Pathological observations on experimental cytomegalovirus infections in pregnancy. Cytomegalovirus injected intramuscularly or intraperitoneally into mice on the 8th day of pregnancy resulted in a significant retardation in foetal growth and a reduced number of offspring surviving until near-term. Pathological changes consisted of inflammatory cell infiltration in the liver, heart, salivary glands and adrenal cortex of affected animals. Necrotic changes were also present in the adrenal cortex in most severely affected animals following intraperitoneal injection of virus. Intraperitoneal infection led to a frank peritonitis and marked splenic necrosis with a degeneration and inflammation of visceral fatty tissue and lymphadenitis. In contrast, intramuscular injection of virus did not produce a peritonitis but the spleen was greatly enlarged with reaction centres and increased numbers of lymphocytes being identified at histology. As a consequence of cytomegalovirus infection in pregnant mice foetal growth impairment and intrauterine death was seen. This was related to the severity of pathological changes in the mothers. In those animals dying or appearing sick at near-term, most foetuses were dead and resorbing. Since murine cytomegalovirus was not identified by inclusion body formation in foetal or placental tissues in this study and virus has not been isolated from these tissues previously, it was concluded that the mechanism for the impaired foetal growth was of an indirect nature and attributable primarily to a severe generalised maternal illness.


Secretory antibody directed against rotavirus in human milk--measurement by means of enzyme-linked immunosorbent assay. Human milk contains antibodies to a variety of enteropathic agents. We utilized the method of enzyme-linked immunosorbent assay to investigate anti-rotavirus secretory IgA in 113 human milk and colostral specimens from a rural area in Guatemala, 32 colostral specimens from an urban area of Costa Rica, and 12 from an urban area of the United States. Anti-rotavirus SCIgA was found in all colostral samples and in 94% of the milk specimans. Both the absolute concentration of anti-rotavirus SCIgA and concentration relative to total SCIgA were highest in colostrum, falling to lower but detectable levels from one week to two years after birth. No significant differences were noted in the results from the specimens from the three different geographic areas. The possible role of this antibody in immunity to rotavirus infections is discussed.


Immunologic studies on hamsters infected with Entamoeba histolytica. The serologic and cell-mediated immune responses of hamsters exposed to 2 strains of Entamoeba histolytica (HM-1 and HM-19) were evaluated by a series of in vitro tests. The pathogenicity of the 2 strains was evaluated in terms of their ability to produce liver abscesses and spleen enlargement. Antibody response was evaluated by the indirect hemagglutination test. The cellular immune response was assayed by increased DNA synthesis by lymphocytes and migration inhibition of macrophages.


Mesoionic xanthine analogs as inhibitors of cyclic AMP phosphodiesterase. Several derivatives of two mesoionic xanthine analogs, mesoionic thiazolo[3,2-alpha]pyrimidine-5,7-diones and mesoionic 1,3,4-thiadiazolo[3,2-alpha]pyrimidine-5,7-diones, were synthesized and evaluated as inhibitors of cyclic AMP phosphodiesterase. A significant number of these compounds demonstrated theophylline-like activity.


Formulation and evaluation of ethiodized oil emulsion for intravenous hepatography. A study was conducted to prepare and evaluate an ethiodized oil emulsion for intravenous administration that would selectively opacify the liver. Several formulations with differing globule size were prepared and compared for their in vivo activity (degree of liver opacification obtained) and stability. Results of studies conducted in rabbits and monkeys revealed that the oil globules that concentrate most in liver were 2.0--3.0 micrometer in diameter. The formulation of choice was stable for 6 months at refrigeration temperature (2--6 degree). In monkeys, this formulation produced an improved diagnostic image of the liver using computerized tomography at a dose of 0.2 ml/kg. The potential use of such emulsions in diagnostic radiology is briefly discussed.


Glutamate current noise: post-synaptic channel kinetics investigated under voltage clamp. 1. Analysis of voltage-clamped noise has been used to investigate the operation of glutamate receptors and associated channels at the locust nerve-muscle junction. Channels opened by glutamate and an agonist have been compared. 2. Glutamate-induced current fluctuations have a power spectrum with a single (1/frequency2) component which fits a simple model for the operation of channels. The form of the spectra for glutamate voltage noise and for 'background' noise has been determined. 3. The single channel conductance was estimated from the spectra, gamma glutamate = 122 +/- 0.4 (S.E.) pS. This estimate is independent of membrane potential and of the amplitude of membrane current change produced by glutamate. 4. The rate constant, alpha, for the closing of glutamate-operated channels depends exponentially on membrane potential, conforming to the equation alpha = approximately alphaeetaVm (approximately alpha = 0.26 +/- 0.014 msec-1, eta = 0.0054 +/- 0.001 msec-1); the duration of the channel lifetime (tau) decreases with hyperpolarization. Membrane potential dependence of alpha reduces as temperature is lowered. 5. For glutamate-operated channels, the temperature dependence of alpha and gamma fits the Arrhenius equation; alpha and gamma decrease exponentially as a function of T-1 (degrees K) with a descrete change in slope at about 6 degrees C, indicating a change in the activaiton energies of the respective rate processes. 6. Spectra of quisqualate-induced current fluctuations have the same form as spectra for glutamate noise. The single channel conductance was estimated from the spectra, gamma quisqualate = 120 +/- 3.9 (S.E.) pS. 7. The rate constnt, alpha, for the closing of quisqualate-induced channels depends exponentially on membrane potential. The duration of the open state for quisqualate channels was 2.2 times longer than for glutamate channels. 8. For glutamate receptors the voltage-sensitivity of the channel life-time is in the opposite direction to that of ACh receptors in vertebrate muscle. Possible explanations for the sharp change in the activation energy of the rate processes associated with the channel are discussed.


The tract of Lissauer and the dorsal root potential. 1. Intersegmental dorsal root potentials (d.r.p.s) have been recorded in the lumbar spinal cord of spinalized cats under Na pentobarbitone anaesthesia, to investigate the spinal cord structures involved in the intersegmental transmission of d.r.p.s. 2. A technique has been developed for restricted surgical isolation of Lissauer's tract between the segments with subsequent histological verification of the extent of the isolation. 3. Section of the ipsilateral dorsal column resulted in an increase in the latency of the intersegmental d.r.p. and a significant reduction in its amplitude. A further reduction in amplitude was achieved by section of the ipsilateral dorso-lateral funiculus. 4. Neither section of Lissauer's tract after the above lesions were performed nor restricted Lissauer's tractotomies abolished intersegmental d.r.p.s; only a small reduction in the amplitude of the d.r.p. was obtained. 5. It is proposed that intersegmental d.r.p.s are produced by the activaiton of a propriospinal system projecting through pathways other than Lissauer's tract and that primary afferent collaterals from the dorsal columns make a major contribtuion to their generation. The contribution made by Lissauer's tract is probably small.


Spontaneous multiquantal release at synapses in guinea-pig hypogastric ganglia: evidence that release can occur in bursts. 1. A study was made of some properties of the spontaneous synaptic potentials recorded in cells of the hypogastric ganglia of guinea-pigs. 2. The distribution of the amplitudes of the spontaneous synaptic potentials arising from a single preganglionic fibre was found to be multimodal, with peaks at roughly integral multiples of a unit peak. 3. It was found that the amplitudes of the larger spontaneous potentials were consistent with them being the result of synchronous or near-synchronous release of two or more unit-sized quanta (multiquantal release). 4. The proportion of multiquantal potentials observed was found to be dependent on the extracellular calcium ion concentration. 5. When the stochastic properties of the spontaneous potentials were examined, it was found that the spontaneous release process was not random and independent but appeared to be clustered. Indeed, the probability of occurrence of a unit spontaneous synaptic potential was greatly enhanced during the 40--60 msec immediately following any given spontaneous synaptic potential. 6. When unit spontaneous potentials were excluded from the analysis, the multiquantal potentials were still found to be clustered although the clustering was less marked than that seen in the over-all process. 7. These results suggested that the multiquantal spontaneous potentials arose from the release of unit quanta in short high frequency bursts and it was found that a mathematical model incorporating such a mechanism could describe the spontaneous quantal release process.


Dopamine receptors in the central thermoregulatory pathways of the rat. 1. Intrahypothalamic injection of either dopamine (10 microgram) or apomorphine (10 microgram) in a dose volume of 1 microliter. caused an almost immediate rise in tail skin temperature and a concomitant fall in core temperature in the conscious rat maintained at an ambient temperature of 17 +/- 1 degrees C. 2. The location of the dopamine-sensitive site was defined more accurately by reducing the dose volume to 0.5 microliter. and injecting dopamine at different points throughout the preoptic and anterior hypothalamic region. 3. The largest mean fall in core temperature (1.13 +/- 0.22 degrees C) was obtained after injection into the preoptic region. Injections with their perimeters more than 0.4 mm rostral or caudal to this site were ineffective. 4. Rats placed 0.65 m below a 250 W infra-red lamp responded to the imposed heat load by vasodilation of the tail skin blood vessels, indicated by an increased tail skin temperature. 5. Bilateral, but not unilateral, injection of either pimozide (0.5 microgram) or haloperidol (2.5 microgram) into the preoptic region significantly reduced the increase in tail skin temperature so that the rats were less able to withstand the imposed heat load. 6. Three serial sections (0.5 mm thick) were prepared from the preoptic anterior hypothalamic region of the rat brain, one anterior, one posterior and one corresponding to the dopamine-sensitive site. 7. Tissue from the middle slice increased its rate of synthesis of 3,5-cyclic AMP in response to addition of dopamine 20 or 100 micron to the incubation medium. The posterior slice was inactive, but the anterior slice had similar activity to the middle slice. 8. The effect of dopamine on the middle slice was specifically blocked by haloperidol (0.1 micron), whereas the effects on the anterior slice were partially blocked by both haloperidol (0.1 micron) and propranolol (0.1 micron). 9. These results indicate that there is within a well defined area of the preoptic region a population of dopamine receptors, which play a part in the transmission of information from warm sensors to heat loss effectors.


Propriospinal neurones as a relay system for transmission of cortico-spinal influences. 1. Synaptic organization and transmission have been studied in the lateral group of short propriospinal neurones of the lumbar and cervical regions of the cat spinal cord. Special attention was paid to their role in the transmission of cortico-spinal volleys. 2. The majority of these neurones are mono- or oligosynaptically excited after pyramidal tract stimulation. Convergence of excitatory actions from rubrospinal and lateral reticulospinal tracts was typical for these cells. Neurones with relatively low-level and delayed effects from segmental afferents are frequent in this population. 3. Temporal summation is important for the transmission of descending vlleys through these neurones. Mutual excitatory and recurrent inhibitory connections are supposed to play a substantial role in their function. 4. Possible participation of the short lateral propriospinal system in the transmission, transformation and re-distribution of corticofugal signals to the segmental spinal mechanisms is discussed.


