Reliability of the serological method of examination as a basis for diagnosis of associated respiratory virus infections. The reliability of the results of serological examination in diagnostics of associated infections was studied on a model of artificially provoked vaccinal infections in humans and in laboratory animals. The effect of administered monopreparations on changes in the level of both homologous and heterologous antibodies was tested. The character of immunological changes following simultaneous administration of two or more respiratory viruses was analysed and the effect of these viruses in diseases of divers etiology was studied. According to the results of experiments on laboratory animals, repeated administration of any of the earlier used respiratory viruses stimulated the accumulation of only homologous antibodies while heterologous antibodies did not increase at all.


[Stich-plaque test--an economic method for quantitative determination of viruses]. An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID 50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity , accuracy, and reproductibility of this method are described and discussed.


The use of early embryo aggregation derived mouse chimaeras. II. The study of disease processes. Early embryo aggregation derived mouse chimeras have proven an extremely valuable tool to study development of disease processes. In this respect chimaeras prove a unique opportunity to study the interaction between cells genetically pre-destined to become diseased and a normal cell population within the same animal. In this situation chimaeras can be studied in view of possible 'correction' of the disease by the provision of a population of normal host cells and/or their products. Various host deficiency states including classic immune deficiency, potentially 'deficient' anemias and tumours have been studied--the subject of the review here.


A unique cytoplasmic structure in papular histiocytoma. Intracytoplasmic granules with a unique and highly complex ultrastructure, have been observed within the cellular infiltrate of the lesions of two patients with papular histiocytosis. Examination of serial sections has enabled construction of a three-dimensional model of these granules, which appear to be composed of an elaborate arrangement of membranes and vesicles. Two hypotheses (endocytosis and exocytosis) are proposed for the mode of formation of these unique cytoplasmic structures.


Clinical and serologic study of four smallpox vaccines comparing variations of dose and route of administration. Basic study and laboratory standardization. The present four-center collaborative study was undertaken in an attempt to define the best vaccine and/or vaccination procedure for use in areas of the world that are free of smallpox. The study was designed to compare the effect of different vaccinial strains, viral concentrations, and routes of administration on the morbidity and antibody response associated with primary vaccination and standard challenge revaccination. Primary vaccinations were performed on 1,585 children; 49.6% of the children were vaccinated by the percutaneous route, and 50.4% received vaccine subcutaneously. The overall age and sex distributions of percutaneous and subcutaneous vaccinees were comparable, but there were marked differences in participants among the four study centers. Vaccines in Kentucky had a greater mean age; the greatest number of Negroid children were enrolled in St. Louis, and more of them were vaccinated by the subcutaneous route; and the dropout rate was much greater in San Diego and Colorado. An analysis of comparative interlaboratory serologic procedures with the use of 20 coded duplicate samples of serum revealed good agreement in the hemagglutination-inhibition test; results of neutralization tests had greater variability of mean titers. On duplicate samples of serum from study participants there was generally good correlation between each of the four study centers and the Center for Disease Control's reference laboratory in titers of hemagglutination-inhibiting antibody. In contrast, 38% of the neutralization titers determined at the four study centers were greater than or equal to 0.67 log10 higher than the respective titers noted at the Center for Disease Control.


Clinical and serologic study of four smallpox vaccines comparing variations of dose and route of administration. Primary percutaneous vaccination. In an investigation of the antigenicity and reactogenicity of four smallpox vaccines, 786 children received primary percutaneous vaccination with vaccine in one of three concentrations, 10(6), 10(7), or10(8) pock-forming units/ml. Dose-response curves indicated that the three licensed vaccines (New York City Board of Health strains grown in calf lymph or chorioallantoic membrane, and the Lister vaccine) had similar potencies, but the CV-1 strain was about 10-fold less infectious. CV-1 also produced smaller skin lesions than the other three vaccines, and the incidence of fever in CV-1 vaccines who developed either "major reactions" or serum antibody was not significantly different from that in children in all vaccine groups with no evidence of "take." Hemmagglutination-inhibiting antibody was consistently seen in individuals who received potent New York City and Lister vaccines, and neutralizing antibody was induced in 82%-85% of children in this group who had some evidence of take (production of hemagglutination-inhibiting antibody or or major reaction). Only 30% of CV-1 vaccines with takes produced neutralizing antibody. Minor complications occurred in all groups, but most often in children who received the New York City strains."


Clinical and serologic study of four smallpox vaccines comparing variations of dose and route of administration. Standard percutaneous revaccination of children who receive primary percutaneous vaccination. A standard challenge with percutaneous smallpox vaccine was administered to 629 children six to 12 months after percutaneous primary inoculation with one of four vaccines (New York City Board of Health strains grown in calf lymph or chorioallantoic membranes, the Lister vaccine, or the CV-1 strain). Of those who had had major reactions on primary vaccination, 8%-21% responded to revasccination with a typical primary-type skin response. In contrast, such a primary-type response occurred in 50% of those who on primary vaccination had developed serum antibody in the absence of major reactions and in 83% of those who had had no serologic or clinical evidence of primary "take." Skin lesions on revaccination tended to be largest in thosewhose primary vaccination was with CV-1, although fever and minor complications were not more frequent. Moreover, even in children who had received CV-1 vaccine, skin responses to challenge vaccine were clearly attenuated when compared with responses of children who had not had takes on primary vaccination. Sizes of lesions and acceleration of skin erytherma after challenge were related in most children to titers of both hemagglutination-inhibiting and neutralizing antibody at the time of revaccination. One month after revaccination, neutralizing antibody was present in 93%-96% of those with takes onprimary vaccination with New York City or Lister vaccines, but only 75% of CV-1 vaccines.


Clinical and serologic study of four smallpox vaccines comparing variations of dose and route of administration. Standard percutaneous revaccination of children who receive primary subcutaneous vaccination. Six months after subcutaneous vaccination with one of four smallpox vaccines, 655 children were challenged with a standard percutaneous smallpox vaccine. Response to reimmunization was characterized by a significant acceleration and diminution of skin response, but not to the degree seen in an equivalent group who had received their primary immunization percutaneously. Fever after revaccination was absent if there had been a "take" with primary subcutaneous vaccination. The overall incidence of minor vaccine-related complications with revaccination was 2-1/2%. The neutralizing antibody response to revaccination was markedly reduced, as compared to that of children who received either one or two successful percutaneous vaccinations. Subcutaneous vaccination followed by percutaneous vaccination is not recommended as a schedule for smallpox immunization, because complications are not avoided, and the incidence and mean titer of resultant neutralizing antibody are low.


Correlation of herpes simplex virus types 1 and 2 with clinical features of infection. Strains (338) of herpes simplex virus (HSV) were isolated in Stockholm during 1965-1974. By immunoelectroosmophoresis it was possible to identify all strains as either HSV type 1 (HSV-1) or 2 (HSV-2). No strains of intermediate antigenic type or with untypable characteristics were found. The antigenic type of HSV was correlated with body site and clinical features of infection. A case of severe, recurrent, abdominal pain in association with HSV-2 infection is described. In one patient with acute aseptic meningitis, both coxsackievirus A9 and HSV-2 were isolated from the same specimen of cerebrospinal fluid. Serology suggested a primary infection with coxsackievirus A9 and a recurrent HSV-2 infection. HSV-1 was isolated from specimens of cerebrospinal fluid. Serology suggested a primary infection with coxsackievirus A9 and a recurrent HSV-2 infection. HSV-1 was isolated from specimens of cerebrospinal fluid from two of four adults with HSV encephalitis.


Abnormalities of cholesterol-phospholipid composition in platelets and low-density lipoproteins of human hyperbetalipoproteinemia. Platelets from most patients with type IIa hyperlipoproteinemia (IIa) aggregate in the presence of lower concentrations of epinephrine and adenosine diphosphate (ADP) than are necessary to aggregate normal platelets. We have observed a comparable functional alteration in human platelets made cholesterol-rich in vitro by incubation in a milieu artificially rich in free cholesterol relative to phospholipid. We therefore examined platelet aggregation and lipid composition of platelets and of plasma low-density lipoprotein (LDL) in 19 individuals with IIa (including three homozygotes), seven normolipidemic individuals with symptomatic, angiographically-documented coronary atherosclerosis (atherosclerosis group), and 23 asymptomatic, normolipidemic subjects (control group). More than 99 percent of platelet cholesterol was unesterified. There was a 7 percent increase in the cholesterol content of whole platelets per mole of platelet phospholipid (C/PL) in IIa as compared to normal controls. This resulted from a 22 percent increase in the C/PL of IIa platelet membranes with no change in the C/PL of the soluble or granule fraction. The C/PL of IIa platelets was 6 percent greater than that of platelets from patients with atherosclerosis. As compared to those of normal controls, IIa platelets aggregated in response to a ninefold lower concentration of epinephrine (p less than 0.001) and a twofold lower concentration of ADP (p less than 0.02). The response of atherosclerosis platelets to these agents was comparable to that of controls. In all groups, there was a negative correlation between the log concentration of epinephrine required to produce complete platelet aggregation and the platelet C/PL (r = -0.06; p less than 0.002). The composition of LDL isolated from the plasma of patients with IIa was characterized by a 39 percent increase in the amount of free cholesterol relative to protein and a 35 percent increase in C/PL, as compared with control LDL. These values were increased 23 and 19 percent, respecitvely, when IIa was compared with the atherosclerosis group. In all groups the C/PL of LDL correlated well with the C/PL of platelets (r = =0.61; p less than 0.001). However, a simple cause-and-effect relationship did not appear to exist since (1) erythrocyte membrane C/PL was not affected and (2) normal platelets or erythrocytes underwent no change in C/PL during 18 hours' incubation in IIa plasma. These studies demonstrate that LDL and platelets in IIa contain an increased amount of free cholesterol relative to its principal solubilizer, phospholipid. In platelets this correlates with an increased sensitivity to aggregating agents. Moreover, the similarity between the functional abnormality in IIa platelets and that previously observed in normal platelets made cholesterol-rich in vitro suggests that the lipid composition of platelet membranes may have a direct effect on the function of platelets in man.


Heterotopic cervical salivary glands.

Heterotopic cervical salivary glands. A four-year-old girl presented with bilaterally symmetrical cutaneous orifices situated at the junction of the middle and lower thirds of the sterno-cleido-mastoid muscle. The preoperative diagnosis was lateral cervical, or branchial, sinuses. At operation the sinus tracts were found to extend only 2 cm. deep to the skin surface, and histological examination revealed mixed salivary glands. The literature is reviewed and the differential diagnosis of sinus orifices in the lower, anterior part of the neck is discussed.


