Sensitivity of a radioimmunoassay method for detection of certain viral antibodies in sera and cerebrospinal fluids. An indirect solid-phase radioimmunoassay (RIA) was applied to titration of serum and cerebrospinal fluid (CSF) antibodies against a variety of viruses including rubella, mumps, measles, herpes simplex, varicella-zoster, and vaccinia. The test used fixed, virus-infected cells as a source of antigen, and conditions for optimal production of viral antigen were determined for each virus-host cell system. In acute, uncomplicated viral infections, sera taken 2 to 5 days after onset generally had low homotypic RIA titers ranging from less than 1:100 to 1:500, whereas convalescent-phase titers ranged from 1:128,000 to 1:512,000. Rubella and measles antibody titers as high as 1:256,000 were demonstrated by RIA in CSF from patients with chronic panencephalitis, whereas homologous antibody titers of 1:4,000 were detected in CSF from acute mumps, herpes simplex, and varicella-zoster virus infections with central nervous system involvement. Some heterotypic antibody was demonstrable by RIA in CSF, but, with the exception of herpes simplex antibody in a mumps virus infection, titers were markedly lower than those to the infecting virus type. RIA generally demonstrated titers at least 1,000 times higher than those obtained by conventional assays such as complement fixation, hemagglutination inhibition, neutralization, and immunofluorescent staining.


Vesicular stomatitis virus plaque production in monolayer cultures with liquid overlay medium: description and adaptation to a one-day, human interferon-plaque. Vesicular stomatitis virus forms discrete, microscopic plaques in stationary cultures of the WISH amnion cell line. Microplaque formation is rapid, reproducible, and easily quantitated, occurs at temperatures ranging from 33 to 40 degrees C, and does not require a semisolid overlay. WISH cells, however, are less sensitive to vesicular stomatitis virus than are chicken embryo, 3T6, or Vero cells. WISH amnion cells also are highly sensitive to the antiviral effects of human interferon, and a quantitative human interferon assay, based on vesicular stomatitis virus plaque reduction in WISH cells, is described. This interferon assay can be performed within 1 day, uses a liquid overlay medium, does not require a vital stain, is as sensitive as other methods that use diploid cell strains, and is performed in a microtiter system.


Extrinsic modulation of human T-lymphocyte E rosette function associated with prolonged hepatocellular injury after viral hepatitis. Defective T-lymphocyte E rosette (ER) function associated with viral hepatitis A and B may be due to mechanisms extrinsic or intrinsic to the target lymphocyte. The extrinsic defect is induced by an immunoregulatory plasma lipoprotein (RIF) and has the capacity to regenerate ER function in vitro. The intrinsic defect is refractory to regeneration and is not associated with RIF. Although both mechanisms occur with high frequency during the acute phase of viral hepatitis they tend to segregate in accordance with progression of hepatocellular injury at later stages of the disease. The extrinsic defect was observed in 7 out of 8 patients with longstanding chronic active hepatitis and in 10 out of 10 patients with unresolved hepatitis 12 wk after the onset of jaundice. In contrast, none of nine patients with resolved hepatitis had extrinsically defective ER function 12 wk after the onset of jaundice whereas eight of them displayed an intrinsic defect of ER function at that time. Among the various viral and liver diseases studied RIF appeared to be specific for hepatitis A and B viral infections. None of 64 sera from a variety of viral infections including Epstein-Barr virus cytomegalovirus mononucleosis with associated hepatitis nor 15 sera from patients with several chronic nonviral liver diseases were positive for RIF. RIF and its associated extrinsic defect in ER function therefore appear to correlate with a particular type of hepatocellular injury initiated by the hepatitis A and B viruses that may have a propensity for persistence and(or) progression to an aggressive form of chronic hepatitis.


Urinary cyclic AMP analyzed as a function of the serum calcium and parathyroid hormone in the idfferential diagnosis of hypercalcemia. Urinary cyclic AMP (UcAMP) appropriate for the serum calcium concentration was determined in normal subjects during the base-line state and during alteration in their serum calcium concentrations by saline and calcium infusions. This was compared to the UcAMP in 76 patients with hypercalcemia and 5 patients with hypocalcemia. In 54 of 56 patients with primary hyperparathyroidism, the UcAMP was inappropriately high for their serum calcium concentration, the 2 exceptions having renal failure. In four patients with vitamin D intoxication, sarcoidosis, milkalkali syndrome, and thiazide-induced hypercalcemia and in five patients with hypocalcemia due to hypoparathyroidism, the UcAMP was appropriately low for their serum calcium concentration. In 16 patients with nonparathyroid neoplasms, 10 had UcAMP levels that were inappropriately high suggesting ectopic parathyroid hormone (PTH)-mediated hypercalcemia and 6 had UcAMP levels that were appropriately low suggesting that their hypercalcemia was due to osteolytic factors other than PTH. Correlations between UcAMP, serum calcium concentration, and carboxyl-terminal immunoreactive PTH suggest that random UcAMP is a sensitive accurate reflection of circulating biologically active PTH. If there is adequate renal function (serum creatinine concentration less than 2.0 mg/dl), a random UcAMP expressed as mumol/g creatinine and analyzed as a function of the serum calcium concentration completely separates patients with PTH and non-PTH-mediated hypercalcemia.


Superoxide production and reducing activity in human platelets. Human platelets contain the cuprozinc (cytoplasmic) and manganese (mitochondrial) forms of superoxide dismutase. Nevertheless, superoxide radicals were detectable in the surrounding medium of metabolically viable platelet suspensions by using two assay systems: cytochrome c and nitroblue tetrazolium. The quantity of superoxide generated by platelets (5 X 10(5) superoxide radicals/platelet per 10 min) was constant and did not increase after aggregation by agents such as collagen and thrombin. The superoxide-generating system was present in the supernate of both aggregated and resting platelets and therefore was not platelet-bound. Platelet superoxide production was unaffected by prior ingestion of aspirin, indicating that the prostaglandin and thromboxane pathways were not involved. Both resting and aggregated platelets exhibited a reductive capacity toward cytochrome c and nitroblue tetrazolium which was unrelated to superoxide production. Furthermore, the aggregation process always resulted in a marked increase in this reduction. The nonsuperoxide reduction associated with aggregation was found to be membrane bound and to decrease with an apparent first order reaction rate (k1 = 0.067 min-1). In addition, accumulative, time-dependent nonsuperoxide-related cytochrome c reduction was also detected. Since there is no superoxide dismutase in plasma, the presence of superoxide radicals in the surrounding medium of platelets may have in vitro significance for platelet and leukocyte concentration and storage and in vivo significance for hemostasis, coagulation, and thrombosis. The nonsuperoxide-related reducing activities may represent a biochemical basis for platelet-blood vessel interactions, with particular reference to blood vessel integrity.


High affinity binding of 125I-angiotensin II to rat glomerular basement membranes. 125I-angiotensin II (AII) specifically bound to rat glomerular basement membrane (GBM). The kinetics of binding were similar to those obtained with the total glomeruli. The apparent dissociation constant was close to 50 pM with both preparations. The number of sites related to the amount of protein was two times greater with GBM than with total glomeruli. Since the amount of GBM protein extracted from a given amount of glomerular protein was about 10%, it was possible to estimate the share of the GBM binding sites for AII as representing 20% of the total number present in the entire glomerulus. Binding studies at equilibrium as a function of 125I-AII concentration and competitive binding experiments suggested either multiplicity of the binding sites or cooperativity in the binding reaction. Degradation of 125I-AII in the presence of GBM was slight and did not increase with time. The difference in the degrees of degradation of 125I-AII was too small to account for the observed difference in binding when the results obtained with GBM and isolated glomeruli preparations were compared. 125I-AII binding to GBM was increased after treatment of these membranes with collagenase, slightly diminished with neuraminidase, and almost completely abolished with trypsin suggesting the proteic nature of the receptor. 125I-AII binding to GBM was diminished after incubation of GBM with anti-GBM antibodies as a result of a decrease in the number of binding sites. 125I-AII binding was even more diminished in preparations of glomeruli isolated from rats passively immunized with anti-GBM antibodies when compared with glomeruli from control animals. This resulted from both smaller affinity for AII and decrease in the number of the binding sites. The present data provides evidence for specific binding sites for AII localized on GBM. This is noteworthy since receptors for polypeptide hormones are currently observed on the surface of cell membranes. These findings also suggest a new physiological role for AII which might involve modification of GBM permeability.


In vivo effect of indomethacin to potentiate the renal medullary cyclic AMP response to vasopressin. In a previous study we demonstrated that indomethacin potentiated the hydro-osmotic action of vasopressin in vivo. It was hypothesized that this action of indomethacin was due to its ability to suppress renal medullary prostaglandin synthesis, since in vitro studies have suggested that prostaglandins interfere with the ability of vasopressin to stimulate production of its intracellular mediator, cyclic AMP. In the present study this hypothesis was tested in vivo. Anesthetized rats undergoing a water diuresis were studied. In a control group, bolus injections of 200 muU of vasopressin caused a rise in urinary osmolality (Uosm) from 124 +/- 6 to 253 +/- 20 mosmol/kg H2O (P less than 0.005). In a group treated with 2 mg/kg of indomethacin the same dose of vasopressin caused a significantly greater (P less than 0.001) rise in Uosm from 124 +/- 7 to 428 +/- 19 mosmol/kg H2O. Medullary tissue cyclic AMP rose from 9.4 +/- 0.9 to 13.4 +/- 1.7 (P less than 0.05) pmol/mg tissue protein after vasopressin administration in animals receiving no indomethacin, while in indomethacin-treated animals there was a significantly greater rise (P less than 0.001) in medullary cyclic AMP from 10.4 +/- 0.9 to 21.6 +/- 2.1 pmol/mg tissue protein in response to the vasopressin injections. In neither control animals nor indomethacin-treated animals were there significant changes in renal hemodynamics, as measured by clearance techniques. Indomethacin, when given alone, had no effect on Uosm or medullary tissue cyclic AMP. Indomethacin did, however, reduce medullary prostaglandin E content from 84.7 +/- 15.0 to 15.6 +/- 4.3 pg/mg tissue. This study has shown that indomethacin, in a dose which suppresses medullary prostaglandin content, potentiates the ability of vasopressin to increase the tissue content of its intracellular mediator, cyclic AMP. Indomethacin caused no demonstrable inhibition of cyclic AMP phosphodiesterase. Therefore, it seems likely that indomethacin enhanced the ability of vasopressin to increase medullary cyclic AMP levels by causing an increased production rather than decreased destruction of the nucleotide. We conclude that this action of indomethacin contributes to its ability to potentiate the hydro-osmotic action of vasopressin in vivo. A corollary to this conclusion is that endogenous medullary prostaglandin E's may be significant physiological modulators of the renal response to vasopressin.


