Separation of antigen-specific lymphocytes. I. Enrichment of antigen-binding cells. Normal mouse spleen cells were fractionated in dishes coated with thin layers of DNP-gelatin or NIP-gelatin, which were insoluble at 4 degrees C. Highly viable cells were recovered from the dishes by melting the gel at 37 degrees C. NIP3- gelatin layers bound approximately 0.1% and DNP4-gelatin layers 0.5% of normal spleen cells. Increasing numbers of low affinity cells were bound with increasing DNP density of the adsorbent. The binding to insoluble DNP-gelatin was hapten-specific since it was inhibited by DNP-lysine, soluble DNP-gelatin or DNP-BSA but not by soluble gelatin or bovine serum albumin (BSA). It was also inhibited by a polyvalent rabbit antimouse Ig. DNP-gelatin was detected on the surface of cells recovered from DNP-gelatin-coated dishes by 125-I-labeled anti-DNP Ig. The cell surface bound DNP-gelatin could be removed by treatment with collagenase. Collagenase treatment did not detectably affect cell viability or surface receptors. More than 90% of DNP-gelatin binding cells were labeled with a polyvalent 125-I-labeled antimouse Ig before or after collagenase treatment under conditions known to label B lymphocytes. Furthermore, the specific antigen-binding capacity of the purified cell populations could be demonstrated after treatment with collagenase. Purified DNP4-gelatin binding cells contained more than 100 times as many DNP-RFC than unfractionated cells. The enrichment of NIP-RFC in the cell population recovered from NIP3 gelatin-coated dishes was more than 200-fold.


Induction of resistance to antibody-mediated cytotoxicity. H-2, Ia, and Ig antigens are independent entities in the membrane of mouse lymphocytes. Mouse spleen or thymus lymphocytes incubated with monospecific H-2 or Ia alloantisera and then coated with a xenogeneic antimouse Ig serum become specifically resistant to the alloantiserum (and complement) they have been incubated with. This so called "lysostrip method" was used to investigate the molecular interrelationships of antigens in the mouse lymphocyte membrane. The results of this investigation confirm that H-2K and H-2D antigens are carried by two distinct populations of molecules. They provide evidence that the Ia antigens move in the membrane independently of both H2-K and H-2D antigens; and finally they demonstrate absence of any physical linkage between Ig receptors in B cells, on the one hand, and Ia, H-2K, and H-2D molecules on the other hand.


Genetic control of immune response. The dose of antigen given in aqueous solution is critical in determining which mouse strain is high responder to poly(LTyr, LGlu)-poly(LPro)--poly(LLys). Antibody response to different doses of (T,G)-Pro--L, given in aqueous solution, was investigated in the high responder SJL and low responder DBA/1 strains by measuring hemolytic plaque-forming cells (PFC) in the spleens as well as hemagglutination titers in the sera. The gene responsible for the difference between the two strains in the response to this antigen, given in complete Freund's adjuvant, has been previously denoted Ir-3. This gene is not linked to the major histocompatibility locus. In the response to the optimal dose (1 mug) of antigen, no difference could be shown between the strains. The peak of the response and the numbers of direct and indirect PFC were similar in both strains in the primary and secondary response. After injection of higher doses (10-100 mug) of antigen, both the direct and indirect PFC responses were lower in the low responder than in the high responder strain. Moreover, the peak of the response occurred earlier in the high responder strain in the primary response to the 10 mu dose of antigen. After administration of a suboptimal dose (0.02 mug) of antigen, the low responder strain produced in the primary response 4-20 times more indirect plaques than the high responder strain. Also the number of direct plaques was higher in the low responder than in the high responder strain. The serum antibody responses to the optimal and higher doses of antigen were parallel to the PFC responses. From inhibition of PFC with free antigen, it was concluded that a similar proportion of cells was producing high and low affinity antibodies to (T,G)-Pro--L in both strains. High and low zone tolerance could be induced in the two strains with (T,G)-Pro--L, but no difference could be shown between the strains. It is suggested that the Ir-3 gene plays a role in the regulation of the balance stimulation and suppression according to the dose of antigen given.


