-embolic complications. If pre-operative hemodilution is used in order to avoid the risks of transfusions of homologous blood, dextran must be used as the diluent. Patients undergoing total hip replacement represent a very high risk group where infusion of dextran is superior to low doses of heparin, but pre-operative hemodilution using dextran is more effective than infusions of dextran. It may thus be concluded that pre-operative hemodilution with dextran would give similar improved results in general surgery. As with all thromboprophylactic agents, side-effects may occur. However, with dextran 60, anaphylactoid reactions have note been described up to the present time in patients undergoing hemodilution before operation.


[Immunological problems in oncology (author's transl)]. The Authors, after a view of the problems of immunology, regarding the most recent discoveries, deal with the conclusions about the immunological reactions in research and clinical oncology. Many specific antibodies, formed as a result of the tumor, have been discovered in the field of research. It is impossible to say the same about human tumors and the probable hypothesis are numerous. The results until now obtained, even if they don't give a definite response, show that the immune defense of the organism has extreme importance for the beginning and the development of tumors, therefore further research could be a contribution to the cure of cancer.


Lymphokine production in primary mixed lymphocyte culture (MLC). III. Alloantigen signals cell-cell interactions involved in migration inhibition factor and immune interferon release. The alloantigen differences stimulation migration inhibition factor (MIF) and immune interferon (i-IF) release in primary one-way murine mixed lymphocyte cultures are reported. Differences at H-2 and Mls coded alloantigens induce both MIF and i-IF release, while the differences at multiple minor histocompatibility antigens induce MIF release only. Both MIF and i-IF release are T-lymphocytes dependent events. However, while T-lymphocytes are sufficient for MIF release, the presence of macrophages is required for i-IF release.


In vivo and in vitro reactions to antigens of Haemophilus influenzae in bronchial obstruction. The frequency of precipitating antibody to heat-labile (H(1--2) and heat-stable (HCW and HCF) antigens of Haemophilus influenzae was determined in patients with asthma, chronic bronchitis, cystic fibrosis and bronchiectasis and compared with that in a control group. This showed that the immune response of asthmatic patients to heat-stable antigens was different from that to the heat-labile antigens. Exposure to antigens of H. influenzae is common in all the disease groups. Skin test reactions having the time course and macroscopic appearance of Type I (immediate) and Type III (late) were obtained after prick and intracutaneous skin testing with HCW antigen in varying concentrations in a group of patients with asthma, chronic bronchitis or cystic fibrosis and in a control group. It is suggested that IgE and short-term sensitizing IgG antibodies may be responsible for the immediate reactions while activation of the alternative pathway of complement by endotoxin contained in HCW could be responsible for the late reactions. HCW antigens were shown to release histamine from non-sensitized human leucocytes; HCW and HCF antigens were shown to release histamine from non-sensitized human lung. None of the antigens tested had an effect on beta-receptors in tracheal preparations. It is proposed that these reactions may contribute to the pathogenicity of H. influenzae in the lower respiratory tract.


[Nucleic acids in the liver of rat treated with "E. coli" O 127 lipopolysaccharide (author's transl)]. The Authors have studied the variations of nucleic acids, proteins and histological aspect in the liver of rats, treated with 1 mg of E. coli ) 127 lipopolysaccharide and killed at 24th, 48th, 72th, 96th hour after the inoculation. The endotoxin determines an increase of nucleic acids and a reduction of hepatic proteic contents. The histological observation shows throw-back phenomenons of the hepatocyte with hyperplasy and hypertrophy of Kupffer cells at 24th hour, while at following intervals the phenomenous of hepatic rigeneration emphasize.