Blocking of catecholamine activation of adenylate cyclase by N, N'dicyclohexyl carbodiimide in turkey erythrocytes. Incubation of erythrocytes or their isolated membranes with N, N'dicyclohexyl carbodiimide (DCC) blocked isoproterenol activation of the adenylate cyclase. Fluoride activation remained unaffected. L-epinephrine and DL-propranolol partially and transiently protected the system against DCC. D-Epinephrine and dopamine did not protect. The enzyme system preactivated by isoproterenol plus Gpp(NH)p was no longer sensitive to DCC. In contrast to the water insoluble DCC, a water soluble carbodiimide acted only at high concentration and blocked fluoride as well as catecholamine activation of the adenylate cyclase. The findings indicate that DCC attacks a group on, or near, the beta-adrenergic receptor and that this group is located in a hydrophobic region of the cell membrane. It is argued that a low nominal concentration of DCC in the aqueous suspension of erythrocytes actually represents a very high concentration of DCC in the hydrophobic region of the cell membranes, near the beta-adrenergic receptor. The reaction with DCC may prove to be a useful tool in future analyses of beta-adrenergic receptor function.


Changes in trout testis cyclic GMP concentration during hormonally induced spermatogenesis. The concentration of cyclic GMP was determined in trout testes (Salmo gairdnerii) during hormonally-induced sermatogenesis. Early in spermatogenesis, a relatively high concentration (1 mumole/kg wet weight testis) of cyclic GMP was present. A sharp, ten-fold, decrease in testis cyclic GMP concentration occurred during the early stages of rapid mitotic activity and testis growth. A further gradual five-fold decrease in cyclic GMP concentration occurred during meiosis and the following transformation of spermatocytes into spermatids and then to mature spermatozoa.


The molecular structure of the free acid of guanosine 3',5'-cyclic monophosphate (cyclic GMP). The structure of the free acid of guanosine 3',5'-cyclic monophosphate has been determined from three dimensional x-ray diffraction data collected with a GE XRD-490 automated diffractiometer. The molecule crystallizes in the orthorhobic space group P212121 with a = 17.953(17), b = 7.530(7), and c = 12.755(12) and Z = 4. The structure was solved by direct methods. The 969 independent data were defined by full matrix least-squares to a final agreement factor, RF, of .050 for FO greater than sigma (FO). The compound exists as a switterion in the solid state. N(7) of the base is protonated. The structure of the free acid is similar to that of the sodium salt. Both have the syn conformation. The puckering of the sugar rings is also similar. Small differences exist in the bond angles of the cyclic phosphate and furanose rings.


Reactions in glass ionomer cements: V. Effect of incorporating tartaric acid in the cement liquid. A description is give of the effect on the ASPA cement reaction of tartaric acid incorporated in the cement liquid. Tartaric acid acts as an accelerator that aids in the extraction of ions from the aluminosilicate glass and facilitates their binding to the polyanion chains. Postgelation hardening is significantly increased. Working time is unaffected possibly because cations are initially present as complexes.


Glass ionomer cements: chemistry of erosion. A three-month study of the chemistry of the water erosion of two forms of ASPA cement has been made. The effect of varying cement consistency and cure time was investigated. The results are discussed in terms of the known chemistry and structure of the cement. The erosion behavior is compared to that of silicate, silicophosphate, and zinc polycarboxylate dental cements. The state of absorbed water and the mechanism of erosion is discussed.


Some characteristics of collagenase activity in gingival crevicular fluid and its relationship to gingival diseases in humans. Collagenase activity in human gingival fluid was measured using a radioactive collagen fibril assay. The activity was positively correlated with the severity of gingival disease. The fluid collagenase seemed to be controlled by alpha 2-macroglobulin, based on its activation by NaSCN, and to be present solely in the extracellular fraction. Examination of the collagen breakdown products by acrylamide gel electrophoresis indicated that the fluid collagenase was of tissue rather than bacterial origin.


Physiological learning theory.

Physiological learning theory. Attention or "concentration" requires control of activity in those excess neurons that are not necessary for the present task. The control is probably not a massive inhibitory suppression but may be a recruiting process, a function of complex perceptual and associative learning that begins with early experience. Inhibition, however, may still be of crucial importance as a sharpener of associative mechanisms, and the child with minimal brain damage may have suffered a selective loss of inhibitory neurons.


Melanotic neuroectodermal tumor of infancy: review of the literature and report of case. A characteristic case of melanotic neuroectodermal tumor of infancy is reported and the literature regarding this lesion is reviewed. The case reported shows the salient features of previously described melanotic neuroectodermal tumors of infancy; it occurs in infants of less than 1 year of age; it most commonly arises from bone (jaws and skull), has distinctive gross and histologic features, is benign, and is cured by total local excision. Since the teeth in the mandible and maxilla are undergoing development about the time that this tumor occurs, it is not surprising to find tooth buds in close proximity to the expanding neoplastic mass. When odontogenic rests are found in association with the lesion, it must be assumed that it is a collision of the two structures and not that the odontogenic epithelium is the origin for the lesion as previously thought. This is especially true since odontogenic epithelium is not found in the tumor when it occurs outside the jaws. This is the type of lesion that will most probably first be noticed by a dentist and should be managed by the oral surgeon. It is important to note that proper advice regarding future dental problems should be given to the parents of the infant. Such possibilities as crowding or absence of the deciduous or permanent teeth, malocclusion, and possible malformation of the remaining maxillary ridge should be explained. Close dental follow-up is mandatory since recurrence is possible.


An immunocytochemical study of TSH beta storage in rat thyroidectomy cells with and without D or L thyroxine treatment. Binding sites to the beta chain of thyroid stimulating hormone (TSH) were localized in pituitaries of thyroidectomized rats. Immunocytochemical staining was observed in hypertrophied TSH cells ("thyroidectomy cells") and primarily located in dilated rough endoplasmic reticulum. Staining was also found on the few secretory granules and on some of the intracisternal granules. Some of the thyroidectomy cells stained intensely, while others exhibited very little staining. When thyroidectomized rats were treated with thyroxine 4 days before death, the TSH cells contained more secretory granules, and the intracisternal granules were larger and more numerous. L-thyroxine was 10 times as potent as D-thyroxine in promoting the build-up of granules. Both types of granules stained intensely.


A new histochemical method for the identification and visualization of both side chain acylated and nonacylated sialic acids. A new histochemical method is described for the differentiation of mucins that utilizes two different Schiff reagents and allows single section identification of side chain O-acylated, and nonacylated, sialic acids in contrasting colors. In the event of mucins containing only one type of sialic acid, it may allow their specific identification (e.g., C7 or C8 side chain O-acylated). It has been shown to be useful in the identification of some metastases from adenocarcinomas of colon (where the primary is potassium hydroxide/periodic acid-Schiff positive) and should prove of great value in the investigation of diseases of the gastrointestinal tract and particularly those of the colon. It should also be valuable in the general field of epithelial mucin histochemistry, particularly for those mucins of the salivary and parotid glands, etc.


Lasar flow cytophotometric immunoperoxidase detection of herpes simplex virus type 2 antigens in infected cultured human cells. Human cells in culture (HEp-2) were infected with herpes simplex virus type 2 (HSV-2) at multiplicities of infection varying from 0.2 to 10, and fixed 6, 12, 18 and 24 hr after infection. Infection-related antigens were detected by an indirect double antibody (peroxidase conjugated goat anti-rabbit to rabbit anti-herpes simplex virus type 2) immunoenzymatic staining reaction that rendered infection-related antigens visible by light microscopy. A corresponding series of laser flow cytophotometric experiments yielded reproducible large-angle (1-19 degrees) laser-light scattering distributions that depended upon multiplicities of infection and the location of the infection-related antigens in the infected cells.


Acrolein as a fixative for enzyme cytochemistry. Since acrolein can penetrate more quickly and deeply into tissue blocks than glutaraldehyde, the possibility of the use of this aldehyde as a prefixative in enzyme cytochemistry was reinvestigated. At low concentrations, acrolein preserves the activities of the enzymes investigated, including those of glucose-6-phosphatase, which is known as one of the most vulnerable to aldehyde fixation; thus, acrolein is usable in enzyme ultracytochemistry. Enzyme activities are also preserved in tissues fixed with acrolein and glutaraldehyde combined. The rapid penetration of acrolein enables fixation in larger tissue blocks and provides greater freedom in specimen selection, especially important advantages when encountering heterogeneous materials as in pathology.


The effect of colostrum-derived antibody on louping-ill virus infection in lambs. The influence of colostrum-derived antibody to louping-ill virus on the course of experimental infection was investigated in lambs. Lambs that had high titres of antibody were refractory to infection. Lambs that had low titres of antibody did not develop a viraemia but either showed an antibody reaction or were sensitized as judged by the immune response, which was typical of an anamnestic response, after rechallenge. Animals that had no antibody 34-20 days before challenge had either no or very slight viraemia, but did develop an antibody response with titres as high as those of control lambs by day 21. Lambs that had been negative for longer periods responded in a similar fashion to controls. These findings are discussed in relation to the occurrence of disease in lambs kept in louping-ill endemic areas. It is concluded that in such areas infections of lambs are likely to be of minor importance as a cause of mortality and of little epidemiological significance.


Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent. Procedures are detailed for the rapid isolation of representative cell membrane antigens with protein A-bearing staphylococci as an adsorbent for IgG antibodies complexed with the antigens. Cell surface membrane proteins were radioiodinated and solubilized in nonionic detergent. Specific antisera were subsequently added and the immune complexes precipitated by addition of the staphylococcal adsorbent and low speed centrifugation. The antigens isolated included surface immunoglobulins from mouse and human lymphocytes, human beta-microglobulin and HL-A alloantigens, mouse H-2 alloantigens, and the murine leukemia virus glycoprotein gp 70. Rabbit, sheep, goat, and mouse antisera were all effective for the specific phase of the precipitation reaction. The surface membrane immunoglobulins of mouse splenic lymphocytes and human peripheral blood lymphocytes differed with respect to class composition and protein A reactivity. Mouse lymphocyte surface immunoglobulins were nonreactive with protein A, whereas a high proportion of human lymphocyte surface immunoglobulins of different classes bound directly to the staphylococci. In sequential immunoprecipitation studies the prior isolation of one antigen had no appreciable effect on the subsequent recovery of another antigen. Adsorption of antigen-antibody complexes is quantitative when protein A sites are provided in excess over antiserum IgG sites, and this obviates the need for equivalence point titrations for optimal precipitation necessary with alternative double antibody techniques.