Clonal nature of the immune response to phosphorylcholine (PC). V. Cross-idiotypic specificity among heavy chains of murine anti-PC antibodies and PC-binding myeloma proteins. Seven mouse myeloma proteins with specificity for phosphorylcholine (PC) were found to share a common antigenic determinant. This group of proteins contained members which differed in genetic origin, heavy chain class, kappa-chain subgroup, individual antigenic determinants and specificity for choline analogues. The cross-idiotypic determinant, VH-PC, was antigenically similar in each of the proteins and was associated with the variable portion of the heavy chain in the region of the antibody combining site. Further studies showed that an indistinguishable determinant was present on IgM anti-PC antibodies isolated from all strains of mice tested regardless of histocompatibility or heavy chain allotype. In view of the finding that this cross-idiotypic determinant was not found on antibodies or myeloma proteins which lacked specificity for PC, the data strongly suggest that a particular heavy chain variable region has been preserved in all mouse antibodies with specificity for PC.


The characterization fo the B-cell repertoire specific for the 2,4-dinitrophenyl and 2,4,6-trinitrophenyl determinants in neonatal BALB/c mice. The (B-cell) repertoire responsive to the DNP and TNP haptenic determinants in BALB/c neonates was analyzed in terms of the specificity of stimulation of neonatal B cells as well as the diversity of specificities available in neonatal populations. The results indicate that the parameters of stimulation of neonatal B cells are similar to those of nonimmune adults, particularly in the exquisitely specific stimulatory process which readily discriminates between haptens as closely related as 2,4-dinitrophenyl (DNP) and 2,4,6-trinitrophenyl (TNP). The clonotypes of monoclonal anti-DNP and anti-TNP antibodies derived from isolated neonatal BALB/c splenic B cells in fragment culture were analyzed by isoelectric focusing. During the first 4 days of neonatal life almost all of the anti-DNP-specific clones were of clonotypes displaying IgM antibodies with pI's of 5.05, 5.25, or 5.55. These could be distinguished from clonotypes responding to TNP which were also predominantly of three distinct pI's, 5.00, 5.15 or 5.40. These clonotypes, which represent the vast majority of the DNP- and TNP-specific antibody capability during the first 4 days of life, represented less than half of the clones by day 6 and were a small  minority by day 9. The observation that individual 1--4-day-old donors had many B cells representative of a given predominant clonotype is evidence for cellular precommitment of specificity and indicates that clones of precommitted B cells exist as the products of normal, antigen-independent, generative processes. The observation of frequently recurring clonotypes in inbred neonates attests to the "germ line" origin of these clonotypes; however, variance in the occurrence of these clonotypes from donor to donor implies a random element in their expression. The finding that several clonotypes occur repeatedly in high numbers early in neonatal development, while other clonotypes occur only sporadically at early times, has been interpreted as a reflection of a sequential ontogenic expression of clonotypes. Thus the DNP- and TNP-specific clonotypes which predominate in neonates may be seen as representative of a total of 5,000-10,000 clonotypes which are expressed as early as the 15th to 17th day of gestation while most clonotypes appear after the 18th day of gestation.


Tumor-secific immunity to chemically induced tumors. Evidence for immunologic specificity and shared antigenicity in lymphocyte responses to soluble tumor antigens. Experiments were designed to explore the apparent paradox that methylcholanthrene-induced tumors of mice evoke tumor-unique transplantation immunity but reveal almost complete cross-reacting antigenicity in tests of lymphocyte behavior in vitro. The approach involved use of tumor membranes solubilized in 3 M KCl, employed both as the stimulating antigen source in a new in vitro proliferation assay of lymphocyte recognition, and as immunogens in vivo. The kinetics of the assay resembled those of in vitro tests of mitogen or specific antigen stimulation in other systems. Lymphoid cell proliferation was assessed in peripheral blood leukocytes, lymph nodes (LN), and spleen over the course of tumor bearing, and in animals immunized by tumor amputation or with the solubilized antigens. The pattern of spread of reactivity was from regional LN to spleen, peripheral blood, and nonregional nodes in each circumstance. An unexplained low antigen dose inhibitory phenomenon was encountered in spontaneously proliferating cell subpopulations taken from some tumor-bearing animals. In vitro responses to some but not all solubilized antigens made from multiple syngeneic tumors were detected in each circumstance. The soluble antigens also induced shared resistance to some tumors. The patterns of spread of responsiveness to syngeneic tumor antigens, the time-course, and relative intensity were most compatible with independent clonal responses to multiple tumor-borne antigens, some but not all of which are shared in any family of syngeneic tumors.