Enzymic synthesis of ether types of choline and ethanolamine phosphoglycerides by microsomal fractions from rat brain and liver. The formation of product by ethanolamine phosphotransferases (EC 2.7.8.1) and cholinephosphotransferases (EC 2.7.8.2) in microsomal fractions from brains and livers of mature rats is increased several fold by 1,2-diacyl-sn-glycerols. With the addition of 1-alkyl-2-acyl-sn-glycerols, we have found an 11-fold increase with brain microsomes and a 20-fold increase with lvier microsomes in the synthesis of choline ether lipids (1-alkyl-2-acyl- and 1-alk-1'-enyl-2-acyl-sn-glycero-3-phosphorylcholines). For the synthesis of ethanolamine ether lipids (1-alkyl-2-acyl and 1-alk-1'-enyl-2-acyl-sn-glycero-3-phosphorylethanolamines), the stimulation of alkylacylglycerols was 7-fold for brain microsomes and 18-fold for liver microsomes. The alkylacyl glycerols (8 mM) also inhibited the synthesis of diacyl phosphoglycerides by 44 to 65%, indicating that the same ethanolaminephosphotransferases and cholinephosphotransferases are utilized for the synthesis of alkylacyl phosphoglycerides and diacyl phosphoglycerides. A desaturation of the alkyl groups may take place in the same reaction mixture. The rate of incorporation of phosphorylcholine into alkenylacyl glycerophosphorylcholines (choline plasmalogens) with alkylacylglycerols, cytidine diphosphate choline, and liver microsomes was 15 nmoles per mg protein per hour. The in vitro synthesis of choline plasmalogens with alkylacylglycerols had not been observed previously. The corresponding rate of incorporation of phosphorylethanolamine into ethanolamine plasmalogens was 10 nmoles per mg protein per hour, a value greater than any of the previously reported values for ethanolamine plasmalogen formation from alkylacyl glycerophosphorylethanolamines.


Analysis of rat serum apolipoproteins by isoelectric focusing. I. Studies on the middle molecular weight subunits. Analytical isoelectric focusing (IEF) has been applied to the study of the apolipoprotein components of rat serum high density and very low density lipoproteins. The apolipoproteins were separated on 7.5% polyacrylamide gels containing 6.8% urea, with a pH gradient of 4-6. The middle molecular weight range apolipoproteins were identified on IEF gels by the use of apolipoproteins purified by electrophoresis on gels containing sodium dodecyl sulfate (SDS). The A-1 protein focused as 4 to 5 bands from pH 5.46 to 5.82; the A-IV protein and the arginine-rich protein each focused as 4 to 6 bands from pH 5.31 to 5.46. The low molecular weight proteins focused from pH. 4.43 to 4.83 and are the subject of a separate communication. Comparisons of the IEF method with SDS gel electrophoresis, polyacrylamide gel electrophoresis in urea, and Sephadex chromatography are also reported. Additional studies were also carried out that tend to rule out carbamylation or incomplete unfolding of the proteins in the presence of urea as the causes of the observed heterogeneity.


Hyperkalemia unresponsive to massive doses of aldosterone and renal tubular acidosis in a patient with chronic interstitial nephritis: clinical and experimental studies. A unique 53-year-old male patient is described in whom aldosterone-refractory hyperkalemia and renal tubular acidosis/RTA/ was due to chronic interstitial nephritis associated with peritubular hyaline deposits in the distal nephron. Hyperkalemia was not caused by an adrenal disorder or acidosis and could not be abolished by interventions enhancing K clearance; saline infusions, high doses of furosemide, cortisone, cortisol, long-acting synthetic ACTH and excessive doses of aldosterone. Glucocorticoids induced a marked decrease in sodium excreting capacity probably by an action on the ascending limb of Henle's loop while aldosterone elicited a paradoxical natriuretic response by unknown mechanism. The results of our experimental studies carried out on the hyperkalemic RTA patient as well as on various control subjects and patients suggest 1. a specific defect in renal K excretion associated with decreased aldosterone responsiveness of the tubules presumably due to the peritubular pathology, and 2. a disturbance in the cellular regulation of K distribution between fluid compartments of unknown origin.


Output of oestrogens, testosterone and their precursors by isolated human adrenal cells as compared with that of glucocorticosteroids. The output of oestrogens, testosterone and their precursors was compared with that of glucocorticosteroids under standardized conditions, in a suspension of isolated human adrenal cells. Cortisol, corticosterone, androstenedione, dehydroepiandrosterone and its sulphate all increased in the same proportions after ACTH stimulation. The response to the logarithm of ACTH concentrations had a sigmoid shape but was fairly linear between 5 and 100 to 1000 muu./ml. The output of dehydroepiandrosterone plus that of its sulphate was of the same order of magnitude as the production of cortisol; the output of free dehydroepiandrosterone averaged half that of the sulphate indicating that the adrenal cortex is capable, under certain conditions, of producing large amounts of the free steroid. The output of androstenedione was very low, on average 35 times lower than that of cortisol, suggesting by extrapolation that the adrenal secretion may not be the main source of androstenedione in vivo or that ACTH is not the unique stimulus to adrenal androstenedione secretion. The output of testosterone was small to negligible and that of oestrogens was practically absent. In three additional experiments the influence of prolactin, prostaglandins, FSH and HCG was explored: no selective stimulation of androgen or oestrogen output was observed except in one experiment in which HCG stimulated adrenal testosterone production.


Testosterone secretion by foetal rat testes in vitro. Testosterone secretion by foetal rat testes (13 1/2-21 1/2 days of gestation) explanted for 3 days in a synthetic medium was measured every 24 h by radioimmunoassay. During the first day of explantation, the foetal testis produced, respectively, 1013 +/- 132, 8734 +/- 1118, 9179 +/- 2185 and 3886 +/- 309 (S.E.M.) pg/testis when explanted at 14 1/2, 16 1/2, 18 1/2 and 21 1/2 days respectively. Testosterone production by 13 1/2-day-old testes was not detectable on the first day of culture, but appeared on subsequent days. Daily testosterone secretion increased on the 2nd and 3rd days of culture in 14 1/2-day-old testes and decreased in older stages. These results suggest that the functional differentiation of the testis is independent of stimulatory factors like gonadotrophins. Dibutyryl cyclic AMP was found to stimulate testosterone production significantly from 14 1/2 days of gestation onwards.


Adrenocortical function in a prototherian mammal, Tachyglossus aculeatus (Shaw). The peripheral plasma concentrations and production rates of corticosterone and cortisol were measured in the conscious, unrestrained echidna (Tachyglossus aculeatus) under basal conditions and during maximal ACTH stimulation. Using Sephadex LH-20 column chromatography and radioligand assay, only cortisol and corticosterone could be detected in the peripheral blood plasma at very low concentrations of 0-07 +/- 0-03 (S.E.M.) mug/100 ml and 0-14 +/- 0-07 mug/100 ml respectively. Two-hourly sampling over periods of 36-48 h disclosed a diurnal periodicity in the combined plasma concentration of these corticosteroids, the high concentrations corresponding to periods of behavioural activity. Marked, short-term fluctuations in plasma corticosteroid concentration were also observed during periods of more frequent (20 min) sampling. Constant rate i.v. infusion of synthetic ACTH increased the plasma concentrations of both steroids to maximal values of 0-42 +/- 0-23 mug cortisol/100 ml and 1-06 +/- 0-56 mug corticosterone/100 ml at infusion rates of 1 i.u. ACTH/kg/h. This is approximately 1/160 of the potency of this ACTH in man. The production rates of corticosterone and cortisol, measured by isotope dilution during constant rate i.v. infusion of 3H-labelled tracers, were only 0-35 +/- 0-21 and 0-56 +/- 0-26 mug/kg/h respectively during saline infusion, and increased to 2-86 +/- 3-47 and 2-74 +/- 2-07 mug/kg/h during the infusion of 1 i.u. ACTH/kg/h. The metabolic clearance rate of cortisol was greater than that of corticosterone and both were depressed by ACTH. Plasma corticosteroid concentrations were increased after surgery during ether anaesthesia and in sick animals with heavy worm infestation. It is concluded that the adrenal cortex of echidnas responds to ACTH stimulation and stress in a similar way to eutherians, but the level of activity is much lower.


Oestrogen-induced uterine protein: presence of associated phosphoprotein phosphatase activity before oestrogen treatment. The phosphoprotein phosphatase (PPPase) activity that is found associated with oestrogen-induced protein(s) is not itself induced by oestrogen. Indeed, this activity was found at the same electrophoretic position (i.e. the position of the oestrogen-induced protein) and in equal amounts relative to the total PPPase activity, whether the cytosol fraction from control or from stimulated uteri was tested.


In-vitro studies of normal human thyroid cells: responses to thyrotrophin and dibutyryl cyclic AMP. Follicular cells isolated from normal human thyroid tissue have been cultured for up to 140 h with bovine thyrotrophin (TSH) or dibutyryl cyclic AMP (DBcAMP). Both compounds induced marked reorganization of the cells into three-dimensional follicular structures, whilst non-supplemented cells assumed a monolayer form. Cultures treated initially with TSH or DBcAMP showed a greater iodide uptake capacity, in comparison with unsupplemented cultures, in which iodide uptake was markedly diminished after 24 h. The release of tri-iodothyronine (T3) and thyroxine (T4) into the medium was determined by radioimmunoassay. Both TSH- and DBcAMP-treated cells showed a significant increase in iodothyronine output compared with unsupplemented control cells. In contrast to the "classical" TSH-induced depression of the T4:T3 ratio in vivo, an increase in the ratio was observed for both TSH- and DBcAMP-supplemented cells in vitro. The ratio was also significantly greater after TSH than after DBcAMP, and possible implications of this findings are discussed.


The structure of the mitochondrial cloud of Xenopus laevis oocytes. The ultrastructure of the mitochondrial cloud (Balbiani body) of the pre-vitellogenic oocytes of Xenopus laevis has been examined using transmission and stereoscan electron microscopy. Examination of conventional thin sections confirm previous observations which suggest that the cloud consists essentially of many thousands mitochondria and numerous small vesicles; larger clouds, in oocytes greater than 200 mum in diameter, contain relatively more vesicles. Using a standard electron microscope at 100 kV very long and coursing arrays of mitochondrial profiles can be detected. The presence of very long mitochondrial elements has been confirmed using a high voltage microscope operating at 500-1000 kV. Stereoscan preparations, isolated from pre-vitellogenic oocytes, lend some support to the view that the mitochondrial cloud amy consist of a mass of long filamentous mitochondria and the possibility that there are large continuous regions of mitochondrial material cannot be ruled out.


