The eyes of mesopelagic crustaceans. II. Streetsia challengeri (amphipoda). In Streetsia challengeri left and right eyes have fused and become a single cylindrical photoreceptor, which occupies the basal half of a forward directed head projection. This unusual compound eye consists of approximately 2500 ommatidia, which are arranged in such a way that the animal has almost circumferential vision, but cannot look ahead or behind. It is thought that the eye operates on light-guide principles, and that the crystalline cones are the major dioptric component. Ommatidia in anterior-posterior rows show a greater overlap of visual fields than dorso-ventrally arranged ommatidia. Cone layer and retinula are separated by a 4 micrometer thick screen-membrane, which contains tiny pigment granules of 0.15 micrometer diameter. Cells of unknown function and origin, containing unusual multitubular organelles, are regularly found near the proximal ends of the crystalline cone threads. The twisted rhabdoms measure 18--20 micrometer in diameter, and consist of microvilli 0.05 micrometer in width, which belong to five retinula cells and which show no trace of disintegration. The position of interommatidial screening pigment, the density of retinula cell vesicles and inclusions, and the narrowness of the perirhabdomal space all suggest that the eyes have been light-adapted at the time of fixation for electron microscopy. The retinula cell nuclei lie on the proximal side of the heavily pigmented basement membrane. A tapetum or basal retinula cells are not developed. It is concluded that the eye optimally combines acuity with sensitivity, and that for distance estimation parallax may be important.


Fine structure of the eyes of orb-weavers, Argiope amoena L. Koch (Aranea: Argiopidae). 2. The anterolateral, posterolateral and posterommedial eyes. The anterolateral eye, the posterolateral eye and the posteromedial eye of the web-building spider, Argiope amoena have been examined by light and electron microscopy. The dioptric apparatus of all three eyes is similar in structure, and consists of a cornea, a lens and a vitreous body. The retina contains monopolar receptor cells, the cell bodies of which are present beneath the vitreous body in all three eyes. Proximal processes of the receptor cells form rhabdoms beneath the cell body layer and then extend toward the first optic glomerulus as an ocellar nerve. Two distinct patterns of retinal organization are present in the three eyes. In one type the rhabdomic layer of the retina is backed by a pigmented layer. In the other type the rhabdomic layer is backed by a tapetal reflecting layer. Rhabdomic structure and cytoplasmic inclusions of the receptor cells differ greatly between the two types. The anterolateral eye possesses a single type of retina with the rhabdoms backed by the tapetum. Both the posterolateral and the posteromedial eye are similar in structure, each possessing beneath the common dioptric apparatus retinae of both types.


The compaction of DNA helices into either continuous supercoils or folded-fiber rods and toroids. We have investigated by electron microscopy the mechanism of DNA compaction and have found that the double helix has the intrinsic potential to direct its own packaging into two distinctly different and mutally exclusive modes. The mode of DNA packaging is determined by the electrostatic charge density and water activity of the immediate microenvironment of the helix. The two basic structures formed by both linear and covalently closed-circular DNA are: a left-handed supercoil characteristic of minimally charge-shielded DNA, and a smooth rod characteristic of fully charge-shielded DNA. We propose that in the supercoil, the double helix is overwound (increased turn-angle), while in the rod, the helix is folded back and forth on itself. Variation of these two basic structures are the beaded fiber of DNA obtained with partially charge-shielded DNA and the toroid formed by the bending of the DNA rod and fusion of its ends in the presence of certain cations. We compare the DNA packaging inside these in vitro generated structures to DNA packaging in chromatin and viral capsids, and conclude that the packaging of DNA brought about by the use of salts and alcohol closely mimics the packaging behavior of the DNA in vivo, where it is usually complexed with histones or polyamines.


Nucleotide sequence of the DNA replication origin for human papovavirus BKV: sequence and structural homology with SV40. DNA and RNA sequencing techniques were used to obtain the sequence surrounding the origin of DNA replication for human papovavirus BKV. The structure is characterized by a true palindrome of 17 residues followed by two sets of symmetrical sequences and a stretch of 20 AT residues. Within the two symmetrical sequences is a segment containing a strong purine bias, 23 of 26 nucleotides. These structures are similar, if not identical, to those found in the region of the SV40 replication, origin. Within the homologous DNA segments, 60-80% of the BKV and SV40 nucleotides are the same. The remarkable similarity of BKV and SV40 sequences containing the origins of DNA replication would appear to confirm our previous suggestion of an evolutionary relationship between the two genomes. In addition, topological similarities between these sequences suggest the possibility of certain structural requirements for bidirectional replication origins in these superhelical DNAs.


Characteristics of cellular RNA related to the transforming gene of avian sarcoma viruses. Nucleotide sequences (sarc) related to the transforming gene of avian sarcoma viruses are present in the DNA and transcribed into RNA in uninfected avian cells (Stehelin et al., 1976b; Spector et al., 1978). Cytoplasmic RNA containing the sarc sequences has a sedimentation coefficient of 30S, is linked to polyadenylic acid and is present in polyribosomes in a form which can be released by treatment with EDTA. Hence the sarc sequences (complexity = 5 x 10(5) daltons) are part of a larger transcriptional unit (approximately 1.8 x 10(6) daltons), and the sarc RNA appears to serve an as yet unknown messenger function. In addition, the sarc sequences in chicken cells are not linked to RNA transcribed from the provisrus of the endogenous virus RAV-O, since RAV-O cytoplasmic RNA has sedimentation coefficients of 32S and 21S. Comparison of sarc-containing RNA in quail embryo fibroblasts and in quail cells derived from a methylcholanthrene-induced fibrosarcoma revealed no differences with respect to size, polyadenylation or concentration of the RNA in nuclei, cytoplasm and polyribosomes; thus the phenotypic differences between these cells are unlikely to be a consequence of alteration in expression of the sarc sequences.


Sites of integration of infectious DNA of avian reticuloendotheliosis viruses in different avian cellular DNAs. The pattern of integration for the infectious DNA of two avian reticuloendotheliosis viruses whose DNA is not inactivated by digestion with the restriction endonuclease, Eco RI was determined. High molecular weight DNA from infected chicken, turkey and pheasant cells was digested with Eco RI, electrophoresed through agarose gels and assayed for infectivity. The same patterns of integration of infectious viral DNA were found for these species of avian cells infected at high or low multiplicities with two reticuloendotheliosis viruses. There were multiple sites of integration in acutely infected cells with concomitant cell death. There was a single site of integration in chronically infected cells with no cell death. There were more integrated infectious viral DNA molecules per cell in acutely infected cells than in chronically infected cells. These results are consistent with the hypotheses that the cell death in the acute phase of infection is a result of the integration of the infectious viral DNA at multiple sites, and that only those cells survive that have the infectious viral DNA integrated exclusively at the single site.


Sterol metabolism. XL. On the failure of superoxide radical anion to react with cholesterol. Superoxide radical anion (O-2) failed to react with cholesterol under a variety of conditions. In some instances products indicative of free radical oxidation by molecular oxygen (O2) were found, but no products of electronically excited (singlet) molecular oxygen (1O2) attack on cholesterol were detected. These results do not support a direct role of O-2 in lipid peroxidation of cholesterol-rich membranes or of the formation of 1O2 from O-2 dismutation.


[Modeling of neuronal plasticity by studies of spatio-temporal behavior of neural structures]. In order to point out spatio-temporal filtering of nervous structures after previous studies of their spatial behaviour, we have built a new model of cell according to synaptic and membrane plasticity (it agrees with properties of integration, memory and synaptic fatigue). This model, intended to be produced in many units, is made with integrated circuits in "thick film" technology.


[Effect of scorpion venom (Androctonus australis) on neuromuscular transmission inhibited by botulinum toxin in the frog]. We have shown that Scorpion venom restores the neuro-muscular transmission inhibited by Botulinum toxin in the Frog. The effectiveness of Scorpion venom was antagonized by excess magnesium.


Assay of vitamins D2 and D3, and 25-hydroxyvitamins D2 and D3 in human plasma by high-performance liquid chromatography. I describe a new assay that is capable of measuring vitamin D2, vitamin D3, 25-hydroxyvitamin D2, and 25-hydroxyvitamin D3 in 2 ml of plasma or serum. Plasma is extracted by the Bligh and Dyer technique [Can. J. Biochem. Physiol. 37, 911 (1959)], the lipid component is fractionated by two high-performance liquid-chromatographic systems based upon adsorption and reversed-phase chromatography, and each of the four vitamin D metabolites is measured by its absorbance at 254 nm. The method has a sensitivity limit of 0.5 mug/liter of plasma. The identity of metabolite peaks was confirmed by mass spectrometry, ultraviolet absorption spectrophotometry, and rechromatography, and there was good correlation (r=0.84) between plasma 25-hydroxyvitamin D as measured by the present method and by a protein binding assay developed in our laboratory. Mean concentrations of vitamin D and 25-hydroxyvitamin D in normal adults (n=25) in December were 2.2 +/- 1.1 (SD) and 16 +/- 3.9 (SD) mug/liter, respectively. 25-Hyroxyvitamin D2 made up 31% of the total 25-hydroxyvitamin D. Patients receiving pharmacological doses of vitamin D had values for vitamin D and 25-hydroxyvitamin D that were 10- to 100-fold normal. This method provides a rapid, reliable physico-chemical assay that appears to have advantages over existing protein binding assays and can be used to measure circulating vitamin D.


An enzymatic triglyceride method that is suitable for long-term population studies. An enzymatic triglyceride method has been shown to be a suitable alternative to the Lipid Research Clinics' extraction/fluorometry method in long-term population studies. Correlation of results obtained with this method by this laboratory (y-axis) and by the Minneapolis Lipid Research Clinic Laboratory (x-axis) during a nine-week standardization period produced a curve with an intercept of -72 mg/liter, a slope of 1.019, and a correlation coefficient of r=0.996 (n=47). During this standardization period certain methodological problems were observed and corrected. An increase in background in certain clinical specimens, caused by spontaneous degradation of NADH, was observed, accurately measured, and taken into account when appropriate.


Vitamin D metabolism.

Vitamin D metabolism. During the past decade, an explosion of information has become available on the metabolism and function of vitamin D which is of great importance to clinicians in the treatment of metabolic bone disease. We have learned that vitamin D is the precursor of at least one hormone and that this hormone carries out functions in calcium and phosphorus metabolism bringing about mineralization of bone on one hand, and the prevention of hypocalcaemic tetany on the other. It may also function in the prevention of such degenerative bone diseases as osteoporosis. An important analogue of this hormone, 1alpha-hydroxyvitamin D3 has been prepared and is used successfully in the treatment of a variety of clinical conditions. This presentation will summarize these findings and their possible implications in these metabolic bone diseases.