The effect of pituitary-adrenal function in the modulation of pain sensitivity in the rat. 1. The relationship between adrenalcortical hormones, adrenocorticotrophic hormone (ACTH), and pain sensitivity was investigated in the rat. Pain sensitivity was assessed by measuring paw-lick and jump latencies in response to being placed on a grid at 55 degrees C.2. Bilateral adrenalectomy increased the sensitivity to pain, but adrenal demedullation had no effect.3. Pain sensitivity was inversely related to the circadian changes in circulating corticosterone and was greater at 7 a.m. than at 7 p.m. However, the same variation in pain sensitivity existed if the adrenals were removed, suggesting that the increase in pain sensitivity after adrenalectomy was not related directly to the levels of corticosteriods.4. The time course of the increase in pain sensitivity after adrenalectomy paralleled that of the changes in circulating ACTH. Adrenalectomy markedly increased pain sensitivity at 9 and 18 days following surgery when circulating ACTH levels were markedly elevated and corticosterone was absent, but not at 3 days following adrenalectomy when ACTH levels were lower and corticosterone was absent.5. Hypophysectomy decreased the sensitivity to pain.6. The results indicate that ACTH can alter pain sensitivity and that the effect of corticosteroids on the sensitivity to pain is an indirect one by virtue of their negative feed-back action on the hypothalamic-pituitary system.


Ultrastructural and cytochemical observations on sporogenesis of Myxobolus sp. (Myxosporida: Myxobolidae) from the common shiner Notropis cornutus. The structure and cytochemistry of spores of Myxobolus sp. from plasmodia which occur in the gill filaments of the common shiner Notropis cornutus were studied by light microscopy and by scanning and transmission electron microscopy. The thin-walled valves of the pyriform spores are thickened in the lateral sutural and apical regions. Mucous material is associated predominantly with the posterior end of many spores. The plasmodium is surrounded by a syncytial wall bounded by 2 membranes. Pinocytotic channels are formed by the inner membrane and numerous dense vesicles are pinched off at the distal ends of the channels. Sporogenesis is initiated by the envelopment of one vegetative cell by another. The larger, enveloped cell divides to form a disporous pansporoblast, which contains 2 pairs of capsulogenic and valvogenic cells and 2 binucleate sporoplasm cells. Each capsular primordium and connecting external tubule gives rise to a polar capsule which houses a helically coiled polar tubule. The apical end of each polar capsule is plugged by a stopper. The valvogenic cells surround the capsulogenic and posteriorly situated sporoplasm cells to form the spore valves. Iodinophilic (glycogen) inclusions were not seen in spores stained with iodine or Best's carmine. A darkly stained band was observed around the posterior region of most spores stained with Best's carmine. In the electron microscope large aggregates of beta glycogen particles were seen in the cytoplasm of sporoplasm cells in mature spores.


11,12-Secoprostaglandins. 5. 8-Acetyl- or 8-(1-hydroxyethyl)-12-hydroxy-13-aryloxytridecanoic acids and sulfonamide isosteres as inhibitors of platelet aggregation. The synthesis of a series of 8-acetyl- (or 1-hydroxyethyl-) 12-hydroxy-13-aryloxytridecanoic acids and their sulfonamide isosteres is described. These compounds are formally derived from members of earlier reported series of modified secoprostaglandins by replacing the omega-butyl chain termini by substituted aryloxy groups. A number of these compounds are potent inhibitors of collagen-induced blood platelet aggregation in guinea pigs on oral administration.


Synthesis, opiate receptor affinity, and conformational parameters of [4-tryptophan]enkephalin analogues. A series of analogues of the opioid peptide enkephalin with tryptophan substituted for phenylalanine in position 4 was synthesized by the solid-phase method. The [Trp4]enkephalin analogues and the corresponding [Phe4]enkephalin analogues displayed nearly parallel affinities in the opiate receptor binding assay throughout the series. In a conformational study fluorescence parameters were measured and intramolecular Tyr-Trp distances were estimated on the basis of resonance energy transfer experiments. No gross conformational differences were observed between analogues with widely differing opiate receptor affinity; however, small but significant changes in the intramolecular distance between the phenol ring and the indole moiety and/or in their relative orientation became apparent in some compounds. Identical intramolecular distances of 9.3 +/- 0.2 angstrom between the two aromatic rings were obtained with [Trp4,Met5]enkephalin, [Trp4,Leu5]enkephalin, and the N-terminal tetrapeptide comprised in the latter two analogues, indicating the existence of folded conformationas in 2 X 10(-5) M aqueous solution and demonstrating conformational analogy between these three peptides. The conformational parameters are discussed in relation to the observed affinities and the putative opiate receptor topography.


Synthesis and biological activities of some uronic acids, uronates, uronamides, and urononitriles of pyrimidine nucleosides. The 5'-hydroxymethylene function of several uracil and cytosine nucleosides has been modified to produce a variety of uronic acids, uronates, uronamides, and urononitriles of 2'-deoxy-beta-D-erythro-pentofuranosyl- and beta-D-arabino-pentofuranosylpyrimidines. In addition, the 5 position in many of these nucleosides has been substituted by a halogen atom. Twenty-one of the 35 compounds synthesized and examined for biological activity have not been previously reported. The purity of the products was measured by a high-pressure liquid chromatographic method. They were then evaluated as potential growth inhibitors of murine Sarcoma 180 cells in culture, of herpes simplex virus type 1 in vitro, and of Streptococcus faecium, a folic acid or deoxythymidine dependent bacterial strain. The ability of these nucleoside analogues to inhibit the phosphorylation of deoxythymidine by herpes simplex virus type 1 encoded pyrimidine deoxyribonucleoside kinase was also investigated and a structure-activity relationship examined.


Potential bis-alkylating agents for cancer chemotherapy. Approaches to the synthesis of 2-sulfonyl-1,4-bis(methanesulfonoxy)butanes. Three general methods for the synthesis of derivatives of the bis-alkylating agent, myleran [1,4-bis(methane-sulfonoxy)butane (1)], are presented. These derivatives contain a lipophilic group attached to the 2 position of 1,4-bis(methanesulfonoxy)butane by means of a sulfonyl function. As examples of the scope of the methods, the synthesis of seven such derivatives (12a-g) is presented. All seven were inactive against L1210 and P388 leukemia in the mouse.


Synthesis and some pharmacological properties of the 23-peptide 15-lysine-secretin-(5--27). Special role of the residue in position 15 in biological activity of the vasoactive intestinal polypeptide. The 23-peptide 15-lysine-secretin-(5-27) [S(5-27)] was synthesized on an insoluble support. The residue in position 15 of secretin, aspartic acid, was replaced by lysine, which occupies that position in the vasoactive intestinal polypeptide (VIP), a member of the secretin family. The resulting analogue showed increased VIP-like activity on smooth muscle preparations and unaltered secretin-like activity on pancreatic juice secretion in the rat. The affinity of the new analogue was high-affinity secretin receptors in acinar cells from guinea pig pancreas was less than that of S(5-27) but was higher than that of S(5-27) for high-affinity VIP receptors in the same cells.


Collagenase-sensitive peptidyl-nitrogen mustards as potential antitumor agents. Attempts to design an agent which would release cytotoxic nitrogen mustards within collagenase-producing tumors led to the synthesis of Cbz-L-Pro-L-Leu-Gly-L-Pro-Gly-NHC6H4N(CH2CH2Cl)2 (10). 10 was cleaved in vitro by bacterial and tumor-associated collagenase as expected at the peptide bond joining L-leucine and glycine to give Gly-L-Pro-Gly-NHC6H4N(CH2CH2Cl)2 which was over six times more toxic, on a molar basis, than 10. In vivo tests of 10 against well-advanced Sarcoma-180 gave disappointing results. The lack of specific antitumor activity may be accounted for by the presence of competing cleavage reactions by collagenases in certain normal tissues.


Inhibition of beef-brain and dog-heart (Na+ + k+) activated adenosine triphosphatase by carbon-3 branched cardenolides. Twenty-two C-3 branched cardenolides were investigated as inhibitors of beef-brain and dog-heart (Na+ + k+) activated adenosine triphosphatase. The synthetic compounds had lower inhibitory strength than digitoxigenin, and there was no indication of an improved safety index. Structure--activity relationships show that increased steric shielding of the 3-OH group results in reduced inhibition.


Photoaffinity labeling of the angiotensin II receptor. 1. Synthesis and biological activities of the labeling peptides. The synthesis and biological activities of analogues of the peptide hormone angiotensin II (AT) for use in photoaffinity labeling and receptor isolation are described. In the modified sequence of AT, Sar-Arg-Val-Tyr-Val-His-Pro-Phe, the aromatic residues Tyr and Phe have been either singly or simultaneously replaced by L-4'-nitrophenylalanine, L-4'-amino-3',5'-diiodophenylalanine, L-4'-aminophenylalanine, L-4'-diazoniumphenylalanine, and L-4'-azidophenylalanine. The peptides were assembled by solid-phase synthesis and the functional groups in position 4 and/or 8 chemically modified. Radioactivity was introduced by catalytic tritiation of the iodinated peptides to form the photolabeling precursors containing L-4'-amino-3',5'-diiodophenylalanine. On rabbit aorta the AT analogues substituted in position 4 showed poor affinities (0--15%), in position 8 high relative affinities (16--118%), and in position 4 and 8 additive effects of simultaneous substitutions. It is also shown that the new Boc derivative of L-4'-amino-3',5'-diiodophenylalanine can be used in peptide synthesis without side-chain protection.


Imidazo[1,2-a]-s-triazine nucleosides. Synthesis and antiviral activity of the N-bridgehead guanine, guanosine, and guanosine monophosphate analogues of imidazo[1,2-a]-s-triazine. The first chemical synthesis of 2-aminoimidazo[1,2-a]-s-triazin-4-one (8), the corresponding nucleoside and nucleotide, and certain related derivatives of a new class of purine analogues containing a bridgehead nitrogen atom is described. Condensation of 2-amino-4-chloro-6-hydroxy-s-triazine (2) with aminoacetaldehyde dimethyl acetal followed by the ring annulation gave the guanine analogue 8. A similar ring annulation of 4-(2,2-dimethoxyethylamino)-s-triazine-2,6-dione (5) gave imidazo[1,2-a]-s-triazine-4,6-dione (9). Direct glycosylation of the trimethylsilyl derivative of 8 with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose in the presence of stannic chloride, followed by debenzoylation, gave the guanosine analogue 2-amino-8-(beta-D-ribofuranosyl)imidazo[1,2-a]-s-triazin-4-one (12b), which on deamination gave the xanthosine analogue 13. Phosphorylation of 12b gave 2-amino-8-(beta-D-ribofuranosyl)imidazo[1,2-a]-s-triazin-4-one 5'-monophosphate (II). The anomeric configuration has been determined unequivocally by using NMR of the 2',3'-O-isopropylidene derivate 10 and the site of ribosylation has been established by using 13C NMR spectroscopy. These compounds were tested against type 1 herpes, type 13 rhino, and type 3 parainfluenza viruses in tissue culture. Moderate rhinovirus activity was observed for several compounds at nontoxic dosage levels.