Complement-mediated antiserum cytotoxic reactions to human chromosome 7 coded antigen(s): immunoselection of rearranged human chromosome 7 in human-mouse somatic cell hybrids. Immunoselection via complement-dependent lysis of human-mouse somatic cell hybrids containing chromosome 7, with antisera reactive to cell surface antigen(s) coded for by chromosome 7, has resulted in growth of somatic cell hybrids containing rearranged human chromosome 7s. Investigation of these hybrids has localized the gene(s) coding for the relevant cell surface antigen(s) to the short arm of human chromosome 7. The simian virus 40 integration site and the gene coding for human beta-glucuronidase appear to be localized to the long arm of chromosome 7 in this hybrid clone.


Thymus-like activities of sulphur derivatives on T-cell differentiation. Levamisole and sodium diethyldithiocarbamate can induce in vivo thymocyte differentiation from precursor spleen cells of nu/nu mice and evoke indirect plaque-forming cells in nude mice immunized with sheep red cells. These sulphur drugs induce in thymusless mice the production of a serum factor which transfer in vivo immune enhancement and in vitro thymocyte differentiation. In vivo treatment with sulphur derivative can substitute for an alleged thymice hormone.


Plasmid modification of radiation and chemical-mutagen sensitivity in Pseudomonas aeruginosa. The R factor pMG2 protects Pseudomonas aeruginosa against the lethal effects of ultraviolet (u.v.) and gamma irradiation, and methyl methanesulphonate and N-methyl-N'-nitro-N-nitrosoguanidine treatment. Enhanced survival occurs in strains of uvr+ rec+ (wild-type) genotype and a variety of uvr rec+ type mutants. No protection occurs in a rec A-type mutant. The plasmid also enhances u.v.-induced mutagenesis. These effects appear to be due to host-cell controlled plasmid-determined DNA repair function(s). Studies on P. aeruginosa strains deficient in DNA polymerase I (polyA) suggest that a plasmid-determined repair resynthesis function may be responsible for increased u.v.-survival and enhanced u.v.-mutability in pMG2-containing bacteria.


Vesicular stomatitis virus: mode of transcription. Recent studies on the mechanism by which the virion-associated RNA polymerase of vesicular stomatitis virus transcribes RNA have revealed several new biological features of general interest. The mode of synthesis of the 5'-terminal cap structure of the mRNAs, the sequential transcription of the genes and the presence of a transcribed "leader" RNA segment are properties which are either not shown by other viruses, or have not yet been described. These features are probably inter-related with the primary transcription process, which itself may be a useful model for future studies on mRNA biosynthesis in eukaryotic systems.


A morphological study of the M-protein of Sendai virus. A purification scheme is described for the M-protein of Sendai virus and an electron microscope study of the isolated protein is presented. The protein exists as subunits of 6 nm in diam., which possess a central hole; the subunits may be dimers of the polypeptide. They are able to form filamentous aggregates which wind around one another to form a helical structure. It is suggested that these filaments may be the form adopted by the protein in the virus, the filaments lying parallel to one another just beneath the virus membrane to form a shell, but that the helical form is likely to be a property only of the isolated protein.


Processing of a pseudorabies virus-induced protein which is glycosylated, sulphated and excreted. Cells infected with pseudorabies virus excrete large amounts of a glycosylated sulphated protein, mol. wt. 89000, into the extracellular fluid. This paper reports the results of studies on the processing of this protein. Glycosylation occurs during, or very soon after, synthesis of the polypeptide chain. After a delay of several minutes the glycoprotein is sulphated; inhibition of glycosylation by high concentrations of glucosamine does not interfere with this process. The glycosylated sulphated polypeptide is then reduced in size from mol. wt. 99000 to 89000, possibly by proteolytic cleavage, and is excreted. Inhibition of glycosylation does not interfere with the excretion of this polypeptide, which is an energy-requiring process.


Aminoacylation of encephalomyocarditis virus RNA. RNA extracted from purified encephalomyocarditis (EMC) virus (EMC-RNA) can be aminoacylated with synthetase praparations from Escherichia coli, beef and rabbit liver. The extent of aminoacylation is between 0-024 and 0-080 moles per mole EMC-RNA and occurs only with serine. Either removal of possible low mol. wt. contaminants with 3 M-sodium acetate nor periodate oxidation of the virus RNA affects its aminoacylation capacity.


A haemagglutination assay for the primate syncytium-forming (foamy) viruses. Preparations of highly concentrated simian foamy virus type 1 (SFV1)agglutinate guinea pig red blood cells. The agglutination occurs both at 22 degrees and 37 degrees C. It is inhibited by specific antisera against SFV1 but also by antisera prepared against several other types of simian foamy viruses.


Effects of pyrimidine derivatives on RNA-dependent RNA polymerase of mengovirus-infected Fogh and Lund (FL) cells. Two newly synthesized pyrimidine derivatives were found to possess antiviral activity against Mengovirus in Fogh and Lund (FL) cells and in a cell-free system. The inhibitory effect on RNA-dependent RNA polymerase of Mengovirus-infected FL cells was assayed using 14C-UTP as precursor. Addition of 50 or 100 muM of the inhibitors in a cell-free system of crude enzyme and nucleoside triphosphate medium for 60 min incubation at 37 degrees C resulted in about 40 to 60% lower counting rates for drug-treated reaction mixtures. The analysis of the polymerase synthesis product (virus RNA extracted from the cell-free reaction mixture and deproteinized by the phenol-SDS method) was carried out by means of agarose-acrylamide gel electrophoresis. The main finding was a reduction of single-stranded Mengovirus RNA (RNase-sensitive and LiCl-precipitable). The rates of synthesis of the replicative intermediate (LiCl-precipitable) and the replicative form of RNA (LiCl-soluble) were not significantly influenced.


Sequential immunofluorescence and infectivity studies on the replication of Herpesvirus saimiri in owl monkey kidney cells. Methods are described for the preparation and authentication of a highly specific antiserum against herpesvirus saimiri (HVS) capsid antigens. The antiserum was used in immunofluorescence tests to follow the development of capsid antigens in HVS-infected owl monkey kidney cells throughout the virus replication cycle in parallel with sequential titrations of virus infectivity in both cells and medium. Fluorescence was detected as a round or oval, bright green area of staining at the centre of the nucleus which was similar in outline to the Cowdry type A inclusion seen in HVS-infected cells stained by haematoxylin and eosin. The first detection of fluorescence towards the end of the eclipse phase of the virus growth cycle, and its abolition by the treatment of infected cultures with cytosine arabinoside confirmed the identity of HVS capsid antigens as late antigens. The failure to detect fluorescence in the cytoplasm of HVS-infected cells has brought to light a conflict between the site of accumulation of virus capsid antigens as determined by immunofluorescence and the finding, by electron microscopy, of cytoplasmic immature particles in intact cells during the early stages of the virus replication cycle. The significance of this discrepancy is discussed in relation to its possible existence for other members of the herpesvirus group.


Protection of mice against encephalomyocarditis virus infection by preparations of transfer RNA. Preparations of bacterial transfer RNA (tRNA), give dose-dependent protection of mice against encephalomyocarditis (EMC) virus infection at up to I mg tRNA per mouse with maximum response when the tRNA is administered around 6 h before infection. Protection occurs with intraperitoneally and intravenously administered tRNA against infections by both these routes. In some experiments significant protection occurs by single treatments of tRNA up to 24 h after infection with virus doses of I X LD100. Some tRNA preparations of eukaryotic origin do not give significant protection. Protection is not a feature of all species of bacterial tRNA; partially purified valine, tyrosine and phenylalanine tRNAs from Escherichia coli are not protective. tRNA treatment does not induce circulating interferon nor does it 'hypo-reactivate' the protective effect of poly (I).poly (C) treatment of mice. Humoral and cell mediated immune responses do not seem to be involved in tRNA mediated protection since first, cytosine arabinoside treatment does not affect protection by tRNA; second, serum from mice treated with tRNA and an EMC vaccine does not protect other mice against infection, and third, mice that survive normally lethal infections as a result of tRNA treatment are generally just as susceptible to re-infection as previously untreated, uninfected mice. Silica treatment abolishes protection of mice by tRNA implying that macrophages are necessary. However, tRNA does not seem to act by clearance of virus particles since vaccination of mice by inactivated EMC virus is not affected by tRNA treatment. These results are considered in relation to the presence of a tRNA-like structure in EMC virus RNA and protection of mice by other single stranded polynucleotides.


Biochemical analysis of a virulent and an avirulent strain of foot-and-mouth disease virus. A comparison has been made of some of the serological and physicochemical properties of a virulent and an avirulent strain of foot-and-mouth disease virus, serotype SAT1. The avirulent strain (SAT1-82) was derived from the virulent strain (SAT1-7) by serial passage in BHK 21 cells. The viruses were indistinguishable in cross-neutralization tests. In immunodiffusion tests a clear spur line was obtained with the SAT1-82 antiserum but not with SAT1-7 antiserum. The major polypeptides of the two viruses were identical when examined by polyacrylamide gel electrophoresis. Hybridization and thermal denaturation experiments failed to distinguish between the RNAs but two-dimensional electrophoresis of the oligonucleotides produced by ribonuclease T1 digestion revealed several differences. Possibly the most significant of these differences was the size of the polycytidylic acid [poly (C)] tract. There were about 170 nucleotides in the poly (C) tract of the SAT1-7 RNA compared with around 100 in the SAT1-82 RNA. Further evidence for this deletion was provided by the slightly different behaviour of the two RNAs when compared by sucrose gradient centrifugation and polyacrylamide gel electrophoresis.


Method for quantitative estimation of thermal thresholds in patients. A quantitative method for the examination of thermal sensibility was applied in 26 normal subjects and in patients with various neurological disorders. The stimulation technique resembled Békésy audiometry: the patient reversed the direction of the temperature change of a thermode whenever warm, cold, or thermal pain thresholds were reached. The resulting temperature curve enables a quantitative description of the subject's thermal sensibility and of the degree of impairment displayed by neurological patients.


Dysautonomia in Parkinsonism: a clinicopathological study. Necropsy studies were done on six patients with idiopathic paralysis agitans, one with multiple system atrophy including features of Parkinsonism, and one control. Autonomic functions had been evaluated during life to a varying degree. Intra-arterial blood pressure studies were carried out on two patients with paralysis agitans (cases 4 and 6) and the one with multiple system atrophy (case 7). Lewy bodies with or without cell loss were seen in the sympathetic ganglia of five cases of paralysis agitans. Three of these had orthostatic hypotension and the severity of the lesion approximately correlated with the degree of hypotension. It is concluded that the lesions of the sympathetic ganglia may play a major role in the production of orthostatic hypotension in idiopathic paralysis agitans.