Long-term therapy with 1alpha-hydroxyvitamin D3 in children with 'pseudo-deficiency' rickets. This investigation confirms that 1alpha-hydroxyvitamin D3 (1alpha-OHD3) is a potent drug for the treatment of patients with pseudo-deficiency rickets (Balsan et al., 1975a; Reade et al., 1975; Prader et al., 1976). 1alpha-OHD3 corrects their intestinal malabsorption of calcium and phosphorus, normalizes their serum calcium and phosphate concentrations and promotes healing of skeletal lesions. This study also shows differences in the needs for 1alpha-OHD3 of children with PDR. Three factors appear to be of importance: familial sensitivity, severity of chronic secondary hyperparathyroidism, and periods of increased growth velocity. Tolerance to long-term 1alpha-OHD3 therapy, at doses varying from 0.5 to 2 microgram/d is excellent. Surveillance of patients should include regular measurements of 24 h urinary excretion of calcium, since hypercalciuria is the first signal of overdosage.


Vitamin D resistant hypophosphataemic osteomalacia: treatment with 1alpha-hydroxyvitamin D3. Ten patients with vitamin D resistant hypophosphataemic osteomalacia are described. They had hypophosphataemia with a decreased tubular reabsorption of phosphate, malabsorption of calcium and phosphorus, proximal myopathy and extensive osteomalacic changes on iliac crest bone biopsy. The plasma alkaline phosphatase and urine hydroxyproline, however, were raised in only some of the patients. Treatment with 1alpha-hydroxyvitamin D3 in high doses rapidly cured the myopathy, increased calcium and phosphorus absorption and retention and healed the osteomalacia. Phosphorus supplements were not required.


1alpha-hydroxyvitamin D3 in the treatment of nutritional and metabolic rickets and osteomalacia. Five patients with nutritional osteomalacia or rickets and six children with rickets unresponsive to physiological doses of vitamin D were treated with 1alpha-hydroxyvitamin D3 (1alpha-OHD3). Patients with nutritional osteomalacia responded to 1--2 microgram/day of 1alpha-OHD3. The most striking findings were rises in plasma calcium and, in one case, a decrease in faecal calcium. In some cases there was a rise in plasma phosphorus, alkaline phosphatase remained unchanged. There was radiological healing. In three patients with cystinosis and one with hypophosphataemia and Barrter's syndrome 2 microgram of 1alpha-OHD3 produced healing of rickets. Plasma phosphate rose on treatment, possibly by a suppression of parathyroid activity. The response to such low doses of 1alpha-OHD3 suggests impaired 1alpha-hydroxylation of 25-hydroxyvitamin D in these patients. A patient with intestinal malabsorption was resistant to high doses of 1alpha-OHD3 by mouth but responded to parenteral administration. A boy with osteopetrosis and the biochemical changes of rickets was resistant to large doses of 1alpha-OHD3 presumably because of failure of osseous response.


An unusual form of galactosemia: studies on erythrocytes and hair roots. An unusual form of galactosemia is described in a 7-month-old boy, characterized by a late onset of the clinical symptoms. A high apparent residual activity of erythrocyte galactose-1-phosphate uridyl transferase (GT) was measured with the spectrophotometric UDP-Glucose consumption test(+/-25% of normal). The residual activity in erythrocyte lysates, determined when the patient was 7, 16 and 22 months old, significantly decreased upon storage and after preincubation with NAD-ase. The radiochemical measurement of GT activity demonstrated a severe deficiency: only a level of +/-1% of normal activity was observed, and no effects of storage or NAD-ase could be demonstrated. GT and galactokinase (GK) activities were measured radiochemically in lysates from hair roots obtained from the human scalp, and it was found that the GT/GK activity ratio is a useful index for the detection of heterozygotes. Erythrocyte and hair root lysates from the heterozygous parents of the patient displayed GT/GK ratios which were intermediate between mutant and normal. Because they offer a simple and fast way to obtain biopsy material, hair roots might become of increasing importance for carrier detection studies.


Double pre-beta lipoprotein in ischaemic heart disease. The presence or absence of a double pre-beta lipoprotein (pre-beta 1+ or pre-beta 1-) peak on agarose gel electrophoresis was recorded in: a) 77 survivors of myocardial infarction, b) their first-degree relatives, and c) 148 controls. Thirty-nine percent of myocardial infarction survivors and 24% of controls had pre-beta 1+ (P less than 0.05). The segregation of pre-beta 1+ in the families of myocardial infarction survivors was consistent with autosomal dominant inheritance. In the control group, fasting serum cholesterol and triglyceride values were higher in the pre beta 1+ subjects, but the differences were not significant. We conclude that the presence of double pre-beta lipoprotein peak on electrophoresis is associated with an increased risk of myocardial infarction.


Blastogenic response of human lymphocytes to human cytomegalovirus. A method was developed for measuring the blastogenic response of human lymphocytes to human cytomegalovirus (CMV). Viral and control antigens were prepared by extracting disrupted infected and uninfected cell cultures with an alkaline buffer. Lymphocytes from ten donors with complement-fixing (CF) antibody exhibited a blastogenic response, whereas cells from ten seronegative donors did not. A relationship between the stimulation index (SI) and the results of neutralization (NT), indirect haemagglutination (IHA) or CF tests was not observed. The maximum blastogenic response occurred after 5 to 7 days of incubation and was usually greater when the cultures were supplemented with homologous plasma instead of sera. The presence of CMV antibody in the supplementary sera did not appear to affect the reactivity of the lymphocytes.


Immunosuppressive effects of lymphocyte (type II) and leucocyte (type I) interferon on primary antibody responses in vivo and in vitro. We have tested the hypothesis that type II interferon (IF), released by immune lymphocytes after in vivo stimulation with tuberculin, has immunosuppressive effects. Mycobacterium bovis (BCG) infected mice injected with tuberculin showed a very intense suppression of antibody response to sheep erythrocytes. Sera containing lymphocyte IF strongly inhibited primary immune responses to sheep erythrocytes in cultures. Addition of macrophages could not counteract the in vitro immunosuppressive effects of lymphocyte IF, suggesting that the main effect is exerted directly on lymphocytes. Sendai virus-induced leucocyte (type I) IF was also shown to have suppressive effects in vivo and in vitro. However, lymphocyte IF was shown to be much more immunosuppressive than a preparation of type I interferon with equivalent antiviral potency. Thus type II IF, as a product of activated lymphocytes, may have a major immunoregulatory role.


Protective activity of secondary effector T cells generated in vitro against ectromelia virus infection in vivo. The anti-viral activity of secondary effector cells generated in vitro against ectromelia virus infection was investigated. Depending upon the order of administration of cells and virus, 2 X 10(6) cells significantly reduce virus titres in recipient mice. Mice injected with lymphocytic choriomeningitis (LCM) virus are not protected by secondary effector cells against ectromelia virus infection and vice versa. The cells conferring anti-viral activity are sensitive to anti-theta and complement treatment, and must share H-2K or H-2D region genes with the recipients in order for significant reduction of virus titres to occur. The possibility of exploiting this approach in clinical medicine by using T cell-mediated mechanisms against certain viral infections is briefly discussed.


Blastogenic response of purified human T-lymphocyte populations to Epstein-Barr virus (EBV). The blastogenic response of T-cell populations to infectious EBV (B95-8 strain) was studied. The addition of virus increased the [3H]thymidine uptake of unfractionated lymphocytes from seropositive and seronegative subjects, and from cord blood, but not from patients with infectious mononucleosis. The same amount of virus evoked a small response in purified T cells from seropositive donors, but not in T cells from the other sources. The addition of autologous B lymphocytes with EBV adsorbed on their surfaces caused a more significant blastogenic response, which was even more pronounced in the presence of mercaptoethanol.


Molecular exclusion electrophoresis of human serum lipoproteins: patterns in control and ischaemic heart-disease populations. 1. Electrophoresis of pre-stained lipoproteins on acrylamide-gel gradients has been carried out on serum from populations of control subjects and patients with ischaemic heart disease. The technique resolves components intermediate in position and, by inference, in size, between very-low-density and low-density lipoproteins. 2. These central band components were found in 37% of a control population but the incidence varied with age and sex, being lowest in young males and highest in elderly males. 3. The incidence of central band components in patients with ischaemic heart disease was 64% (males) and 71% (females), and the difference between these figures and those for matched control subjects was highly significant. The intensity of central band components in the group with ischaemic heart disease was significantly greater than in the control group. 4. The presence and intensity of central bands show positive correlation with serum cholesterol and triglyceride values, but many patients showing the phenomenon have normal lipid values. Of patients with ischaemic heart disease 31% showed central band components and had normal lipid values.


Cutaneous pain without tumor: a manifestation of occult synovioma. A thirty-four year old male presented with severe spot tenderness for eighteen months, situated in the right upper anterior thigh. The skin was normal. No tumor, swelling, or inflammation was found. On the basis of a clinical diagnosis of glomus tumor, the area was biopsied and a microscopic synovioma was documented in the fascia just deep to the subcutis. From the literature, it is apparent that approximately 20 percent of the patients with this cancer go through a pretumor phase characterized by only sustained pain or tenderness. By detecting the lesion in this phase, the dermatologist can contribute substantially to an increased cure rate, which has lingered at about 50 percent for at least ten years.


Biopsy of the nail area.

Biopsy of the nail area. It is as simple to biopsy around the nail as it is at any other site, and there are multiple lesions that deserve biopsy. Biopsy of the nail bed or nail matrix is avoided unless there is possible malignant change. Biopsy sites in the nail bed usually heal satisfactorily, but biopsy of the matrix is more likely to produce permanent deformity.


Therapy of parotid tumors.

Therapy of parotid tumors. A therapy schedule is presented for the surgical treatment of parotid tumors, based on the experience gained from more than 600 operations for parotid tumors. The individual surgical techniques (lateral parotidectomy, total parotidectomy with preservation or resection of the facial nerve, with or without reconstruction of the nerve) are selected depending on the histologic findings in the tumors. The fact is stressed that in a tumor disease the lateral parotidectomy constitutes the minimal operation. The different tumors can be divided into four groups. The operative treatment of tumors of the facial nerve in its extratemporal course is particularly mentioned since clinically these growths usually appear to be parotid tumors.


[Ectopic production of chorionic gonadotropin in apudomas (author's transl)]. Two pheochromocytomas, two carcinoid tumours of the ileum and one pancreatic vipoma showed APUD-system properties and formed electron-dense secretory granules indicating synthesis and storage of proteohormones. Besides catecholamines, 5-hydroxytryptamine and vasoactive intestinal polypeptide, all five tumours contained immunoreactive chorionic gonadotropin and also its free, protomeric beta chain as a sign of de-differentiation. Plasma of the tumour patients also contained immunoreactive chorionic gonadotropin and the free protomeric beta chain, both of which are useful tumour markers.