Synthesis and prostaglandin-like activity of 2-(trans-3-hydroxy-1-octenyl)-3-indoleheptanoic acid. The synthesis of 2-(trans-3-hydroxy-1-octenyl)-3-indoleheptanoic acid (1) is described. The title compound appeared to show a weak prostaglandin-like activity in two different systems. It contracted rat stomach fundus strips and guinea-pig ileum preparations only at concentrations about 10(3)- and 10(2)-fold higher, respectively, than PGE1. Moreover, it stimulated adenylate cyclase from rat liver plasma membrane, but the relative potency was 4--5 X 10(2)-fold lower than the natural compound. The title compound showed also a certain degree of PGE1 antagonism.


Jejunal microbial flora of southern indian infants in health and with acute gastroenteritis. The microbial flora of the jejunal lumen of 28 infants with acute gastroenteritis was compared with that of a group of 10 normal infants. The jejunum of control subjects harboured an "oral" type of flora and in a few instances enterobacteria in small numbers. The concentrations of all but one of the groups of organism were higher in the patients than in controls, and the differences were of statistical significance for enterobacteria and lactobacilli. In eight subjects, the same pathogen was identified in the jejunum and the stool. In six subjects with rotavirus infection, there were almost no Gram-negative aerobic rods in the jejunum. The possible role of other Gram-negative aerobic rods in producing gastroenteritis is discussed. It is suggested that studies of jejunal flora are of considerable importance in assigning an aetiological role to bacteria in the causation of acute gastroenteritis.


Conditions leading to the establishment of the N (a gene dependent) and A (a gene independent) transformed states after polyoma virus infection of rat fibroblasts. Infection of normal rat fibroblasts (FR 3T3) with the early tsa mutant of polyoma virus may lead to either the A or the N phenotype, tsa-A transformants, originally derived by agar selection, are not temperature dependent for maintenance of the transformed phenotype, whereas tsa-N transormants revert at high temperature to normal growth control. A transformants did not result from an independent cellular mutation selected in agar medium, but rather from a transformation process distinct from that leading to the N state. It occurred in both liquid and agar media when the infected cells were maintained under growth-restricting conditions, such as absence of anchorage and contact inhibition at confluency. N transformation occurred in cells maintained in active growth after virus infection (sparse cultures on a solid substratum). Physiological conditions during a critical period after virus infection thus appear to be a crucial parameter of the transformation process.


Detergent-inhibited, heat-labile nucleoside triphosphatase in cores of avian myeloblastosis virus. Endogenous DNA synthesis was studied in isolated core particles of avian myeloblastosis virus. It was found that cores contained an enzymatic activity which rapidly converted the added nucleoside triphosphates to diphosphates (but not further) at 0 degrees C, thus inhibiting DNA synthesis. This triphosphatase probably originates from the viral membranes. In the cores the enzyme is completely inactivated by low concentrations (0.02%) of Nonident P-40. Also, the enzyme is very thermolabile and denatures rapidly at 38 degrees C.


Differential sensitivity of normal and transformed human cells to reovirus infection. Normal and simian virus 40-transformed WI-38 cells exhibited a differential sensitivity to infection with type 3 reovirus. A progressive decrease in viability began 24 to 36 h after infection of transformed cells terminating in complete lysis of cultures by 96 h. Normal cells were productively infected and continued to produce and release virus for as long as 14 days after infection, but exhibited no detectable cytopathology. Inhibition of cellular DNA synthesis began 15 to 18 h after infection in transformed cells before development of cytopathology. No inhibition of DNA synthesis was detected in infected normal cells. No significant differences were noted in the adsorption or early replication characteristics of reovirus in normal and transformed cells. Virus replication and host cell DNA synthesis in normal and transformed human cells were compared to reovirus-infected L-929 mouse fibroblast cells.


Autoregulation of adenovirus type 5 early gene expression II. Effect of temperature-sensitive early mutations on virus RNA accumulation. The kinetics of accumulation of early virus RNA in the cytoplasm of KB cells infected at 40.5 degrees C by wild-type (WT) adenovirus type 5 and a temperature-sensitive "early" mutant, H5ts125 (ts125), were compared by hybridization of unlabeled RNA in solution to the (3)H-labeled l strand of Ad5 DNA HindIII restriction endonuclease fragment A. In the presence of 1-beta-d-arabinofuranosylcytosine, A(l) RNA accumulated in WT-infected cells for 9 h and then decreased in concentration to 6% of the 9-h concentration by 18 h. In ts125-infected cells, A(l) RNA accumulated for 12 h and then remained at the same concentration for at least 6 h thereafter. The concentrations of virus RNA from the four early transcription regions of the genome were measured at 15 h in cells infected at 40.5 degrees C in the presence of 1-beta-d-arabinofuranosylcytosine by: (i) ts125 and WT; (ii) two other ts early mutants, ts107 and ts149; and (iii) a revertant of ts125. The revertant and ts149, a mutant from a different complementation group than ts125, both accumulated all early virus cytoplasmic RNA species in amounts similar to, or less than, WT. However, both ts125 and ts107, independently isolated mutations in the 72,000-molecular-weight (72K) DNA-binding protein gene, accumulated cytoplasmic early RNA in excess of that found in WT infection. This pattern of RNA accumulation with the mutants and WT virus was the same in the nuclei as in the cytoplasm at 40.5 degrees C. At 32 degrees C, however, the abundance of nuclear virus RNA from all four early regions was the same in cells infected by either ts125 or WT. Differences in the relative abundance of nuclear RNA from the four early regions were observed in cells infected at 40.5 and 32 degrees C, but were not dependent upon the infecting virus genotype. These results are consistent with autoregulation of early gene expression by the 72K protein and support the hypothesis that the 72K protein either decreases the rate of early virus transcription or increases the rate of virus RNA degradation in the nucleus.


Does simian virus 40 DNA integrate into cellular DNA during productive infection? Late after infection of permissive monkey cells by simian virus 40 (SV40), large amounts of SV40 DNA (30,000 to 220,000 viral genome equivalents per cell) can be isolated with the high-molecular-weight fraction of cellular DNA. Hirai and Defendi (J. Virol.9:705-707, 1972) and Hölzel and Sokol (J. Mol. Biol. 84:423-444, 1974) suggested that this SV40 DNA is covalently integrated into the cellular DNA. However, our data indicate that the high-molecular-weight viral DNA is  composed of tandem, "head-to-tail" repeats of SV40 DNA and that very little, if any, of this viral DNA is covalently joined to the cellular DNA. This was deduced from the following experimental findings. The size of the SV40 DNA associated with the high-molecular-weight cellular DNA fraction is greater than 45 kilobases, based on its electrophoretic mobility in agarose gels. In this form the SV40 DNA did not produce heteroduplex structures with a marker viral DNA (an SV40 genome with a characteristic deletion and duplication). After the high-molecular-weight DNA was digested with EcoRI or HpaII endonucleases, enzymes which cleave SV40 DNA once, more than 95% of the SV40 DNA migrated as unit-length linear molecules and, after hybridization with the marker viral DNA, the expected heteroduplex structures were easily detected. Digestion of the high-molecular-weight DNA fraction with restriction endonucleases that cleave cellular, but not SV40. DNA did not alter the electrophoretic mobility of the polymeric SV40 DNA, nor did it give rise to molecules that form heteroduplex structures with the marker viral DNA. Polymeric SV40 DNA molecules produced after coinfection by two physically distinguishable SV40 genomes contain only a single type of genome, suggesting that they arise by replication rather than by recombination. The polymeric form of SV40 DNA is highly infectious for CV-1P monolayers (6.5 X 10(4) PFU per  microgram of SV40 DNA), yielding virtually exclusively normal, covalently closed circular, monomer-length DNA. Quite clearly these cells have an efficient mechanism for generating monomeric viral DNA from the SV40 DNA polymers.


Recombinants between herpes simplex virus types 1 and 2: analyses of genome structures and expression of immediate early polypeptides. Recombinants between temperature-sensitive mutants of herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) were constructed. Using restriction endonucleases, we analyzed the genome composition of 17 intertypic recombinants and detected crossovers in every region of the genome. The virion DNA of one recombinant appeared to be largely "frozen" in two of the four possible genome arrangements of HSV. Knowledge of the genome structures of recombinants enabled us to physically map immediate early polypeptides. We present evidence that the immediate early polypeptide Vmw IE 110 of HSV-1 and its functionally equivalent polypeptide, Vmw IE 118, of HSV-2 may map in the repetitive sequences bounding the long unique region of HSV.


DNA of Epstein-Barr virus. IV. Linkage map of restriction enzyme fragments of the B95-8 and W91 strains of Epstein-Barr Virus. The arrangement of EcoRI, Hsu I, and Sal I restriction enzyme sites in the DNA of the B95-8 and W91 isolates of Epstein-Barr virus (EBV) has been determined from the size of the single-enzyme-cleaved fragments and from blot hybridizations that identify which fragments cut from the DNA with one enzyme contain nucleotide sequences in common with fragments cut from the DNA with a second enzyme. The DNA of the B95-8 isolate was the prototype for this study. The data indicate that  (i) approximately 95 X 10(6) to 100 X 10(6) daltons of EBV (B95-8) DNA is in a consistent and unique sequence arrangement. (ii) Both termini are variable in length. One end of the molecule after Hsu I endonuclease cleavage consists of approximately 3,000 base pairs, with as many as 10 additional 500-base pair segments. The opposite end of the molecule after Sal I endonuclease cleavage consists of approximately 1,500 base pairs, with as many as 10 additional 500-base pair segments. (iii) The opposite ends of the molecule contain homologous sequences. The high degree of homology between the opposite ends of the molecule and the similarity in size of the "additional" 500-base pair segments suggests that there are identical repeating units at both ends of the DNA. The arrangement of restriction endonuclease fragments of the DNA of the W91 isolate of EBV is similar to that of the B95-8 isolate and differs from the latter in the presence of approximately 7 X 10(6) daltons of "extra" DNA at a single site. Thus, the size of almost all EcoRI, Hsu I, and Sal I fragments of EBV (W91) DNA is identical to that of fragments of EBV (B95-8) DNA. A single EcoRI fragment, C, of EBV (W91) DNA is approximately 7 X 10(6) daltons larger than the corresponding EcoRI fragment of EBV (B95-8) DNA. Digestion of EBV (W91) DNA with Hsu I or Sal I restriction endonucleases produces two fragments (Hsu I D1 and D2 or Sal I G2 and G3) which differ in total size by approximately 7 X 10(6) daltons from the fragments of EBV (B95-8) DNA. Furthermore, the EcoRI, Hsu I, and Sal I fragments of EBV (W91) and (B95-8) DNAs, which are of similar molecular weight, have homologous nucleotide sequences. Moreover, the W91 fragments contain only sequences from a single region of the B95-8 genome. Two lines of evidence indicate that the "extra" sequences present in W91 EcoRI fragment C are viral DNA and not cellular. (i) The molecular weight of the "enlarged" EcoRI C fragment of EBV (W91) DNA is identical to that of the EcoRI C fragment of another isolate of EBV (Jijoye), (ii) The HR-1 clone of Jijoye has previously been shown to contain DNA which is not present in the B95-8 strain but is present in the EcoRI C and Hsu I D2 and D1 fragments of EBV (W91) DNA (N. Raab-Traub, R. Pritchett, and E. Kieff, J. Virol. 27:388-398, 1978).