Studies on cyclic AMP in different compartments of cerebrospinal fluid. Adenosine 3', 5'-monophosphate (cAMP) was measured in the CSF of 42 patients undergoing radiological investigation, neurosurgical procedures, or investigation of hepatic coma. The concentration of cAMP was significantly higher in ventricular CSF than in lumbar CSF. Premedication with pentobarbitone plus promethazine increased cAMP in lumbar CSF. There was no difference in cAMP concentration in lumbar CSF obtained before or after injection of air or after the administration of diazepam during lumbar pneumoencephalography. Lumbar CSF cAMP concentration was significantly increased in patients in hepatic coma. The concentration of cAMP in the lateral ventricle was not affected by general anaesthesia or by the presence of a complete block of the aqueduct of Sylvius. There was no decrease in lumbar CSF cAMP in patients with a complete stenosis of the aqueduct of Sylvius, partial blocks of CSF flow at the cervical level, or a complete block at the lower thoracic level. The concentration of cisternal CSF cAMP was similar to that of lumbar CSF. These results suggest that (1) there is a ventriculolumbar gradient in the concentration of cAMP but of insufficient magnitude to be detected by mixing of lumbar and ventricular CSF during pneumoencephalography, (2) lumbar CSF cAMP concentration is not dependent on brain as a source of this nucleotide; the source of this nucleotide may be largely derived from the spinal cord, (3) premedication may affect the concentration of cAMP in lumbar CSF cAMP, (4) the formation of cAMP is unimpaired in hepatic coma.


Anticonvulsant prolongation of survival in adult murine lymphocytic choriomeningitis. II. Ultrastructural observations of pathogenetic events. Because previous ultrastructural studies of murine lymphocytic choriomeningitis (LCM) had revealed only mononuclear cell infiltration with no cytopathology of target cells in the choroid plexus, ependyma, and leptomeninges, diazepam treatment was used to prolong survival for characterization of late pathogenetic events. Mice which were treated with diazepam and sacrificed 8, 9, and 10 days after intracerebral inoculation with LCM virus showed an increasing amount of inflammatory infiltration into choroid plexuses, leptomeninges, Virchow-Robin spaces, and ependyma. Mononuclear cells, lymphocytes, and polymorphonuclear (PMN) leukocytes increased in number as compared with terminally infected mice sacrificed 7 days after inoculation. Ultrastructurally, choroidal epithelial cells showed cytopathological changes varying from dilated endoplasmic reticulum through necrosis. Greater numbers of PMN leukocytes, macrophages, and activated macrophages and fewer undifferentiated mononuclear cells were seen in choroid plexuses of the drug-treated survivors. Virions and larger, more numerous arenavirus inclusions were present in choroid plexus and ependyma. Ultrastructurally the leptomeningitis was characterized by large numbers of activated macrophages. Choroidal epithelial necrosis appears to be the in vivo correlate of T-cell-mediated cytotoxicity in vitro.


The ethnic distribution of primary central nervous system tumors: AFIP, 1958 to 1970. A retrospective ethnic study was made of 16,311 cases of primary central nervous system (CNS) tumors seen at the Armed Forces Institute of Pathology (AFIP), Washington, D.C., from 1958 to 1970. Results showed a considerably higher Caucasian:Negro (C:N) case ratio (13.7:1) than the C:N population ratio (8.4:1), indicating a higher relative frequency of primary CNS tumors in American Caucasians as compared to American Negroes. The glioma was significantly more frequent in Caucasians than in Negroes (p less than 0.005). In contrast, Negroes had an excess of the pituitary adenoma as compared to Caucasians (P less than 0.01). The proportional frequencies of the meningioma and the nerve sheath tumor were also higher in Negroes than in Caucasians. When this pattern of the tumor distribution of American Negroes was compared to that of African Negroes (a composite African series), the preponderance of the pituitary adenoma and the meningioma and the relative paucity of the glioma in the Negro race as compared to Caucasians were again confirmed. The differences in the relative frequency and the tumor distribution between American Negroes and Caucasians and the considerable similarity of the tumor distribution between American and African Negroes emphasize the importance of genetic factors in the development of at least some primary CNS tumors.


235 cases of excessive daytime sleepiness. Diagnosis and tentative classification. A series of 235 consecutive patients refferred to the Stanford University Sleep Disorders Clinic with the complaint of excessive daytime sleepiness (EDS) were investigated extensively. A satisfactory final diagnosis involving a consistent syndrome or pathogenic process was made in all but 7 patients. In the course of this work a variety of tests, including prolonged polygraphic monitoring of multiple variables and CSF measurements before and after probenecid ingestion, were utilized. Different syndromes were confirmed (harmonious hypersomnia, subwakefulness syndrome); the definitions of others were clarified and extended (narcolepsy, drug dependency, periodic hypersomnia associated with menstruation, upper airway sleep apnea in children). Two new entities were tentatively identified (narcolepsy with sleep apnea, the neutral state syndrome). Narcolepsy and upper airway sleep apnea accounted for the majority of the cases (199). A strategic schema utilizing specific categories and frequency of occurrence in the case series is presented to improve the diagnosis of the complaint of excessive daytime sleepiness by the practicing physician. This case series was analysed in order to develop tentatively a meaningful nosology.


Intracisternal A and bar-shaped particles in murine neuroblastoma C 1300. Transplantable murine neuroblastoma C 1300 was studied ultrastructurally at varying time intervals ranging from 2 hours to 40 days before and after X-irradiation. Following X-irradiation, 2000 and 4000 rads in a single dose, the uniformly small tumor cells became progressively enlarged multinucleated and degenerated, starting at one to two days. At five to seven days, the uni- and multinucleated giant cells predominated over the small tumor cells, while the giant cells progressively disappeared therafter and the small tumor cells predominated over the giant cells at 10 to 14 days. The giant cells contained abundant subcellular organelles and the X-irradiated tumor cells apparently continued to produce the organelles until they degenerated. Two types of cytoplasmic particles, intracisternal A and bar-shaped, were observed in the tumor cells. The intracisternal A particles occurred in almost all non-irradiated tumor cells though their number varied considerably from cell to cell, while they were observed less frequently in the radiation-induced giant cells probably due to a dilution effect rather than an actual numerical decrease. The bar-shaped particles, hitherto undescribed in the neuroblastoma, were 23 nm in diameter, variable in length and occasionally tubular. They occurred only in degenerating cells regardless of X-irradiation but were encountered more frequently in irradiated tumors than in non-irradiated ones. It is suggested that they may represent an unknown degenerative product of cytoplasm and/or nucleus rather than virus particles, despite their morphological resemblance to certain virus particles.


Elevated parainfluenza virus type 1 antibody in patients with subacute sclerosing panencephalitis. Parainfluenza virus hemagglutination inhibition (HI) antibodies were determined 3 times in the sera of 9 patients with subacute sclerosiing panencephalitis (SSPE) and 20 healthy controls matched for age and place of residence. Serum antibody against parainfluenza virus type 1 was significantly elevated in SSPE patients as compared with controls, whereas antibodies against type 2 and 3 were found to be in normal ranges. Higher titres of parainfluenza virus type 1 antibody might depend on: (1) dual viral infection, (2) cross-reaction between antigens of SSPE virus and parainfluenza virus type 1, and (3) non-specific activation of latent virus type 1 genome. The latter explanation seems to be particularly interesting since the parainfluenza type 1 antibody titres remained constant despite the clinical progression. This finding is comparable to the elevated titres against Epstein-Barr virus of adenovirus which have been found occasionally in this disease.


Functional organization of catfish retina. 1. The basic organization of the biphasic (or concentric) receptive field is established in the bipolar cells as the result of an interaction between two signals, one local representing the activity of a small number of receptors, and the other integrating (19, 20) or global (28) coming from the S space or a lamina formed by the horizontal cells (8, 14, 22, 29). 2. Bipolar-ganglion cell pairs are segregated into two types; A (on center) and B (off center) pairs. A depolarization of a bipolar cell produces spike discharges from ganglion cells of the same type and a hyperpolarization depresses their discharges. I haven't detected any cross talk between the types A and B pairs. Bipolar and ganglion cells must be interfaced by the classical chemical synapses, the only such kind in the catfish retina. 3. Horizontal and type N neurons form two lateral transmission systems, one distal and the other proximal (19, 20). Signals in the lateral systems are shared by the two receptive-field types and are not excitatory or inhibitory in themselves; it is incumbent upon the postsynaptic neurons to decide the polarity of the synaptic transmission. The horizontal cell participates directly in the formation of biphasic receptive fields of bipolar cells by providing their surrounding, whereas type N neuron seems to modify the receptive-field organization established in the bipolar cells. 4. Type N neurons are amacrine cells because they do not produce spike discharges (2, 18, 21) and because they influence the activity of both A and B receptive fields. 5. The function of the type C neuron is as unique as its structure (21) and is not fully clear as yet. It is not a conventional amacrine cell as the type N appears to be, nor is it a classical ganglion cell which forms either a type A or B receptive field (2). 6. Type Y neurons are a class of ganglion cells which forms either a type A or B receptive field.


Neuropathy in hepatic disorders. A clinical, electrophysiological and histopathological appraisal. The present study deals with 30 patients with cirrhosis of the liver and 12 patients with infective hepatitis who were studied clinically, neurophysiologically and histopathologically for the presence of neuropathy. Simultaneously, 13 healthy individuals were evaluated as controls. Clinical evidence of neuropathy was found in 63.3% of the patients with hepatic cirrhosis and in 16.6% of the patients with infective hepatitis. In hepatic cirrhosis, the conduction velocities were abnormal in 33.3% and histopathological demyelination was found in 80% of the patients. In infective hepatitis, on the other hand, altered nerve conduction velocities were found in 41.6% and segmental demyelination in 75% of the patients. Our data reveal that peripheral nerve involvement is seen both in chronic and acute liver disorders. The neuropathy in hepatic cirrhosis is unrelated to diabetes, alchoholism or portacaval shunt and may be due to unknown metabolic abnormality or to toxins. In infective hepatitis, the neuropathy may either be due to some acute metabolic derangement or may be purely viral in origin.


Physiological responses of human adults to foods containing phosphate additives. An experiment was conducted to assess the physiological effects of a diet rich in foods cotaining phosphate additives. During a 4-week control period, eight adults were fed a balanced diet free of phosphate additives providing approximately 95 g protein 0.7 g Ca and 1.0 g P per day. During a subsequent 4-week period, food items containing phosphate additives were substituted for counterpart items devoid of added phosphates. This diet contained 0.7 g Ca and 2.1 g P per day. The introduction of foods containing phosphate additives was associated with intestinal distress, soft stools or mild diarrhea. These symptoms subsided in six subjects but occurred intermittently throughout the experimental period in the other two subjects. The high-phosphorus diet induced increases in serum phosphorus and urinary phosphorus and decreases in serum calcium and urinary calcium. Hydroxyproline excretion in the urine was increased and cyclic AMP excretion was elevated in six of the eight subjects. These changes are analogous to those seen in experimental animals fed high-phosphorus diets which were shown to be due to enhanced parathyroid activity (secondary hyperparathyroidism). The use of phosphate food additives is discussed with respect to their possible stimulating effect on adult bone resorption.