The involvement of cyclic nucleotide metabolism in the initiation of lymphocyte proliferation induced by mitogens. Murine lymphocytes incubated with specific mitogens were examined for alterations in levels of intracellular cyclic AMP and cyclic GMP. Mitogenic concentrations of LPS elevate cyclic GMP levels; however, DxSO4, Poly I:C, PPD, Con A, and PHA show little immediate effect on cyclic GMP levels. Higher concentrations of all of these mitogens consistently elevate cyclic AMP levels. Exogenous cyclic GMP stimulates DNA synthesis in homozygous nude (nu/nu) spleen cultures but not in murine thymocyte cultures. Both LPS and cyclic GMP enhance the mitogenic response of thymocytes in the presence of suboptimal concentrations of Con A. Exogenous cyclic AMP inhibits mitogenic responses to both LPS and Con A; however, these inhibitory effects are partially reversed when cyclic GMP is also added to cultures. The mitogenic response of nu/nu spleen cells to LPS can be inhibited by high concentrations of Con A. This inhibition by Con A is also partially reversed by cyclic GMP, indicating that Con A may exert its effect by elevating intracellular levels of cyclic AMP. Thus, while lymphocytee mitogens may affect intracellular levels of both cyclic AMP and cyclic GMP, three effects show a consistent pattern. 1) Exogenous cyclic GMP is either mitogenic for B cells or agents that elevate cyclic GMP levels enhance the mitogenic response of T cells to Con A. 2) Increasing cyclic AMP levels inhibit cell proliferation induced by both B and T cell mitogens. 3) The inhibitory effects of cyclic AMP or cyclic AMP-elevating agents on mitogen-induced proliferation can be partially reversed by the presence of cyclic GMP. The opposing effects of these two cyclic nucleotides may reflect either their involvement in the initiation of cell proliferation or thei modulating effects on cell cycle events after the initiation process has been completed.


Chemical treatment of macrophages increases their responsiveness to migration inhibitory factor (MIF). The response of guinea pig macrophages to migration inhibitory factor (MIF) is altered by several chemical treatments. Treatment of macrophages with the diazonium salt of sulfanilic acid (5 x 10(-6) to 4 x 10(-4) M) significantly increases the response of these cells to MIF. Treatment with acetic anhydride also augments the response of these cells to MIF. The latter finding suggests that alteration of amino, hydroxyl, or sulfhydryl groups is involved in this phenomenon. Treatment of macrophages with sodium periodate (2 x 10(-5) to 10(-3) M) which is known to oxidize cis-glycols and with hydroxylamine (2 x 10(-5) to 2 x 10(-3) M), which reacts with carbonyl groups also increases response to MIF. The following experiments suggest that the significant alteration occurs at the level of the cell surface. Incubation of macrophages with the diazonium salt of sulfanilic acid at 4 degrees C, at which temperature pinocytosis is largely inhibited, is sufficient to increase the MIF response. The activity of the cytoplasmic enzyme aspartate aminotransferase, which in homogenates is susceptible to inactivation by low concentrations of the diazonium salt of sulfanilic acid, is not decreased when intact macrophages are incubated with high concentrations of the diazonium salt of sulfanilic acid. Cumulatively, these findings suggest that modification of different functional groups on the macrophage surface causes the same physiologic effect.


Immune interactions with cells infected with herpes simplex virus: antibodies to radioiodinated surface antigens. Lactoperoxidase-catalyzed radioiodination was used to study reactions between surface antigens and antibodies on BHK-21 cells infected with HSV-1 and HSV-2. Isolation of iodinated surface antigens was achieved by indirect immune precipitation of Triton X-100 disrupted cells with antisera to HSV and IgG. Analysis of immune precipitates by polyacrylamide gel electrophoresis (PAGE) revealed at least 10 antigens, ranging in m.w. from 35 x 103 to 160 x 103 daltons. Antigens were detectable on cell surfaces as early as 2 hr post-infection. Electrophoretic patterns of surface antigens induced by HSV-1 were similar to those induced by HSV-2. In both cases the major portion of activity was associated with glycoprotein(s) in the range of 115 x 103 to 130 x 103 daltons. A reduced amount of radioactivity was obtained if cells were reacted with anti-HSV sera before disruption with Triton X-100, suggesting that less surface antigen was accessible to HSV antibody applied directly to intact cells.


Histamine modulation of eosinophil migration. Preincubation of eosinophils with 10(-5) M or higher concentrations of histamine inhibited the eosinophil chemotactic response to endotoxin-activated serum whether by using the nucleopore filter assay and counting the cells migrating through the filter, or by using the Zigmond-Hirsch assay and counting the cells at each 10-mum interval. When the H2-receptor sites on the eosinophils were blocked by metiamide, the inhibitory capacity of histamine was prevented. Preincubation of eosinophils with 10(-6) M histamine increased the number of responding eosinophils to endotoxin-activated serum and this enhancement was blocked by an H1-receptor antagonist. Isoproteronol and aminophylline inhibited eosinophil movement and increasing concentrations of dibutryl cyclic AMP inhibited eosinophil migration. Concentrations of histamine that consistently resulted in inhibition of eosinophil movement stimulated an increase in cyclic AMP that was prevented by blocking the H2-receptor but not the H1-receptor. Thus, histamine-dependent inhibition of the eosinophil chemotactic response to other agents is mediated through the H2-receptor and is associated with an increase in the intracellular level of cyclic AMP whereas histamine dependent enhancement of eosinophil migration to other agents appears to be mediated through the H1-receptor. Eosinophils behave as a heterogeneous population as assessed by the ability of histamine to augment or inhibit cell migration. This may reflect differences in H1 to H2 receptor density or cell responsiveness to receptor stimulation. The chemoattractant activity of histamine itself is not influenced by H1 or H2 receptor antagonists, thus it is possible that an eosinophil has a third type of histamine receptor.


Reversal of SV40 tumor-mediated suppression of spleen cell cytotoxicity by antibody. Spleen cells obtained from hamsters bearing PARA-7 tumors greater than 1.0 cm were not reactive in microcytotoxicity assays unless preincubated overnight. The events occurring during in vitro incubation which lead to reversal of tumor-mediated suppression of cellular immunity were investigated. After 24 hr of incubation, supernatants overlying spleen cells from tumor-bearing hosts contained a factor which blocked cytotoxicity of simian virus 40 (SV40)3-sensitized spleen cells at the PARA-7 target cell level but not at the effector cell level. The preparations did not mediate antibody-dependent cellular cytotoxicity (ADCC). Opposite results were obtained in assays of culture medium overlying spleen cells from hosts with a tumor burden less than 0.1 cm. Although ADCC activity was present, no significant blocking was detectable. Treatment of inactive spleen cells with anti-hamster gamma-globlin in the presence of complement (anti-HGG + C) prevented activation and formation of blocking factor but did not impair the cytotoxic activity of already activated cells. Addition of SV40 antiserum to anti-HGG + C-treated cells led to effector cell activation, whereas heterologous virus-immune sera did not. Control studies established that the antibody-mediated recovery of cytotoxicity was not due to arming. Further studies showed that PARA-7 tumor antigen extract blocked at the effector cell level, not at the target cell level. Addition of PARA-7 extract to spleen cell supernatants mediating ADCC resulted in formation of a factor which blocked at the target cell level but not at the effector cell level. These data are compatible with the following interpretation. Spleen cell unresponsiveness is due to antigen blockade. Recovery of cytotoxicity occurs because antibody synthesized during the incubation period promotes elution of antigen from the effector cell surface. Thus, activation is accompanied by the generation of tumor antigen-antibody complexes.


Inhibition of the mitogenic response to lipopolysaccharide (LPS) in mouse spleen cells by polymyxin B. The addition of low doses of the cationic polypeptide antibiotic, polymyxin B (PB), to cultures of mouse spleen cells inhibits lipopolysaccharide-(LPS) induced DNA synthesis but not that stimulated by PPD, PHA, or Con A. Inhibition is stoichiometric; the mitogenic response is suppressed by 50% at a weight ratio of PB:LPS of 0.055 to 1. Furthermore, PB-LPS complexes have a much reduced mitogenic capacity. These complexes inhibit the mitogenic response of spleen cells to unmodified LPS but not to PPD, Con A, or PHA. The inhibitory activity of PB is less effective when added after LPS is mixed with responding cells, achieving 50% inhibition when addition is made at 4 to 6 hr. Time course experiments indicate that partial inhibition is a reflection of a lower rate of DNA synthesis. Thus, PB inhibition of LPS mitogenesis apparently occurs as a result of formation of PB-LPS complexes with reduced mitogenic capacity. Specific inhibition by the complexes of mitogenesis induced by native LPS suggests that the inactive complex may bind to B cells but is unable to trigger them.


Immune response to acute otitis media in children III. Implications of viral antibody in middle ear fluid. With the technique of radioimmunodiffusion and indirect FA staining, IgA antibody to measles, mumps, rubella, and polio-1 were determined in serum and middle ear fluid (MEF) of 103 patients with otitis media. The occurrence of IgA viral antibody in MEF and its absence in simultaneously drawn serum was used as an indicator of local antibody production. Of the 401 assays performed, 41 instances of IgA antibody exclusively in MEF were found. Only four of these occurred in specimens from unimmunized patients and were directed against rubella virus. Of the 37 remaining cases, 16 each were directed against measles and polio-1 and four and one, respectively, for mumps and rubella viruses. The mean specific IgA titers were from 8- to 17-fold higher in MEF from immunized individuals than in the unimmunized and persisted for at least 9 to 19 months after immunization. The data thus indicate 1) specific immunologic sensitization of the middle ear mucosa can be achieved by parenteral as well as oral routes of immunization, and 2) specific immunologic memory exists in the middle ear mucosa. These two factors are crucial to feasibility of immunization against acute otitis media.


Effect of selective complement deficiency on the rate of neutralization of enveloped viruses by human sera. The capacity of human sera genetically deficient in selective complement (C) components to enhance neutralization of enveloped viruses was examined by kinetic plaque reduction assays. Vaccinia virus, a DNA virus, and vesicular stomatitis virus (VSV), an RNA virus, were studied. Exogenous rabbit: or human antibody to vaccinia virus, and guinea pig or human antibody to VSV were provided in limiting, C-dependent concentrations. IgG antibodies predominated in most of the antisera employed. C5-deficient and C6-deficient human sera consistently supported normal rates of neutralization of either virus; this effect was heat-labile. C4-deficient human serum did hot exceed heat-inactivated serum in any neutralization assay. C1r-deficient serum displayed slight heat-labile neutralizing capacity against vaccinia but none against VSV. C2- and C3-deficient sera consistently exhibited measurable but clearly subnormal rates of neutralization. Two fresh agammaglobulinemic sera failed to inactivate either virus in the absence of added antibody. These results confirm and extend earlier evidence, based on neutralization of herpes simplex and Newcastle disease viruses in the presence of early (IgM) antibody and functionally pure guinea pig C components or C-deficient animal sera, that the late-acting components C5-C9 are not required for C-dependent neutralization. Data on four enveloped viruses now agree that this function is mediated by C1-C3, although C1 plus C4 appear to have some neutralizing capacity. This requirement for C1-C3 is overcome, however, in the presence of higher antibody cohcentrations, suggesting that the contribution of the C system to viral neutralization in vivo may be chiefly in the early phase of infection when antibody is limited.