The in vitro induction of immunological tolerance in the B lymphocyte by oligovalent thymus-dependent antigens. B-cell tolerance has been induced by oligovalent thymus-dependent antigens in an entirely in vitro system. Dissociated spleen cells from congenitally athymic (nu/nu) mice were preincubated for 24 h with 0.1 -- 1 mg/ml of either fowl gamma globulin (FGG) of DNP-human gamma globulin (DNP-HGG). After washing, the cells were tested for the ability to mount in vitro, thymus-independent responses against FGG and DNP. A state of specific responsiveness to either FGG or DNP was thus demonstrated. Features of this wholly in vitro system that paralleled previous findings on the in vivo induction of B-cell tolerance in nu/nu mice were the kinetics, 24 h being required for tolerance induction in either case, the abrogation of tolerance induction by the presence of POL both in vivo and in vitro, and finally the observation that in neither case was there a requirement for the antigens to be deaggregated. It was shown that DNP-(Fab) 2 fragments prepared from HGG induced DNP-specific tolerance indicating that the Fc piece was not required for tolerance induction in this in vitro system. DNP-bovine serum albumin was less effective than DNP-HGG or DNP-(Fab)2. Preincubation with subtoxic concentrations of DNP-lysine of DNP-epsilon-capric acid had only a marginal effect on DNP responsiveness. Since nu/nu mice, lacking in detectable T-cell function, were used as spleen cell donors, this work provides further evidence that B-cell tolerance to thymus-dependent antigens can be induced without the participation of T cells. It is suggested that B-cell tolerance to thymus-dependent antigens occurs when the antigen in a sufficient concentration and over a sufficient period of time has direct access to the B cell. This contact with antigen must be in the absence of an additional influence provided either by adjuvants like endotoxin or POL, or by activated macrophages, which may be stimulated by activated T cells; otherwise not tolerance but B-cell activation will occur.


Tolerance induction in B lymphocytes but thymus-dependent antigens. T cells may abrogate B-cell tolerance induction by prevent an antibody response. Thymus-dependent protein antigens such as fowl gamma globulin (FGG) and dinitrophenylated-human gamma globulin (DNP-HGG), readily induced tolerance of the B cell in the absence of T cells even when these antigens were not deaggregated. However, when the same doses of antigen were given in the presence of T cells, the B-cell population was shown to be protected from tolerance induction, especially when the antigen was not in a deaggregated form. In this case, there was in fact evidence of a priming effect, manifest in both the B-cell and T-cell populations. The priming effect on the B-cell population was demonstrated by an increased response of mice pretreated with DNP-HGG, upon challenge with DNP conjugated to a heterologous carrier. The priming effect on the T-cell population was evident in a helper effect demonstrated in vitro. However, when euthymic mice which had been pretreated with large doses of FGG or DNP-HGG were challenged with the homologous carrier, the results were different. In this case, there was a profound suppression of the response against the carrier or the hapten on that carrier. Suppressor activity was also demonstrated in vitro and was shown to be sensitive to treatment with anti-theta-serum plus complement. Additionally it was shown that the effector phase of the suppression had a definite nonantigen-specific component. Thus, in pretreated euthymic mice, provided the homologous carrier was present, the response to a heterologous carrier was also suppressed. To account for the observation that nondeaggregated antigens can induce B-cell tolerance in athymic mice, but B-cell priming and T-cell-mediated suppression in euthymic mice, it is proposed that B-cell tolerance occurs when antigen at some critical dose interacts with the B cell in the absence of some second signal. This second signal is normally provided by the macrophage, probably with the assistance of the T cell, and its effect is to divert the result of the interaction of the B cell with antigen towards immunization and away from tolerance induction. When a large dose of an antigen that tends to form aggregates is given to an animal possessing functional T cells, both T-dependent helper and T-dependent suppressor activities are generated, thus accounting for a situation where the B-cell population is immunized, but B-cell activation is suppressed in the presence of the original carrier.