Relative activities of some metabolites and analogs of cholecalciferol in stimulation of tibia ash weight in chicks otherwise deprived of vitamin D. Nine metabolites and analogs of cholecalciferol (CC) were tested for ability to increase tibia ash weight in chicks otherwise deprived of vitamin D. All of the compounds promoted bone mineralization in a linear log dose-response relationship. The maximal response obtained for any compound was an approximate doubling in bone ash weight compared to vehicle-treated controls. Relative potencies, based upon the calculated ash weight doubling dose, were as follows: 1 alpha, 25-(OH)2-CC = 1 alpha-OH-CC greater than CC greater than 25-OH-CC greater than 24R, 25-(OH)2-CC = 1 alpha,24R, 25-(OH)3- CC greater than 5,6-trans-25-OH-CC greater than 1 alpha, 24S, 25- (OH)3-CC greater than 5,6-trans-CC greater than 24S, 25-(OH)2-CC.


Partial characterization of molecular species of retinol-binding protein found in tubular proteinuria due to chronic cadmium poisoning in the rabbit. A study was conducted to characterize the molecular species of retinol-binding protein (RBP) isolated from the urine of rabbits chronically poisoned with cadmium. The RBP species, identical with regard to both molecular size (approximately 20,000) and immunoreactivity, were separated into four fractions by means of polyacrylamide gel electrophoresis (PAGE), which yielded two holo-RBP (H2 and H1) and two apo-RBP (A2 and A1) species. The urinary excretion ratio of these fractions (H2 : H1 : A2 : A1) was found to be about 70 : 6 : 21 : 3. No distinct difference of amino acid composition between holo- and apo-RBP was observed. An additional species of apo-RBP (designated An) was also isolated from theaged holo-RBP (H2) by isoelectric focusing in gel. Using the separated molecular species of rabbit RBP as well as of human RBP, their interactions with human prealbumin (PA) were examined both by human PA-Sepharose affinity chromatography, and by gel filtration on Sephadex G-100 after in vitro incubation of the RBP with human PA. Purified rabbit holo-RBP exhibited almost the same binding ability to human PA as did human RBP. Retinol within RBP molecule enhanced the affinity to PA, resumably through the change of tertiary structure, although the presence of retinol was not essential for the protein-protein interaction.


Synthesis and biological activity of 5,6-trans-vitamin D3 in anephric rats. A modified procedure for the purification of 5,6-trans-D3 from a reaction mixture of vitamin D3 with iodine and its biological activity in sham-operated and anephric rats are described. When a solution of vitamin D3 in n-hexane was reacted with small quantities of iodine under influence of visible light, the reaction mixture gave four or five spots, including vitamin D3 and 5,6-trans-D3, on a thin layer chromatogram. After purifying the mixture by alumina column chromatography, a colorless oil from the separated 5,6-trans-D3 fractions was crystallized from n-hexane and snow white crystalline 5,6-trans-D3 was obtained. The purification method was thought to be better than the previously reported methods (14-16) because it gave good yield without the procedure of esterification using expensive p-phenylazobenzoyl chloride. A dose of 25 mug of 5,6-trans-D3 obtained thus gave significant elevation on intestinal calcium transport in both sham-operated and anephric rats, whereas the dose did not give positive effects on serum calcium levels in anephric rats. Serum phosphorous levels were extremely elevated by nephrectomy, but in both sham-operated and anephric rats they were unaffected by the administration of 25 myg of 5,6-trans-D3.


The effects of ascorbic acid on diphtheria toxin and intoxicated HeLa cells. Ascorbic acid (vitamin C) prevented diphtheria toxin from inhibiting the incorporation of [U-14C]-alanine into trichloroacetic acid precipitable material in HeLa cells. Ascorbic acid did not exhibit an effect on the adenosine diphosphate-ribosylation of amino acyl transferase II nor did it separate fragment A from fragment B in "nicked" toxin. A non-specific reducing agent, para-methylaminophenol sulfate, exhibited an effect on HeLa cells very similar to the results of ascorbic acid. Citric acid, a tricarboxylic acid, had no effect on HeLa cells.


An immunopathologic study of interstitial nephritis associated with experimental canine adenovirus infection. The localisation of canine adenovirus in the renal tubules induces a marked cellular response in the interstitium. A number of mechanisms appear to be involved in the genesis of the lesions. Direct lytic damage to tubular epithelial cells by the virus is probably augmented by immunological factors. The production of local anti-viral antibody by plasma cells in the vicinity of virus-containing cells may result in the formation of toxic antigen-antibody complexes while the accumulation of large populations of macrophages suggests that cell-mediated hypersensitivity is also operative.


Differential effects of the isomers of tetramisole on adrenergic neurotransmission in cutaneous veins of dog. Clinical observations indicate that dexamisole and levamisole, the isomers of tetramisole, cause mood elevation. Their effects on smooth muscle cells and adrenergic nerves were investigated in strips of dogs' saphenous veins. Dexamisole (2.5 X 10(-6) to 4 X 10(-5) M) augmented the contractile response to norepinephrine but depressed that to tyramine; cocaine inhibited the augmentation of the norepinephrine response. Levamisole (10(-5) M) did not alter the response to norepinephrine, but augmented that to tyramine. At 1.6 X 10(-4) M dexamisole, more than levamisole, depressed the responses to norepinephrine, tyramine and acetylcholine. Activation by K+ ions was not affected by the isomers. Preparations, incubated with 3H-norepinephrine, were mounted for superfusion, tension recording and determination of 3H-norepinephrine and metabolites in the superfusate. Dexamisole and levamisole augmented the 3H-norepinephrine overflow during nerve stimulation; levamisole decreased the efflux of deaminated metabolites. During tyramine-induced contractions, dexamisole depressed and levamisole augmented the efflux of 3H-norepinephrine; they reduced the appearance of metabolites. The increases in 3H-norepinephrine caused by the isomers during nerve stimulation were not seen after phenoxybenzamine. Dexamisole, more than levamisole, inhibited tissular uptake of 3H-norepinephrine. Levamisole, more than dexamisole, inhibited monoamine oxidase activity in vein homogenates. These interferences with release and disposition of norepinephrine may be related to the antidepressant properties of the tetramisole isomers.


The effect of tolbutamide on contractility and cyclic adenosine 3':5'-monophosphate concentration in the intact beating rat heart. The effect of tolbutamide on contractility and the concentration of cyclic adenosine 3':5'-monophosphate (cyclic AMP) in ventricular muscle was examined in the intact beating rat heart. The hearts were perfused in a nonrecirculated (Langendorff) fashion. Bolus injections of tolbutamide caused an increase in cardiac contractility. This increase in contractility was markedly inhibited when bovine albumin (3 g/100 ml) was present in the perfusing fluid. The increase in contractility caused by tolbutamide was not preceded by or associated with any change in the concentration of cyclic AMP in the ventricular muscle. Further studies utilizing a simultaneous injection of norepinephrine and tolbutamide demonstrated no significant effect of this combination on the concentration of cardiac cylcic AMP produced by an injection of norepinephrine alone. Our findings suggest that in the intact beating rat heart the positive inotropic effect of tolbutamide is not mediated via an increase in the concentration of cardiac cyclic AMP and that tolbutamide does not significantly potentiate the effect of catecholamines on cardiac cyclic AMP concentration.


Effects of carbamazepine on the water permeability and short-circuit current of the urinary bladder of the toad and the response to vasopressin, adenosine 3',5'-cyclic phosphate and theophylline. Carbamazepine has been reported to decrease urine output and increase urinary concentration in patients with diabetes insipidus. The effects of the drug on the osmotic water permeability of the bladder of the toad, Bufo marinus, were studied. Carbamazepine had no effect on osmotic water flow when present in the serosal or mucosal bathing media. The submaximal or maximal increase in osmotic water flow caused by vasopressin was inhibited by serosal carbamazepine concentrations as low as 0.01 mM. Higher concentrations of carbamazepine in the mucosal solution also inhibited the submaximal antidiuretic hormone (ADH) response. The response to 2 mM cyclic AMP was inhibited by 0.5 mM serosal carbamazepine. Carbamazepine did not affect the response to 20 mM theophylline. Significant inhibition of the ADH response occurred only after preincubation of the bladders with the drug for at least 1 hour. The inhibition was reversed by rinsing the drug from the bladders before ADH was added. The mechanism of action of carbamazepine in diabetes insipidus remains obscure. In the toad bladder, the drug neither directly increases osmotic water flow nor potentiates the response to vasopressin.


Spontaneous subminature end-plate potentials in mouse diaphragm muscle: evidence for synchronous release. 1. Miniature end-plate potentials (min.e.p.p.s) were recorded from small muscle cells of mouse diaphragms. Min.e.p.p. amplitude histograms showed successive peaks which were integral multiples of the smallest peak. The smallest potentials (submin.e.p.p.s) averaged 0-3-0-6mV and the mean of the larger min.e.p.p.s averaged 3-7 mV, depending on the muscle cell diameter. There was a positive correlation between time-to-peak and min.e.p.p. amplitude. Time-to-peak of the submin.e.p.p.s fell slightly below the regression line through the larger min.e.p.p.s. 2. Sometimes min.e.p.p. amplitude distributions changed spontaneously such that the mean of the major mode min.e.p.p.s decreased twofold during which time the mean of the submin.e.p.p.s did not change. Spontaneous decreases were most pronounced during low frequencies of release (10/min) achieved at 32 degrees C. 3. Small changes in temperature (2 degrees C steps in the range 32-40 degrees C) greatly altered the number of peaks of min.e.p.p. amplitude histograms without noticeably changing the position of the submin.e.p.p. peak. At 32 degrees C submin.e.p.p.s composed 5-20% of the histograms and the amplitude of the major mode peak was twelve to fifteen-times that of the submin.e.p.p.s. Over-all bell-shaped distributions were obtained at 37 degrees C which showed up to eight peaks with the major peak at the fourth to sixth peak. Temperatures slightly above 37 degrees C gave a flat distribution with the mean amplitude at the third peak. Min.e.p.p. amplitude histograms were initially skewed (mostly small min.e.p.p.s) after a 40 degrees C heat challenge. 4. Two to eight-times the normal concentration of Ca2+ in the saline reversibly increased the min.e.p.p. frequency and also decreased the mean of the major mode min.e.p.p.s (two to nine-times) without noticeably changing the mean of the submin.e.p.p.s. 5. Botulinum toxin A, 10(5) X intraperitoneal median lethal dose (10(5)I.P.LD50)/ml., almost abolished min.e.p.p.s in 30-90 min. The relative proportion of submin.e.p.p.s increased and the mean of the major mode min.e.p.p.s usually did not change during the initial decrease in frequency. Major mode min.e.p.p.s essentially ceased after 200-1000 were generated and remaining min.e.p.p.s of some cells showed skewed distributions with three small peaks that were integral multiples of the submin.e.p.p. peak. Smaller min.e.p.p.s were more resistant to block than the larger min.e.p.p.s and, although frequencies were low, small min.e.p.p.s were recorded after 4 hr of botulinum toxin incubation. 6. Colchicine (5 X 10(-4)M) within minutes reduced the major mode min.e.p.p.s by half (mean of major peak reduced to sixth or seventh peak). Additional colchicine (10(-3)M reduced the major mode min.e.p.p. amplitude to a fifth of that of control (mean of major mode min.e.p.p.s at the third peak) with no change in position of the submin.e.p.p. peak. Min.e.p.p. amplitudes slowly recovered to half control values after washing. 7...