[Clinical studies of the antiarrhythmic efficacy of disopyramide phosphate (Norpace) (author's transl)]. The antiarrhythmic action of Norpace was investigated in three clinical studies. 1. A single blind placebo controlled study in 10 centers confirmed the antiarrhythmic action of Norpace in 120 patients with ventricular or supraventricular cardiac arrhythmias. It was shown by means of continuous ECG recordings that Norpace produced a statistically significant reduction in the frequency of both ventricular and supraventricular extrasystoles. Paroxysmal ventricular tachycardia was prevented in 70% of the patients. 2. In a double blind comparative study of Norpace and quinidine sulphate a similar antiarrhythmic action against ventricular and supraventricular extrasystoles was shown in both patient groups. In contrast, the tolerance of Norpace was markedly superior to quinidne sulphate. 3. An acute study of the action of Norpace was also conducted. Therapeutic plasma levels were attained within 2 1/2 hours following an initial single dose and within 60 to 120 minutes when this dose was doubled. Orally administered disopyramide phosphate has therefore proved effective in both the short and longer term treatment of various types of supraventricular and ventricular arrhythmias. Its antiarrhythmic properties are very similar to those of quinidine sulphate. The dosage of disopyramide phosphate should be sufficiently high and must be individualized. The dosage generally lies between 400 and 800 mg daily and should be given in 4 divided doses at intervals of 6 hours. To achieve rapid therapeutic effects an initial dose of 2 x 150 mg is recommended. This method of administration will produce effective therapeutic plasma levels within 1 to 2 hours. Due to possible negative inotropic effects it is necessary to digitalize patients with congestive heart failure and caution should be exercised in poorly compensated patients. Observed side effects were mainly of the anticholinergic type.


Prevention of neural tube malformation by genetic counselling, and prenatal diagnostic surveillance. The aetiology and genetics of the spectrum of dysraphic neural tube malformations from anencephaly through encephalocele, myelocele and meningocele to complicated spina bifida occulta, is outlined. The risk of recurrence, the methods and problems of genetic counselling are discussed. All women known to be at risk for neural tube malformation should be counselled carefully and offered prenatal diagnostic surveillance for any future pregnancy. There is no longer any need for a couple able to accept termination, to go through with a pregnancy in fear of recurrence.


[Effects of DDT and aldrin on swine cell line IB-RS-2]. Swine cells IB-RS-2 were exposed to 0,1-100 microgram/ml medium of DDT and aldrin for 48 hours. The data obtained showed that both compounds decreased cell growth. This effect depends on the concentration and on the time of cell exposition to the insecticides. These compounds also decreased the levels of cellular protein, RNA and DNA. The degrees of changes with aldrin were less than with DDT.


[Significance and limits of alpha-fetoprotein levels in the prenatal diagnosis of malformations of the central nervous system]. Amniotic fluid or/and serum alpha-foeto protein (AFP) determination is used as a test-system for screening of neural tube defects. The physiologic basis of this screening are described taking into account the evolution of AFP concentration in foetal and maternal blood, and in amniotic fluid. As for all the biologic screening systems, the acquired experience reveals a lack of sensibility and specificity. First the authors analyse the analytic and biologic problems which have an effect upon sensibility; then, they consider the mechanisms which explain the specificity lack showing itself in some foetal malformations. Practically, interpretation of AFP results requires necessarily familial story and echography results. Taking into consideration the different problems concerning AFP, a programme to utilize the test-system is presented for prenatal diagnosis of malformations of central nervous system.


[The present work concern the diagnostic significance and utility of enzymology in clinical practice (author's transl)]. To a short treatment concerning the main concepts of this discipline, follows the description on the enzymatic patterns typical of the most frequent diseases of various organs and apparatus, and a review of the literature about the main features of the enzymes of the greatest use and interest for internal medicine.


[Chemotherapy of non-microcellular disseminated bronchial cancer (author's transl)]. Fifty-one patients suffering from non-microcellular bronchial cancer with metastasis were divided at random into two groups: 33 patients received BACFON polychemotherapy, 18 patients did not receive chemotherapy. There was no global significant difference between the two groups (total mean survival time : 22 weeks with good therapy and 18 without), but a significant difference could be observed if there was an early response (M.S.T. : 38 weeks) compared to no response (M.S.T. : 15 weeks). Similar results were obtained when radiotherapy was associated (M.S.T. : 34 weeks) compared to chemotherapy without radiotherapy (M.S.T. : 22 weeks).


On the occurrence and prognosis of aphasia in patients with cerebral infarction. Of 338 consecutive patients with cerebral infarction, aphasia in the acute phase was found in 96. Of these, 18 had totally recovered from aphasic speech disturbances when leaving the hospital. The patients were examined 2-8, in average 6 years after the onset of the illness. 39% of them had died, 27% still suffered from aphasic speech disturbances, while 32% were recovered from aphasia. In addition, disturbances in writing, reading, and/or calculation were observed in more than the half of the survivors. Of patients over 65 years of age 56% were dead compared with 32% of younger patients. Aphasia was observed relatively more in geriatric patients, and the recovery of them was poorer.