In vitro induction of cell-mediated immunity to murine leukemia cells. I. Optimization of tissue culture conditions for the generation of cytotoxic lymphocytes. Cytotoxic lymphocytes (CTL) to allogeneic and syngeneic murine leukemia cells were generated in vitro in "one way" mixed lymphocyte-tumor cell cultures carried out under a variety of conditions. In an attempt to define the optimal culture conditions for sensitization, the following parameters were analyzed: culture vessel, culture volume, responder: stimulator (R:S) cell ratio, lymphoid cell density, source of lymphoid cells, duration of culture, fetal calf serum (FCS) concentration, 2-mercaptoethanol (2-ME) concentration, concentration of amino acids, and buffering system. Of the many variables examined, of particular importance were the R:S cell ratio, the cell density, the FCS concentration, the presence of 2-ME, and the time of harvesting. These factors exerted various effects, both quantitatively and qualitatively, on the magnitude and kinetics of the responses induced in microcultures (5 X 10(6) lymphocytes) and macrocultures (25 X 10(6) lymphocytes). Moreover, optimal sensitization in syngeneic cultures required different conditions than those for allogeneic cultures. Cytotoxic activity was assessed in vitro by a quantitative 51Cr-release assay. The sensitized lymphocytes demonstrated the characteristics ofT lymphocytes and reacted specifically with the sensitizing leukemia cells.


The development of adrenocorticotrophin-sensitive adenylate cyclase activity in the foetal rabbit adrenal: a correlated biochemical and morphological study. Proliferation of the smooth endoplasmic reticulum, the site of some hydroxylating steroidogenic enzymes in foetal adrenocortical cells, is the first major change in the process of their differentiation into steroidogenic tissue. This was observed in our ultrastructural studies on foetal rabbit adrenals to begin at about day 19 of development. Morphological changes in the mitochondria, the site of production of other steroidogenic enzymes, occurred at about day 24. The elongated or rod-shaped forms of the earlier stages became flattened and rounded by this time, while the cristae were transformed from a flattened lamellar type of the earlier stages to the tubulo-vesicular form of the adult. Other changes observed included an increase  in microvilli and in cell size, with a concomitant increase in thickness of the gland. Adenylate cyclase activity in foetal adrenal homogenates was assessed in response to sodium fluoride (NaF) and ACTH. All preparations responded to NaF. While good responses to ACTH were observed at days 24, 27, 28 and in the neonate, there was a lack of any significant response in the day 19 gland. Foetal ACTH was depressed by administration of cortisol, and the effects of this treatment on both the morphological changes and adenylate cyclase activity was reassessed. The response of foetal adrenals to ACTH was depressed by this treatment and differentiation of the mitochondria was arrested. These results suggest a circumscribed period for the development of ACTH-sensitive adenylate cyclase coinciding with the time at which final differentiation of the mitochondria is completed. Furthermore, both the differentiation of the mitochondria and the development of ACTH-sensitive adenylate cyclase in the foetal adrenal may be dependent on foetal ACTH secretion.


Effect of adrenergic blockade and dexamethasone on the aldosterone response to brain stimulation in the rhesus monkey. The increase in aldosterone and plasma renin activity (PRA) observed after stimulation of extrahypothalamic sites within the brain of the rhesus monkey was prevented by the prior administration of the beta-adrenergic blocking agent propranolol. alpha-Adrenergic blockade by phentolamine had no inhibiting effect. Propranolol only partially reduced the response of aldosterone to lateral hypothalamic stimulation in spite of inhibition of PRA; a partial reduction in aldosterone was also obtained from this site after dexamethasone treatment without any effect on PRA. It was concluded that the increase in aldosterone observed after extra-hypothalamic stimulation was mediated mainly through the renin-angiotensin mechanism whereas in the case of the hypothalamus, release of ACTH was also a contributory factor.


Pregnant mare serum gonadotrophin: ratio of follicle-stimulating hormone and luteinizing hormone activities measured by radioreceptor assay. Specific radioreceptor assays for FSH and LH, which employ tissue receptors from rat testis and highly purified human FSH (LER 1575-C) and LH (Hartree IRC-2, 24/6/69) as standards, have been developed to determine the FSH-like and LH-like activities in pregnant mare serum gonadotropin (PMSG). Measurements of FSH and LH concentrations in the serum of six pregnant Pony mares showed that the ratio of these two activities did not vary significantly between mares and remained constant between days 40 and 80 of gestation with a value of 1-45 +/- 0-04 (S.E.M.). The FSH:LH ratio of PMSG produced by cultured equine trophoblast cells was found to be 0-72 +/- 0-03 (S.E.M.) and that of partially purified serum extracts of PMSG 1-08 (range 0-87-1-30).


The signal from fruiting body and conus tips of Dictyostelium discoideum. Tips from fruiting bodies and conuses were transplanted into interphase fields of Dictyostelium discoideum amoebae. Progressively increasing concentrations of beef-heart phosphodiesterase added to the fields significantly decreased the chemotactic range of the responding amoebae. The findings suggest that the tip secretes c-AMP. We also find that the chemotactic range is independent of the size of the tip implying that the tip may produce a regulating gradient.


Cell aggregation and sexual differentiation in pairs of aggregation-deficient mutants of Dictyostelium discoideum. A diffusible aggregation-stimulating factor (ASF) is released from a series of aggregation-deficient mutants. Biochemical markers indicate that these ASF-donor mutants are blocked at a later step of cell differentiation than an ASF-requiring mutant. ASF is able to bridge the initial block of differentiation in the latter mutant, such that development proceeds up to aggregation and even further. ASF is most probably neither identical with cyclic-AMP phosphodiesterase nor with an inhibitor of this enzyme which both are released from donor strains. In certain combinations of aggregation-deficient mutants, macrocysts, the sexual stages of Dictyostelium, are formed. Also, motile giant cells believed to be zygotes are observed in these mutant combinations. The gamone known to be released from one mating type and to induce macrocysts in the other, is probably not identical with ASF since this factor is produced by mutants derived from either one of both mating types.


The neuronal basis of a sensory analyser, the acridid movement detector system. II. response decrement, convergence, and the nature of the excitatory afferents to the fan-like dendrites of the LGMD. No dendritic spikes occur in the input fan of the lobular giant movement detector (LGMD) neurone. The action potentials are initiated at the point of thickening of the axon, which therefore represents the site of convergence of the retinotopic projection in the MD system. Previous work has shown that the site of decrement in response to repetitive visual stimulation is distal to this point. No change in spiking threshold in the LGMD could be demonstrated, and decrement in the number of LGMD action potentials is completely explained by the observed decrement of EPSPs recorded in the LGMD input dendritic fan. Possible postsynaptic mechanisms which might affect EPSP amplitude are excluded experimentally or shown to be improbable. Latency measurements during electrical stimulation in the second chiasma (which produces a decrementing EPSP in the fan) indicate that the pathway from the chiasma afferents to the LGMD fan is probably monosynaptic. By exclusion, the site of decrement appears to be located at the presynaptic terminal of that synapse. Generalization of habituation of the response to ON and OFF stimuli is demonstrated, showing that the presynaptic neurone at the labile synapse is an ON/OFF unit. The greater part of the previously described sensitivity gradient on the retina, relative to the MD response, appears to be explicable by the geometry of the LGMD fan and of the retinotopic projection. We conclude that the LGMD is fed by a homogeneous population of ON/OFF units running in the second optic chiasma, which form labile synapses on the input fan.


Properties of a symmetric pair of serotonin-containing neurones in the cerebral ganglia of Planorbis. 1. There is a bilaterally symmetric pair of large serotonin-containing neurones in the cerebral ganglia of Planorbis corneus. 2. In some animals these neurones are connected by a non-rectifying electrotonic synapse, and fire in synchrony even at prolonged high frequency. In other animals the neurones are not coupled, and fire independently except when driven by common input. Occasionally the coupling is weak. 3. Both coupled and non-coupled serotonin neurones have processes in the major nerve trunks of both buccal ganglia. 4. Synapses are made with many neurones in the buccal ganglia. The serotonin neurones can initiate firing in several motoneurones and thus produce movements of the buccal mass. 5. During spontaneous feeding cycles the input and firing pattern of the serotonin neurones do not bear any obvious relation to the movements of the buccal mass. 6. The data suggest that the serotonin neurones are modulatory cells, altering the level of excitability of buccal ganglion neurones.


Identification of a cell-surface antigen selectively expressed on the natural killer cell. We have studied the cell-surface phenotype of natural killer (NK) cells of NZB and B6 mice which react to an MuLV+ lymphoid tumor. (a) NK cells do not express Thy1, Ly2, or Ig surface markers. (b) NK cells express an antigen recognized by C3H anti-CE antiserum ('anti-Ly1.2 antiserum'). Inasmuch as NK activity of spleen cells from B6 and B6/Ly1.1 congenic strains were both equally sensitive to C3H anti-CE antiserum, the NK antigen is distinct from Ly1.2. This point was confirmed by the observation that alphaNK activity was removed by absorption of C3H anti-CE antiserum with spleen cells from either B6 or B6/Ly1.1 congenic strains. Absorption of C3H alphaCE serum with BALB/c thymocytes and spleen cells (which are Ly1.2+NK-) removed anti-Ly1.2 activity and left anti-NK activity intact. This absorption step could be circumvented by inserting the BALB/c genotype into the recipient immunized to CE cells (i.e., (C3H X BALB/c)F1 alphaCE spleen cells). This antiserum, provisionally termed 'anti-NK', defines a new subclass of lymphocytes which may play a central role in the immunosurveillance against tumors.


The in vitro effects of Bordetella pertussis lymphocytosis-promoting factor on murine lymphocytes. I. Proliferative response. The lymphocytosis-promoting factor of Bordetella pertussis is a potent mitogen for murine lymphocytes in vitro. The stimulatory response was not the result of specific antigen stimulation. Spleen and lymph node cells were responsive, whereas normal thymocytes were unresponsive. However, DNA replication was induced in cortisone-resistant thymocytes by lymphocytosis-promoting factor (LPF). Bone marrow cells were not stimulated by LPF.