The mechanism of tolerance induction in thymus-derived lymphocytes; I. intracellular inactivation of hapten-reactive helper T lymphocytes by hapten-nonimmunogenic copolymer of D-amino acids. Treatment of a p-azobenzoate (PAB) derivative of a copolymer of D-glutamic acid and D-lysine (D-GL) induced a profound state of unresponsiveness to PAB-reactive helper T lymphocytes generated in PAB-mouse gamma globulin (MGG)-primed mice. This unresponsiveness in T lymphocytes was specific for PAB-reactive cells, since the bacterial alpha-amylase-, keyhole limpet hemocyanin-, or ovalbumin-primed helper T lymphocytes were not suppressed by PAB-D-GL treatment. Taking advantage of the relative ease with which PAB-D-GL can induce specific unresponsiveness to helper T lymphocytes in an animal previously primed with PAB-MGG, it was possible to approach certain questions concerning the mechanisms of tolerance-induction and the fate of tolerant helper T lymphocytes in the PAB-D-GL model by utilizing a classical adoptive cell transfer systemmelimination of the possibility of carry-over of the tolerogen with cells or of the generation of suppressor cells as the result of PAB-D-GL treatment as an explanation of the suppression of helper T-cell activity strongly inplicates the existence of a central intracellular mechanism of specific tolerance on the helper T-cell level. The possibility that suppression of the activity of PAB-reactive helper T lymphocytes by PAB-D-GL reflects simple blocking of surface receptor molecules on T lymphocytes was ruled out as it was found that the helper activity of PAB-reactive cells was minimally suppressed even when PAB-D-GL was directly exposed in vitro to helper T lymphocytesmmoreover, the most conclusive evidence on te the tolerant state induced by in vivo exposure of primed T cells to PAB-D-GL. It appears, therefore, that specific tolerance induced by PAB-D-GL' TO PAB-reactive helper T lymphocytes is an example of irreversible inhibition of T-cell reactivity to antigen, reflecting yet to be determined events at the intra- and subcellular levels.


Cell-mediated lympholysis of trinitrophenyl-modified autologous lymphocytes. Effector cell specificity to modified cell surface components controlled by H-2K and H-2D serological regions of the murine major histocompatibility complex. Splenic lymphocytes from four C57BL/10 congenic resistant mouse strains were sensitized in vitro with trinitrophenyl (TNP)-modified autologous spleen cellsmthe effector cells generated were incubated with 51-Cr-labeled unmodified or TNP-modified spleen or tumor target cells, and the percentage of specific lympholysis determined. The results obtained using syngeneic-, congenic-, recombinante, and allogeneic-modified target cells indicated that TNP modification of the target cells was a necessary but insufficient requirement for lympholysis. Intra-H-2 homology either between modified stimulating cells and modified target cells or between responding lymphocytes and modified target cells was also important in the specificity for lysis. Homology at the K serological region or at K plus I-A in the B10.A and B10BR strains, and at either the D serological region or at some other region (possibly K) in the B10.D2 and C57BL/10 strains were shown to be necessary in order to detect lympholysis. Experiments using (B10itimes C57BL/10)F1 responding lymphocytes sensitized and assayed with TNP-modified parental cells indicated that the homology required for lympholysis was between modified stimulating and modified target cellsmthe possibility is raised that histocompatibility antigens may serve in the autologous system as cell surface components which are modified by viruses or autoimmune complexes to form cell-bound modified-self antigens, which are particularly suited for cell-mediated immune reactions. Evidence is presented suggesting that H-2-linked Ir genes are expressed in the TNP-modified autologous cytotoxic system. These findings imply that the major histocompatibility complex can be functionally involved both in the response potential to and in the formation of new antigenic determinants involving modified-self components.