Absence of circularly permuted and largely redundant sequences in the genome of visna virus. A previous study of the infectivity of visna virus proviral DNA suggested that the genetic information of the virus is distributed over at least two of the RNA subunits. Because the genetic complexity of visna virus corresponds to the size of one subunit, this result may imply that sequence redundancies exist within each subunit. In the present article we have examined this question by constructing a map of the large RNase T1-resistant oligonucleotides of the viral genome. Our principal results are as follows: (i) all 36S RNA subunits have the same genetic content regardless of their polyadenylic acid [poly(A)] content; (ii) the poly(A) tract is present at the 3' end of the molecule; and (iii) the recoveries of 19 large RNase T1-resistant oligonucleotides from poly(A)-tagged RNA fragments of various sizes demonstrate that the oligonucleotides are organized in the same linear order within all subunits. Our results, therefore, exclude the existence of large sequence redundancies in the genome of visna virus.


Characterization of Rous sarcoma virus src gene products synthesized in vitro. The cell-free synthesis of three major proteins from virion RNA of nondefective Rous sarcoma virus (RSV), but not from RNA of transformation-defective deletion mutants, has been observed. The apparent molecular weights of these transformation-specific proteins are approximately 60,000 (60K), 25K, and 17K. Tryptic maps of methionine-containing peptides revealed the 17K, 25K, and 60K proteins to be overlapping in sequence. However, only partial homology was observed between the 17K, 25K and 60K proteins synthesized from Schmidt-Ruppin strain, subgroup D, RSV RNA and those synthesized from Prague strain, subgroup B, RSV, RNA. About half of the methionine peptides in the Schmidt-Ruppin strain, subgroup D, 60K protein were shared with the Prague strain, subgroup D, 60K protein, and the rest were distinct to each. The virion RNAs coding for the 60K, 25K, and 17K proteins were found to be polyadenylated and to sediment with maximal mRNA activity at about 23, 19 to 20, and 18S, respectively. In addition, transformation-specific proteins with molecular weights of 39K and 33K were observed by in vitro synthesis. These proteins are also related to the 60K, 25K, and 17K proteins and were synthesized from polyadenylated RSV RNA of approximately 21 to 22S. RNase T1-resistant oligonucleotides were analyzed in parallel, and the src-specific oligonucleotides were found to be first present in equimolar amounts in those gradient fractions sedimenting at 21 to 22S. Our data suggest that synthesis of the 60K protein is initiated near the 5' terminus of the src gene, whereas the 39K, 33K, 25K, and 17K proteins are initiated internally in the src gene. All of these proteins appear to be initiated independently, but they may have a common termination site.


Ends of the RNA within Sendai virus defective interfering nucleocapsids are not free. Sendai virus defective interfering nucleocapsids, isolated from infected cell cytoplasm by equilibrium banding in CsCl gradients, contain only the viral N protein. Neither end of the genomic RNA within these nucleocapsids is accessible to RNase digestion.


Selective degradation of newly synthesized nonmessenger simian virus 40 transcripts. By pretreating simian virus 40-infected BSC-1 cells with glucosamine, [(3)H]uridine labeling of both cellular and viral RNA can be halted instantaneously by addition of cold uridine. We have studied the fate of pulse-labeled viral RNA from cells at 45 h postinfection under these conditions. During a 5-min period of labeling, both the messenger and nonmessenger regions of the late strand were transcribed. After various chase periods, nuclear viral species which sediment at 19, 17.5, and 16S were observed. Nuclear viral RNA decays in a multiphasic manner. Of the material present at the beginning of the chase period, 50% was degraded rapidly with a half-life of 8 min (initial processing). This rapidly degraded material was that fraction of the late strand which did not give rise to stable late mRNA species. Forty percent was transported to the cytoplasm, and 10% remained in the nucleus as material which sedimented in the 2 to 4S region. These 2 to 4S viral RNAs had a half-life of 3 h, and hybridization studies suggest that they are in part coded for by the late-strand nonmessenger region and are derived from the initial nuclear processing step. Another part is coded for by the late-strand messenger region and may be generated by some subsequent nuclear cleavages of 19S RNA into 17.5 and 16S RNAs. Transport of nuclear viral RNA into the cytoplasm was detected after a 5-min pulse and a 7-min chase. The maximum amount of labeled viral RNA was accumulated in the cytoplasm after a 30-min to 1-h chase. At least two viral cytoplasmic species were observed. Kinetic data suggest that 19S RNA is transported directly from the nucleus. Whether cytoplasmic 16S is formed by cleavage of 19S RNA in the cytoplasm is not clear. The half-lives of cytoplasmic 19 and 16S RNAs can be approximated as 2 and 5 h, respectively.


Identification of nucleotide sequences which may encode the oncogenic capacity of avian retrovirus MC29. The retrovirus strain MC29 induces a variety of tumors in chickens, including myelocytomatosis and carcinomas of the kidney and liver. In addition, the virus can transform cultures of embryonic avian macrophages and fibroblasts. We have characterized the genome of MC29 virus and have identified nucleotide sequences that may encode the oncogenic potential ofthe virus. MC29 virus can replicate only with the assistance of a related helper virus. The defect in replication is apparently a consequence of a deletion in one or more viral genes: the haploid genome of the MC29 virus has a molecular weight of ca. 1.7 X 10(6), whereas the genome of the helper virus MCAV has a molecular weight of ca. 3.1 X 10(6). Although MC29 virus transforms fibroblasts in culture, its genome has no detectable homology with the gene src that is responsible for transformation of fibroblasts by avian sarcoma viruses. We prepared radioactive single-stranded DNA complementary to nucleotide sequences present in the genome of MC29 virus but not in the genome of MCAV (cDNA(MC29)). If they are contiguous, these sequences (ca. 1,500 nucleotides) are sufficiently complex to encode at least one protein. Homologous sequences were not detectable in several strains of avian sarcoma viruses or in an endogenous virus of chickens. Our findings confirm and extend recent reports from other laboratories and lead to the conclusion that MC29 virus may contain a previously unidentified gene(s) that is capable of transforming several distinct target cells. The evolutionary origins of this putative gene and its location on the viral genome can be explored with cDNA(MC29).


Modulation of mouse mammary tumor virus production in the MJY-alpha cell line. Implantation of the mouse mammary tumor virus (MMTV)-producing mammary tumor cell line MJY-alpha into isogeneic mice elicited both humoral and T-cell responses against MMTV virion antigens. The carcinosarcomas which developed from the implanted cells showed a significant decrease in MMTV synthesis, compared with cells remaining in culture, which was detectable as early as 7 days after implantation and for five transplant generations. Electron microscopic examination of thin sections of the tumors revealed that intracytoplasmic A particles, budding particles, and cell-free MMTV B particles were all affected. However, immunofluorescence assays of tumor sections demonstrated the presence of MMTV viral antigens in the cells. Cell cultures initiated from first-, third-, and fourth-generation tumors were morphologically identical to the original in vitro cell line, although virus production was barely detectable. Analysis of the cultures by electron microscopy revealed a significant increase in MMTV virions after in vitro passage 3. Polypeptide profiles obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of virions purified from these cultures were identical to MMTV. Immunodiffusion demonstrated the cross-reactivity between these virions and MMTV particles obtained from mouse milk. In vitro treatment of MJY-alpha cell cultures with rabbit anti-MMTV antiserum resulted in a reduction of extracellular MMTV virions, as well as alterations in their sodium dodecyl sulfate-polyacrylamide gel electrophoretic polypeptide patterns.


Physical mapping of herpes simplex virus-induced polypeptides. Analysis of the polypeptides induced by 29 herpes simplex virus type 1/type 2 intertypic recombinants and correlation of the data with the crossover points in the recombinant DNAs have enabled the map positions of many polypeptides to be deduced. These include 25 polypeptides which label with [35S]methionine, 11 which label with [32P]orthophosphate, and 4 which label with [14C]glucosamine. Together with the data of Preston et al. (J. Virol., in press) on the mapping of five immediate-early polypeptides, the results show that representatives of four groups of proteins--immediate-early, late, phosphorylated, and glycosylated--map in both long and short regions. The functional organization of the herpes simplex virus genome does not therefore restrict any of these four groups to either the long or the short region.


Replication-defective ecotropic murine leukemia viruses: Detection and quantitation of infectivity using helper-dependent XC plaque formation. Clones 8A and NP-N, which appear to be infected with replication-defective variants of murine leukemia virus, produce particles which do not form plques in the XC test. These particles formed XC plaques when amphotropic murine leukemia virus, which is XC negative, was added to the assay plates. This phenomenon can be used as a quantitiative infectivity assay for these replication-defective murine leukemia viruses.


Chromatin pattern consisting of repeating bipartite structures in WI-38 cells infected with human cytomegalovirus. A novel chromatin structural pattern displaying bipartite and oblate ellipsoid structures orderly arranged along a fiber axis has been observed in WI-38 cells infected with human cytomegalovirus. This chromatin type coexists with chromatin fibers showing conventional nucleosomes. Each bipartite-oblate structure is 40 nm in length, about four times as long as an ordinary nucleosome, and the number of these structures per micrometer (11/micrometer) is clearly less than that of typical cellular nucleosomes (32/micrometer).


Translation of type C viral RNAs in Xenopus laevis oocytes: evidence that the 120,000-molecular-weight polyprotein expressed in Abelson leukemia virus-transformed cells is virus coded. The genomic RNA of Abelson leukemia virus (AbLV) has been purified and translated in Xenopus laevis oocytes. The primary AbLV-specific protein synthesized is a polyprotein corresponding in molecular weight and immunological properties to a previously described p15 and p12 containing 110,000- to 130,000-molecular-weight polyprotein expressed in AbLV-transformed cells. In contrast, translation of woolly monkey sarcoma virus genomic RNA resulted in symthesis of a 55,000-molecular-weight polyprotein consisting of woolly helper virus p30, p15, and p12. These findings demonstrate the value of the X. laevis oocyte in vitro system for studies of translational products of replication-defective transforming viruses and establish the virus-coded nature of the nonstructural component of the 110,000- to 130,000-molecular-weight polyprotein expressed in AbLV-transformed cells.