[Cerebral damage and cytomegalovirus infection (author's transl)]. Serological tests were performed on 58 children with microcephalus and 223 children with cerebral damage but no microcephalus (all patients were living in the Munich region). In the microcephalus group 38% had antibodies against cytomegalovirus, while in healthy controls the incidence was only 18%. Mean titre levels were significantly higher in microcephalic children. Of eleven microcephalic children seven were cytomegalovirus excreters: all of the latter had severe cerebral defects. On the other hand, in patients with cerebral damage but no microcephaly there was no difference serologically or with regard to virus excretion from healthy controls. The authors suggest, in conformity with opinions expressed by others, that there is a definite connection between--usually unrecognised--cytometalovirus infection and certain forms of cerebral damage. They may be atypical and symptom-poor diseases in which CNS signs appear only in the course of time.


Metabolic effects of some antibilharzial drugs inhibition of citrate oxidation by Bilharcid. Bilharcid inhibited citrate oxidation and the inhibition was decreased by time. The inhibition is not restored or reduced by increasing citrate concentration. NAD stimulated citrate oxidation in the presence of Bilharcid. Cysteine slightly stimulated citrate oxidation, while ATP had no effect. Addition of citrate before Bilharcid protected the enzyme against the inhibitory action of the drug.


PCBs and environmental contamination.

PCBs and environmental contamination. A survey is given on reported PCB-residues and accumulation in various environmental media including water and aquatic environment, flora, animals, food, and humans. Model studies with technical PCB, as well as with pure individual components, are presented from various working groups. As examples, metabolic studies and photochemical experiments are discussed. According to our present knowledge, a major metabolic pathway in animals and plants is hydroxylation, often followed by methylation or conjugation. By UV-irradiation, however, oxygenation, dechlorination and chlorination, polymerization and isomerization may occur.


Overproduction disease in man due to enzyme feedback resistance mutation. Purine overproduction in gout due to excessive activity of mutant feedback-resistant phosphoribosylpyrophosphate synthetase. Physiologically superactive phosphoribosylpyrophosphate (PRPP) synthetase, due to feedback resistance mutation, was found in a family with excessive purine production, gout and uric acid lithiasis. The superactivity of the mutant enzyme was manifest in the propositus' erythrocytes and cultured fibroblasts, in increased generation, content and metabolic availability of PRPP, leading in the fibroblasts to acceleration of the rate of purine synthesis de novo. One of the propositus' two siblings was similarly affected, but the propositus' father, his second brother and four sons, were all clinically and biochemically normal. The mother was clinically normal and normouricemic, but hyperuricosuric. Cultured fibroblasts from her skin exhibited variability in PRPP content and availability and in the rate of purine synthesis de novo. The mother's cultures were found to contain a mosaicism of two cell populations, one with normal and the other with mutant PRPP synthetase, indicating an X-linked pattern of inheritance of the PRPP synthetase abnormality in this gouty family.


The effects of fasting on glucose-6-phosphatase of mouse liver and kidney. Glucose-6-phosphatase levels were measured in livers and kidneys of control mice and mice fasted for 24, 48, and 72 h, respectively. The enzyme was assayed with and without deoxycholate supplementation. When assayed in the absence of deoxycholate, significant increases in liver enzyme levels was observed after fasting for 48 and 72 h, respectively. For the kidney enzyme, a significant increase was observed only after 48 h fasting. When the enzyme was assayed in the presence of 4.8 mmol/l deoxycholate, fasting-induced increases of liver enzyme levels were further potentiated, but no significant differences were observed in kidney enzyme levels between fasted and control mice.


Evidence for the participation of cytosolic protein kinases in membrane phosphorylation in intact erythrocytes. The effects of adenosine 3':5'-monophosphate (cyclic AMP) on the phosphorylation of membrane proteins in intact rabbit and human erythrocytes were investigated. The addition of cyclic AMP to intact human or rabbit erythrocytes results in an increase in the incorporation of ortho[32P]phosphate into several membrane protein components which are known to serve as substrates for the cyclic-AMP-dependent protein kinases. Thus this increase in protein phsophorylation is probably due to the activation of either soluble or membrane-bound cyclic-AMP-dependent protein kinases. Incubation of human erythrocytes in the presence of ortho [32P]phosphate and cyclic AMP also leads to the phosphorylation of a membrane protein component, band 7, which has not been previously detected in the autophosphorylation of isolated ghosts. Since rabbit erythrocyte membranes do not contain any cyclic-AMP-dependent protein kinase, the results suggest that cytoplasmic kinases also play a role in the phosphorylation of membrane proteins in intact cells.


A universal lipid exchange protein from rat hepatoma. The postmicrosomal protein fraction from rat hepatoma 27 adjusted to pH 5.1 stimulates phospholipid exchange between rat liver microsomes and mitochondria with higher rates and in a less specific way than the corresponding fraction from rat liver. A phospholipid exchange protein has been purified to homogeneity from the hepatoma pH-5.1 supernatant by gel filtration on Sephadex G-75 and ion-exchange chromatography on carboxymethylcellulose. The isolated protein had a molecular weight of 11200 as determined by electrophoresis on polyacrylamide in the presence of dodecyl sulfate and of 11168 as calculated from the amino acid composition. Isoelectric focusing showed a single band at pH 5.2. in the assay system rat liver microsomes leads to mitochondria the protein exhibits a complete lack of substrate specificity transferring all the major microsomal phospholipids to about the same extent. The possible role of the isolated phospholipid exchange protein in the chemical dedifferentiation of hepatoma cell membranes is discussed.


A hydrogen-deuterium exchange study of the amide protons of polymyxin B by nuclear-magnetic-resonance spectroscopy. 1. Proton magnetic resonance spectra at 270 MHz of polymyxin B, a cationic oligopeptide antibiotic, show the influence of the inorganic counteranion present in solution. 2. Hydrogen-deuterium exchange rates for the amide protons are of two types, depending on whether the anion is monovalent or polyvalent. Polyvalent anions catalyse the acid-catalysed reaction more than the monovalent anions. 3. The structure in solution was monitored using the proton signals of the amides, the phenylalanine aromatic protons, and the leucine methyl and gamma-CH protons in several polymyxin salts. The temperature coefficients of the chemical shifts of the N-H protons are used to identify two beta turns in the cyclic ring of polymyxin B. The variation in chemical shift of the N-H protons, the aromatic protons and the leucine protons are correlated with anionic size and electronegativity.


Quantitative studies on competitive ligand binding to bovine serum albumin by use of the spin label 5-doxyl dodecanoic acid. The binding of the spin label 5-doxyl dodecanoic acid to bovine serum albumin in phosphate buffer at pH 7.4 was studied by electron spin resonance spectroscopy. Free label and label bound to serum albumin could be quantitatively measured and evaluated from the superposition spectra of these two species with no previous separation. The efficiency relative to the spin label as competitors for binding to serum albumin was studied with salicylic acid and some fatty acids of medium length. The results were represented both by the stoichiometric model involving equilibrium constants Ki, by binding isotherms constructed from the Ki values, and by a purely graphical representation of the experimental data points without connection with any special binding model.


Calf-spleen nicotinamide-adenine dinucleotide glycohydrolase. Properties of the active site. The interaction between the nicotinamide adenine dinucleotide binding domain of calf spleen NAD glycohydrolase and its ligands has been studied. The use of competitive inhibitors, structurally related to different portions of the NAD molecule (i.e. adenosine and nicotinamide moieties), revealed the considerable importance of the binding between the pyrophosphate linkage and probably an arginyl residue of the active site. This interaction allows the positioning of the substrate in a conformation which permits catalysis to occur. The binding between the 2'-hydroxyl of the adenosine moiety and a residue of the active site, which exists in NAD-linked dehydrogenases, is probably missing in the calf spleen NAD glycohydrolase, based on the inhibition by salicylates, 2'-deoxyadenosine 5'-monophosphate and the hydrolysis of the 2'-deoxyadenosine analogue of NAD. The NAD glycohydrolase could be completely inactivated by 2,3-butanedione, an arginyl-modifying reagent. The reaction followed pseudo-first-order kinetics and the modification was found to be reversible. Woodward's reagent K, a reagent for carboxyl residues, partially inactivated the enzyme, which resulted in a change of the NAD glycohydrolase kinetic parameters Km and V. The inactivation rate was complicated by a parallel decomposition of the reagent.


Adenosine-3':5'-monophosphate-binding proteins from bovine kidney. Isolation by affinity chromatography and limited proteolysis of the regulatory subunit of protein kinase II. Affinity chromatography on cyclic AMP columns allowed a two-step isolation of the cyclic-AMP-binding proteins from bovine kidney cytosol. An AMP-binding protein (apparent molecular weight approximately 60 000) and large amounts of a low affinity binding protein ('P35'; apparent subunit size approximately 35 000) were obtained in practically pure form besides the high affinity binding proteins of the R type. Among the R proteins the dimer R2 of the regulatory subunit of protein kinase II (apparent subunit size approximately 54 000) represented the bulk material. Small amounts of monomer, of higher aggregates, and of a protein 'P49' (subunit size approximately 49 000) presumably identical with the regulatory subunit of protein kinase I were also detected. The R protein fraction of kidney also contained a high affinity binding protein of smaller size (designated as R'; molecular weight approximately 37 000) which appeared to be derived from protein R2 of protein kinase II by limited proteolysis. At all stages of purification, R protein and its aggregates could be quantitatively transformed into R' protein (or a closely related polypeptide) by several proteases including the relatively unspecific proteinase K. The degradation product exhibited unchanged cyclic-AMP-binding capacities but had largely lost the ability to inhibit the catalytic subunit C of protein kinase, to be phosphorylated by C, and to form a dimer. Preliminary experiments indicate that protein R' may be a natural component of kidney tissue.


Structural homology of cytochromes c.

Structural homology of cytochromes c. Cytochromes c from many eukaryotic and diverse prokaryotic organisms have been investigated and compared using high-resolution nuclear magnetic resonance spectroscopy. Resonances have been assigned to a large number of specific groups, mostly in the immediate environment of the heme. This information, together with sequence data, has allowed a comparison of the heme environment and protein conformation for these cytochromes. All mitochondrial cytochromes c are found to be very similar to the cytochromes c2 from Rhodospirillaceae. In the smaller bacterial cytochromes, Pseudomonas aeruginosa cytochrome c551 and Euglena gracilis cytochrome c552, the orientation of groups near the heme is very similar, but the folding of the polypeptide chain is different. The heme environment of these two proteins is similar to that of the larger bacterial and mitochondrial cytochromes. Two low-potential cytochromes, Desulfovibrio vulgaris cytochrome c553 and cytochrome c554 from a halotolerant micrococcus have heme environments which are not very similar to those of the other proteins reported here.