The in vitro effects of Bordetella pertussis lymphocytosis-promoting factor on murine lymphocytes: II. Nature of the responding cells. The mitogenic response of murine lymphocytes to the lymphocytosis-promoting factor of Bordetella pertussis has been shown to be due to activation of T cells. The selectivity of responsiveness to LPF with respect to the population of T cells which is stimllated, differs from that of PHA as well as Con A, and the surface receptors are different. A population of adherent cells, which does not appear to consist of macrophages or other phagocytic cells, is required for the T-cell response.


Isolation of variants in phagocytosis of a macrophage-like continuous cell line. We have isolated cloned variants in phagocytosis from a cloned continuous murine macrophage-like cell line, J 774.2. A selection procedure against Fc receptor-mediated phagocytosis was devised using IgG-coated SRBC containing a toxic drug, tubercidin, as the lethal agent. A series of variant clones deficient in Fc receptor-mediated phagocytosis were isolated. Such variants occurred at low frequency (approximately 6 X 10(-5)), were stable, and appeared to possess Fc receptors. The degree to which they were defective in phagocytosis of IgG-coated SRBC varied from clone to clone, yet all clones, were able to phagocytize latex particles. The phagocytic defect in some variants could be corrected by the addition of 8 Br-cAMP, in others, the drug was without effect. It is likely, therefore, that different variants are defective in several distinct steps critical to Fc receptor-mediated phagocytosis.


Hypercalcemia.

Hypercalcemia. Hypercalcemia is a potentially life-threatening metabolic disorder which may be effectively treated once its presence is recognized and its probable cause determined. The family physician should be aware of the various clinical circumstances in which hypercalcemia occurs and the appropriate initial therapy for patients who are symptomatic at the time of diagnosis. This paper provides a clear approach to the pathogenesis, diagnosis, and management of this problem.


Enhanced resistance of mice to infection with Langat (TP21) virus following pre-treatment with Sindbis or Semliki forest virus. Significant protection to heterologous i.c. challenge with the flavovirus Langat occurred after a single i.c. injection of avirulent strains of the alpha viruses Semliki Forest or Sindbis given 1 day to 5 weeks before challenge. Some protection also occurred after an i.p. infection with these viruses. We consider that the protection afforded by the alpha viruses is due to interference with the multiplication of Langat virus and is related to the maximum level of brain infectivity reached in the alpha virus infection.


Changes in transcription of endogenous type-C virus genome during mouse liver regeneration. In the present study the extent of endogenous, type-C virus genome transcription in normal and regenerating mouse liver was analysed by using the technique of nucleic acid hybridization. The RNA preparations from regenerating liver tissues collected at various intervals following partial hepatectomy, and from normal liver samples of BALB/c mice, were hybridized to 3H-DNA complementary to 60 to 70S RNA of an endogenous, N-tropic virus, released spontaneously from BALB/c mouse cells in culture. Although partial transcription of the endogenous virus genome can be clearly detected in normal liver, a significant increase in the level of virus-specific RNA synthesis in the regenerating liver, in comparison to normal liver, is apparent, following partial hepatectomy. This increase in virus-specific RNA synthesis attains its highest level just before the level of DNA synthesis in the regenerating liver reaches its maximum. These observations may indicate a qualitative or quantitative change in the endogenous type-C virus genome transcription pattern in hepatocytes, in response to partial hepatectomy and suggest that this change in the transcription pattern and the initiation of cell proliferation, in regenerating livers, are probably sequential and related events.


An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. Cells infected with herpes simplex virus type 1 (HSV-1) DNA by the calcium phosphate precipitation technique produce virus which leads to the formation of plaques (Graham, Veldhuisen & Wilkie, 1973). In the study reported here we show that treatment of cell monolayers with dimethyl sulphoxide (DMSO) solutions after infection with DNA-calcium phosphate complexes leads to a considerable increase in the number of plaques obtained. The conditions for this enhancement of infectivity have been optimized for baby hamster kidney (BHK) cells, and increases in plaque numbers of over 100-fold have been obtained. The treatment appears to increase the proportion of cells which respond to DNA infection by initiating plaque formation, and results in a large increase in the measured specific infectivity of HSV-1 DNA. DMSO causes similar (but quantitatively different) responses in various other cell lines infected with HSV-1 DNA. BHK cells infected with either virus particles, or virus DNA by the DEAE-dextran technique (Laithier & Sheldrick, 1975), do not exhibit this massive enhancement following exposure to DMSO.


The effects of herpes simplex virus type 1 on cellular DNA-dependent RNA polymerase activities. An investigation of the activity of nuclear RNA polymerase following infection of LS cells with HSV-1 shows a decline in both major activities. This effect is not entirely due to inhibition of cellular protein synthesis, and the effect of alpha-amanitin-sensitive RNA polymerase is mediated by a protein(s) synthesized in the infected cell. Changes in the properties of this RNA polymerase activity include a reduction in the relative UTP/GTP incorporation ratio and an increased sensitivity to inhibition by actinomycin D, indicating that RNA polymerase II is involved in virus transcription.


Polyadenylated virus-specific RNA in baby hamster kidney cells, transformed by polyoma virus. RNA from a clone of polyoma virus-transformed hamster cells was fractionated by chromatography on oligo(dT)-cellulose. The proportion of virus-specific RNA in the polyadenylated and non-polyadenylated fractions was determined by hybridization of the labelled RNA with excess purified polyoma DNA, immobilized on filters. Seventy to 80% of the virus-specific RNA in both polysomal and total cell RNA was found in the polyadenylated fraction. Since it has been shown previously that more than 65% of the total virus-specific RNA is restricted to the nucleus in this cell line, these results indicate that a high proportion (at least 53%) of the nuclear virus-specific RNA is polyadenylated. The sedimentation profile of total polyadenylated virus-specific RNA in dimethyl sulphoxide was comprised mainly of a broad band with a median sedimentation coefficient about 26S (relative to 28S rRNA). This profile was similar to that of total nuclear, and not cytoplasmic, virus-specific RNA. To estimate the intramolecular proximity of virus-specific sequences to poly(A), total RNA was subjected to limited thermal scission to an average mol. wt. similar to that of mRNA. The RNA remaining attached to poly(A) was then isolated, using oligo(dT)-cellulose. It was found that 65% of the virus-specific RNA that was originally attached to poly(A) was released by the thermal scission. Most of the virus-specific sequence within polyadenylated RNA molecules therefore must have been located at some distance from the polyadenylated 3'-terminus. This observation, together with the results of sedimentation analysis, can most simply be explained by postulating the existence of 'hybrid' RNA molecules containing a host-specified sequence located between a virus-specific sequence and the 3'-terminal poly(A).


The location of the ploy(C) tract in the RNA of foot-and-mouth disease virus. Fragments of foot-and-mouth disease virus RNA of decreasing size, containing the 3' poly(A) sequence have been prepared by alkali treatment and sucrose gradient centrifugation followed by oligo(dT)-cellulose affinity chromatography. Polyacrylamide gel electrophoresis of the ribonuclease T1 resistant oligonucleotides from these polyadenylated fragments has enabled us to locate the position of some of the longer oligonucleotides on the RNA. In particular the poly C tract has been shown to be near the 5' end of the RNA; The possible function of the poly(C) tract is discussed in the light of these findings.


An antigenically atypical strain of variola virus. The Kuwait-5-67 strain of variola virus, although indistinguishable from variola virus in its other properties, behaved atypically in immuno-diffusion tests with antivaccinia serum. The precipitation line pattern produced was similar to that of cowpox virus but lacked one of the main precipitation bands given by all other strains of variola, alastrim and vaccinia viruses tested. Trypsin treatment of cowpox virus preparations revealed the missing precipitation line but did not do so with the Kuwait-5-67 strain. The antivaccinia serum absorbed by Kuwait-5-67 strain lost its ability to interact with this strain, but still gave precipitation lines with variola, vaccinia and cowpox viruses.


Comparison of the activities in inhibition of haemagglutination by different togaviruses for human serum lipoproteins and their constituents. The 3 major lipoprotein fractions of human serum were investigated for their ability to inhibit the haemagglutination of 24 different togaviruses including rubella virus and arboviruses from 5 antigenic groups. All viruses were inhibited by the 3 fractions and this was most evident with low and very low density lipoproteins; inhibitory activity appeared to be associated with the lipid part of the lipoproteins. When artificial dispersions of the lipids cholesterol, cholesteryl ester and triglyceride were emulsified with egg lecithin three categories of inhibitory activity were observed which were independent of virus antigenic grouping -- category 1, no activity; category 2, activity confined to cholesterol and category 3, activity for cholesterol and to a lesser extent the other lipids.


Amino acid composition of the poliovirus capsid polypeptides isolated as fluorescamine conjugates. Poliovirus was treated with fluorescamine and disrupted with sodium dodecyl sulphate (SDS). The polypeptides were isolated by electrophoresis in a polyacrylamide gel (SDS-PAGE) and their amino acid compositions determined. The compositions of VP1, 2 and 3 were very similar but different from that of VP4.


Isolation of the matrix (membrane) protein of vesicular stomatitis virus by gel filtration in guanidine hydrochloride. The L, N and M proteins of vesicular stomatitis virus (VSV) were resolved from each other by gel filtration in the presence of 6 m-guanidine hydrochloride. Amino acid analysis for purified M protein of VSV showed that its chemical composition differed from those of influenza and SV5 M proteins.


Reactivation of herpes simplex virus infection by ultraviolet light and possible involvement of prostaglandins. Herpes simplex infection in the mouse ear was used to investigate whether various treatments would reactivate the disease. Immunosuppressive drugs failed to induce clinical signs of reactivation but irradiation of the skin of the originally infected ear with ultraviolet light or injection of prostaglandin E2 or PBSA into this site, caused reactivation of infection. This was detected by the appearance of infectious virus in the skin 2 to 3 days after these treatments. The results are discussed in relation to the mechanism of herpes reactivation in man.


[Quantitative study of alpha-1-antitrypsin in an ethnic group of Ivory Coast: the Abrons]. A logarithmic-normal distribution of the concentrations of alpha1AT in 578 Abrons in Ivory Coast was found. A significant, negative logarithmic regression of alpha1AT rates according to age is demonstrated. alpha1AT rate = 0,376 (log age) + 3,003 The quantitative analysis by immunoprecipitation of Technicon only does not permit a definitive conclusion as to the presence of allele Z.


Visual pathways, acuity dominance, and visual half-field asymmetry. The effects of the visual pathways and acuity dominance on visual half-field (VHF) recall of verbal stimuli were investigated with 36 right-handed male undergraduates, with the use of the Hines-Satz short-term memory paradigm under monocular conditions. Scores for stimuli transmitted via the left temporal and right nasal pathways were significantly greater than those for stimuli transmitted via the left nasal and right temporal pathways. No other significant differences were found for either the pathways or acuity dominance variables. The results were hypothesized to reflect the primary influence of hemispheric asymmetry of function on the VHF recall task.