Parainfluenza type 3 in a transplant unit. We observed an epidemic of parainfluenza type 3 viral infection in a renal transplantation service among 16 recent transplant recipients. To assess the effect of infection on transplant function, we retrospectively compared the infected patients with a matched, uninfected control group. In addition to symptoms of a mild upper respiratory tract infection associated with the epidemic, there was an increase in the frequency of acute rejection episodes during the period of infection in the infected group. Nevertheless, outcome in terms of patient and graft survival at six months was not affected when compared with the survival rates of the control group.


Herpes simplex virus type I from a patient with radiculoneuropathy. Herpes simplex virus type I was isolated from the CSF of a patient with atypical lumbosacral pain. The features of this case are unusual and important in light of the current understanding of herpes-simplex-virus-associated neurologic disease.


Effects of cadmium ion on the Na,K-ATPase of microsomes obtained from frog skin. We have developed a new simple procedure for separating the epidermis from the dermis of the bullfrog (Rana catesbeiana) (freezing and scraping off the skin), and extracting microsomal fraction from the epidermis. The effects of Cd on the ATPase activity of the microsomal fraction was investigated by varying Na and/or K concentrations in the reaction medium. The higher the concentration of Cd, the larger the decreases were found in both activities of ouabain-sensitive ATPase and ouabain-insensitive ATPase in the range of Cd concentration from 10(-7) M to 10(-3) M. The activity of ouabain-sensitive ATPase (Na, K-ATPase) was completely inhibited by 1 mM Cd. The amount of Cd uptake into the whole skin was about 150 micrometer/kg wet weight. This concentration of Cd was sufficient to inhibit the Na, K-ATPase activity of the microsomes. The mechanisms underlying the three types of Cd effects on the frog skin were discussed in relation to the Na, K-ATPase activity.


Differential effects of D 600 on release of catecholamines by acetylcholine, histamine, tyramine and by cyclic AMP from canine adrenal medulla. The isolated canine adrenal glands were perfused retrogradely with Locke's solution, and the catecholamine contents of the effluents were measured by the trihydroxyindole-fluorimetric method. Stimulation of the glands by acetylcholine, histamine, tyramine and cyclic AMP caused an increase in release of catecholamines from the glands. Introduction of D 600 of the perfusion medium reduced release of catecholamines in response to acetylcholine, and this reduction was overcome by raising calcium ion concentrations of the perfusion medium. Similarly, D 600 reduced release of catecholamines in response to histamine. The release of catecholamines evoked by tyramine was also inhibited by D 600, although to a lesser degree than the release by acetylcholine. In contrast, D 600 was entirely ineffective on the catecholamine release in response to cyclic AMP. D 600 had no effect on the spontaneous catecholamine output. From these results it was concluded that release of catecholamines from adrenal chromaffin cells by acetylcholine and histamine, and by tyramine in part requires the entry of calcium ions across the cell membane, whereas that by cyclic AMP does not.


Enhanced plating efficiency of herpesvirus DNA on herpes simplex virus type 1 DNA-transformed cells. Partial resistance of hamster cells transformed by sheared herpes simplex virus (HSV) DNA to superinfection by intact HSV could be overcome by transfection with viral DNA's. HSV DNA plated more efficiently on HSV type 1 DNA-transformed hamster cells than on spontaneously transformed hamster or rabbit cells or on hamster cells transformed by other DNA viruses.


Congenital transmission of murine leukemia virus from wild mice prone to the development of lymphoma and paralysis. Maternal congenital transmission of infectious murine leukemia virus, primarily via milk, was the major route of virus spread in Lake Casitas (LC) wild mice and in crosses of LC mice with uninfected wild and laboratory mice. An indirect extrachromosomal male transmission in utero of LC virus also readily occurred in matings of viremic LC males with C57L females but apparently not with other uninfected wild or NIH Swiss females. Both amphotropic and ecotropic classes of LC murine leukemia viruses were potentially transmissible by congenital and venereal epigenetic means and could induce the same two diseases, lymphoma and paralysis, that occurred naturally in LC wild mice. Lymphoma and paralysis both failed to occur in uninfected LC mice or their hybrid progeny that escaped congenital infection.


A rapid neutralization test for antibodies to bovine leukemia virus, with the use of rhabdovirus pseudotypes. Two rhabdoviruses, vesicular stomatitis (type Indiana) and Chandipura viruses, formed pseudotype particles with envelope antigens provided by bovine leukemia virus (BLV). The pseudotypes are infectious for calf, human, mink, and rat cells, but the most sensitive indicator proved to be the Vero cells. Infectivity of the pseudotypes was increased by DEAE-dextran present during adsorption. Sera of spontaneously infected cattle contained high titers (some over 1/10,000) of antibodies neutralizing the pseudotypes, whereas sera of cattle from uninfected herds possessed no neutralizing activity in 1/10 dilution. The neutralization of these pseudotypes can serve as a rapid and sensitive test for the detection of antibodies in the cattle infected with BLV.


Venereal herpes: a nursing overview.

Venereal herpes: a nursing overview. Nursing problems associated with venereal herpes are examined. Clinical manifestations, as well as specific effects on the pregnant woman, fetus, and neonate, are discussed. Additional discussion focuses on prevention and therapy and the relation of venereal herpes to cervical cancer. A list of nursing implications is appended; it includes assessment and intervention, patient teaching, nursing education, and public policy.


[Cyclic nucleotides in the blood plasma in arterial hypertension]. The reaction of cyclic nucleotides in blood to an orthostatic position, furosemid administration, and submaximal bicycle ergometry load was studied in 20 healthy individuals and in 50 patients with hypertensive disease. A special group was composed of 25 patients the disease in whom was marked by crises. It is shown that a walk of one hour and intravenous infusion of 40 mg of furosemid caused an increase in the blood cAMP but did not change the level of cGMP. During a bicycle ergometry load the levels of cGMP and cGMP increased to an equal measure and returned to their initial values 30 min after its cessation. In patients with arterial hypertension the cGMP system becomes most functionally mobile, whereas the cAMP level, if it increases, does so at a later term after the effect of the stimulus begins. The increased cGMP level is maintained for quite a lengthy period of time after the load is discontinued. In patients suffering from hypertensive disease with crises the cAMP/cGMP ratio, which is reduced at rest also, diminishes still more in an orthostatic position and particularly during a crisis. The role of changes in the metabolism of cyclic nucleotides and their sensitivity to regulatory factors in the pathogenesis of arterial hypertension is discussed.


[The judgement of collagen metabolism by hydroxyproline in plasma and urine and plasma collagenic peptidase under different diet (author's transl)]. In 34 subjects--coal workers with pneumoconiosis, patients with collagenic diseases and healthy subjects--the daily urinary excretion of hydroxyproline (HP) and the plasma concentrations of HP and of collagenic peptidase have been measured during one week under HP-free and HP-enriched diet. It is shown that a 24-hour period of HP-free diet is necessary and sufficient before collecting urine or plasma for diet independent measurements of total or non-protein bound HP. Concentrations in plasma correlate well with the amounts excreted in urine. Collagenic peptidase is independent from the diet and well suited for assessment of the activity of the metabolism of collagene. There are no differences between healthy subjects and coal workers with pneumoconiosis.


Enterotoxemia in rabbits.

Enterotoxemia in rabbits. The presence of Clostridium perfringens Type E iota toxin was confirmed from the cecal contents of 23 of 46 rabbits which died of enteritis complex. The most consistent lesions observed were hemorrhage and edema in the cecum. Rabbit toxicity tests showed the toxic cecal contents were lethal for young rabbits unless incubated with Clostridium perfringens Type E antiserum.


A fatal enteric syndrome in Mystromys albicaudatus (white tailed rat) following topical antibiotic treatment. Heavy death losses (59%) occurred in adult Mystromys 3--14 days after muscle biopsies were taken from their rear legs. Clinical signs included anorexia, depression, and rough hair coat. Predominant necropsy findings were hrmorrhagic typhlitis and colitis. Results of studies evaluating the roles of the anesthetic and the tropical antibiotic ointment suggested that the deaths were caused by ingestion of the topically applied antibiotic.


Inhibition of the binding of low-density lipoprotein to its cell surface receptor in human fibroblasts by positively charged proteins. A group of proteins and polyamino acids with positively charged domains were shown to inhibit the binding of 125I-LDL to its receptor on the surface of human fibroblasts. The list of inhibitory proteins included platelet factor 4 (which has a cluster of lysine residues at its carboxyl terminus), two lysine-rich histones, poly-L-lysines of chain length greater than 4, and protamine. These proteins were effective in the concentration range of 5--10 microgram/ml. Two other positively charged proteins, lysozyme and avidin, did not inhibit 125I-LDL binding. Kinetic studies suggested that protamine was not acting simply as a competitive inhibitor with regard to the LDL receptor. In light of previous data showing that polyanions such as heparin and polyphosphates also inhibit 125-I-LDL binding to its cell surface receptor, the current findings suggest that charge interactions are important in this binding reaction. In a related series of studies, a number of glycoproteins and their asialo derivatives as well as a number of sugar phosphates failed to inhibit 125I-LDL binding to its receptor in fibroblasts.


The program of Friend cell erythroid differentiation: early changes in Na+/K+ ATPase function. Treatment of Friend erythroleukemia cells with several different chemical agents causes an early decrease in the 86Rb+ influx mediated by Na+/K+ adenosine triphosphatase (ATPase). These agents, which induce Friend cells to differentiate, include dimethylsulfoxide (DMSO), ouabain, hypoxanthine, and actinomycin D. The magnitude of the early decrease in 86Rb+ influx correlates with the proportion of cells in cultures of inducible Friend cell clones which later go on to synthesize hemoglobin. Compounds which do not incude differentiation in these cells, such as xanthine, exogenous hematin, and erythropoietin, do not cause a change in 86Rb+ influx. A change in the intracellular K+ ion concentration does not occur during induction by DMSO because, although there is a decrease in K+ content per cell soon after induction, there is a parallel decrease in cell volume. These results and previous observations from this laboratory are discussed in terms of the posible involvement of the Na+/K+ ATPase in Friend cell differentiation.


Mechanism of action of choleragen.