Congenital generalized lipodystrophy with insulin-resistant diabetes. Hyperglycemia, glucose intolerance, hyperinsulinemia and resistance to exogenous insulin were found in a 10-year-old Japanese boy diagnosed as having congenital generalized lipodystrophy. Studies on insulin receptors of circulating mononuclear leucocytes indicated that insulin-resistant diabetes combined with congenital generalized lipodystrophy may be due to disturbance of insulin binding to membrane receptors. No insulin-binding antibody or antibody that impairs insulin-receptor binding was found. Plasma glucagon showed an exaggerated response to L-arginine before treatment. After treatment with a controlled diet and an oral sulfonylurea (500 mg/day) for 4 weeks, there was improvement in the plasma glucagon response to L-arginine. Improvement in the hyperglycemia, hyperinsulinemia and acanthosis nigricans was also observed. On the other hand, on completion of a 7-day high-fat diet, a marked increase in serum free fatty acids, triglycerides and beta-lipoproteins was observed. The total plasma post-heparin lipolytic activity during the high fat diet was within the normal range. However, the level of protamine-inactivated activity was 3 times that of the control.


Congenital lactic acidosis due to pyruvate carboxylase deficiency: absence of an inhibitor of TPP-ATP phosphoryl transferase. Two children are described who suffered from episodes of metabolic acidosis and progressive mental and motor deterioration. The patients showed periodic elevation of blood lactate, pyruvate and alanine, which was accompanied by vomiting, hypotonia or convulsions. The concentrations of lactate and pyruvate in cerebrospinal fluid were found to be increased. Liver biopsies revealed a decrease in pyruvate carboxylase activity and normal pyruvate decarboxylase activity. No inhibitor of TPP-ATP phosphoryl transferase was detected in urine from the patients. These findings suggest that congenital lactic acidosis due to pyruvate carboxylase deficiency is probably a different disease entity from Leigh's encephalomyelopathy. A possible mechanism of brain damage caused by a defect in pyruvate carboxylase is postulated.


Recurrent multiple cranial neuropathies. A report of 12 patients with recurrent multiple cranial neuropathy is presented. In most cases the palsies a accompanied a mild respiratory infection. Both motor and sensory cranial nerves were affected, the facial nerve and optic nerve being the most common. The symptoms were self-limited in course and steroid therapy seemed to hasten the recovery in the cases it was used. Only in a few cases a noticeable neurological deficiency remained. Aetiological aspects of the syndrome are discussed.


Use of trypsin for isolation of islets of Langerhans in the rat. A new technique for pancreatic islet isolation, based on trypsin administered into the pancreatic duct system and a reduced amount of collagenase for digestion of the removed and chopped pancreatic tissue, yielded viable islets as judged by the metabolic response of 27 inbred, streptozotocin-diabetic rats after intraportal transplantation of the islets: all recipients of greater than 240 islets normalized their blood glucose, plasma insulin, urine volume and urinary glucose. The number of islets isolated was the same as with the conventional collagenase technique.


Modification of theophylline-induced lipolysis in human fat cells after trypsination. Trypsin-treatment of human fat cells results in the potentiation of the lipolytic response and the cAMP accumulation induced by theophylline (5 . 10(-4) M) but not of those induced by theophylline (5 . 10(-3) M). The amount of cAMP formed after exposure to theophylline (5 . 10(-3) M) plus norepinephrine (5 . 10(-6) M) remains, however, 2.6 fold higher in trypsin-treated human fat cells than in the control ones.


Search for sex-dependent and gestation-induced changes in choline and ethanolamine phosphorylating activities. Choline kinase and ethanolamine kinase of liver, brain and kidney had nearly the same activity in 4-month-old male and virgin female mice. Ethanolamine kinase activity was almost doubled in the liver and brain of mice in advanced pregnancy compared with the virgin, while choline kinase activity was unaltered.


Effect of low density lipoprotein on proteoglycan synthesis by aorta cells in culture. Increasing the content of human serum low density lipoprotein in the growth medium led to greater incorporation of 35S-sulfate into proteoglycan (mostly into dermatan sulfate) by primary aorta cells but did not affect similar incorporation by fibroblast cells. These results suggest a mechanism which can explain the increased deposition of lipid in aorta due to hyperlipidemia.


The metabolism of methadone by cultured mammalian cells. Rat hepatoma tissue culture cells and mouse leukemic cells were found to metabolize [1-3H] methadone to at least 2 unidentified radioactive compounds. These results suggest that cultured cells may be useful models for studying methadone metabolism by specific cell types.


Effect of sodium octanoate on leucine incorporation into protein of rat liver slices and of Yoshida ascites hepatoma cells. 7.38 X 10(-4) M octanoate does not significantly modify leucine incorporation into protein of rat liver slices, while in hepatoma cells a 19% inhibition has been noted. 3.69 x 10(-3) M octanoate reduces leucine incorporation to about the same extent (71-76%) in both liver slices and hepatoma cells.


Evidence for possible emergence of immunochemically distinct nucleotide-peptides in malignant transformation. Antibodies against nucleotide-peptides of beef heart do crossreact with nucleotide-peptides of other beef organs, but with nucleotide-peptides of Ehrlich tumor cells. Antibodies against the latter do not crossreact with nucleotide-peptides of normal organs, but do so with nucleotide-peptides of a rat hepatoma.


[Effect of prostaglandin F2alpha on inhibitory and excitative processes in the sensory nerve endings of the cornea]. In concentration of 0.002 to 0.025 per cent prostaglandin F2alpha potentiated the anesthetic action of dicain and decreased the mitigating effect of acetylcholine in regard to dicain-induced anesthesia. These facts prove that prostaglandin F2alpha can in certain conditions attenuate the process of excitation and intensity that of inhibition in the sensory endings of the cornea.


Identification and regulation of alpha- and beta-adrenergic receptors. Direct radioligand binding methods for studying the alpha- and beta-adrenergic receptors have been developed over the past several years. These techniques use radioactively labeled adrenergic antagonists and agonists to identify the receptors in appropriate membrane fractions from catecholamine-sensitive tissues. In the case of the beta-adrenergic receptors, confident receptor identification has been aided by the close correlation of binding data with data on adenylate cyclase activation. Such direct binding studies are providing new insights about the molecular characteristics and regulatory properties of the receptors.


Possible involvement of cerebroside sulfate in opiate receptor binding. Cerebroside sulfate (CS) appears to fulfill most of the structural requirements of a hypothetical opiate receptor. It possesses many of the properties that are thought to be necessary for the identification of an "opiate receptor," exhibiting high affinity and stereoselective binding to a number of narcotic drugs. Although these properties are insufficient to establish identity of the receptor, it is highly significant that the affinity of this binding can be correlated with the analgetic potency of these drugs in both man and rodents. CS is an endogenous component of brain tissue, and a partially purified opiate receptor from mouse brain has been found to be CS. Other experiments indicate that reduced availability of brain CS decreases the analgetic effects of morphine and this is accompanied by a reduction in number of binding sites, suggesting that the interaction of opiates with CS observed in vitro may also have importance in vivo. CS was also found to be a component of the opiate receptor after marking with 125I-labeled diazosulfanilic acid. The possibility that CS or the SO4-2 group of this lipid may be the "anionic site" of the opiate receptor should be considered.


Histochemical localization of opiate receptors and opioid peptides. It is possible to localize opiate receptors by histochemical methods. They appear in high densities in anatomical areas associated with physiologic functions altered by opiates. They appear to mediate inhibitory responses; some of them, in certain regions could be involved in axo-axonic synapses. The immunohistochemical studies as well as the electrophysiologic results are compatible with the view that the enkephalins are the endogenous substrates for the opiate receptors.


Leydig and Sertoli cell function in normal and oligospermic males: a preliminary report. Sertoli and Leydig cell function was evaluated in patients with oligospermia. Testicular biopsy specimens were incubated in vitro with follicle-stimulating hormone (FSH) or luteinizing  hormone (LH), and the amount of cyclic adenosine 3':5'-monophosphate (cyclic AMP) generated was measured. Cyclic AMP has been shown to be intracellular mediator for the pituitary gonadotropins FSH and LH. The amount generated by the Sertoli cells response to FSH, and by the Leydig cells in response to LH, was contrasted with that in age-matched, normal control subjects. The Leydig cell response was similar in both groups. However, the amount of cyclic AMP generated by the Sertoli cells from oligospermic males was much less when compared with normal control subjects. This finding suggests a possible metabolic abnormality in the Sertoli cells of patients with oligospermia which might, in turn, alter spermatogenesis.


The association between oral contraception and hepatocellular adenoma--a preliminary report. Women with long-term use of oral contraception (OC) are at increased risk of developing a serious, though nonmalignant, liver tumor--hepatocellular ademona (HCA)--according to a case-control study conducted by the Center for Disease Control (CDC) in collaboration with the Armed Forces Institute of Pathology (AFIP). The tumor is sometimes fatal, deaths usually being due to sudden rupture and hemorrhage. This study suggests that, in addition to long-term OC use, a women's age and the hormonal potency of the OC she uses affect her changes of developing HCA. Women 27 years old and older who have used OC with high hormonal potency for 7 or more years are at the greatest risk.


Steroidogenesis in isolated adrenal cells: excitation by calcium. Calcium salts were found to replace ACTH in inducing steroidogenesis in isolated adrenocortical cells from rats. This Ca-specific stimulation occurred when the cation was presented to the cells in the presence of phosphate and carbonate as a counter-ions under conditions which favoured the formation of colloidal calcium. Colloid generation and stabilization was facilitated by the use of calcium buffers and gelatin. Stable soluble or sparingly soluble calcium complexes were inactive. The preparation of cells and metastable calcium solutions is described in detail. The Ca trigger was sensitive to Ca deprivation or inhibitors of Ca transport and could be replced by Sr. The relative role of Ca and cyclic AMP as second messengers is discussed.


Effect of cordycepin on CRF stimulation and steroid inhibition of ACTH secretion by rat pituitary cells. Isolated pituitary cells prepared from adrenalectomized rats secrete ACTH in response to CRF, and this response is inhibited by corticosterone. Both the stimulation of release by CRF and the inhibition of release by corticosterone are antagonized by cordycepin (3'-deoxyadenosine). Inhibition of CRF-stimulated secretion by cordycepin is apparently not related to inhibition of RNA synthesis, since high doses of actinomycin D do not affect ACTH secretion. More likely, cordycepin's inhibition of secretion stems from its inhibition of adenylate cyclase activity. Inhibition of corticosterone action by cordycepin is qualitatively similar to that previously reported actinomycin D. This effect of both drugs is probably due to inhibition of RNA synthesis. Significantly, a low dose of cordycepin has a greater inhibitory effect on corticosterone action than on total cellular RNA synthesis. Cordycepin is reported to preferentially inhibit messenger RNA synthesis, and low dose preferentially inhibits appearance of cytoplasmic RNA in pituitary cells. These data suggest that corticosterone-induced RNA is a cytoplasmic (messenger) RNA.