Community mental health nursing: a social network approach. This paper describes an intervention technique that the psychiatric nurse may find useful in community mental health settings. This technique is called social network intervention. It attempts to combine the best of traditional clinical psychotherapeutic method with the advantages of a social systems approach for working with the patient in psychiatric distress. The technique is based on the assumption that the solution to a variety of human predicaments lies within the collective instrumental and affective resources of a patient's social network. The vehicle for accomplishing this is the "Network Session" during which a mental health clinician convenes the patient and his/her social network to help plan and resolve the difficulty. A Case Report demonstrating the use of the technique is included.


[Results of concentrated irridiation in glioblastomas, astrocytomas and brain tumours of the adult (author's transl)]. Concentrated irradiation (two series of 1,800 rads in 2 sessions and 3 days) separated by a rest period of 3 to 4 weeks has been used to treat 214 adult patients with brain tumors between 1965 and 1972. These included 108 glioblastomas, 18 astrocytomas and 88 brain metastases from different origins. Early side effects are moderate or inexistent provided synthetic ACTH or corticoid steroids have been given several days before irradiation. Survival is related, at least, in the first 2 groups of primary tumor to local failure. For brain metastases, death could be related to others reasons. In spite of the fact that comparison is made with a previous series of 121 cases it seems that this type of irradiation gives the same survival rates. Moreover it has specific advantages as reducing duration of hospitalization and the number of treatment sessions.


[Interstitial pneumonia in the immunodepressed child (author's transl)]. The authors stress the value of radiological examinations in the diagnosis of interstitial pneumonia in children with immuno-depression secondary to chemotherapy and/or radiotherapy. X-rays play a capital role in the early diagnosis of these conditions and may be of vital interest in some cases, in particular pneumocystosis which is curable. The diagnosis is based upon the discovery of minor signs of radiological interstitial syndrome, which in one third of cases is limited to a decrease in lung volume. This syndrome will be associated sooner or later with signs of alveolar involvement. The authors subsequently describe the underlying background, frequency, immune state, the circumstances of diagnosis and features of the aetiological diagnosis.


The metalloenzymic nature of collagenase-like peptidase of the rat testis. Collagenase-like peptidase, an enzyme degrading synthetic collagenase substrate (PZ-pentapeptide), was purified from rat testes and its properties were examined. Its activity was strongly inhibited by chelating agents, such as EDTA and 1,10-phenanthroline. By chelation and exhaustive dialysis it was possible to obtain this enzyme in its inactive, metal-free form. The activity of the metal-free enzyme was partly recovered by treatment with zinc or manganese ions, while a combined zinc and manganese treatment resulted in complete recovery of enzyme activity.


Amniogenic bands associated with facial dysplasia and paresis. A case of the amniogenic band syndrome is described with typical digital constriction bands amputations and syndactyly. The amniotic bands were demonstrated macroscopically and histologically, thus establishing the diagnosis. Facial dysplasia and unilateral facial paresis were attributed to undefined mechanical effects following amniotic rupture. In addition, there was evidence of partial umbilical cord compression which did not appear to have affected fetal well-being.


11,12-Secoprostaglandins. 1. Acylhydroxyalkanoic acids and related compounds. The synthesis is described of a series of acylhydroxyalkanoic acids which embody structural modifications of that class of secoprostaglandins which are formally derived from the natural substances by scission of the cyclopentane ring between carbon atoms 11 and 12. These analogues have been tested for their ability to stimulate cAMP formation in the mouse ovary, a characteristic action of the (E)-prostaglandins, and for their ability to bind to the rat lipocyte prostaglandin receptor. Certain members of the series that most closely resemble the prostaglandins in structure (e.g., 8-acetyl-12-hydroxyheptadecanoic acid) markedly stimulate cAMP formation at concentrations in the pharmacological range and show a significant affinity for the prostaglandin receptor. Conversely, these compounds are not substrates for prostaglandin 15-hydroxydehydrogenase which catalyzes a major reaction in the biological deactivation of the prostaglandins.


11,12-Secoprostaglandins. 2. N-acyl-N-alkyl-7-aminoheptanoic acids. A series of N-acyl-N-alkyl-7-aminoheptanoic acids has been prepared and evaluated for their ability to mimic the natural prostaglandins in certain biological systems. These compounds can be regarded as 8-aza-11,12-secoprostaglandins and, indeed, most of them stimulate cAMP formation in the mouse ovary assay, just as is observed with the natural prostaglandins. Selected compounds from this series also have been studied and shown to have prostaglandin-like effects in vivo.


Irreversible enzyme inhibitors. 200. Active-site-directed inhibitors of deoxycytidine kinase. Forty-three pyrimidine derivatives, mainly containing the 4-aminopyrimidine system, have been prepared and evaluated as inhibitors of deoxycytidine kinase. The most effective inhibitors were 2-alkylthio-4-aminopyrimidines and 1-alky-1-cytosines. The best inhibitors in both groups were those with large alkyl substituents, which indicate that hydrophobic bonding is occurring, possibly in the same area adjacent to the active site.


Inhibition of squalene synthetase by farnesyl pyrophosphate analogues. The pyrophosphates of the following farnesol analogues have been synthesized: 2-methylfarnesol; 7,11-dimethyl-3-ethyl-2,6,10-dodecatrien-1-ol; 3-demethylfarnesol; 4-methylthiofarnesol; 7,11-dimethyl-3-iodo-2,6,10-dodecatrien-1-ol; 7,11-dimethyl02-iodo-2,6,10-dodecatrien-1-ol; 7,11-dimethyldodeca-6,10-dien-2-yn-1-ol; phytol; 3,7,11-trimethyl-2-dodecen-1-ol; 3,7,11-trimethyldodecan-1-ol; and geraniol. The double bonds in all the above compounds were in the E configuration, except phytol, which was a 7:3 mixture of 2E and 2Z isomers. Each of the pyrophosphates inhibits the incorporation of labeled farnesyl pyrophosphate into squalene by a yeast enzyme preparation. Free alcohols and monophosphates are inactive. The analogues, listed in order of decreasing inhibitory strength, are, by kinetic analysis, competitive or mixed inhibitors. Irreversible inhibition is not observed. The results suggest that binding to the enzyme is primarily mediated by the pyrophosphate moiety assisted by relatively nonspecific lipophilic interactions. Decreasing the chain length and saturating double bonds severely reduces binding, while substitution at the 2,3, and 4 positions, and lengthening of the chain, is well tolerated.


Synthesis and antiviral activity of certain thiazole C-nucleosides. A general reaction of glycosyl cyanides with liquid hydrogen sulfide in the presence of 4-dimethylaminopyridine to provide the corresponding glycosylthiocarboxamides is described. These glycosylthiocarboxamides were utilized as the precursors for the synthesis of 2-D-ribofuranosylthiazole-4-carboxamide and 2-beta-D-ribofuranosylthiazole-5-carboxamide (23). The structural modification of 2-beta-D-ribofuranosylthiazole-4-carboxamide (12) into 2-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)thiazole-4-carboxamide (15), 2-beta-D-ribofuranosylthiazole-4-thiocarboxamide (17), and 2-(5-deoxy-beta-D-ribofuranosyl)thiazole-4-carboxamide (19) is also described. These thiazole nucleosides were tested for in vitro activity against type 1 herpes virus, type 3 parainfluenza virus, and type 13 rhinovirus and an in vivo experiment was run against parainfluenza virus. They were also evaluated as potential inhibitors of purine nucleotide biosynthesis. It was shown that the compounds (12 and 15) which possessed the most significant antiviral activity were also active inhibitors (40-70%) of guanine nucleotide biosynthesis.


6-aminopyrazolo[4,3-c]pyridin-4(5H)-one, a novel analogue of guanine. Treatment of dimethyl alpha-ethoxymethylidineacetonedicarboxylate with hydrazine gave methyl 3-(methoxy-carbonylmethyl)pyrazole-4-carboxylate which, upon ammonolysis and dehydration, afforded methyl 3-(cyanomethyl)pyrazole-4-carboxylate. This compound, when heated with liquid ammonia, gave 6-aminopyrazolo[4,3-c]pyridin-4(5H)-one, a new guanine analogue, which did not possess any of the potent antiviral activity shown by 6-aminoimidazo[4,5-c]pyridin-4(5H)-one (3-deazaguanine).


Intercellular communication and tissue growth: VII. A cancer cell strain with retarded formation of permeable membrane junction and reduced exchange of a 330-dalton molecule. A cancer (hepatoma) cell strain is described in which the formation of junctional membrane channels is abnormally slow. The development of electrical junctional coupling following the establishment of contact between these (reaggregated) cells is at least 15 times slower than that between their normal counterparts; and junctional transfer of fluorescein eventually develops, but only in about 5 per cent of the contacts (as against 100 per cent normally). This deviant membrane behavior is interpreted as a retardation in the process of accretion of junctional membrane channels. Its possible etiological role in defective growth regulation is discussed.


Virus isolation test for feline leukemia virus. A method was developed for detecting feline leukemia virus (FeLV) infection in cats by coculturing separated blood lymphocytes and platelets with clone 81 cells carrying a murine sarcoma genome but free of leukemia virus (S+L-). FeLV caused cell transformation that was easily visible 3-12 days after infection. This method is simple, sensitive, and easily read; it has the advantage of directly detecting infectious FeLV rather than viral antigens.


A carcinogenicity assay of Mirex in Charles River CD rats. The long-term administration of 50 and 100 ppm of Mirex in the diets of male and female Charles River CD rats was associated with a spectrum of liver lesions, from foci or areas of cellular alteration and neoplastic nodules to hepatocellular carcinoma. Statistically significant numbers of neoplastic nodules were observed in the livers of male rats receiving the high dose. Neoplastic nodules and hepatocellular carcinomas were not observed in control rats.