Mechanism of action of choleragen. Choleragen exerts its effect on cells through activation of adenylate cyclase. Choleragen initially interacts with cells through binding of the B subunit of the toxin to the ganglioside GM1 on the cell surface. Subsequent events are less clear. Patching or capping of toxin on the cell surface may be an obligatory step in choleragen action. Studies in cell-free systems have demonstrated that activation of adenylate cyclase by choleragen requires NAD. In addition to NAD, requirements have been observed for ATP, GTP, and calcium-dependent regulatory protein. GTP also is required for the expression of choleragen-activated adenylate cyclase. In preparations from turkey erythrocytes, choleragen appears to inhibit an isoproterenol-stimulated GTPase. It has been postulated that by decreasing the activity of a specific GTPase, choleragen would stabilize a GTP-adenylate cyclase complex and maintain the cyclase in an activated state. Although the holotoxin is most effective in intact cells, with the A subunit having 1/20th of its activity and the B subunit (choleragenoid) being inactive, in cell-free systems the A subunit, specifically the A1 fragment, is required for adenylate cyclase activation. The B protomer is inactive. Choleragen, the A subunit, or A1 fragment under suitable conditions hydrolyzes NAD to ADP-ribose and nicotinamide (NAD glycohydrolase activity) and catalyzes the transfer of the ADP-ribose moiety of NAD to the guandino group of arginine (ADP-ribosyltransferase activity). The NAD glycohydrolase activity is similar to that exhibited by other NAD-dependent bacterial toxins (diphtheria toxin, Pseudomonas exotoxin A), which act by catalyzing the ADP-ribosylation of a specific acceptor protein. If the ADP-ribosylation of arginine is a model for the reaction catalyzed by choleragen in vivo, then arginine is presumably an analog of the amino acid which is ADP-ribosylated in the acceptor protein. It is postulated that choleragen exerts its effects on cells through the NAD-dependent ADP-ribosylation of an arginine or similar amino acid in either the cyclase itself or a regulatory protein of the cyclase system.


[Sudden deafness and cytomegalievirusinfection of adults (author's transl)]. By means of virologuc--serologic examination methods it was possible to find in four patients with sudden deafness a cytomegaloinfection, whereby the complement-fixation reaction was determined with the CMV antigen. With the usual therapeutical measures all cases with an, in part high-degree, labyrinthine deafness improved or normalized simultaneously with the serologically ascertained subsiding of the virus infection. The paper discusses the pathogenesis of viruscaused sudden deafness and tries to explain the benign nature of central complications in cytomegaloinfections.


[Giant cell tumor of tendon sheath in association with synovialchondroma. An unusual tumor-combination at the temporomandibular joint (author's transl)]. A tumor at the temporomandibular joint is reported, which so far has not yet been described. Histologically it turned out to be a combination of synovialchondroma and giant cell tumor of tendon sheath. By reason of the microscopic picture and the course of 2 1/2 years without relapse it is a benign tumor. In postoperative x-rays of the mandible a thickening and sclerosis as well as cystic areas of increased translucence at the place of removal of the tumor were found, which had already existed preoperatively in a lower degree.


[Myxoma of the mastoid with destruction of the facial nerve (author's transl)]. A case of Myxoma of the mastoid with initial facial spasme followed by palsy of the nerve is reported. After removing the tumor the nerve has been reconstructed from the secound knee until the stylomastoid foramen. The probable origine of the tumor is discussed.


Differences in the phospholipid, cholesterol, and fatty acyl composition of 3T3 and SV3T3 plasma membranes. An analysis of the phospholipid, cholesterol, and phospholipid fatty acyl composition of isolated plasma membranes of 3T3 and SV3T3 mouse mebryo cells has been performed. The results show that the plasma membrane of SV3T3 cells contain relatively less phosphatidylethanolamine and sphingomyelin and more cholesterol than 3T3 plasma membranes. The fatty acyl composition of individual phospholipid classes as determined by gas liquid chromatography also showed differences between 3T3 and SV3T3 plasma membranes. The plasma membranes of SV40 transformed 3T3 cells contain: (a) a higher percentage of 18:1 and less 20:3 and 20:4 in phosphatidylethanolamine; (b) a higher percentage of 18:1 in phosphatidylserine; and (c) a higher percentage of 18:2 and 20:4 in phosphatidylinositol.


[Experimental silicosis. II. Fibrogenic effect of natural amorphous silica]. Fibrogenic effect of two natural amorphous silica dusts-diatomite from deposits near Leszczawka (Poland) and silica earth (from the USA) was tested on the rats. Pneumoconiosis was produced by intratracheal introduction of 50 mg dust, at a single dose. Fibrogenic effect was assessed after 3, 6 and 9 months after introduction of dust. Analysis of diatomite dust carried out with X-ray diffraction method showed the presence of quartz in a quantity not exceeding 5%, while in silica earth dust from the USA no silica critalline structures were found. Development of fibrogenic changes in lungs of the rats assessed with hydroxyproline (collagen) increase and weight increment of wet lungs was small. The increment of the indices was twice higher as compared with their increment found in the lungs of control animals but lower than in the rats which were given cristalline silica dusts. In histopathological examinations no progressive lung fibrosis was found though some signs of destruction and necrobiosis in some cells were noticed.


[Treatment of African vesico-vaginal fistulae in a rural hospital (author's transl)]. The author reports his own experience of this difficult surgery in a rural african hospital. He describes various types of fistulae and 3 technics which gave him 10 recoveries in 19 patients. Complications, rules of treatment and nursing are considered.


Bioassay of TSH using dog thyroid cells in monolayer culture. The cAMP response to TSH stimulation in dog thyroid cells in monolayer culture was adapted as a means to assay TSH bioactivity. Of a variety of polypeptide hormones examined, only TSH and LH stimulated thyroid cell cAMP generation, but stimulation by LH probably represented contamination with TSH. Serum was found to be a potent, noncompetitive inhibitor of the thyroid cell cAMP response to TSH stimulation. The inhibitor(s) present in serum was nondialyzable and, on Sephadex G-200 gel filtration, eluted over a wide range between the void volume and the albumin peak. Because of this inhibitory effect of serum, partial purification of TSH from serum was necessary, and was achieved using Sephacryl S-200 gel filtration. Columns were calibrated with human TSH as measured by radioimmunoassay and bovine TSH as measured by bioassay. Intraassay variation of the thyroid cell cAMP response to a 100 microU/ml TSH standard was 9.2%. TSH standards were added to human serum following which the TSH was extracted by gel filtration and assayed for bioactivity. With TSH standards of 100 microU/ml intra-assay variation was 19.9%; and with TSH standards of 25 microU/ml, interassay variation was 34.5%. TSH bioactivity was demonstrable in serum from 21 of 25 patients with primary hypothyroidism, 4 of 14 normal subjects and none of 3 patients on exogenous thyroxine replacement. Although a positive correlation was observed between serum TSH bioactivity and immunoactivity, numerous individual samples displayed a dichotomy between the two measurements. This study provides new evidence that immunoassayable TSH in the serum of patients with primary hypothyroidism is not necessarily synonymous with TSH bioactivity.


[Children death rates in a rural village of Ivory Coast (author's transl)]. A 10 years statistical study with a global death rate of 63 per 1.0000 and its changes according to the age groups.


The role of free fatty acids in the regulation of lipolysis by human adipose tissue cells. The effect of added fatty acid on lipolysis and cyclic AMP concentration of human adipose tissue cells was studied. The addition of sodium oleate decreased the lipolytic response of adipocytes to 10(-7) M isoproterenol. Inhibition was detectable with the lowest quantity of oleate added, 0.2 mM, and was progressively greater with increasing quantities of added fatty acid. Palmitic and linoleic acids were as effective as oleic acid in suppressing isoproterenol-stimulated lipolysis. Suppression of cyclic AMP formation was detectable within one minute after the addition of oleate. Cyclic AMP formation, suppressed by accumulated fatty acids, could not be stimulated by the addition of fresh isoproterenol. However, after the accumulated fatty acids were removed by buffer change, cyclic AMP formation was stimulated by fresh isoproterenol. These findings are consistent with the view that fatty acids are physiologically significant regulators of lipolysis.


Electron microscopic studies of tumor viruses. II. Entry and uncoating of Epstein-Barr virus. Entry of Epstein-Barr virus into human lymphoblastoid cells (Daudi cells) was studied by electron microscopy. At the site of viral attachment, two distinct interactions conducive to penetration of the virus occurred between the viral envelope and cell membrane, namely, (i) simultaneous dissolution of both the envelope and cell membrane, presumably resulting in passage of viral capsids into the cytoplasm and (ii) dissolution confined to the cell membrane with resulting penetration of enveloped virus. In the latter case envelope dissolution appears to occur subsequently in the cytoplasm with release of capsids. Fusion of the viral envelope with the cell membrane was not observed. The capsids exhibited two distinct structural forms--one dense, the other translucent or light in appearance. The former disrupted near the cell membrane with release of viral cores into the cytoplasm whereas the light capsids containing dense cores migrated toward the nucleus and accumulated in the perinuclear region. Apparently the process of releasing deoxyribonucleic acid (DNA) from the light capsid is slowed down or prevented in Daudi cells. A hypothesis is presented concerning the manner in which these two types of capsids initiate infection.


[A serological survey of rubella among pregnant women in Abidjan (author's transl)]. A serological survey about the prevalence of the rubella hemagglutination inhibiting antibodies among the pregnant women in Abidjan during the year 1975 has shown an average rate of seroconversion of 84 p. 100. This rate depends on age and increases from 70 to 80 p. 100 between 15 and 35 years old. A regression line is fitted, and the goodness of fit is tested. These rates are in accordance with what is presently observed in America, in Europe and in Australia.


[Value of indirect fluorescent tests for on the spot prospection of human African trypanosomiasis (author's transl)]. The equipment of a laboratory making indirect antibody-fluorescent tests on dried blood blots in the heart of the trypanosomiasis focus of Bouaflé (Ivory Coast) gave good results. On a visited population of 1444 people, 13 cases were diagnosed by immunofluorescence tests; 8 were already diagnosed on the spot, trypanosome found in gland juice; 5 were diagnosed following the results of immunofluorescence tests. Reduction of time between sampling and immunological results makes easier the search of suspects. Based on these observations, the writers emphasize the great value of the use of immunofluorescence technics on the spot and the establishment of a regional mobile immunofluorescence team for supplementing the work of local prospection teams.


Preparation and efficacy of an inactivated subunit vaccine (NFUIBHK) against type 2 Herpes simplex virus infection. A vaccine against Herpes simplex virus infection was prepared by Nonidet NP 40 and formalin treatment of a type 1, infected-cell extract; virus particles were removed by ultracentrifugation over sucrose. These procedures were not detrimental to the antigenic quality of the vaccine preparation. The vaccine afforded significant protection to experimental type 2 genital herpes virus infection in mice, as adjudged by clinical observations, cytopathological change, and virus yields.