Thyrotropin receptors in adipose tissue, retro-orbital tissue and lymphocytes. Thyrotropin (TSH) receptors on retro-orbital muscle and fat have been implicated in the pathogenesis of Graves' exophthalmos and it has been suggested that TSH has a direct effect on human fat metabolism. We have therefore investigated the interaction of biologically active 125I-labelled TSH with membranes prepared from human adipose, retro-orbital and thyroid tissue. Since lymphocytes contain receptors for several polypeptide hormones, TSH binding to lymphocyte membranes was also studied. We were unable to demonstrate TSH receptors in adult human adipose tissue, retro-orbital muscle and fat, or peripheral blood lymphocytes. In contrast, adult and neonatal guinea pig adipose tissue membranes showed similar TSH binding characteristics to guinea pig thyroid membranes.


Selected properties of [3H]prostaglandin E1 binding to dispersed bovine luteal cells. Selected properties of [3H]prostaglandin (PG) E1 binding to collagenase dispersed bovine luteal cells were studied and compared with those observed in luteal plasma membranes. [3H]-PGE1 specific binding to a relatively homogeneous population of luteal cells was a rapid (K1 = 4.2 X 10(5) M-1 .sec-1), reversible (K-1 = 3.9 X 10(-3) sec-1), saturable and specific process at 38 degrees C. The binding was homogeneous with an apparent dissociation constant of 2.4 nM and 1.8 X 10(5) receptors per cell. The presence of increasing amounts of unlabeled PGs inhibited [3H]PGE1 binding in a dose-dependent manner. The potency order for this inhibition of binding was: PGE 2 greater than PGE1, (15S)-15-methyl-PGE2 methyl ester greater than PGF2alpha greater than PGF1alpha greater than other PGs, PGE, PGF metabolites and PGF analogs. Other than the homogeneous nature of [3H]PGE1 binding and the greater effectiveness of PGE2 compared to PGE1 in cells, the rest of the properties of [3H]PGE1 binding to cells were in excellent agreement with those observed in plasma membranes.


Properties of solubilized nuclear triiodothyronine binding proteins. Rat liver nuclear proteins bind 3,5,3'-triiodo-L-thyronine (T3) to essentially one class of sites (Ka approximately 1 X 10(10) M-1). Gel filtration and sucrose gradient centrifugation studies show a main T3 binding component with a Stokes radius of 33 A and a sedimentation coefficient of 3.5S, and variable amounts of high molecular weight binding components, most of them being reversible aggregates of the main component formed during storage. But the uniqueness of the nuclean T3 binding proteins (NTBP) cannot be ascertained from polyacrylamide gel electrophoresis data. During storage in the absence of reducing agents, NTBP aggregate and rapidly lose their ability to bind T3; T3--NTBP complexes also aggregate and progressively dissociate. This can be reversed by dithiothreitol. Bound T3 could temporarily stabilize the binding site but cannot protect NTBP against general conformational changes which follow the oxidation of their essential --SH group(s). NTBP are DNA binding proteins with probably a relative independence of their DNA and T3 binding sites: they bind T3 to the same class of high affinity sites whether complexed or not with DNA; bound T3 is not a prerequisite for DNA binding.


Regional pancreatic concentration and in-vitro secretion of canine pancreatic polypeptide, insulin, and glucagon. The regional concentrations and in-vitro secretions of canine pancreatic polypeptide (cPP), insulin, and glucagon were studied. CPP is found predominantly in the uncinate process of the dog pancreas, whereas insulin and, more markedly, glucagon predominate in the body and tail of the pancreas. In-vitro secretion studies of pancreatic pieces indicate that dibutyryl cyclic AMP (dcAMP) alone can stimulate cPP release whereas glucose and arginine alone have no effect. Arginine, but not glucose, potentiates this stimulant effect of dcAMP. These studies suggest that the cAMP generating system may play a role in regulation of cPP secretion.


Idiopathic granulomatous hepatitis with a prolonged course: effect of corticosteroid therapy. The effect of corticosteroid (or ACTH) therapy on 4 patients with idiopathic granulomatous hepatitis is described. All patients presented with spiking fever and chills and none had jaundice. Only 1 patient had an enlarged tender liver and 3 had splenomegaly. The erythrocyte sedimentation rate was increased in all cases while the white blood cell count was typically normal. Impairment in liver function was insignificant and consisted of a mild elevation of SGOT and alkaline phosphatase activities and prolonged prothrombin time. All patients presented a diagnostic challenge. The diagnosis was established by routine liver biopsies in 3 cases and by laparotomy in the 4th. The etiology could not be established. All patients reacted dramatically to prednisone (or ACTH) after failure of other therapeutic regimens. The disease has, however, been present for 5 years in 1 patient and 10 years in another, Relapses occur after cessation of therapy.


The metabolic fate of high density lipoprotein (HDL) in the diabetic rat. Diabetic rats, pre-fed and maintained on a sucrose-rich diet, have marked hyperlipoproteinaemia, with an increase in both very low density (VLDL) and high density lipoproteins (HDL). HDL obtained from both diabetic and non-diabetic rats and labelled with 125I or 135I was injected simultaneously into diabetic and non-diabetic rats. The half life of the two HDL preparations was similar in both diabetic and non-diabetic rats and ranged between 11.4 to 12.0 hours. A-I apolipoprotein had a disappearance rate parallel to the whole HDL, in contrast to the apo-C peptides which had a faster rate of removal. Although the fractional catabolic rate (FCR) of HDL preparations was slower in the diabetic rats, there was a 16% increase in the calculated synthetic rate (SR) of HDL-protein. These observations could explain the increased plasma HDL levels in the sucrose-fed, streptozotocin-induced diabetic rat.


Cell mediated immunity in experimental influenza and parainfluenza infection. Cell mediated immunity (CMI) to Sendai and Kunz viruses, determined by a 51Cr release assay of cytotoxicity, has been shown to develop during the course of these infections. This virus specific response is mediated by sensitised T cells, and takes place against a background of innate cell mediated immunity. The mediator of this innate cytotoxicity exhibits none of the characteristics of T, B or K cells, but bears close resemblance to the NK cell which is spontaneously cytotoxic for certain tumour cells. Removal of the virus specific CMI by T cell depletion produces a moderate immunosuppressive effect. The nature of this effect and the role of CMI in these infections are discussed.


[Effect of mutations for adenylate cyclase (cya) and the cyclic adenosine monophosphate receptor protein (cgp) on the gene expression of nucleoside catabolism in Escherichia coli]. The effect of cya and crp mutations on the expression of the activity of nucleoside catabolizing genes has been studied in Escherichia coli. It is found that cya and crp mutants lose their ability to grow on nucleosides as carbon sources in spite of the preservation of the basal levels of nucleoside catabolizing enzymes, found in cell-free extracts of cya and crp mutants. It is shown that cya and crp mutations completely release the influence of the regulatory gene cytR on the activity of uridine phosphorylase (udp gene) and thymidine phosphorylase (tpp gene). On this ground it is assumed that the cytR gene product acts at the level of promotors of the corresponding structural genes, causing their insensitivity to the positive action of cAMP--CRP complex. The same data concerning the effect of cya and crp mutations on cytR regulation have been reported [8], but these authors favoured the hypothesis that the cytR gene product is a repressor protein, which binds to the specific operator.


Experimental infection of pheasants with Lednice (Yaba 1) virus. Young pheasants (24-hour- and 7-day-old) are capable of producing viraemia and antibodies after experimental infection with Lednice virus even after low doses of virus (0.7-0.9 log mouse LD50/g). Viraemia lasts approximately 3 days, but the titres of virus in the blood are low. The possible role of young pheasants in the circulation of virus in nature is discussed.


[A new topographic method for localising the venous angle of the deep cerebral veins on the lateral phlebographic phase (author's transl)]. A simple topographic method has been developed for the localisation of the venous angle on the lateral cerebral phlebogram. The principle depends on an angular measurement and an estimation of proportions which are statistically independant of each other. The method was tested on 103 phlebograms of adult patients with supratentorial spaceoccupying lesions. It was compared with conventional measurements; a number of advantages were found for using our method and its accuracy is as good as the other, most precise, methods.


[Electroencephalography in clinical research into psychoses (author's transl)]. At present EEG investigations in heterogenous acute schizophrenic and psychotic syndromes suggest changes in the regulating, reticulo-thalamo-cortical, functional system. One may further assume that cerebral functional changes consist of a dissociation of normally closely linked functional systems. In favor are findings in functional psychoses ("Funktionspsychosen"), also the dissociation of usually coordinated systems in normal psychologic changes during going to sleep and REM stages of dreams. A further thalamohippocampo-reticulo system which might act in parallel to the activating reticular system has become important in EEG research into psychoses. Changes here probably in part of focal EEG phenomenon localized in the temporal region. Here, too, EEG findings in schizophrenic syndromes requiring further proof emerge.


Carbon and hydrogen metabolism of xylitol and various sugars in human erythrocytes. 1) Erythrocytes are able to metabolize D-ribose, D-xylitol, D-xylulose, D-fructose and D-glucose; the rates of metabolism increase in that order from 2430 to 26200 ng atom C/ml packed cells per 120 min of incubation. 2) The utilization of the carbon of these substrates and its recovery in the products were found to be in balance, when the change in the 2,3-bisphosphoglycerate concentration was taken into account. 3) The metabolic rates strongly affected the 2,3-bisphosphoglycerate level. Without addition of substrate the decomposition rate of this intermediate was found to be 1030 nmol/ml packed cells per 120 min. 4) The net decrease of the 2,3-bisphosphoglycerate concentration and the conversion of this compound into lactate provides a NAD regeneration system which enables the red blood cells to utilize xylitol. 5) The rate of carbon metabolism via the pentose phosphate cycle is determined by the NADPH requirement of the erythrocytes which was found to be 160 nmol/ml packed cells per 120 min under the experimental conditions employed.


Factors involved in the modulation of cell proliferation in vivo and in vitro: the role of fibroblast and epidermal growth factors in the proliferative response of mammalian cells. The control of proliferation of mesoderm-derived cells by EGF and FGF has been examined taking, as an example, the vascular endothelium. The mechanisms by which cell proliferation can be brought to a stop in vivo and in vitro have been reviewed.