Possible carcinogenic effects of cedar shavings in bedding of C3H-Avy fB mice. C3H-AvyfB female mice were tested on bedding consisting only of low-resin pine and on bedding of pine plus cedar shavings. Tumor occurrences were similar in both groups of animals, with a slightly lower incidence and slightly higher average age for mammary gland tumors in the females on bedding of pine plus cedar. From these results, the high incidence of cancer in the C3H-AvyfB strain could not be attributed to the routine use of cedar shavings in the bedding material.


Detection of complement-dependent lytic antibodies in sera from feline leukemia virus-infected cats by the chromium-51 release assay. Sera from feline leukemia virus-infected cats lysed FL74 cells in the presence of an appropriate complement source; lysis was detected with the use of the 51 Cr release method. Serum complement from both rabbits and guinea pigs mediated lysis. Partial correlation of results was obtained from preliminary comparisons of lysis and immunofluorescence tests with the use of FL74 cells as targets.


Transplantable murine salivary gland carcinoma (myoepithelioma). I. Biologic behavior and ultrastructural features. Three salivary gland adenocarcinomas that arose spontaneously in female BALB/c mice, originally being maintained for plasmacytoma (MOPC) investigations, were studied; only one of the original hosts was MOPC-bearing. The initial carcinomas clearly originated from the salivary gland, not the breast, and did not resemble plasma cell lesions. The submandibular gland was the site of origin in each instance. Lesions were identical to those described previously in strains A and C mice as myoepitheliomas, with intracellular fibrils positive by phosphotungstic acid-hematoxylin (PTAH) stain. Each cell line (A, B, and C) of the salivary gland carcinomas was serially transplanted for 28, 29 and 14 generations, respecitvely. The lesion was easily transplanted, achieved almost 100% takes, and uniformly killed the host in 6-12 weeks, with metastasis to the peritoneum and liver. A myeloproliferative reaction characterized by leukemoid changes regularly occurred during the growth of the tumors. Each transplant generation was identical to its original salivary gland precursor by light and electron microscopy. In their growth pattern, the tumors resembled sarcomas with large areas of spindle cells. Ultrastructurally, the neoplastic cells contained the organelles of acinar and possibly ductal cells and exhibited varying degrees of synthetic and secretory activity, characterized by abundant free ribosomes and granular ergastoplasm, with prominent canaliculi containing abundant secretory material. Numerous desmosomes were seen, as well as coarse filaments and fine fibrils in most tumors cells, particularly in cells exhibiting lesser degrees of secretion. The filaments most likely represented the PTAH-positive intracellular fibrils observed by light microscopy. All tumors had numerous cells with resting features.


Vesicular stomatitis virus-infected L1210 murine leukemia cells: increased immunogenicity and altered surface antigens. Homogenates of L1210 cells infected in vitro with vesicular stomatitis virus (VSV) were immunogenic agains a tumor graft of 100 times the LD50 dose of L1210 cells" whereas those of uninfected cells were not. The immunogenicity of intact X-irradiated L1210 cells was distinguishable from that of VSV-infected cell homogenates on the basis of the susceptibility of immunogenicity to experimental procedures used in preparation of the immunogenic homogenates: Homogenization of intact X-irradiated cells or their infection with VSV prior to irradiation led to loss of immunogenicity. In addition, uninfected cell homogenates were not made immunogenic nor was the immunogenicity of VSV-infected cell homogenates eliminated by X-irradiation. At the time of tumor challenge, sera from mice that were effectively immunized with VSV-infected cell homogenate showed a high VSV-neutralizing titer but no complement-dependent cytotoxicity for L1210 cells. Quantitative absorption studies demonstrated that VSV infection led to a marked reduction in L1210 surface antigens recognized by cytotoxic alloantibody; spatial association between these antigens and VSV antigens was not demonstrable on VSV-infected cells. Antigens recognized by heterologous antiserum to L1210 cells were also reduced following VSV infection.


Incidence patterns of spontaneous tumors in BN/Bi rats. The occurrence of tumors in 236 female and 74 male BN/Bi rats that were allowed to live out their normal lifespans was described. For most neoplasms, the risk of a rat dying with a specific tumor increased with age. Some tumors, however, had a peak incidence; therefore, animals surviving these periods were at less risk than were their younger cohorts. The number dying with metastatic cancers was greatest in females over 30 months of age. Unexpectedly, males had a peak risk period at 25-30 months, so that males that died in older age groups had less risk of dying with a metastatic tumor. Lesions were often multiple, even in rats dying at a young age. Surprisingly, the percent dead with multiple tumors did not increase significantly with age. Some animals in every age group died without a tumor, and the percentage without neoplasms did not decrease, even in the older age groups.


Transient virus expression during murine leukemia induction by X-irradiation. Most X-irradiation-induced thymomas in C57BL/6 mice are virus-free when assayed by immunofluorescence for the gs antigen (gsa) of murine leukemia virus (MuLV). Virus was induced transiently in bone marrow cells and later appeared in thymus cells. Six to 7 weeks post irradiation, thymocytes and bone marrow cells were MuLV gsa-negative and remained negative for the lifetime of most animals, whether or not they contracted overt leukemia. During the period when MuLV gsa-positive bone marrow cells were found, XC-positive syncytia-producing bone marrow cells were also found. Virus information was expressed, therefore, for a limited duration, long before any signs of leukemia in the animals were evident. MuLV gsa-positive  thymocytes taken from mice 4 weeks after X-irradiation were cocultivated with a series of indicator cells. B-tropic virus, in addition to a xenotropic virus, was isolated from these cells. Ecotropic virus was not found in normal mouse thymocytes, in irradiated thymocytes a few days after termination of the X-irradiation sequence, or in most primary thymomas. All thymocytes produced only xenotropic virus in the cocultivation assays. Expression of the ecotropic virus was, therefore, transient, as assayed by immunofluorescence, XC syncytia formation, and virus isolation from MuLV gsa-positive thymus cells.


Inhibition of dimethylnitrosamine-induced strand breaks in liver DNA and liver cell necrosis by diethyldithiocarbamate. Diethyldithiocarbamate (DEDTC) prevented dimethylnitrosamine (DMN)-induced strand breaks in liver DNA and liver cell necrosis in male Wistar rats. In contrast, DEDTC did not inhibit the fragmentation of liver DNA caused by several other chemical carcinogens (N-hydroxy-2-acetylaminofluorene, 3-hydroxyxanthine, aflatoxin B1, N-acetoxy-2-acetylaminofluorene, methyl methanesulfonate, methylnitrosourea, and methylazoxy-methanol acetate), whether or not they required metabolic activation. Aminoacetonitrile exerted an action similar to that of DEDTC. The inhibitory effect was transitory, lasting at least for 4 hours, and protection for longer than 4 hours required multiple administrations of DEDTC. DEDTC also inhibited the serum clearance of DMN, methylation of liver DNA, and oxidative demethylation of DMN in the in vitro hepatic microsomal system prepared from either male Wistar rats or from hamsters. Interference of the metabolism of DMN appeared to be the mechanism by which DEDTC arrested DMN-induced biochemical and biologic effects.


Separation of cells containing R-type virus-like particles from a simian virus 40-induced hamster tumor cell line. Cells derived from a simian virus 40-induced hamster fibrosarcoma were separated into two distinct cell bands of differing buoyant densities. The lighter cell band or fraction (F1) had a buoyant density range of 1.025-1.032 g/ml and comprised 3.8% of the total cells applied to the gradient, whereas the heavier cell fraction (F2) had a buoyant density range of 1.054-1.074 g/ml and comprised 95.3% of the total cells applied. Both cell fractions were tumorigenic and did not differ greatly in cell type, viability, mitotic index, or their ability to incorporate [3H]thymidine. However, ultrastructurally, the F1 cells contained R-type virus-like particles within dilated intracisternal spaces and exhibited cytoplasmic vacuoles. In the F2 cells, few detectable R-type particles and cytoplasmic vacuoles were revealed by electron microscopy. The F2 cells demonstrated a twofold greater ability to incorporate [14C]protein hydrolysate into proteins than did the F1 cells.


A continuous line of Rous sarcoma virus-transformed chick embryo cells. A continuous line of Rous sarcoma virus (RSV)-transformed chick embryo cells was established. The cells, designated RTAZ-1, which constitute the only known line of continuously growing RSV-transformed cells of chick embryo origin, grow rapidly, display uniform morphology, and perpetually release large amounts of RSV (Rous-associated virus, type 1). RTAZ-1 cells display a heteroploid chromosome complement with 92-94 chromosomes characteristic of chicken cells. Molecular hybridization studies demonstrated that DNA from these cells hybridized with normal chick DNA but not with human (KB) cell DNA, thus providing additional evidence of the chicken origin of RTAZ-1.


Inhibition of hepatocarcinogenesis by adrenocorticotropin in aflatoxin B1-treated rats. We examined whether hormones would modify the carcinogenic action of aflatoxin B1 (AFB1). Four groups of inbred Fischer rats received AFB1, 125 mug per animal, weekly per os. In three of the groups, certain hormones were administered simultaneously: One group received 1 U growth hormone (GH) sc weekly, another was given 4 U adrenocorticotropin (ACTH) weekly, and a third received 0.5 U insulin weekly sc. AFB1, ACTH, and insulin were given for 20 weeks; GH was given for only 10 weeks. The control group did not receive hormone adjuvant. In each group, 4 animals were killed at 7, 14, 21, 28, and 35 weeks; the remaining rats were killed at 77 weeks. Their livers were carefully examined and samples prepared for light and electron microscopy. Animals receiving AFB1 and ACTH failed to exhibit hepatocellular carcinoma. On the other hand, malignant lymphoma appeared at 56 weeks in 3 of the 6 surviving males on this regime. AFB1, alone or when given with insulin or GH, caused hepatocellular carcinoma in all animals; in these, lymphoma was not observed. Lymphoma comprised two cell types, each with similar neclear characteristics but differing in their nucleocytoplasmic ratios and in the amount and distribution of cytoplasmic organelles. Alterations leading to hepatocellular carcinoma were examined at various stages of development. "Basophilic hyperplasia" reflected an increase in free ribosomes. "Hyperplastic nodules" were composed of hepatocyte aggregates with characteristics similar to those encountered in the earlier stage. Both the "neoplastic nodules" and hepatocellular carcinomas were formed by cells containing large, "smooth fingerprints" and free ribosomal aggregates. These features supported the concept that AFB1 impairs ribosomal binding to endoplasmic reticulum membranes. The failure of ACTH-treated animals to develop hepatocellular carcinoma was ascribed to the effect of adrenal cortical stimulation upon membrane-polysome binding.