[Intramedullary topography of the motor neurons of the brachial plexus of the rat. Preliminary study]. The exact localisation of the motor pools of anterior horn providing the different nerves of the brachial plexus remains unclear. The data obtained have been furnished by different methods: Nissl methods furnish cytoarchitectonic data -- Golgi methods furnish dendro-architectonic data. The topographical data were obtained by the chromatolytic modifications of the moto-neurons after a peripheral nerve injury. The recently used method of the retrograde transport of HRP permits to get information about the exact localisation and the dendritic extent of the neurons of a motor pool related to a particular nerve. We have studied the localisation of the nerves of the brachial plexus in the cervical enlargement of the spinal cord. We have determined the exact extent of the different motor pools in relation to each nerve. It seems that two different regions may be recognized in the anterior horn: a ventro lateral one in relation with the girdle muscles and a dorso lateral region related with the more distal muscles of the anterior limb.


[Myelin].

[Myelin]. The lipid and protein composition and the metabolic turnover of myelin in the C.N.S., are briefly reported. For a better understanding, it seemed useful to introduce some historical data on the discovery of myelin and to give a morphological description at structural and ultrastructural level.


Efflux of magnesium and potassium ions from liver mitochondria induced by inorganic phosphate and by diamide. Addition to rat liver mitochondria of 2 mM inorganic phosphate or 0.15 mM diamide, a thiol-oxidizing agent, induced an efflux of endogenous Mg2+ linear with time and dependent on coupled respiration. No net Ca2+ release occurred under these conditions, while a concomitant release of K+ was observed. Mg2+ efflux mediated either by Pi or low concentration of diamide was completely prevented by EGTA, Ruthenium red, and NEM. These reagents also inhibited the increased rate of state 4 respiration induced both by Pi and diamide. At higher concentrations (0.4 mM), diamide induced an efflux of Mg2+ which was associated also with a release of endogenous Ca2+. Under these conditions EGTA completely prevented Mg2+ and K+ effluxes, while they were only partially inhibited by Ruthenium red and NEM. It is assumed that Mg2+ efflux, occurring at low diamide concentrations or in the presence of phosphate, is dependent on a cyclic in-and-out movement of Ca2+ across the inner mitochondrial membrane, in which the passive efflux is compensated by a continuous energy linked reuptake. This explains the dependence of Mg2+ efflux on coupled respiration, as well as the increased rate of state 4 respiration. The dependence of Mg2+ efflux on phosphate transport is explained by the phosphate requirement for Ca2+ movement.


The messianic idea and messianic delusion. The messianic delusional syndrome repeats an historical prototype that manifests itself in each patient with individual changes. The syndrome expresses a serious impairment of identity and reflects a social, cultural and religious reality through generations. The regularities of its clinical features comprise a delusional system, centered on the patient's conviction that he has been chosen by God for a special and intransferable mission. The patient has special powers for carrying out this mission. He is a savior and announces resurrection. His delusions have a clear symbolic character. For the patient's social group, the messianic idea is an attempt at annulling the effect of oppression or persecution that have become unbearable for the individual. They represent a flight from the human sphere and an attempt to be God. The patient's behavior is in consonance with this purpose; it expresses itself, on the one hand, through preaching repentance and compassion and, on the other hand, the patient gives up his earthly links and replaces them by parental relations with God. In the above-mentioned context, the author analyzes the different elements of the religious conception in the Christian, Moslem and Jewish religions, and the way each of them expresses itself in the general symptomatology.


Reappraisal of prick test to tree and weed antigens. Although 3+ or 4+ prick test reactions to olive tree or Russian thistle are valid, the RAST values to these antigens are significantly lower than those in patients with similar reactions to grass antigens, suggesting that nonimmunologic factors are partially responsible for the skin test reactions to the tree and weed antigens.


Is dementia primarily a subcortical disease? Appraisal of recent biochemical observations. Recent studies have shown some morphological, structural, metabolic, and biochemical alterations in cortical and subcortical areas of brain of patients with Dementia. In this work we have studied Choline Acetyltransferase activity in cortical and subcortical areas and we present some data that shown a involvement of subcortical structures in Demented brain.