Susceptibility of inbred and outbred mouse strains to Sendai virus and prevalence of infection in laboratory rodents. Sendai virus is one of the more prevalent and serious virus infections of rodents. Infection was found in 66% of the mouse, 63% of the rat, 83% of the hamster, and 44% of the guinea pig colonies examined. Twenty-four inbred and outbred strains of mice were tested for their sensitivity to lethal Sendai virus infection. The 129/J mice tested were approximately 25,000-fold more sensitive than SJL/J mice; however, both mouse strains were similarly permissive in support of viral replication in their lung tissues. Histopathological studies revealed that whereas lesions in both sensitive and resistant mice were qualitatively similar, the lesions in the more sensitive 129/J mice appeared earlier, were much more extensive, and persisted longer than in the resistant SJL/J mice. These results suggest that the observed variance in sensitivity is not the result of a genetic restriction on virus infection and replication but rather is the result of a physiological factor(s) possibly related to some aberration of strain difference in the humoral or cell-mediated immune response.


Extraction of skin test activity from Coccidioides immitis mycelia by water, perchloric acid, and aqueous phenol extraction. Water, perchloric acid extracts, and fractions of partially defatted whole mycelia of Coccidioides immitis contained delayed skin test activity when tested in C. immitis-infected guinea pits. Aqueous phenol extraction of these fractions resulted in partitioning of activity between aqueous-soluble and phenol-soluble fractions; activity was found to be water soluble after removal of phenol by extensive dialysis. Highest specific activity skin test antigen was invariably found in the phenol-soluble phase, water-soluble fraction. Material of equivalent activity could also be extracted directly from the defatted mycelia. Skin test active fractions contained glucose, mannose, 3-O-methylmannose, glucosamine, and amino acids.


Varicella-zoster virus: isolation and propagation in human melanoma cells at 36 and 32 degrees C. Cell lines derived from human malignant melanoma tumors are susceptible to infection with varicella-zoster virus (VZV). Within 5 days after inoculation of vesicular fluid, cytopathic changes appeared in melanoma cell monolayer cultures that were incubated at either 36 or 32 degrees C. The VZV isolates at the two temperatures were serially propagated by passage of trypsin-dispersed infected cells. A plaque assay was developed utilizing melanoma cell monolayers overlaid with nutrient medium containing carboxymethylcellulose. By this assay method, the growth cycle of a VZV isolate propagated at 36 degrees C was studied and compared with that of another VZV isolate grown at 32 degrees C. With equivalent infected-cell inocula at a ratio on one inoculum cell to eight uninfected cells, the yield of cell-free virus at an incubation temperature of 32 degrees C was slightly higher than at 36 degrees C, although the peak occurred 60 h, rather than 36 h, postinfection. It was also found that the titer of low-passage VZV propagated at 36 degrees C was 0.5 to 1 log higher when assayed at 32 degrees C rather than at 36 degrees C.


Enhanced resistance against encephalomyocarditis virus infection in mice, induced by a nonviable Mycobacterium tuberculosis oil-droplet vaccine. Female C57B1/10 mice injected intravenously (i.v.) with nonviable Mycobacterium tuberculosis Jamaica cells associated with oil-droplet emulsions (WCV) were highly resistant to the i.v. injection of encephalomyocarditis virus (EMCV). Resistance to infection (87% survival) was detected from 1 week to at least 12 weeks after injection of WCV. Mice vaccinated i.v. also were resistant to intraperitoneal, subcutaneous, or intramuscular virus challenge, but were not resistant to intracranial challenge. Mice vaccinated intraperitoneally also were resistant to virus infection, whereas WCV administered intramuscularly or subcutaneously did not protect mice from virus injected by any route. Less than 50% of WCV mice that survived virus challenge possessed serum anti-EMCV-neutralizing antibody (less than 1:10), and none had detectable (less than 1:10) serum interferon. Interferon was not detected in sera of WCV mice from 4 to 144 h after i.v. injection of EMCV. Studies concerning the effects of WCV on EMCV infection suggest that mice may be protected by mechanisms that inhibit early viral replication and spread of virus to the central nervous system.


Changes in liver and L-cell plasma membranes during infection with Coxiella burnetii. Changes in plasma membrane proteins of guinea pig liver and L-929 cells were studied during infection with Coxiella burnetii. Polypeptide species resolved by disc polyacrylamide gel electrophoresis with sodium dodecyl sulfate showed quantitative but no qualitative differences between uninfected and infected samples. When the O'Farrell technique of isoelectric focusing, followed by sodium dodecyl sulfate-slab gel polyacrylamide gel electrophoresis, was employed, additional polypeptides were resolved. Livers and L cells were labeled with [3H]-glucosamine. Infected livers incorporated less [3H]glucosamine in the membrane proteins than uninfected material, presumably indicating lower glycoprotein levels. Infected L-cell membranes incorporated greater amounts of [3H]glucosamine, and also were labeled to a greater extent than uninfected membranes, employing the [125I]lactoperoxidase technique. Uninfected L cells showed a greater agglutinability with concanavalin A than did infected cells. Infected livers had much greater levels of cyclic adenosine 3',5'-monophosphate. The data indicate changes in plasma membranes as a result of infection. Possible physiological consequences of membrane changes are discussed.


Phytohemagglutinin enhancement of dengue-2 virus replication in nonimmune rhesus monkey peripheral blood leukocytes. Phytohemagglutinin treatment of peripheral blood leukocytes from dengue nonimmune monkeys enhanced dengue-2 virus replication. Enhancement was due primarily to an increase in the number of infected cells. Destruction of mononuclear phagocytes with silica did not significantly inhibit virus replication in phytohemagglutinin-treated cultures. Pokeweed mitogen, concanavalin A, and streptolysin O stimulated increased deoxyribonucleic acid synthesis in monkey leukocytes but did not enhance virus replication. None of the mitogens significantly affected virus replication in cultures of dengue-immune monkey peripheral blood leukocytes.


Macrophages and age-dependent resistance to hepatitis induced by herpes simplex virus type 2 im mice. An age-dependent increase in the resistance of BALB/c mice to induction of focal necrotic hepatitis by herpes simplex virus type 2 was demonstrated. In 3-week-old mice inoculated intraperitoneally with virus, numerous necrotic foci developed in the liver. As the mice matured, the number of lesions declined until the age of 8 weeks, when no further increase in resistance appeared. Corresponding to this, the virus titers of livers and spleens of 3-week-old mice were higher than in 8-week-old animals throughout the infection, and the infection was apparently terminated in these organs of the adult mice by day 5. In vitro infection of peritoneal macrophages from 3-week-old and 8-week-old mice showed that this age-related resistance was concomitant with an increased restriction of virus replication in peritoneal macrophages from adult mice. Since, furthermore, the resistance of adult mice could be abolished by intravenous inoculation of the macrophage-toxic agent silica before infection, and since adoptive transfer of 2 X 10(6) syngeneic macrophages from adult mice to young ones conferred to the latter a resistance comparable to that of the adult mice, it is concluded that macrophage maturation is responsible for the age-dependent resistance seen in this infection.


Requirement for macrophages for interferon to be effective against encephalomyocarditis virus infection of mice. Suppression of macrophages in mice by treatments with silica or auro-thiomalate (Myocrisin) reduced production of serum interferon by polyriboinosinic acid:polyribocytidylic acid by 85 to 90%, indicating that this double-stranded polynucleotide caused interferon production primarily in macrophages. Suppression of macrophages in mice by silica or Myocrisin treatment did not significantly affect the susceptibility of mice to encephalomyocarditis virus, although at virus doses around 20 times the 50% lethal dose they died about 48 h earlier. Macrophage interferon protected mice from encephalomyocarditis virus infection at much lower doses than fibroblast interferon, and treatment of mice with silica or Myocrisin abolished the protection conferred by macrophage interferon, whereas these treatments had a much smaller effect on the protection afforded by fibroblast interferon. The requirement for macrophages for interferon to be effective in mice can explain why macrophage suppression can cause normally nonlethal viruses to kill adult mice.


Variable effect of encephalomyocarditis virus on host defense mechanisms. A number of viruses have been shown to suppress a variety of host defense mechanisms. To further define the effect of a viral infection on host resistance, a number of parameters were examined during the course of a lethal encephalomyocarditis virus infection of mice. The peripheral lymphocyte count, the induction of interferon in spleen cells by Newcastle disease virus, and the proportion of B cells in the spleen were all decreased or suppressed. In contrast, the responsiveness of spleen lymphocytes to phytohemagglutinin, the delayed-type skin hypersensitivity to dinitrofluorobenzene, and the proportion of T cells in the spleen remained normal. These results indicate that a viral infection may have divergent effects on different parameters of host resistance and emphasize the need to examine several functions before drawing conclusions about the effect of viral infections on host defense mechanisms.


Screening of human brain tumors for SV40-related T antigen. A series of 39 human brain tumors has been screened for the presence or absence of SV40-related T antigen by the direct and indirect immunoperoxidase methods. Two tumors of ependymal origin (malignant ependymoma, choroid plexus papilloma) revealed markedly positive nuclear staining for T antigen both in vivo and in vitro. The relationship of these tumors to their experimental counterparts inducible by recent human papovavirus isolates is discussed.


Type-C oncovirus isolate from human leukemic bone marrow: further in vitro and in vivo characterization. Rabbit corneal cells transformed by a putative human type-C helper virus pseudotype of the mouse sarcoma virus produce large amounts of transforming and non-transforming viruses. The virions are antigenically related to the woolly monkey (simian) sarcoma-leukemia type-C oncovirus. Typical sarcoma virus lesions developed in newborn rats injected with virus-producing rabbit cells. Cells producing only the putative type-C helper viruses as a result of exposure to a high dilution of transforming virus stock induce lymphosarcomas after inoculation into newborn rats.


Increased immunogenicity of low-antigenic rat tumors after superinfection with endogenous murine C-type virus in nude mice. Four chemical carcinogen-induced and two polyoma-virus-induced rat tumors were repeatedly passaged through nude mice. A methylcholanthrene-induced tumor in BDIX rats (MBDB) and a polyomavirus-induced tumor in Wistar/Fu rats (PW41) became infected with endogenous mouse virus (EMV), as judged by the expression of murine C-type virus-associated gp71, p30 and p12 antigens on their cell surface. Two ethylnitrosourea-induced tumors in BDIX rats (290T and GE3A) were exposed in vitro to the supernatant of EMV-infected PW41. Subsequently, 290T but not GE3A converted to murine gp71, p30 and p12 positivity. All these successfully infected rat tumors (EMV-MBDB, EMV-PW41 and EMV-290T) became less transplantable to and more rejectable in otherwise susceptible syngeneic rats. To compare the immunogenicity of the virus-infected and non-infected tumors, syngeneic rats were immunized three times with irradiated cells, and challenged with the non-infected tumor. Wistar/Fu rats immunized with irradiated EMV-PW41 showed no improvement in PW41 rejection, compared to rats immunized with irradiated, non-infected cells. On the other hand, BDIX rats immunized with EMV-MBDB or EMV-290T rejected MBDB or 290T, respectively, with no cross-immunity, while the rats immunized with irradiated but non-infected tumors showed no significant rejection. These results indicate that EMV infection augmented the immunogenicity of non-immunogenic or only low-immunogenic rat tumors.