Spinal evoked responses recorded from the epidural space in normal and diseased humans. In 22 subjects, including normal subjects and patients with radicular or spinal cord lesions, the authors studied the spinal evoked responses recorded extradurally after stimulating mixed limb nerves. The responses obtained are discussed with particular reference to the clinical value of the changes in amplitude and latency of the spinal evoked potential with particular lesions.


Carnitine-palmityl-transferase deficiency. An increasing number of cases of myopathy due to disordered lipid metabolism have recently been recognised and these appear to fall into two contrasting clinical and biochemical categories. Some patients present with steadily prograssive proximal weakness which sometimes responds to steroid therapy and which is due to carnitine deficiency in skeletal muscle. In the second category, patients usually present with muscle cramps on exertion then followed by myoglobinuria without established proximal myopathy and their symptoms are associated with deficiency of carnitine palmityl transferase in skeletal muscle. In this paper we report a patient who has well-documented carnitine palmityl transferase deficiency, whose symptoms were triggered by violent exercise after fasting and in whom there was variable histochemical and ultrastructural evidence of lipid accumulation in muscle biopsy samples. Development of the patient's symptoms was suppressed by a high carbohydrate diet.


[Multiple systemic atrophies, mental retardation, neurogenic amyotrophy and congenital bone fragility. A new neuro-generative disorder]. Five cases of a congenital neurological disorder are reported. Four patients, born after a breech delivery, belong to one sibship while the fifth patient is the only child in another family. The clinical features include quadriplegia, amyotrophy, a peripheral neuropathy, severe mental retardation and a subluxation of the hips. X-rays reveal diffuse osteoporosis and multiple spontaneous fractures. Autopsies in 3 patients showed multiple system atrophies involving the spinal cord and the cerebellum, coarse cerebral gyri and a marked reduction in volume of the white matter. These various pathological features are compared with the lesions found in a few other cases reported in the literature, none of which can be considered to be identical to the ones described. It is therefore felt that the condition under discussion represents a new syndrome to be classified, at least temporarily, within the group of multiple system atrophies.


Fatal systemic carnitine deficiency with lipid storage in skeletal muscle, heart, liver and kidney. A fatal case of systemic carnitine deficiency is reported. The patient suffered from slowly progressive muscle weakness since early childhood. After the age of 17 years her weakness progressed more rapidly until her death at the age of 20. A pregnancy during the last year of the patient's life was followed by rapid deterioration in her condition. An episode of renal insufficiency occurred at the age of 17 years and hepatomegaly, increased BSP dye retention and intermittent ketoacidosis were present during the last month of her life. Biopsy and autopsy specimens of muscle showed a lipid storage myopathy. Type 1 fibers were selectively severely affected, and many Type 1 fibers were atrophic. Abundant large mitochondria, some also containing abnormal inclusions, were also present in the muscle fibers. At autopsy there was marked accumulation of sudanophilic lipid deposits in all hepatocytes, in the renal tubular epithelial cells, and a patchy increase of lipid material was found in the myocardial fibers. There was marked carnitine deficiency in the patient's liver as well as muscel, while the carnitine palmityltransferase activities in these tissues were abnormally high. The basic metabolic abnormality is assumed to be a defect in carnitine biosynthesis.


Hereditary hypertrophic neuropathy in the trembler mouse. Part 1. Histopathological studies: light microscopy. The Trembler mouse suffers from a dominantly inherited chronic hypertrophic neuropathy. Quantitative light-microscopic studies have been performed on Trembler peripheral nerves from birth to adulthood, and compared with the findings in age-matched control mice. Teased fibres of Trembler nerves contained virtually no myelin, nodes of Ranvier were difficult to identify, and supernumerary Schwann cells were prominent. Results of quantitative studies performed on the dorsal root ganglia and spinal cord of Trembler mice did not differ from those in controls. The density of myelinated fibres in the case of Tremblers was reduced at all ages when compared with controls, and there was a predominant loss of large diameter fibres.


Hereditary hypertrophic neuropathy in the trembler mouse. Part 2. Histopathological studies: electron microscopy. The Trembler mouse suffers from a dominantly inherited hypertrophic neuropathy. Electron microscopy, including a quantitative analysis of myelination was performed on the nerves of Trembler mice from birth to senility and compared with the findings in control mice. Axons in adult Trembler nerves were thinly myelinated and were surrounded by very few myelin lamellae which in turn were often uncompact circumferentially and longitudinally. Schwann cell cytoplasm was copious and had a normal content of organelles. Well-developed "onion-bulb" formations which consisted of thinly myelinated axons surrounded by empty membrane configurations were frequently seen. The initiation of myelination was studied. The diameter distribution of promyelin fibres of control and Trembler sciatic nerve at ages day 2, j, and 7 was calculated Myelination in Trembler nerves commenced on axons of larger diameters than controls. The effectiveness of myelination was studied by relating the number of turns of myelin to the axon area of control and Trembler sciatic nerves from age 2 days to adult mice. At all ages Trembler axons were less well myelinated than controls and the difference was more pronounced with age. Schwann cell activity was examined by relating the area of the Schwann cell cytoplasm to the area of the axon it invests. The relative amount of Schwann cell cytoplasm decreased progressively in control axons with age and as the axon became better myelinated. By contrast, that of Tremblers did not undergo a similar reduction as the animal matured and the relative amount of Schwann cell cytoplasm was markedly increased in adult Tremblers when compared with controls. The periodicity of control and Trembler compact myelin was compared. The myelin period of Trembler mouse was significantly greater than that of controls. The defect in Trembler peripheral nerves was considered to be that of dysmyelinogenesis. The Schwann cell was active but ineffective in the synthesis, compaction and maintenance of myelin.


Abducens nerve paralysis due to giant aneurysm in the medial carotid canal. Case report. A case of a giant aneurysm located in the medial part of the carotid canal is reported. The patient had isolated abducens nerve paralysis that completely resolved after common carotid ligation in the neck.


Myocardial uptake (rabbit) of six 99mTc-tagged pharmaceuticals and 85Sr after vasopressin-induced necrosis. A new rapid method for producing myocardial necrosis in rabbits was developed, using percutaneous intramyocardial injection of vasopressin in peanut oil. The 15-min procedure resulted in a mortality rate of 15% and a success rate among surviving animals of 50%. When the lesions were 24 hr old, strontium-85 and a technetium-99m-tagged agent were injected intravenously simultaneously, and the animals were killed 1,6, and 24 hr later for tissue radioassay. Strontium-85 failed to accumulate appreciably in the lesions. Three bone-seeking technetium complexes (pyrophosphate, methylene diphosphonate, and imidodiphosphonate) produced lesion-to-normal myocardial ratios of 6,5, and 14, respectively, at 1 hr, and 20,30, and 33 at 6 hr. The ratios for 99mTc-glucoheptonate were only 2 at 1 hr and 4 at 6 hr, while the ratios of 99mTc-acetylcysteine and 99mTc-citrate were even lower.


Electron microscopic observations of "keratin pools" in chronic hyperplastic oral mucosa. "Keratin pools," previously characterized clinically and histochemically in the superficial epithelium of chronic hyperplastic oral mucosa, were studied by light and electron microscopy. These occured as small beaded and larger coalescent masses which varied in metachromasia. Ultrastructurally, the "keratin pools" consisted of electron-dense, amorphous or finely-granular material developing and coalescing, chiefly as extracellular deposits. The "pools" frequently possessed a layered arrangement alternating with cells having distinct tonofilaments, desmosomes, and definite cell membranes. Occasional bands of filamentous-like material, possibly representing tonofilament bundles, were observed in some "pools."


A postaxial polydactyly-dental-vertebral syndrome. Three patients with postaxial polydactyly and other abnormalities of the hands and feet, hypoplasia and fusion of the vertebral bodies, and dental abnormalities are reported. Two were sisters born to normal unrelated parents; the other patient was the male offspring of a consanguineous marriage. We suggest that this constellation of abnormalities represents a recessively inherited syndrome.


Acid hydrolase activity during growth and encystment in Acanthamoeba castellanii. The activity and sedimentation of acid phosphatase (APase), acid deoxyribonuclease (DNase), and acid ribonuclease (RNase) were investigated throughout growth and encystment in Acanthamoeba castellanii. The activities/mg protein of all 3 hydrolases are high in young cultures and decrease to constant levels in postlog cells. The RNase activity/ameba decreases 50% during growth, whereas the activity/cell of both APase and DNase remains constant. The percent sedimentation at 20,000 g of all 3 enzymes gradually increases from about 40% in midlog to a plateau of 80% in postlog cells. During encystment, the sedimentation behavior of RNase differs from that of APase and DNase. Encystment is characterized by a differential decrease in the activity/cell of the 3 hydrolases, with RNase decreasing most rapidly and APase least rapidly. APase is unique in that a transient increase of its specific activity is noted during encystment, even though its activity/cell is decreasing.


Evidence for species-specific substrate-site-directed inactivation of rabbit adenylate kinase by N6-(6-iodoacetamido-n-hexyl)adenosine 5'-triphosphate. Adenosine 5'-triphosphate (ATP) derivatives bearing iodoacetylamino-n-alkyl substituents [(CH2)nNHCOCH2I] on N6 were synthesized as potential ATP-site-directed irreversible inhibitors of adenylate kinases from rabbit, pig, and carp muscle. When n was 5 no enzyme was progressively inhibited (inactivated) by 1 mM inhibitor under the test conditions (6 h at 0 degrees); when n was 6 the rabbit enzyme was 76% inactivated by 0.79 mM inhibitor whereas the pig and carp enzymes were unaffected by 2.76 mM inhibitor; when n was 7, 1 mM inhibitor inactivated 14% of the rabbit enzyme and did not inactivate the pig and carp enzymes; when n was 8, all enzymes were inactivated 11-15% by 1 mM inhibitor. No inactivation occurred when the iodine of the hexamethylene analogue was replaced by hydrogen. The selective effect occured also in mixtures of the rabbit and pig enzymes and evidence could not be found that the hexamethylene analogue was activated by the rabbit enzyme or deactivated by the pig and carp preparations. The species-specific inactivation in concluded from various lines of evidence to be ATP-site-directed and is attributed to alkylation of an amino acid residue of the rabbit enzyme which in the pig and carp enzymes is absent, inaccessible, or less reactive. These and previous studies with several other enzymes provide evidence that substrate-site-directed agents capable of bonding covalently to an amino acid residue outside the substrate site can be designed to exert species-specific or tissue-specific irreversible inhibition of target enzymes.