Minor metabolism a xanthene dye by African giant rat (Cricetomys gambianus, Waterhouse). In a continuation of our previous report (Adekunle, 1978), we have carried out a comparative study on the metabolism of eosin (2, 4, 5, 7-tetrabromofluorescein), a xanthene, in 2 mammalian species, the Albino rat (Rattus norvegicus) and the African giant rat (Cricetomys gambianus, Waterhouse). The latter shows a minor metabolism of this dye compared with the former. The findings are discussed in relation to the disposition of Erythrosine BS (tetraiodofluorescein) reported by other workers.


Characterization of stem cells and progenitors of hemopoiesis by cell sorting. Cell sorting has been used as a method for characterizing hemopoietic stem cells and progenitors. Fluorescent antibody-surface labels and changes in fluorescence polarization induced by in vitro stimulation with potential hemopoietic regulators were used. As detected by significant enrichment of CFU-S (pluripotent stem cells) in fluorescence-activated cell sorting, some CFU-S bear 'unique antigens' recognized by rabbit anti-human brain sera, human anti-human sperm sera, and 129 anti-F9 serum, but not A . TH anti-A . TL (Ia) ascites. Significant changes in fluorescence polarization induced by in vitro stimulation of mouse bone marrow with potential hemopoietic regulators were also observed; further, progenitors of human T-lymphocyte colonies were observed to exhibit a significantly decreased mean polarization value after short-term stimulation with PHA-LCM (phytohemagglutinin-stimulated leukocyte conditioned medium).


Fetal hemoglobin: in vivo stimulation by D-thyroxine in adult marmosets. Reactivation of fetal hemoglobin gene expressions was studied in new-world monkeys (marmosets). After a series of intraperitoneal injections of D-thyroxine, fetal hemoglobin production was increased in three experimental animals. Control animals were unaffected by the injection of the solvent. Because the percentage of cells containing fetal hemoglobin exceeded the total percentage of fetal hemoglobin, it was concluded that thyroid hormone influenced the synthesis of fetal hemoglobin rather than the development of a population of fetal cells.


[Alpha fetoprotein titers in HBsAg positive hospital personnel]. A survey was carried on the sera secured from the Ankara University Hospital for HBsAG. Out of 23 HBsAg positive sera, four sera was demonstrated containing high levels of Alpha-feto-protein, by haemagglutination test (Mochida). The cases were being followed up and within one year, second testing also proved high levels. One of them suffered from hepatitis two years ago and working at gynecology clinic as nurse.


Developmental evaluation in congenital esotropia. Eleven patients with congenital esotropia were tested with the Bayley Scales of Infant Development (BSID) before surgery and at two and six months after surgery. Eight patients completed all three tests and no obvious changes were found between the first and last test. Of the eight patients, two were considered developmentally normal by the BSID; three had significantly delayed mental and motor scores; one showed significant mental delay; and one had motor delay. Medial rectus muscle recessions were done bilaterally on all patients. Four required a second operation. Orthophoria was obtained in six patients.


Nucleic acids quantitation in polyacrylamide disc gels with a modified double beam spectrophotometer. Quantitation of RNA and protein bands separated in polyacrylamide disc gel electrophoresis is time consuming and not reliable when performed after staining processes. The best theoretical approach is the use of absorption spectra of the samples. The present paper reports the construction of an efficient and inexpensive adapter to perform direct scanning of 100 mm gels in a Unicam SP-800 double beam spectrophotometer. Conditions for gel casting and electrophoresis were established for RNA. A linear response between 0.25 micrograms and 15 micrograms per individual peak was found with a quantitative limit of detection of 0.25 micrograms. Spectral analysis of each band can be performed allowing identification of samples as nucleotides and detection of contaminants.


IgG in human cornea stained by various techniques. Four eyes were divided into portions which were prepared by various techniques for immunofluorescence microscopy. One portion was frozen and processed by cryostat cutting, a second was fixed in alcohol-acetic acid and embedded in paraffin, and a third was soaked in saline for 2 days to elute IgG and was then processed by the alcohol-acetic acid technique. All tissues received stain for IgG by either the direct (one-layer) or indirect (two-layer) technique. In all instances IgG was demonstrated in the same areas of the cornea. We conclude that the presence of IgG in human cornea is not dependent on the method used for fixation or processing of the tissue.