Immunological study of two stocks of Moloney sarcoma virus producing regressor and progressor tumors in C57BL/6 mice. The immune response to progressor murine sarcoma virus (P-MuSV)2 and regressor murine sarcoma virus (R-MuSV) was analyzed by the radioisotopic footpad assay (FPA) for delayed hypersensitivity (DH) and by the membrane immunofluorescence assay for anti-tumor antibodies. In mice injected with P-MuSV, the footpad response remained at a low level for 7 weeks, while membrane immunofluorescence antibody titers reached their maximum levels after 5 weeks. By contrast, in mice injected with R-MuSV, the footpad response rose to a maximum by 4 weeks and membrane immunofluorescence antibody titers reached a maximum after 4 weeks. The adoptive footpad response reached its highest level in 2 weeks. A most striking difference between the P-MuSV and the R-MuSV systems was the lower immunogenicity of tumor cells induced by the P-MuSV as measured by the FPA. Considerable differences between the two stocks were also observed in oncogenic activity and tumor growth patterns. The P-MuSV contained one-tenth the amount of helper Moloney leukemia virus found in R-MuSV. Addition of more helper Moloney leukemia virus to the P-MuSV did not produce regressor tumors.


Induction of EBNA precedes the first cellular S-phase after EBV-infection of human lymphocytes. Using a combination of immunofluorescence and autoradiography, we studied the appearance of EBNA and DNA synthesis in cord-blood lymphocytes after infection with EBV derived from the B95-8 cell line. EBNA appeared between 12 and 25 h after addition of the virus. DNA synthesis was detected in EBNA-positive cells approximately 20 h after the appearance of EBNA. This shows that EBNA induction precedes the first cellular S-phase and suggests that the cells have not yet entered the cell-division cycle when EBNA appears. Little, if any, of the total DNA synthesis induced at this stage can be attributed to EBV-mediated immunologic stimulation.


Seroepidemiological studies of human papilloma virus (HPV-1) infections. At least two groups of human papilloma viruses can be distinguished serologically: on the one hand HPV 1-3, which are closely related but differ in the restriction enzyme pattern of their DNA, and on the other hand HPV-4. The age distributions of patients with warts induced by HPV 1-3 or by HPV-4, respectively, differ markedly. HPV 1-3 predominates between 5 and 15 years of age, whereas HPV-4 could be isolated more often between the ages of 20-25 years. The large number of HPV-1-3-induced warts in children is paralleled by a high percentage of HPV-1 antibody-positive sera in the same age group (about 50%). With increasing age the percentage of HPV-1 antibody-positive sera decline gradually. This pattern of seroreactivity was compared to that of patients with various papillomas and with several malignant tumors. There is no evidence to suggest a link between HPV 1-3 and condylomata acuminata, laryngeal papillomas or any of the malignant tumors tested.


Two E-rosette-forming lymphoid cell lines. Two E-rosette (spontaneous rosette with sheep red blood cells)-forming lymphoid cell lines were established. One (HPB-ALL) was derived from a young male Japanese patient with acute lymphoblastic leukemia (ALL), and the other (HPB-MLT) was from a 62-year-old female Japanese patient with a leukemic T-lymphoid malignancy. Formation of E rosettes, absence of any immunoglobulin determinants, absence of EBNA (Epstein-Barr virus associated nuclear antigen) and very limited stimulating ability in mixed lymphocyte culture, were characteristics mostly identical with those of so far established T-cell-derived lymphoid cell lines, MOLT, CCRF-CEM, CCRF-HS-B2, RPMI-8402 and JM. Only HPB-MLT, however, has been derived from an aged patient with T-lymphoid malignancy.


Immune response of the draining and distal lymph nodes during the progressive grwoth of a chemically-induced transplantable rat hepatoma. The immunological and histological changes occurring in the lymph node draining the site of a progressively growing intramuscular tumour (D192A) implant were monitored during a 4-week time course. Cell-mediated cytotoxicity against hepatoma-D192A and 15-day rat embryo cell targets, was detected with cells derived from the draining "lumbar" lymph node 4 days after tumour implantation and persisted up to the 2nd week of tumour growth, decreasing rapidly during the 3rd week. The observed lymph-node anergy demonstrated in cytotoxicity tests correlated with the histological findings, in that an initial marked paracortical (T-dependent) response also declined towards the end of the 3rd week of tumour growth. The B-dependent cortex showed active lymphocyte follicles in the 2nd week of the time course, and plasma-cell production continued until the experiment was terminated. These changes were shown to occur with the progressive increase in lymph-node mass. Serum antibody specific for the developing tumour was detected during the latter stages of tumour growth. The immunological and histological changes displayed were out of phase with those shown by the draining lymph node.


Relationship between Moloney MSV tumor resistance and endogenous virogene expression in AKR mouse strain and its hybrids. We have examined the induction of Moloney-MSV tumors in AKR and other mouse strains in relation to endogenous virus expression. All virus-free strains so far tested were tumor-susceptible, while AKR was resistant. The selectivity of MSV tumor resistance, characteristic of AKR mice, was associated with AKR virogens segregation in first backcross to MuLV-negative/MSV-susceptible mice, and in a few second backcross families. Stronger evidence that AKR ecotropic virogene expression is the major, but not the sole, determinant of M-MSV tumor resistance in AKR mice was obtained in strains partially congeneic for akv-2 viral gene and in recombinant inbred lines derived from an original cross between AKR and C57L mice. These mice or lines, in which inheritance of endogenous viral genes had occurred, at the same time inherited the ability to show strong resistance to tumor induction by an exogenous oncogenic virus. This finding suggests that ecotropic endogenous viruses can exert a beneficial effect in their hosts providing some protective functions. Although the association between AKR-MuLV and M-MSV resistance is definite, the mechanisms by which MuLV+ mice are refractory to M-MSV tumors have still to be elucidated.


Natural cell-mediated cytotoxicity in rats. III. Effects of immunopharmacologic treatments on natural reactivity and on reactivity augmented by polyinosinic-polycytidylic acid. Treatment of rats with high doses of hydrocortisone, X-irradiation, or cyclophosphamide had a suppressive effect on natural cytotoxicity in vivo. However, when rats were given poly I:C after any of these agents, the levels of NK activity were similar to those in normal rats which had been given poly I:C alone. To explain these findings, we have postulated that a population of pre-NK cells, resistant to hydrocortisone, cyclophosphamide and X-irradiation was induced by poly I:C to become cytotoxic NK cells. Treatment of rats with silica, in doses that had no effect on proliferative responses of host lymphocytes to Con A in vitro, sharply diminished NK activity. With this agent, the boosting effect of poly I:C, although still detectable, was diminished. Since there is little if any indication that NK cells are phagocytic, these data suggest that phagocytes may play a role in maintaining high levels of NK activity in vivo and, further, may be involved in the mechanism by which natural cytotoxicity is boosted by poly I:C. Adult thymectomized rats had easily detectable levels of natural reactivity and the response to poly I:C was unimpaired, indicating a lack of thymic dependence for boosting as well as for spontaneous levels of NK activity.


B-tropic oncornavirus production by BALB/c methylcholanthrene-induced sarcoma cells. Six oncogenic cell lines were established from primary sarcomas that had been induced in thymectomized, aged, BALB/cSt mice by the injection of 3-methylcholanthrene. Of these lines, three produced B-tropic murine leukemia virus (MuLV), i.e. MuLV that would patently infect fibroblasts of BALB/c origin but not those from NIH Swiss mice. One of these lines, M-138, produced large quantities of virus, permitting the isolation of milligram quantities of B-tropic MuLV. Neonatal BALB/c mice injected with the M-138 MuLV developed high plasma levels of the species-specific virion antigen p30, but during 9 months of observation failed to manifest clinical signs of leukemia. Plasma p30 levels of NIH Swiss mice (non-permissive host) that were inoculated also as neonates with the same virus preparation, remained undetectable by radioimmunoassay. Xenotropic MuLV, i.e. that which infects cells of species other than the species of origin, was released by one of the three remaining cell lines. Two of the six original lines have remained free of detectable oncornavirus production. Our results suggest that cell lines that are stable sources of large quantities of B-tropic MuLV may be easily obtainable by the described manipulations. Equally important, however, is the fact that these data re-emphasize the danger that is inherent in assuming that a cell line is virus-free simply because it was isolated from a chemical carcinogen-induced neoplasm


E.s.r. of free radicals in aqueous solutions of substituted pyrimidines. Reactions of OH radicals with methyl and ethyl derivatives of uracil, cytosine and thymine in aqueous solutions have been investigated. Photolysis of H2O2 was used to generate OH radicals and the radicals on the base derivatives were spin-trapped using t-nitrosobutane and identified with the help of e.s.r. spectroscopy. Addition of OH radicals was found to take place predominantly to the C(5)--C(6) double bond of the bases. H-abstraction from the methyl group occurred in the N(1) methyl derivatives of uracil, cytosine and thymine. Radicals formed by H-abstraction from the methyl group were also detected for 3-methyluracil, thymine, 1-methylthymine and 1-ethylthymine. Introduction of a methyl or ethyl group at the N(1) position of uracil, cytosine and thymine causes an increase in the C(6) proton coupling and a decrease in the N(1) splitting for radicals formed by OH addition at the C(5) position.


Expression of human prostatic acid phosphatase in a pancreatic islet cell carcinoma. The immunologic specificity of human prostatic acidphosphatase has been established by several previous investigations as well as in this study. An apparent exception to this specificity was observed--a case of pancreatic islet cell carcinoma metastasized to the liver produced acid phosphatase that was immunologically indistinguishable from the prostatic acid phosphatase. In this case, the possibility of prostatic involvement was convincingly ruled out by clinical follow-ups and by postmortem pathologic studies. Highly purified prostatic acid phosphatase and this tumor acid phosphatase exhibited very similar Km values and identical molecular weights. Immunochemical analysis of the two enzymes using antiprostatic acid phosphatase sera showed that enzymes are antigenically identical. The implications of our observation are discussed in relation to clinical application of immunoassays for prostatic phosphatase in the future and to the molecular basis of human acid phosphatase polymorphism.