Characterization of proteins of human papilloma viruses (HPV) and antibody response to HPV 1. The serological characterization of virus isolates from verrucae vulgares and plantar warts revealed that HPV 1 and HPV 4 are present in about 50% of these warts with HPV 1 being more prevalent, especially in plantar warts. Parallel to the high incidence of HPV 1 infections, about 50% of non-selected young adults contained antibodies against HPV 1. Only HPV 4 particles, however, reacted with serum from a patient with epidermodysplasia verruciformis when tested by immuno-electron microscopy. An examination of HPV 1 proteins indicated that the major structural proteins VP2 and VP3 are trypsin sensitive. Tryptic degradation leads to distinct polypeptides with molecular weights between 37,000 and 23,000 which may be correlated to minor protein components of HPV 1 preparations. HPV 1 histone-like proteins, which co-migrate with purified cellular histones in SDS gel electrophoresis were analyzed in an acetic acid urea system. It was shown that H3- and H4-like proteins differ from cellular histones. The reason for this difference and its meaning are discussed.


Attempts to isolate virus from human trigeminal and facial ganglions. Herpes virus hominis type 1 was isolated from the trigeminal ganglion (ganglion semilunare, gasservian) in three out of 20 randomly selected autopsies. Two of the three patients had been treated with immunosuppressive or cytostatic agents. Clinical signs of herpes infection were not observed during the previous 6 months. No virus was isolated from the facial ganglion (geniculate ganglion) in the same 20 cases. The findings are discussed in relation to the viral etiology of acute peripheral facial palsy.


Comparison of five diagnostic methods for the detection of rotavirus antigens in calf faeces. Immunoelectroosmophoresis, complement fixation, immunofluorescence on inoculated cell cultures, electron microscopy, and an enzyme-linked immunosorbent assay were compared for the detection of rotavirus in faecal samples from calves. Rotavirus particles could be detected in 39 out of 98 faecal samples by electron microscopy. Immunofluorescence, complement fixation, and immunoelectroosmophoresis detected rotavirus antigens in 39, 42, and 30 samples, respectively. The enzyme-linked immunosorbent assay demonstrated rotavirus antigens in 49 faecal samples. The assay is not only sensitive but also simple to perform and suitable for large-scale testing.


Nonspecific inhibitors of coronavirus OC43 haemagglutination in human sera. Antibodies against the human coronavirus OC43 in human sera were measured by haemagglutination inhibition (HI), complement fixation (CF), and radial diffusion-haemolysis in gel (HIG) techniques. The apparent HI titres in a fraction of sera with no antibodies detectable by the two other methods were found to be reduced considerably after treating the sera with phospholipase C (PLC). The PLC treatment also reduced the apparent HI titres in some sera containing variable amounts of CF and/or HIG antibodies, but did not affect antibody determinations by the two latter methods. These results suggest that false positive results can be obtained in the OC43 HI test unless the sera are treated with phospholipase C before the assay.


Application of a solid-phase radioimmunoassay and immune electron microscopy for hepatitis A in diagnosis and research. With crude virus suspensions from stool and antibodies from hepatitis-A patients, a solid-phase radioimmunoassay (RIA) for detection of hepatitis virus A (HVA) had antibodies against hepatitis virus A (anti-HVA) has been developed. Examples for the application of this test are demonstrated. Virus particles from the stools of the two patients were further characterized. Serologically, they were identical or very similar to the MS-1 strain. Isopycnic CsCl-gradient centrifugation of both strains revealed two peaks, but the particles of different densities did not differ in size or serologically. A modification of the RIA was also useful for determination of IgM antibodies in patients' sera fractionated by sucrose-density centrifugation. The application of the RIA method for serologic epidemiology is demonstrated by a comparison of anti-HVA prevalence in German and non-German women residing in Germany.


Epstein-Barr virus early antigen titer by immunofluorescence in microplates. A new semi-automated method based on microtiter system. For the titration of antibodies directed against Epstein-Barr Virus early antigen (EBV-EA) we describe a method in microplates with the Microtiter System (Cooke Engineering Co., Alexandria, Va). Using this technique, which allows rapid epidemiological investigations, we have titered anti-EBV-EA antibodies in different groups of patients and controls, and particularly in group which had recent contact with infectious mononucleosis. In this group the percentage of individuals having antibodies directed against the EBV-EA antigen was significantly higher than in the group of controls.


The proteins of varicella-zoster-virus.

The proteins of varicella-zoster-virus. [35S] methionine-labelled Varicella-Zoster-Virus (VZV) was partly purified by sucrose density gradient centrifugation. Enveloped virus, checked by electron microscopy, was analyzed by high resolution polyacrylamide gel electrophoresis. A polypeptide pattern of at least 31 proteins could be identified on the fluorogram. The molecular weight of the proteins, estimated by co-electrophoresis with marker proteins, ranged from 18,000 to 240,000 daltons. Immunoprecipitation using a specific human antiserum showed at least 14 proteins.


HBsAg-negative chronic active hepatitis related to hepatitis B virus. Numerous cases of chronic hepatitis have been shown to be closely associated with persistent infection with hepatitis B virus (HBV). A group of 100 patients suffering from chronic active hepatitis (CAH) was investigated for HBV serologic markers. Of these, 35 patients were HbsAg-positive; in 26 HBsAg-negative subjects, anti-HBc were detected using counterimmune electrophoresis and complement-fixation tests. These data suggest that chronic liver disease in patients who were only anti-HBc-positive might be related to persistent infection with hepatitis B virus. Epidemiological clinical and histopathological data were different when we compared CAH patients who were HBsAg-negative, but anti-HBc-positive, with HBsAg-positive CAH patients. A sequence is proposed leading from HBsAg-positive to HBsAg-negative CAH, cirrhosis, and hepatoma in temperate areas, according to a model similar to the one described in intertropical Africa.


Physical characterization of deoxyribonucleic acids of different human papilloma viruses (HPV). Different types of human papilloma viruses (HPV) had been identified by the use of bacterial restriction endonucleases (Gissmann et al., 1977). The DNAs of HPV 1, 2, and 3 showing a similar restriction enzyme pattern can be purified in sufficient quantities to permit further characterization. These viruses appear not to be identical, as far as molecular weight (4.9 X 10(6) d), buoyant density (1.700 mu/ml), and sedimentation values of the different DNA components (Co I: 22--23S; Co II: 17S; Co III: 16S) are concerned. HPV 4 DNA, which is entirely different from HPV 1--3 in its restriction enzyme pattern, does not hybridize with cRNA transcribed in vitro from HPV 1 DNA. The cRNA was shown to represent the whole viral genome.


Genetic susceptibility to wild and vaccine polio virus: genotypes and their frequency. The frequency of proposed genotypes which predispose to poliomyelitis, is tabled. Susceptibility is due to a gene or linkage group of genes with a frequency of 2% for the homozygote and 24% for the heterozygote. Two subgroups are identified where a second gene might make the persons susceptible to vaccine strains of virus. Cutter vaccinees might form a third group with increased susceptibility under special circumstances. The age at which genetic susceptibility changes to phenotypic susceptibility may be modified by physical factors such as those due to congenital syphilis and Salvarsan, and thalidomide.


Protection of guinea pigs against primary and recurrent genital herpes infections by immunization with live heterologous or homologous Herpes simplex virus: implications for a herpes virus vaccine. Guinea pigs, inoculated intravaginally with HSV type 2 develop a primary and spontaneously recurring genital herpes with clinical symptoms resembling genital herpes in humans. The effect of a previous infection with either HSV type 1 or HSV type 2 on the clinical course of the intravaginal challenge infection was studied. Prior infection with either subtype of HSV moderated considerably the acute disease induced by intravaginal HSV type 2 challenge. No effect was, however, observed on the development of the chronic phase of the challenge infection. The significance of these studies for the possibility of vaccination against recurrent herpes infections is discussed.


Stimulation of human lymphocytes by cytomegalovirions and dense bodies. Lymphocytes from healthy adult individuals were examined for their ability to incorporate thymidine in the presence of cytomegalovirus (CMV) and dense bodies. It was found that lymphocytes from donors with antibodies to CMV were stimulated to incorporate thymidine-14C both by preparation of CMV and CMV dense bodies. Lymphocytes from CMV seronegative donors did not respond. Dilution experiments and the dose-response curve indicate that the stimulation induced by the dense body preparation was not caused by the small amounts of contaminating CMV particles. These results indicate that in healthy human adults there is a correlation between CMV seropositivity and in vitro lymphocyte transformation, induced by either CMV or by dense bodies.


Primary chemotherapeutic management of unresectable and metastatic hepatoblastoma in children: report of four cases. Four children presenting with unresectable hepatoblastomas and one with metastatic disease are reported. Following initial biopsy all were treated with chemotherapy which included Adriamycin. Three of the four children showed a significant reduction in tumor size, and in two, delayed resection of the primary lesion was possible. Chemotherapy including Adriamycin represents effective initial cytoreductive therapy for childhood hepatoblastoma, thereby reducing the morbidity and mortality associated with the extensive hepatic resection usually required for an untreated lesion.


Prevention of brain extension in small-cell carcinoma of the bronchus. In small-cell carcinoma of the bronchus, extension to the brain is common. The role of prophylactic whole-brain irradiation and combination chemotherapy using CCNU in the prevention of brain extension is examined in the light of recent studies.


Treatment of oat-cell carcinoma of the lung in the community. Nine patients with "limited" (thoracic) small-cell lung carcinoma were treated with combined chemotherapy and radiotherapy (involved field and prophylactic whole brain). Induction drugs were Vincristine, Adriamycin, and Cyclophosphamide; and Cyclophosphamide, Vincristine, and CCNU for maintenance. Eight out of nine patients (89%) continue NED at a median follow-up time of 12 months. This group is compared to 21 patients with more extensive disease and 6 patients with limited disease using various other regimens. Median survival in the extensive group was 5 months and 8 months for the limited-diseased group treated with single agents. Intensive combination chemo-irradiation therapy is safe and highly effective in patients with limited oat-cell carcinoma of the lung, and leads to prolonged disease-free survival.


Specific effect of zinc ions on DNA polymerase activity of avian myeloblastosis virus. The effect of selected cations on DNA synthesis by DNA-polymerase of avian myeloblastosis virus (AMV) was studied. Zinc ions at low concentration (0.2mM) in the assay system enhanced the activity about 2 x fold and at higher concentration (2.0 mM) inhibited the activity completely. In contrast, addition of lithium and potassium salts produced inhibitory effects in this ionic concentration range. Replacement of K+ ion had an inhibitory effect on the activity.


[alpha1-antitrypsin deficiency. Clinical and morphological aspects during childhood (author's transl)]. In many cases so called neonatal hepatitis of unknown origin nowadays is recognized as a manifestation of alpha1-antitrypsin deficiency. Out of 12 patients with Pi-type ZZ, 5 were diagnosed because of cholestatic jaundice, 2 because of hepato-splenomegaly in the first trimenon, and 3 by family examination. We believe that the affection may be due to a perinataly acquired cytomegalic inclusion disease in one case, in another to a congenital rubella infection. The latter child died at the age of one year because of an esophageal hemorrhage. Over a mean observation time of 3 years the other patients are doing well and show no signs of portal hypertension. The very different course of the hepatopathy is demonstrated. Common bacteria or toxins which do not usually lead to an illness may be realisation factors just as "classical" causes of hepatitis. Up to now it is not known how these factors influence the course of alpha-antitrypsin deficiency.