Iodo-bis(quaternary ammonium) salts. Potential cartilage-selective x-ray contrast agents. Bis(quaternary ammonium) compounds in which one or both quaternary nitrogens bear iodinated benzyl moieties and the charged centers are separated by 2, 4, 6, or 10 methylene units have been synthesized and evaluted for (a) binding to cartilaginous material, (b) radiocontrast characteristics, and (c) in vitro pharmacological effects. Also prepared was 1,5-diiodo-2,4-bis(beta-trimethylammonioethyl)benzene diiodide, an analogue in which the iodinated aryl group lies between the charged nitrogen centers. Biological studies show that these compounds bind to cartilage but at relatively slow rates and with low persistence, resulting in low and transient levels of radiopacity. In common with simpler bisquaternary compounds, these compounds block synaptic transmission.


Design of substrate-site-directed inhibitors of adenylate kinase and hexokinase. Effect of substrate substituents on affinity on affinity for the adenine nucleotide sites. Nineteen derivatives of adenosine 5'-phosphate (AMP) bearing acylaminomethyl, acetoxy, or alkylaminomethyl substituents on the phosphate-ribose bridge (5' and O-5' positions) of AMP together with 2',3'-O-ethylidene, 2',3',-O-isopropylidene, and 2',3'-di-O-acetyl derivatives of AMP have been synthesized. Their substrate and/or competitive inhibitor properties with pig rabbit muscle AMP kinases indicate that all the substituents except 2',3'-O-ethlidene with the pig enzyme permitted binding of AMP at its enzymic site. Determination of enzyme-inhibitor dissociation constants showed that several compounds with substituents on the ribose-phosphate bridge bind as well or better than AMP. The affinity is ascribed in part to interaction between substituents and a lipophilic region of the enzymes adjacent to the ribose-phosphate bridge in the enzyme-AMP complexes. The enzyme-inhibitor dissociation constants reveal a structural dissimilarity between the pig and rabbit enzymes in the vicinity of the lipophilic region. The substrate and inhibitor properties of eight ATP derivatives gave evidence that affinity of ATP for its substrate site on the AMP kinases is compatible with acetyl- or chloroacetylaminomethyl groups at the phosphate-ribose bridge or with 2',3'-O-ethylidene or isopropylidene residues. The yeast hexokinase-ATP complex tolerated an acetylaminomethyl group at C-5' or a benzoylaminomethyl group adjacent to O-5'. The present findings regarding substituent tolerance could be used in the design of adenine nucleotide site-directed irreversible inhibitors.


Purity of staphylococcal delta-haemolysin obtained by three different procedures. Three methods at present available for the purification of staphylococcal delta-haemolysin were compared as to the purity and identify of the product obtained. None yielded a pure preparation of delta-haemolysin; one of the three preparations did not contain demonstrable delta-haemolysin when tested electrophoretically, but it contained deoxyribonuclease, penicillinase, phosphatase and alpha-haemolysin. The second preparation had delta-haemolysin activity and was free of alpha-haemolysin, but it contained lipase, egg-yolk factor, esterase, deoxyribonuclease, penicillinase, phosphatase and hyaluronidase. The third preparation contained all of the products mentioned above, except phosphatase, and it also contained alpha-haemolysin, staphylokinase, lysozyme and caseinase. These findings are discussed with special reference to the requirement for criteria of purity in work with staphylococcal products.


Various degrees of susceptibility of different stocks of rats to N-2-fluorenyldiacetamide hepatic carcinogenesis. Outbred Osborne Mendel, Japanese, Wistar, NIH Black, and Sorague-Dawley male rats 12 weeks of age ingested 0.025% N-2-fluorenyldiacetamide in a semisynthetic diet Sprague-Daeley and NIH Black male rats were most susceptible to the development of carcinomas and cirrhosis of the liver and also had the highest incidence of metastases. More and larger carcinomas per liver and more poorly differentiated and undifferentiated carcinomas were found, as well as more advanced cirrhosis, Japanese and Wistar male rats were susceptible, but less so, to hepatic carcinogenesis and cirrhosis. These rats had fewer and smaller hepatic carcinomas per liver, and the neoplasms were well differentiated. By contrast, Osborne Mendel male rats were least susceptible to hepatic carcinogenesis and cirrhosis.


Persistence of immunoglobulin and complement components C4 and C3 bound to guinea pig tumor cells. Guinea pig hepatoma cells (line-10) growing as ascites were studied for the presence of immunoglobulin, C4, and C3 (components of complement) on their surfaces. Immunoglobulin, C4, and C3 content increased with length of time spent in the peritoneal cavity. The persistence of these factors bound in vivo or in vitro was also determined. Complement-fixing (CF) activity of cellbound antibody disappeared from the surface more rapidly at 37 degrees C than at 4 degrees C; the continued presence of cellbound immunoglobulin in non-complement-fixing form could be demonostrated at either temperature. No CF activity was released into the medium. C4 and C3 were released into the medium and could be demonstrated in the medium by immunochemical methods.


Herpesvirus saimiri-induced malignant lymphoma in rabbits. Of 9 New Zealand White rabbits inoculated at multiple sc and im sites with a single dose of Herpesvirus saimiri (HVS), 2 developed malignant lymphomas 40-50 days post inoculation. At least 1 animal developed a terminal leukemic phase of the disease. HVS was isolated from the oral and conjunctival swabs and blood and tissue lymphocytes, but not from monolayer cell cultures derived from kidney or lung tissues of the diseased animals. The inoculated rabbits developed low titers of neutralizing antibodies against the virus. Antibodies against HVS specific early and late antigens were not detected in the sera of 7 animals that failed to develop clinical disease, but were detected in the serum of the 1 rabbit with lymphoma. The immunologic response of rabbits to HVS infection was compared to similar responses in infected nonhuman primates.


Carcinogenicity of Camellia sinensis (tea) and some tannin-containing folk medicinal herbs administered subcutaneously in rats. In an attempt to correlate the high incidence of esophageal carcinoma in natives of certain places with their habit of using herbaceous folk medicines, we performed bioassays of several plant extracts and the fractions prepared from them. Fourteen extracts and fractions from 6 plants were injected sc into NIH Black rats. The tannin fractions from Quercus falcata pagodaefolia, Diospyros virginiana, and Camellia sinensis were very active and produced tumors at the injection site in 66% or more of the treated animals. Tannin fractions from 3 other plants and total aqueous extracts from 5 to 6 tested plants were also tumorigenic rats. The induced tumors were malignant fibrous histiocytomas similar, if not identical, to those encountered in humans. The experiment indicated a possibility of induction of tumor in man by the tested plant materials.


Nucleic acids from subcellular fractions of N-nitrosodiethylamine-induced hepatoma in mice. During eight successive isologous passages of hepatoma induced in male C3HA mice by N-nitrosodiethylamine, no common features of tumor progression were observed, although both the mitotic pattern and ploidy differed from generation to generation. These additional cytologic criteria allowed the biochemical examination of material least changed due to tumor progression. Tumor nDNA's were characterized by greater actinomycin D (AD)- and acridine orange (AO)-binding abilities than were normal nDNA's; this could have resulted from a higher proportion of double-stranded regions in tumor DNA. Isolated tumor deoxyribonucleoprotein had both lower template activity in an RNA polymerase system and fewer AD- and AO-binding sites, when compared with the activity and sites from normal mouse liver. RNA-DNA hybridization data with the above-mentioned findings showed that in hepatoma, part of the nuclear genome was repressed. Also, RNA "new classes" appeared and a certain proportion of nuclear genes controlling mitochondrial protein biosynthesis were derepressed in tumor mitochondria. The hybridization of mitochondrial RNA (mtRNA) and DNA revealed new classes of pulse-labeled RNA's in in vitro-incubated liver mitochondria that were absent from intact cell organelles; the hybridization properties of in vivo- and in vitro-formed hepatoma mtRNA's were similar. Competition and hybridization experiments demonstrated that in tumor mitochondria in vivo, some new classes of RNA existed. Hepatoma mitochondrial mRNA had a higher metabolic stability than did normal mRNA.


Elevated adenosine 3',5'-cyclic monophosphate levels in human and animal tumors in vivo. Cyclic AMP (cAMP) levels were measured in several animal and human tumors and in normal human tissues. Malignant tissues did not appear to be deficient in cAMP.


Adenosine 3',5'-cyclic monophosphate levels in x-ray-induced small-bowel adenocarcinoma in the rat. X-ray-induced adenocarcinomas in the small bowels of outbred Lewis Brown Norway and Holtzman rats contained significantly lower concentrations of adenosine 3',5'-cyclic monophosphate than did normal rat intestinal tissue. No significant differences were observed between the intracellular concentrations of the nucleotide in the normal rat small bowel and those occurring in normal-appearing intestinal tissue exposed to tumor induction conditions.


Nonimmune and immune surveillance. I. Growth of tumors and normal fetal tissues grafted into newborn mice. Growth of various fetal tissues and transplantable tumors in syngeneic newborn and adult mice [BALB/c, DBA/2, and (CBA X C57BL/6J)F1] was compared. Fetal skin, a mixture of all fetal tissues, and tumors were transplanted. The tumors arose spontaneously [hepatomas, mammary gland adenocarcinoma (MGAC)] or resulted from malignant conversion of ectopic transplants either of fetal tissues (urinary bladder carcinoma, adenocarcinoma of small intestine, stomach sarcoma) or of adult animal tissues (ovarian carcinoma) in the syngeneic system. The growth of fetal skin transplants and teratomas, which developed after transplantation of minced tissue from 18- to 20-day and 12- to 14-day fetuses, was considerably inferior in newborn syngeneic recipients, as compared with similar transplants in adults. Inhibition of tumor growth observed in newborn animals was manifested in prolongation of latent period before tumor node appearance and in slowing of growth rate of developed tumors. One of six tumors studied (MGAC) grew at the same rate in newborn and adult recipients. It was suggested that a special type of cellular and/or humoral mechanisms controlling tumor growth exists in newborns. The activity of such factors was conceivably based on fetal tumor antigens as targets. We assumed that weakly antigenic and strongly antigenic tumors behaved differently in respect to nonimmune and immune surveillance mechanisms.


Natural occurrence of tumors in mouse strains with different xenotropic and ecotropic endogenous viruses. Natural tumor incidence and type C virus expression in HIH Swiss and BALB/c mice were investigated. The BALB/c mice showed a moderate incidence of lymphoreticular tumors containing infectious ecotropic virus. A second class of lymphoreticular tumors occurred with approximately the same incidence in both strains; though infectious virus could not be isolated from it, it contained the antigens of a common xenotropic virus. These xenotropic viral antigens were found in all NIH Swiss and BALB/c tumors examined and in normal hematopoietic tissues as well. The relationship between different classes of endogenous viruses and tumor development was discussed.