H-2 compatability requirement for T-cell-mediated lysis of target cells infected with lymphocytic choriomeningitis virus. Different cytotoxic T-cell specificities are associated with structures coded for in H-2K or H-2D;. Use of syngeneic, allogeneic, F1, AND H-2 recombinatn mice has shown that animals injected with lymphocytic choriomeningitis (LCM) virus generate T cells which are cytotoxic for H-2K or H-2D compatible, but not H-2 different, virus-infected target cells. Three separate lines of evidence are presented which indicate that these immune T cells are sensitized to "altered-self," the self antigens involved being coded for in the H-2K or H-2d regions. Firstly, cytotoxic activity associated with mutuality at H-2D iy, lysis mediated by immune T cells from F1 or H-2 recombinant mice is specifically inhibited only by presence of unlabeled, virus-infected cells that are H-2 compatible with the targets. Thirdly, LCM-immune F1 and H-2 recombinant T cells inoculated into irradiated, virus-infected recipients proliferate only to kill target cells that are H-2 compatible with both the donor and the recipient. All of these experiments establish that there is a dissociation of T-cell activities between parental haplotypes in F1 mice, and between H-2K and H-2D in recombinants. It would thus seem that there are at least two specificities of tlcm-immune T cells in homozygotes, associated with either H-2K or H-2D, and four specificities in F1 hybrids. The significance of these findings, with respect both to gene duplication and to the marked polymorphism in the H-2 system, is discussed.


Correlative study of exfoliative cytology and histopathology of oral carcinomas. Cytology smears from 325 oral carcinomas were studied; the false-negative rate was 37%. Cancers of the floor of mouth exfoliated most malignant cells and those of the gingiva exofoliated the least. There was no correlation between the histologic grading, location, sex of the patient, and the result of the cytology smear. Oral exfoliative cytology should only be used as an adjunctive measure and is not a substitute for biopsy.


Spasmolytic constituents of Cedrus deodara (Roxb.) Loud: pharmacological evaluation of himachalol. Himachalol has been identified as the major antispasmodic constituent in the wood of Cedrus deodara. The pharmacological studies of himachalol on various isolated smooth muscles (guinea pig ileum, rabbit jejunum, rat uterus, and guinea pig seminal vesicle) and against different agonists (acetylcholine, histamine, serotonin, nicotine, and barium chloride) indicated spasmolytic activity similar to that of papaverine. It was a more potent antagonist of barium chloride-induced spasm of guinea pig ileum than papaverine but less effective in reverting a similar spasm of rabbit jejunum and had no relaxing effect alone. In the conscious immobilized cat, intragastric administration of himachalol or papaverine (100 mg/kg) produced equal inhibition of carbachol-induced spasm of the intestine, lasting about 2 hr, but himachalol had a faster onset of action. Himachalol was devoid of spasmolytic effect on the bronchial musculature of guinea pig but was 3.3 times more potent than papaverine in antagonizing epinephrine-induced contraction of the guinea pig seminal vesicle. Intravenous injection of himachalol (3-10 mg/kg) in the cat produced a dose-dependent fall in blood pressure and an increased femoral blood flow.


Cytologic changes induced in rat livers cells by short-term exposure to chemical substances. All the hepatocarcinogens administered to rats markedly increased the mitotic rate in the liver. Except for alpha-benzene hexachloride and nitrosobutylurea, the nonhepatocarcinogens did not appreciably increase the mitotic rate. Cytogenetic analyses on the ploidy rate and chromosome abnormalities evidence qualitative differnces among mitotic liver cells in animals treated with hepatocarcinogens and chromosome changes in liver cells observed after the administration of hepatocarcinogenic substances may have some relation to an essential process in hepatocarcinogenesis.