Chromosome banding patterns of two transplantable pituitary tumors induced in rats by 2,4,6-trimethylaniline. The chromosomal constitution of a mammosomatotrophic and an inactive transplantable pituitary tumor induced in rats by 2,4,6-trimethylaniline was studied by the Giemsa-banding techniques. The inactive tumor line 7315i evolved from the active tumor line 7315a in one of the early transfers. A comparison was also made with chromosomal patterns of a transplantable rat hepatoma (7316A) induced by the same chemical carcinogen. Pituitary tumor line 7315a had a pseudotriploid complement with 63 chromosomes, whereas the inactive line 7315i was hypodiploid, with a dominant stemline of 36 chromosomes. The stemline chromosome number of the hepatoma was in the hypotetraploid region. Giemsa banding revealed that all chromosomes of the normal rat complement were present in both pituitary tumors. Four abnormal chromosomes were detected in the active tumor line and three in the inactive line. Both tumor lines contained a single minute chromosome. One common marker, a deleted chromosome No.1, was found in both pituitary tumors. This common marker chromosome was, however, not present in the hepatoma. The stemline karyotype of the hepatoma contained seven different markers, but none of them was identical to the abnormal chromosomes of the pituitary neoplasms. The findings suggested that the abnormal karyotypes of lines 7315a and 7315i could reflect the multisecretory activities of these neoplastic pituitary cells but that chromosomal localization of the secretory defect awaits exact genetic mapping of the rat chromosomes.


Induction of malignant vascular tumors of the liver in guinea pigs treated with 2,2'-dihydroxy-di-n-propylnitrosamine. The effect of chronic administration of 2,2'-dihydroxy-di-n-propylnitrosamine (DHPN) was studied in randombred guinea pigs. DHPN, dissolved in olive oil, was injected sc into 40 animals at a dose of 250 mg/kg body weight/week for 30 weeks, and the animals were observed until their death or termination of the experiment at the end of 40 weeks. Of the 32 guinea pigs that survived more than 20 weeks of DHPN treatment, 23 developed angiosarcoma of the liver between 22 and 40 weeks. Metastases to lungs, spleen, and peripancreatic lymph nodes were observed in 8 animals. Other tumors included hepatocellular carcinoma (1 animal), cholangiocarcinoma (1 animal), chronic myeloid leukemia (1 animal), acinar cell adenoma of pancreas (1 animal), and acinar cell carcinoma of pancreas (1 animal). In addition, megalocytic change of hepatic cells with intranuclear inclusions, pelliosis hepatis, and cholangiomatous lesions were also encountered frequently in the livers.


Horizontal transmission of feline leukemia virus under experimental conditions. Thirty-seven specific-pathogen-free (SPF) cats ranging from newborn to 1 year were inoculated with the Rickard strain of feline leukemia virus (FeLV). Each inoculated cat shared a cage with a control SPF cat for 40 weeks post inoculation. After 4-5 weeks, 20 of the inoculated cats became group-specific antigen (gsa)-positive; the other 17 remained gsa-negative but developed virus neutralizing and feline oncornavirus cell membrane-associated antigen antibody titers. Seventeen of the control cats in contact with the gsa-positive cats developed evidence of FeLV infection 4-18 weeks after virema was detected in their inoculated cage mates. Of the control cats in contact with inoculated cats that remained gsa-negative, none developed evidence of FeLV infection. Data indicated that the gsa-positive state in cats inoculated with FeLV correlated with the capacity for horizontal transmission of the virus.


Frog lysozyme. IV. Isozyme of lysozyme and the Lucké renal adenocarcinoma. The normal kidney of the frog Rana pipiens contains eight isozymes of lysozyme. Three of these, however, were not present in five Lucke renal adenocarcinomas. No isozymes were found to be unique to the tumor kidneys. Of the five isozymes detected in tumor kidneys, only one was uniformly present, a second was present in four of five tumors, and the appearance of the other three was highly variable, although no tumor kidney contained all of them. The uniform absence of the three isozymes and the variable distribution of the other five among the tumors attested to the metabolic uniqueness of this tumor and was consistent with a prior hypothesis concerning the role of lysozyme in the etiology of this herpesvirus-induced tumor.


Herd health status of animals exposed to polybrominated biphenyls (PBB). The health status of 16 herds of dairly cattle exposed to low levels of polybrominated bipenyl (PBB) was compared with that of 15 control herds. Milk production of the contaminated herds was not significantly changed in 1972, 1973, and 1974 and was not significantly different from that of control herds in the same years. Mortality of adult cows and calves, the percentages of cows culled from the herds because of old age and low production, disease, or sterility, and the general health conditions were similar in the two groups. Serum concentrations of calcium, glucose, and cholesterol in contaminated herds were significantly different from those of the control herds, but the relationship to PBB exposure needs further investigation.


Biological responsiveness to cholinesterase inhibition: a test for exploring the developmental maturity of the cholinergic system. The acute mortality caused by two irreversible inhibitors of cholinesterases [diisopropylfluorophosphate (DFP) and diethoxyphosphorylthiocholine, 217 MI-phospholine iodide] has been investigated on chick embryos at different stages of development. The results demonstrate that the above compounds do not show any acute lethal action when administered before the 9th day of incubation; on the other hand, the administration is regularly followed by death after the 9th day of incubation. The doses are comparable to those causing death in hatched chicks. It has also been observed that no appreciable difference exists in DFP uptake from the yolk before and after the 9th day of incubation and that drug-induced cholinesterase inhibition is of the same order of magnitude at any developmental stage; the compound pyridine-2-aldoxime methanesulfonate (2-PAM) was a good antidote against DFP acute lethality. It seems likely that between the 8th and the 9th day of incubation the target system of organophosphorus inhibitors, that is, the cholinesterase enzymatic system, reaches a new point of maturation.


Effects of neonatal hypothyroidism on cerebral and cerebellar synaptosome development. The effect of hypothyroidism on cerebral and cerebellar synaptosome development has been studied. Neonatal hypothyroidism was induced following addition of 0.3% propylthiouracil to the diet of nursing mothers. Maturation profiles of total synaptosome fraction and specific activities of lactate dehydrogenase, Na+K ATPase, cytochrome c oxidase, and protein were obtained from days 6 to 32 on synaptosomes isolated from Ficoll-sucrose gradients. The greatest changes were found in the total activities of enzymes isolated from the cerebellum. Hypothyroidism induced a retardation of LDH and cytochrome c oxidase in cerebellar synaptosomes, but no change in corresponding specific activities. Maximum rates of 14C-leucine incorporation into cerebellar synaptosome protein was found at 16-20 days, after which a rapid decline occurred to adult levels at 32 days. In neonatal hypothyroidism, synthesis was significantly reduced at 8 and 14 days, but reached control levels or above at 21--32 days. In the cerebrum, maximum rates of 14C-leucine incorporation into synaptosome protein were identified at 8--12 days in normal with a rapid drop to adult levels at approximately 20 days. In neonatal hypothyroidism, peak activities were identified at 14 days and increased activities over control were noted at 14, 20 and 30 days. These observations demonstrate the sensitivity of the developing cerebellar synaptic apparatus to neonatal hypothyroidism, with a protraction in the peak levels of synaptosome protein synthesis in cerebrum and cerebellum.


Drug and immuno-diagnostic resistant amoebic liver abscess in Ibadan: an elucidation of a possible mechanism. A thick fibrous walling of amoebic abscess of the liver is very rare. Using as a basis the case of drug and immunodiagnostic resistant amoebic liver abscess in an adult Nigerian, an account is given of the possible mechanisms for such therapeutic and diagnostic failure. A theory of blood-abscess cavity barrier is postulated and suggested as a mechanism for the constant failure to detect antibodies to E. histolytica in the small percentage of cases of amoebic liver abscess (Morris, Powell and Elsdon-Dew, 1970). A study of the rate of diffusion of amoebic excretion/secretion through a thick fibrous tissue or artificial membrane of comparable properties is strongly advocated.


Endobronchial adenomata in children.

Endobronchial adenomata in children. Two cases of carcinoid adenomata of the bronchus in children are added to 25 previously reported cases of bronchial adenomata in children in the literature. Carcinoid tumors represent close to 50 per cent of bronchial adenomata in children, with cylindromas (adenoid cystic) forming 13 per cent and mucoepidermoids two per cent. Over 1/3 of bronchial adenomata have metastases at the time of diagnosis. No cases of carcinoid syndrome have been reported in children. The treatment of choice is lobectomy with sleeve resection or, failing that, pulmonary resection. The overall five-year survival time varies from 50 to 80 per cent and is enhanced by surgical resection.


Spontaneous mutations affecting the host range of the B77 strain of avian sarcoma virus involve type-specific changes in the virion envelope antigen. Previously it was shown that the host-range gene of the Bratislava strain of avain sarcoma virus (B77 virus) spontaneously mutates with a very high rate. The wild-type B77 virus called B77 virus-II, mutates either to virus that efficiently infects duck cells (B77 virus-III) or to virus that does not mutate to the ability to infect duck cells (B77 virus-I) (Zarling and Temin, 1976). No significant differences in either the virion envelope glycoproteins or other major virion proteins were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, pseudotypes of B77 virus-I with proteins of a transformation-defective mutant of B77 virus-III formed foci efficiently in duck cells. An alteration in the envelope protein of B77 virus-I was demonstrated by experiments in which B77 firus-I was fused into duck cells with UV-irradiated Sendai virus and formed foci. Neutralization experiments further demonstrated that B77 virus host-range mutants have altered type-specific envelope antigens. Thus, the spontaneous mutations in the host-range gene of B77 virus involve changes in the type-specific virion envelope antigen.


Steroid induction of mouse mammary tumor virus: effect upon synthesis and degradation of viral RNA. Steroid hormones have been demonstrated to induce in tissue culture the production of mouse mammary tumor viral (MMTV) RNA, proteins, and particles 10-fold compared with constitutive levels. However, previous data of increased viral RNA levels did not distinguish between an increased rate of viral-specific RNA synthesis and a slower rate of viral RNA degradation. According to the recently developed assay of Coffin et al. (1974) for measuring rates of viral RNA synthesis, short-term labeling experiments of a mouse mammary tumor cell line indicate that the glucocorticoid hormone dexamethasone stimulates a 3-fold increase in the synthesis of MMTV-specific RNA within 10 min after the addition of hormone and that stimulation of RNA synthesis reaches 5- to 10-fold within 30 to 60 min, while the synthesis of Moloney leukemia virus-specific RNA in the same cell is unaffected by steroids. The decay rates of pulse-labeled and accumulated MMTV RNA in the presence or absence of dexamethasone show this RNA to have a half-life of greater than 8 h. In addition, hormone-stimulated MMTV RNA appears to have an increased rate of decay compared to basal MMTV RNA, thus ruling out an increased stability of MMTV RNA in the presence of steroid hormones as the basis for increased RNA levels. Thus, the magnitude, rapidity, and specificity of hormone action on MMTV RNA synthesis indicate a primary effect upon transcription.