Immunologic identification of Epstein-Barr virus early antigen in a P3HR-1 cell extract. We extracted the Epstein-Barr soluble antigen (EBSA) from the P3HR-1 human lymphoid cell line, which carries the Epstein-Barr virus (EBV), after P3HR-1 cells were activated with 5-iodo-2'-deoxyuridine. EBSA was identified as the early antigen (EA) complex by immunodiffusion and blocking immunofluorescence tests with high-titered human antiserum to EA. The identity of the EA complex was confirmed with antiserum to EA prepared in rabbits and adsorbed to assure its immunologic specificity. The EA was partially purified on Sephadex G-200. Further characterization by immunoelectrophoresis showed that the EBSA moved rapidly toward the anode. This indicated a highly negative charge. EBSA appeared to be distinct from the previously described complement-fixing S-antigen. It was demonstrable only in EA-producer lymphoid cell lines and reacted specifically with human antiserum to EA, whereas the S-antigen was found in all EBV-carrying lymphoid cell lines and reacted with all EBV antibody-positive human antisera.


Suppression of the avian sarcoma virus genome in 8-azaquanine-resistant, transformed, hamster cells. The avian sarcoma virus genome (Schmidt-Ruppin strain) in transformed hamster cells resistant to 8-azaquanine [Ha(SR)AG-50] was strongly suppressed. The suppression was genetically stable and could not be overcome by attempts at induction with 5-iodo-2'-deoxyuridine. Fusion of hamster cells, which had suppressed virus genome, with chicken Rous-associated virus (RAV-1)-preinfected cells easily rescued the sarcoma virus. The rescued virus had envelope properties of RAV-1, as determined by viral interference, virus neutralization, and plating on genetically resistant chicken cells. By repeatedly cloning the rescued virus, we determined that virus recombined in the rescue experiment and that the recombinant virus had the envelope properties of helper virus used for its rescue. Cells with suppressed avian sarcoma virus genome were suitable for preparation of different recombinant viruses.


Nephroblastoma in the rat: histology of a spontaneous tumor, identity with respect to renal mesenchymal neoplasms, and a review of previously recorded cases. The histology of a spontaneously occurring neoplasm of the rat kidney conforming to a classification of nephroblastoma is described and compared with that of N-nitrosodimethylamine-induced renal mesenchymal tumors. This rat nephroblastoma was an encapsulated epitheloid neoplasm with a uniform histologic pattern. Clumps of densely crowded, hyperchromatic cells frequently associated with central, well-differentiated ducts were supported by a less cellular, interconnecting stroma of loose areolar or mature fibrous connective tissue. Neoplastic cells were organized into primitive, ill-defined tubular formations. The neoplastic cell component strongly resembled metanephrogenic blastema. In contrast, the renal mesenchymal tumor was nonencapsulated and consisted of a heterogeneous mixture of connective tissue elements including fibroblast-like spindle cells, smooth muscle, and embryonic mesenchyme that engulfed and sequestered preexisting renal tubules and glomeruli. The separate morphologic identities and apparently unrelated existence of rat nephroblastoma and renal mesenchymal tumor were stressed. The rat nephroblastoma morphologically resembled the malignant epithelial component of human Wilms' tumor, whereas rat renal mesenchymal tumor appeared to have counter-parts in the mesenchymal component of Wilms' tumor and in congenital mesoblastic nephroma (leiomyomatous hamartoma) of infancy. The histologic descriptions of previously recorded occurrences of spontaneous and experimentally induced rat neoplasma classified as nephroblastoma or its synonyms were reevaluated in comparison to the present case. In all but four instances, in which sufficient histologic detail was provided in previous reports, a consistent histologic pattern emerged for this neoplasm in the rat.


Antipyretic and antiviral action of vitamin A in Moloney sarcoma virus- and poxvirus-inoculated mice. Six week-old male CBA/J mice fed a commercial powdered laboratory chow or the same chow supplemented with vitamin A palmitate (150,000 U/kg) were inoculated with either the Moloney strain of murine sarcoma virus (M-MuSV) or poxvirus. Central body temperature was measured daily. Both viruses elicited fevers, but the fevers were less pronounced and of shorter duration in the mice ingesting the vitamin A-supplemented diet. Palpable M-MuSV-induced tumors appeared later, were less frequent, grew more slowly, and were resorbed sooner in the mice fed the vitamin A supplement. Similarly, in these mice the appearance of pox lesions was delayed, their numbers reduced, and their disappearance hastened.


Feline leukemia virus infection: age-related variation in response of cats to experimental infection. Sixty-seven specific-pathogen-free cats of various ages (newborn, 2 wk, 1 mo, 2 mo, 4 mo, and 1 yr) were inoculated ip with either the Rickard (R) or the Kawakami-Theilen (KT) strain of feline leukemia virus (FeLV). Susceptibility to FeLV was judged by induction of a) FeLV group-specific antigens (gsa) in leukocytes, b) FeLV-related disease, c) antibody to feline oncornavirus-associated cell membrane antigen (FOCMA), and d) virus-neutralizing (VN) antibody. Susceptibility to FeLV-decreased with age. Persistent viremia and FeLV-related disease developed in 100% of cats inoculated as newborns, in 85% of cats inoculated at 2 weeks to 2 months of age, and in 15% of cats inoculated at 4 months or 1 year of age. Cats susceptible to FeLV leukemogenesis became persistently FeLV gsa-positive (viremic) at 4 weeks post inoculation and thereafter and produced little or no FOCMA or VN antibody. Cats that resisted leukemogenesis by FeLV all developed persistent FOCMA and VN titers and never became FeLV gsa-positive. The disease in inoculated cats was influenced by virus strain; FeLV-R induced predominantly thymic lymphosarcoma, whereas FeLV-KT caused fatal nonregenerative anemia without concurrent neoplasia.


Cylindroid lamella-particle complexes in lymphoma cells by northern pike (Esox lucius). Cells of the malignant lymphoma of northern pike of Canada contained cylindrical structures with central cores of cytoplasmic elements and walls composed of lamellae in a spiral arrangement, separated by ribosome-like particles or by membranes. These cylindroids were similar, if not identical, to structures in the cells of certain human lymphomas and leukemias. In cross and longitudinal sections, the lamellae showed a pattern of light and dark bands resulting in an approximately 95 A-period that has not been described in the human cylindroids.


Transmission of murine leukemia virus (Scripps) from parent to progeny mice: a comparison of assay systems. Transmission of murine leukemia virus (MuLV) from parent to progeny C3H/St and C57BL/St mice was examined by four assay systems: 1) recovery of infectious NB-tropic MuLV from spleen cultures, 2) the radioimmunoassay for p30 antigenemia, 3) morphologic examination for lymphoma development, and 4) the indirect fluorescent antibody technique for antinuclear antibodies (ANA). Transmission of MuLV (Scripps) occurred in 90-100% of C3H/St and C57BL/St progeny nursed by mothers with p30 antigenemia. All assays except ANA were equally sensitive for the determination of MuLV transmission in C3H/St mice, but the incidence of transmission in C57BL/St mice was determined only by assays of their cultured spleens for MuLV. Incidences of ANA were increased in all generations of C57BL/St mice compared with controls; the route of transmission of MuLV (Scripps) was not a factor. Only C3H/St mice infected by virus transmitted from parent to progeny developed ANA. Infectious MuLV was invariably recovered from spleens cultured from mice with p30 antigenemia, which was present in all mice that developed lymphoma. NB-tropic MuLV was also recovered after prolonged cultivation from spleens of 75% of C57BL/St progeny mice that did not develop p30 antigenemia. These suggested that MuLV (Scripps) could exist either as a productive persistent or nonproductive latent infection in C57BL/St mice.


Two-way selection of a stock of Swiss albino mice for mammary tumorigenesis: establishment of two new strains (SHN and SLN). Two mouse strains (SHN and SLN) with high and low incidences, respectively, of mammary tumors were established from the same basal stock of Swiss albino mice that were unrelated in origin to other mouse  strains with mammary tumors.  In the breeders, mammary tumor incidence and average age of the mice when they developed mammary tumors were 97.2 % and 6.6 months for SHN, and 5.57% and 10.1 months for SLN, respectively...


In vitro susceptibility of mink lung cells to the mouse mammary tumor virus. Lung cells from mink embryos were infected in vitro with a purified mammary tumor virus isolated from RIII mouse milk. Specific virus antigen at the cell surface was detected by membrane immunofluorescence; B-type virions budding from the cell membrane were seen by electron microscopy. Nucleic acid hybridization confirmed replication and specificity of the virus produced.


Evaluation of Moloney murine sarcoma and leukemia virus complex as a model for airborne oncogenic virus biohazards: survival of airborne virus and exposure of mice. Aerosols of the Moloney murine sarcoma virus (MuSV-M) and leukemia virus (MuLV-M) complex (MuSV-M/MuLV-M) were generated from refluxing atomizers and then aged in rotating drums at 21 degrees C holding temperature with relative humidities ranging from 25 to 76%. The MuSV-M and MuLV-M aerosolized from the same tumor extract preparation survived almost equally at the four humidity levels. Both viruses remained viable in the airborne state for at least 2 hours after aerosolization. When mice were exposed to airborne MuSV-M/MuLV-M, no macroscopic lesions were observed in lungs or other tissues examined during the 2-month postexposure period. On the basis of this study, MuSV-M was determined unsuitable as a "model system" in which a simple aerosol dose response could be used for biohazard evaluation of oncogenic virus aerosols.


Adenocarcinoma of the kidney. II. Enzyme histochemistry of renal adenocarcinomas induced in rats by N-(4'-fluoro-4-biphenylyl)acetamide. Activities of a broad spectrum of enzymes were studied histochemically in renal adenocarcinomas induced in young male F344 rats by chronic dietary administration of the carcinogen N(4'-fluoro-4-biphenylyl)acetamide. Enzymes included were: dehydrogenases of glucose-6-phosphate, lactate, succinate, malate, and alpha-glycerophosphate; peroxidase (catalase); glucose-6-phosphatase; alkaline and acid phosphatase; Mg2+ ATPase; 5'-nucleotidase; and aminopeptidase. Levels of enzyme activity were estimated visually and scored from 0 (not detectable) to a maximum of 5 (intense). Comparison of estimated activity for each enzyme was made between small neoplastic nodules (stage III tumors) and large adenocarcinomas (stage IV tumors) and between tumors and portions of normal proximal tubules in parenchyma of kidneys from untreated control rats. The results, which revealed nearly identical levels of activity for most enzymes in both stages III and IV tumors, suggested similar metabolic and biologic behavior of these lesions. However, when data for tumors were compared with data for normal proximal tubules, striking differences were observed consistent with: 1) a marked shift of energy metabolism from oxidative to glycolytic production of ATP, with a corresponding reduction in mitochondrial respiration; and 2) simplification of plasma membrane specializations that were possibly associated with a reduction or loss of transport function. These findings were compared with other histochemical, biochemical, and ultrastructural studies of renal adenocarcinomas in rats and man.