[Diagnosis experiences in 188 patients with primary liver cancer]. In Austria primary carcinoma of the liver (plc) is no rare malignancy. From 1973 and 1979, in our department, 188 patients with plc had been registered. By separate evaluation of the results of alpha-1-fetoprotein (AFP)-determination and 67-gallium-liverscintigraphy, a diagnostic accuracy of 81.6%, resp. 78. % could be demonstrated. By combination of both methods, a correct diagnosis of plc was obtained in 95%. Based on the promising results of lobectomies performed in patients with plc, therapeutic nihilism is not longer justifiable.


[Initial experiences with a new treatment protocol in otorhinolaryngologic cancers]. A new combined modality treatment in advanced squamous cell carcinoma of the head and neck, is presented. It consists of surgery, or radiotherapy, followed by adjuvant chemotherapy with MTX and CF rescue, cis-DDP, Bleomycin, Cyclophosphamide, given sequentially. Toxic effects were not very severe and did not cause an interruption of therapy.


[Hematologic diseases and opsonic alpha 2-macroglobulin. A correlative study]. We have investigated the opsonic alpha 2 Macroglobulin in Patients with various haematological diseases by means of turbidimetry. Patients with a poor prognosis tended to have lower levels. Therapeutic implications are discussed.


[Clinical significance of the radioimmunological determination of beta-thromboglobulin and platelet factor 4]. Recently new radioimmunoassay methods have been established to measure plasma concentrations of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4), platelet release products which are set free when platelets aggregate. Plasma concentrations of beta-TG and PF4 were investigated in disorders with increased thromboembolic risk. Extremely high concentrations of these platelet proteins were found in patients with venous thrombosis, pulmonary embolism, polycythemia vera, and chronic renal failure. Moderately increased beta-TG and PF4 levels were observed in patients with peripheral vascular disease, coronary artery disease, chronic rheumatoid arthritis, multiple myeloma, and diabetes mellitus. These data indicate, that plasma concentrations of beta-TG and PF4 are useful parameters for the evaluation of the "in vivo" platelet activity. By using these new methods for clinical applications special blood sampling conditions have been taken into account; moreover one has to consider that the plasma levels of the platelet "release products" are dependent from renal function.


Separation of the microvillous (maternal) from the basal (fetal) plasma membrane of human term placenta: methods and physiological significance of marker enzyme distribution. Plasma membranes from normal, full-term human placental trophoblast have been isolated by a new procedure. The method depends upon isopycnic zonal centrifugation using linear sucrose/Ficoll density gradients. Enrichment of plasma membrane marker enzymes with respect to trophoblast homogenate is found in two distinct peaks (designated B and D) of the fractionated effluent recovered from the rotor. Fraction B is enriched with membrane-bound alkaline phosphatase and 5'-nucleotidase, but not with (Na+, K+)-ATPase of F(-)-stimulated adenylate cyclase. It is suggested that this material is derived from the maternal-facing microvillous plasma membrane. Fraction D, enriched with (Na+, K+)-ATPase, F(-)-stimulated adenylate cyclase and, to a smaller extent, with 5'-nucleotidase and alkaline phosphatase is, by exclusion, proposed to be derived from the fetal-facing basal plasma membrane. Both plasma membrane fractions are shown to be free of appreciable contamination, using specific markers for endoplasmic reticulum, mitochondria, nuclei and lysosomes. The separation of the two membrane fractions is shown to depend both upon these membranes forming closed vesicles during homogenization and upon the buoyant densities of such vesicles differing in such a way that microvillous plasma membranes band at a lower density than basal plasma membranes. No separation of the membranes is achieved in gradients in which the vesicles are collapsed.


Protection of primate lung from stored-blood perfusion by peptidase inhibition and target membrane stabilization. Microaggregate filtration and separation of the cellular components of stored autologous blood have not changed the functional and morphological damage in primate lungs from stored-blood perfusion. Pharmacologic prevention of these "shock lung" changes was attempted with trasylol and with methylprednisolone pretreatment. In an in situ primate lung perfusion model, fresh and stored blood was perfused in untreated baboons and in groups of seven animals each pretreated with Trasylol or methylprednisolone. Pulmonary damage from stored-blood perfusion was evident by pulmonary edema, increase in pulmonary vascular resistance, decreases in effective compliance and arteriovenous pO2 gradient as well as by morphological criteria. Some protection was afforded by methylprednisolone and, to a lesser extent, trasylol pretreatment against this damage in primate lung form and function induced by stored-blood perfusion.