Surface ATPase activity at cell-cell contacts in hepatic parenchymal cells and in cAMP-treated hepatoma cells in monolayer culture. Histochemical investigation shows that ATPase activity is located intensively on the surface of cell contacts in hepatoma cells grown in confluent monolayer culture. Dibutyryl cyclic AMP and theophylline-treated hepatoma cells which exhibit contact-inhibited growth show the absence of localization of intense ATPase activity at cell-cell contacts. However, after removal of these additives the activity fully recovers to the intense level of control cells, when the release of cells from contact inhibition of growth occurs. Cultured hepatic parenchymal cells in monolayer have little or no ATPase activity at their surface immediately after contacts are established, and again after growth to a confluent state. In a different type of hepatoma cell which is less malignant and forms a small tissue mass or island, cell surface ATPase activity at cell-cell contacts in an island is very weak or scarcely detected even when cells are not treated with dibutyryl cyclic AMP and theophylline.


Estimating Va/Q distributions from inert gas data with an enforced smoothing algorithm. The accuracy of a noniterative smoothing algorithm without a nonnegativity constraint for estimating continuous Va/Q distributions from inert gas retentions was evaluated. Simulated retention measurements computed from three assumed distributions were processed by the algorithm. Original and recovered distributions were compared with the root-mean-square (rms) difference. With noise-free retentions, the rms differences were minimum with zeta = 0 (no smoothing). With simulated 3% noise, the minima occurred when 0.01 less than zeta less than 0.10. Seven trials with independent simulated errors showed a large variation in the recovered distribution and the rms difference. All recovered distributions were qualitatively different from the originals especially at low and high Va/Q values. Negative fractional flows existed in all distributions that were not grossly different from the originals. We believe that this method which did not include a nonnegativity constraint, is inappropriate for processing inert gas retentions.


Autonomic blockade and coronary catecholamines and cyclic AMP in exercising man. To assess in man the effects of autonomic blockade on the response of catecholamines in the coronary circulation to dynamic exercise, arterial and coronary sinus catecholamine concentrations were measured in six patients during supine cycling exercise, following atropine 1.8 mg and oxprenolol 0.2 mg/kg iv. Although arterial concentrations did not increase significantly, coronary sinus catecholamine concentrations increased from 2.54 +/- 0.59 nmol/1 at rest 4.44 +/- 1.3 nmol/1 during exercise (P less than 0.05; one-tailed test) and were associated with a small increase in heart rate and coronary sinus cyclic AMP concentrations from 9.4 +/- 0.7 nmol/1 (rest) to 11.6 +/- 1.1 nmol/1 (exercise) (0.05 greater than P greater than 0.01). Although autonomic blockade may have increased catecholamine release, this was not reflected in an increased efflux of catecholamines from the heart, because similar increases in coronary sinus catecholamine concentrations occurred in the absence of autonomic blockade.


Adrenocortical function in response to myocardial necrosis in exercise-trained rats. Plasma corticosterone concentrations and in vitro adrenal secretion of corticosterone was determined in exercise-trained rats. Virgin, male rats, 100 days of age, were trained for an 11-wk period by treadmill running. Following the training program, rats were subjected to two subcutaneous injections of l-isoproterenol 24 h apart and killed 24 h after the second injection. All exercise-trained rats survived isoproterenol treatment, while 44% of the control rats died. Plasma corticosterone concentrations were elevated only in exercise-trained rats treated with isoproterenol. Control rats treated with isoproterenol had plasma corticosterone concentrations similar to that in control and exercise-treated rats given placebo injections. Exercise training reduced adrenocortical responsiveness to ACTH in vitro, but isoproterenol treatment increased in vitro responsiveness to ACTH in exercise-trained and control rats. Total unstimulated corticosterone secretion rates in vitro were similar. The reason for better rat survival in exercise-trained rats is unknown; however, improved energy metabolism, depressed aldosterone secretion, or both are suggested as reasons for the better survival of exercise-trained rats.


Thermally induced peripheral blood flow changes in chickens. Changes in wattle blood flow (Q) and vascular resistance (R) were examined during both local heating of the wattle and/or general body heating of unanesthetized, male White Leghorn chickens. Q increased and R decreased during both local and general heating. Termination of heating reversed the response. Beta stimulation with isoproterenol markedly reduced R and increased Q in normothermic birds, suggesting the presence of beta receptors in the wattle vasculature. Alpha blockade with phenoxybenzamine also resulted in pronounced vasodilatation, suggesting tonic alpha-sympathetic tone in the wattle vasculature under normothermic conditions. Neither cholinergic blockade with atropine sulfate nor beta-adrenergic blockade with propranolol altered the vascular response to general heating when administered near its peak. Release of alpha-sympathetic tone is believed to subserve the response to general heating.


Adaptation of the rat myocardium to endurance training. The purpose of this study was to assess cardiac adaptation to endurance training in rats. After 11 wk of progressive treadmill exercise (1 h/day), gastrocnemius cytochrome c oxidase activity was 38% higher (P less than 0.01) in the trained (n = 20) as compared to control (n = 20) rats. Cardiac Mg2+-stimulated myofibril ATPase activity (0.308 +/- 0.012 vs. 0.324 +/- 0.006 micrometer.mg-1.min-1) did not change nor was there any change in myofibril protein concentration (60.0 +/- 1.12 vs. 59.9 +/- 0.85 mg.g-1). The isolated left ventricular papillary muscle showed no significant change in time-to-peak tension (TPT) or half-relaxation time. Tension output, however, was significantly increased with training, 2.2 +/- 0.3 vs. 1.5 +/- 0.1 g.mm-2 (P less than 0.025). Furthermore, when the papillary preparations were perfused with 0.5 mM lanthanum (La3+) to displace membrane-bound Ca2+, the time course for tension decay was significantly prolonged in the trained muscles (P less than 0.001). We conclude that endurance running of this type does not necessarily increase myofibril ATPase activity or the time course of the isometric twitch of rat papillary muscle. However, tension output per unit area does increase and this appears to be due to a greater amount of Ca2+ being made available to the contractile apparatus.


Rotavirus as a cause of diarrhea in pigs. A rotavirus (reovirus-like agent) was associated with diarrheal diseases occurring in 1- to 4-week-old suckling pigs in 8 herds and in weaned pigs in 2 herds. Transmissible gastroenteritis virus was also detected in 2 of these herds, as was enteropathogenic Escherichia coli in 5 herds. Morbidity was generally greater than 80% in pigs of the affected age group within these herds, and mortality from diarrhea ranged from 7 to 20%. The disease due to rotavirus in suckling pigs appeared similar to the syndrome commonly referred to as milk scours, white scours, or 3-week scours. Diarrhea and villous atrophy, resembling that seen in transmissible gastroenteritis, occurred in naturally infected pigs and in gnotobiotic pigs experimentally infected with rotavirus. Diagnosis was accomplished by immune electron microscopy of intestinal contents and by immunofluorescent staining of enterocytes. A massive infection of enterocytes with rotavirus was demonstrated by immunofluorescence, which helps explain the pathogenesis of this disease. The apparent rarity of clinical rotaviral infections in suckling pigs greater than 7 days old is probably due to the acquisition of passive immunity from immune sows.


Infectious agents associated with fetal and early neonatal death and abortion in swine. Laboratory examination of specimens from 824 porcine abortions over a 6-year period produced 320 (38.8%) diagnoses. Viruses were involved in 22% and bacteria in 16.5% of all cases. Enterovirus was the virus isolated most often (10.9%), followed by parvovirus (4.9%), reovirus (4.4%), pseudorabies virus (1%), and adenovirus (0.8%). Leptospirosis was the bacterial infection most commonly diagnosed (9.8%), with numerous miscellaneous bacterial and fungal infections making up the remainder (7.0%) of the infectious causes.


Cyclic AMP regulation of the hexose phosphate transport system in Escherichia coli. Synthesis of the hexosephosphate transport system in Escherichia coli required the cyclic AMP-receptor protein regulatory complex. The apparent Km value for hexosephosphate activity was affected by the level of phosphate in the uptake environment.


Sequence of b cytochromes relative to ubiquinone in the electron transport chain of Escherichia coli. A ubiquinone-deficient mutant, carrying mutations in two genes affecting ubiquinone biosynthesis, has been used, in comparison with a normal strain, to determine the sequence of some of the components of the electron transport chain of Escherichia coli. The amounts of cytochromes reduced during aerobic steady-state conditions were estimated by comparing low-temperature difference spectra of normal or ubiquinone-deficient membranes with either D-lactate or reduced nicotinamide adenine dinucleotide as substrate. From the amounts of cytochromes reduced it was concluded that ubiquinone functions at two sites, one site  being between the dehydrogenases and cytochromes and the second site being after cytochromes b562 and b556 but before cytochromes b558, d, and o. The scheme proposed is discussed in relation to the Mitchell protonmotive ubiquinone cycle.


Mapping and characterization of the nad genes in Salmonella typhimurium LT-2. An ampicillin enrichment technique was used to isolate 39 nicotinic acid-requiring mutants of Salmonella typhimurium LT-2. Using interrupted-mating and transductional mapping procedures, three loci, designated nadA, nadB, and nadC, were identified. These loci mapped at 33, 82, and 6 min, respectively, on the S. typhimurium linkage map. The arrangement of the loci on the Salmonella linkage map corresponded closely to the nadA, nadB, and nadC loci on the Escherichia coli K-12 linkage map, indicating that the de novo pathway to nicotinamide adenine dinucleotide and the genes governing the enzymes involved in this pathway in S. typhimurium are very similar to those in E. coli. Evidence is also presented which indicates that the product of the nadC locus in S. typhimurium LT-2 is the enzyme quinolinic acid phosphoribosyltransferase. All nadC mutants of S. typhimurium secreted between 2 and 8 mumol of quinolinic acid per 100 ml of secretion medium. In addition, none of the nadC mutants isolated were able to grow in 10(-3) M quinolinic acid, whereas all nadA and nadB mutants of S. typhimurium grew well in the presence of quinolinic acid. Transductional crosses between nadB mutants provided evidence suggestive of more than one locus in the nadB region.


Biological properties of islets-activating protein (IAP) purified from the culture medium of Bordetella pertussis. The biological activities were studied of a new protein, islets-activating protein (IAP), purified from the culture medium of Bordetella pertussis. Rats injected intravenously with 1 microgram of purified IAP exhibited markedly enhanced insulin secretory responses to glucose, glucagon, epinephrine, and sulfonylureas over a period from 3 to 10 days after the injection. The degree and duration of the enhancement were proportional to the dose of IAP; the maximal effect induced by 1-2 microgram of IAP persisted for as long as 2 months. There was a highly significant correlation between the enhancement of insulin secretion and suppression of epinephrine hyperglycemia over a wide range of doses of IAP, indicating that suppression of epinephrine hyperglycemia resulted from hypoglycemic action of insulin secreted in response to epinephrine challenge. Additional actions of IAP were observed in mice; mice treated with higher doses of IAP showed symptoms were observed when lower doses of IAP were injected into mice. Thus, it is concluded that IAP is a protein primarily possessing a unique action to potentiate insulin secretory responses of experimental animals to nutritional and hormonal stimuli.