[Dose-effect of beta-sitosterin in type IIa and IIb hypercholesterolemias (author's transl)]. In a trial with 59 outpatients with hypercholesterolemia the effect of two different doses of 5.28 and 10,56 g Beta-Sitosterol was compared to placebo. Each treatment phase lasted six weeks. Serum cholesterol decreased on the average under Beta-Sitosterol by 10--13% (p less than 0.05) in correlation with LDL. The triglycerides were not affected; there was a slight increase in the HDL/LDL quotient. The optimal daily dose of Beta-Sitosterol was about 6 gm; the double dose was no more effective.


Perturbations in simian virus 40 DNA synthesis by ultraviolet light. Perturbations of Simian Virus 40 (SV40) DNA replication by ultraviolet (UV) light during the lytic cycle in permissive monkey CV-1 cells resemble those seen in host cell DNA replication. Formation of Form I DNA molecules (i.e. completion of SV40 DNA synthesis) was more sensitive to UV irradiation than synthesis of replicative intermediates or Form II molecules, consistent with inhibition of DNA chain elongation. The observed amounts of [3H]thymidine incorporated in UV-irradiated molecules could be predicted on the assumption that pyrimidine dimers are responsible for blocking nascent DNA strand growth. The relative proportion of labeled Form I molecules in UV-irradiated cultures rapidly increased to near-control values with incubation after 20 or 40 J/m2 of light (0.9--1.0 or 1.8--2.0 dimers per SV40 genome, respectively). This rapid increase and the failure of Form II molecules to accumulate suggest that SV40 growing forks can rapidly bypass many dimers. Form II molecules formed after UV irradiation were not converted to linear (Form III) molecules by the dimer-specific T4 endonuclease V, suggesting either that there are no gaps opposite dimers in these molecules or that T4 endonuclease V cannot use Form II molecules as substrates.


The enkephalins. Peptides with morphine-like activity. Two penta-peptides with morphine-like properties have recently been found in the nervous system and gastrointestinal tract. These peptides known as enkephalins are unevenly distributed in the brain, the highest concentration being in the globus pallidus. The enkephalins are found to have analgesic properties and are also anti-tussive, anti-diarrheal and able to stimulate release of prolactin and growth hormone. Synthetic analogues of the enkephalins containing specific D-amino acid substitutions have been developed and show a wide spectrum of activities.


Site of naloxone-precipitated opiate withdrawal dissociates from that at which apomorphine reinitiates this phenomenon. The site of action of naloxone to induce jumping in morphine tolerant/dependent rats appears to be dissociated from structures where apomorphine initiates its action to reinduce jumping in previously withdrawn animals. These findings suggest that dopaminergic pathways do not directly affect the neuronal circuit involved in withdrawal jumping behavior, but instead exert a facilitatory influence on neurons that become supersensitive during the state of withdrawal. Thus, an increased response to apomorphine during naloxone-precipitated opiate withdrawal does not necessarily imply a specific supersensitivity of the dopaminergic system.


[Benign childhood epilepsy with spike activity in the area of the precentral gyrus]. The authors report 18 cases of benign childhood epilepsy with spike activity in the Rolandic area. These cases were isolated from a group of 103 children with partial epilepsy. In agreement with the observations of other authors the benign course of this form of epilepsy is stressed. The isolation of this type of epilepsy is necessary in view of its practical management, treatment and prognosis. Neuroradiological and neurological examinations failed to demonstrate any evidence of organic brain disease in this epilepsy. Focal EEG changes with spike activity in the frontal or temporal area regressed gradually until complete normalization of the record. The authors agree with the view of other authors that the diagnosis of this epilepsy in typical from does not require radioneurological contrast investigations.


Steroid hormones in ovarian vein and cyst fluid of a virilizing stromal tumor. A 23-year-old female with clinical signs of excessive androgen and estrogen production was found to have a gonadal stromal tumor. The clinical symptoms correlated well with steroid hormone values in ovarian vein blood and ovarian cyst fluid of the ovarian tumor. Ovarian vein blood draining the ovarian tumor measured as follows: estrone, 162 pg/ml; estradiol, 1688 pg/ml; progesterone, 9.6 ng/ml; testosterone, 9775 ng/100 ml; and dihydrotestosterone, 2905 ng/100 ml. The ovarian cyst fluid produced higher estrogen and lower androgen values compared with ipsilateral ovarian vein blood. Peripheral levels of immunoreactive testosterone ranged from 210 to 244 ng/100 ml. It is concluded that excessive androgen and estrogen production may arise from a gonadal stromal tumor that has been previously classified as a purely virilizing ovarian tumor.


Colposcopic examination of the cervix of cebus monkeys. A total of 410 colposcopic examinations were performed on 188 female cebus monkeys that were under study to determine the oncogenic potential of herpes simplex virus type 2 in this genus. A split-cone vaginal speculum was developed that permitted good observation of the vaginal cervix in the cebus monkey. The cervical anatomy of cebus monkeys was found to differ from that of humans in that the surface of the animal cervix was more papilliform, with thinner squamous epithelium, and the squamocolumnar junction lay within the endocervical canal. Therefore, the ability to detect abnormalities in the cervical epithelium by colposcopic examination in the cebus monkey was restricted to vascular changes in the squamous epithelium. After 100 examinations, several vascular patterns were distinguishable and interpretations of these patterns were compared with cytologic findings on the same animals. Findings by both cytology and colposcopy were mild in nature; no carcinoma was detected. Colposcopic and cytologic findings correlated at a level of 84%. More abnormalities were detected with colposcopy than with use of cytologic techniques.


The value of levamisole in a Wilms' tumor animal model. The effect of Levamisole was studied in an animal model of Wilms' tumor. No tumoridical effect of Levamisol could be documented in this tumor model, and no effect was shown on prevention of tumor, when Levamisole was given before tumor implantation. In previous experience with Wilms' tumor model, a good correlation between the human and animal tumor was found in regard to treatment with different drugs. The fact that Levamisole had no effect on our animal model and its reported immunosuppressive effect at some doses, should be considered in planning clinical trials.


Quartz fibers as templates for biopolymers. The polymerization of silica in water solution to form quartz fibers proceeds by a dehydration process, analogous to condensation polymerization in organic high-polymers, in which monomeric Si(OH)4 groups unite through Si--O--Si bonds with the elimination of H2O. The resulting fibers are structurally polar along the direction of elongation, are enantiomorphous, and generally show stereospecific twisting around the direction of elongation. In these regards the fibers are analogues of biopolymers such as RNA and DNA. Quartz also possesses specific adsorptive relations to a wide range of organic substances including monomer amino acids, short-chain polypeptides, and proteins. These involve hydrogen-bonding between (OH) or silanol groups on the surface of the quartz with active side-groups on the organic molecules, and in part are epitaxial through dimensional coincidences in the interface. Geochemical evidence indicates that quartz was deposited in the early Precambrian ocean either by direct crystallization from seawater or by recrystallization of amorphous silica. What is of interest is the possible role of quartz fibers as a template and co-polymer in the passage of biomonomers in the pre-biotic ocean to the long-chain biopolymers such as nucleic acids and proteins that are involved in life processes.


Degradation of biochemical activity in soil sterilized by dry heat and gamma radiation. The activities of three enzymes present in soil, phosphatases, urease, and decarboxylase, were monitered as indicators of the loss of biochemical information occurring when soil was sterilized by dry heat (0.08% relative humidity), gamma radiation, or a combination of both. More enzymatic activity was retained in soil sterilized by a long exposure to dry heat at relatively low temperature (8 weeks at 100.5 degrees C) than by a shorter exposure to a higher temperature (2 weeks at 124.5 degrees C). No enzymatic activity was detectable in soil sterilized by an even higher temperature (4 days at 148.5 degrees C). Soil sterilized with 7.5 Mrads of radiation retained much higher enzymatic activity than with heat sterilization. Combining sublethal doses of heat radiation effectively sterilized the soil and yielded enzymatic activities higher than those of soil sterilized by dry heat alone but lower than those of soil sterilized by radiation.


The role of interferon in virus infections and antibody formation. Virus-type interferon is capable of suppressing the antibody response both in vivo and in vitro. Interferon induced by antigen or mitogen (immune interferon), primarily in T-lymphocytes, also appears capable of suppressing the antibody response. Cyclic AMP has a regulatory effect on the induction of both virus-type and immune interferon. The immunosuppressive effects of virus-type and immune interferon may differ at the cellular or subcellular level.


Calcification within metastatic pulmonary nodules from synovial sarcoma. A case of calcified metastatic lung deposits from synovial sarcoma of the leg is reported. Calcification within metastatic lung nodules from synovial sarcoma has not been previously reported. The differential diagnoses of calcified lung metastases are presented.


Calcium and sodium transport processes in patients with cystic fibrosis. I. A specific decrease in Mg2+-dependent, Ca2+-adenosine triphosphatase activity in erythrocyte membranes from cystic fibrosis patients. Calcium-ATPase activity (Mg2+-dependent Ca2+-ATPase, ATP phosphohydrolase, EC 3.6.1.3) in erythrocyte membrane preparations from cystic fibrosis (CF) patients was greatly reduced compared to erythrocyte membranes from control subjects. The Km for calcium was found to be similar in the two groups; however, the Vmax, the maximal rate of activation of the Ca2+-ATPase, is reduced by 50% in the erythrocyte membrane preparations of the CF patients (P less than 0.001). In contrast, the Mg2+-ATPase activity of erythrocyte membranes from CF patients was unchanged compared to the control subjects. No difference in the Na+,K+-ATPase activity in erythrocyte membranes from CF patients compared to control patients could be observed. This indicates that the Ca2+-ATPase activity noted in CF erythrocytes is not part of a generalized membrane or membrane-bound enzyme alteration. It remains to be determined whether this alteration in Ca2+-ATPase activity is directly related to a defect in calcium transport in these cells and is a generalized phenomenon in CF present in cell types more directly involved in secretion.


Reye's syndrome: preservation of mitochondrial enzymes in brain and muscle compared with liver. The activities of five mitochondrial enzymes tested in liver from patients with Reye's syndrome were measured. Citrate synthase, glutamic dehydrogenase, succinic dehydrogenase, pyruvate carboxylase, and pyruvate dehydrogenase were all outside of the range shown by control samples and well below them in activity. The activity of two extramitochondrial enzymes, glucose-6-phosphatase, which is a microsomal enzyme, and fructose-1,6-diphosphatase, which is a soluble enzyme, were in the normal range in samples from Reye's syndrome patients. In both muscle and brain the activities of the mitochondrial enzyme, citrate synthase, glutamic dehydrogenase, and succinic dehydrogenase were all within the control range. Pyruvate dehydrogenase was found to be normal in muscle from these patients.