Maltotetraose as a substrate for enzyme-coupled assay of amylase activity in serum and urine. The use of maltotetraose ss a new substrate for the enzyme-coupled determination of amylase activity in biological fluids was developed by Beckman Microbics. We evaluated a manual and a centrifugal analyzer version of the method in comparison with two commonly used manual starch-dye amylase techniques: Roche Amylochrome and Pharmacia Phadebas. Both maltotetraose amylase procedures proved to be rapid and precise, and results correlated satisfactorily with the starch-dye methods for serum and urine samples.


On the potential variation in the gramicidin channel. A simple model for the gramicidin ion pore is analyzed in which the potential energy of the ion in the pore can be resolved into three parts: a series of equal potential barriers, the contribution from the applied potential, and a parabolic variation leading to either a potential maximum or a potential minimum at the center of the membrane. For low ion concentrations, the particular case corresponding to a potential minima 2kT below the surface potential predicts experimentally observed behavior.


Na+ interacting with gramicidin D. A nuclear magnetic resonance study. 23Na nuclei in a milk-white emulsion composed to nonionic surfactant and higher alcohol in saline were characterized by single values of T1 and T2 and a single Larmor frequency. In the presence of small amounts of gramicidin D (Dubos), the relaxations of 23Na were greatly accelerated, and the transverse relaxation was a sum of two decaying exponentials. But only a single T1 was observed; it was roughly equal to the slow T2. The slow T2 accounted for about 40% of the total resonance intensity. The relaxation rates increased linearly with the increase of the gramicidin concentration. The absorption signal consisted of a narrow and a broad line, both centered at the same frequency. The present results suggest that nuclear magnetic resonance spectroscopy is a useful tool for studying the nature of ion-permeable channels of biological membranes, even when the channel has no ionizable groups.


An effect of ultraviolet light on RNA and protein synthesis in nondividing human diploid fibroblasts. Nondividing human diploid fibroblasts maintained in medium containing 0.5% calf serum do not survive when exposed to low doses of UV (254 nm). The extent of killing is dose and strain dependent. DNA excision repair-proficient cells are more resistant than excision repair-deficient cells. Results of measurements of the effect of UV on RNA and protein synthesis in repair-proficient and -deficient (XP12BE) cells are reported. UV causes an immediate and equal depression of the RNA synthesis rate in both kinds of cells. A recovery to control rates was observed only at low (5 J/m2) doses in repair-deficient cells and at higher doses (20 J/m2) in repair-proficient cells. No recovery was observed at doses that cause substantial reductions in survival (greater than 5 J/m2 for XP12BE; greater than 40 J/m2 for repair-proficient populations). No initial effect on rate of protein synthesis was detected at doses less than 20 J/m2. However, in XP12BE populations, a decreased rate first evident at 15-30 h post-UV and before any cell degeneration and loss was observed for doses as low at 7 J/m2. This delayed effect was not observed in repair-proficient populations. The results are consistent with the hypothesis that the lethal action of UV in nondividing cells is one on DNA that leads to an inhibition of required protein synthesis by preventing RNA transcription.


Light-scattering studies of the monomer-dimer states of gramicidin A. Measurements of the intensity and autocorrelation function of ligh quasielastically scattered by gramicidin A indicate that the molecules exist as dimers in methanol and dioxane and monomers in dimethyl sulfoxide when solute concentrations range from 18 to 50 mg/ml. This gives further evidence that gramicidin A can dimerize in low-polarity media such as the hydrocarbon part of the membrane bilayers.


A reinterpretation of the infrared linear dichroism of oriented nucleic acid films and a calculation of some effective partial changes on the ribose phosphate backbone. In this paper we show, based on symmetry considerations, that structural information cannot be obtained from the linear infrared dichroism of the dioxy vibrations of the phosphate group of nucleic acids. Consequently, the discrepancies between the results of x-ray structure measurements and linear dichroism measurements are not meaningful. The linear dichroism measurements are instead important for a calculation of transition dipole moments that involve both the vibrations of all the atoms of the nucleotide and their charges. Independent information on either the atomic displacements contributing to a given vibration or the atomic charges permits a refinement of the unknown quantities. Based on the molecular dynamics calculations of Prohofsky et al., atomic charges of DNA are calculated to reproduce the observed linear dichroism results. Some of the resulting charges are unexpected and may reflect the inadequacy of the molecular dynamic calculation.