Palliative urinary diversion for pelvic malignancy. Palliative urinary diversion was done in 47 cases for ureteral obstruction secondary to advanced pelvic malignancy. The average survival time was 5.3 months, with only 50 per cent of the patients alive at 3 months and only 22.7 per cent alive at 6 months. After the diversion 63.8 per cent of the survival time was spent in the hospital. Patients with carcinoma of the prostate fared better than those with other sites of tumor origin, which may reflect the natural history of this tumor.


Clinical applications of a new uroflowmeter. A transportable, easy to handle and accurate uroflowmeter was designed by using standard laboratory equipment. The satisfactory results of assessment of the diagnosis and effects of treatment for relieving obstruction of the lower urinary tract are described.


Rational use of albumin and plasma substitutes. Salt-poor human albumin (25 g/dl) is a safe and effective colloid for increasing intravascular volume. It is usually given together with saline solutions at a final concentration of 5 g albumin per 100 ml infusion volume. Although relatively nontoxic, albumin costs about $120 per liter, more than ten times the cost of artificial macromolecular colloids used as "plasma substitutes". Hydroxyethyl starch (6 g/dl NaCl) is a new polysaccharide volemic colloid similar in effect to dextran-70; 70% of an infused volume remains intravascular at 3 hours and 30% at 24 hours. It is nonallergenic, causes no anaphylactoid reactions, and interferes with coagulation less than dextran-70 or dextran-40. Bleeding may be observed when doses of more than 1500 ml are given without blood replacement, especially in thrombocytopenic patients. The effects of dextran-70 and dextran-40 on inhibiting thrombogenesis and facilitating blood flow in small vessels are due largely to hemodilution; the relative efficacy of hydroxyethyl starch for these purposes has not yet been established. Hydroxyethyl starch is a safe and effective colloidal solution for replacement of lost blood and augmentation of blood volume.


Immunoperoxidase labeling of subacute sclerosing panencephalitis virus in hamster acute encephalitis. A hamster neuroadapted strain of subacute sclerosing panencephalitis (SSPE) virus induces in the newborn animal an acute encephalitis with ultrastructural features similar to those observed earlier in measles encephalitis and characteristic of a productive viral infection. The direct immunoperoxidase technique, using labeled SSPE gamma-globulin, was applied to formaldehyde-fixed thick sections. These were either chopped after perfusion of the animal or cryocut after quick freezing of the tissue and then fixed. The latter procedure showed good correlation with light microscopic immunoperoxidase or fluorescent antibody staining on paired thin cryostat sections, whereas the former procedure resulted in easier handling of the sections and better ultrastructural preservation, especially when the lysine-periodate-formaldehyde fixative was used. Under electron microscopy the plasma membrane of the nerve cells was often labeled on its inner side in areas where nucleocapsids accumulate to form buds. The membrane antigen appeared to be more focal than in productive SSPE infection in vitro. Within the cytoplasm, most of the tubular inclusions were stained with the label located on the external granular coat of the nucleocapsid. Nonstructural antigen was also detected in the rough endoplasmic reticulum membrane (often on the side of a tubular inclusion), in the cytoplasmic matrix, and sometimes in the nucleus and on the nuclear membrane of giant cells. It thus seems that SSPE gamma-globulin reacts with viral proteins close to the site of synthesis. Because of the high specificity and reproducibility of this method for the detection of intracellular viral antigens, its use is recommended for further studies of conventional and nonconventional viral central nervous system diseases in which an immune response is present.


Surgery of congenital heart disease assessed by radionuclide scintigraphy. Intravenous injection of 99mTc-labeled macroaggregated albumin (MAA) followed by lung and whole-body scintigraphy results in data from which magnitude of right-to-left shunting and distribution of pulmonary arterial blood flow can be calculated. This information is useful in assessing the functional capacity of a surgical systemic-pulmonic anastomosis. Malfunctioning anastomoses do not significantly reduce right-to-left shunting and may cause unilateral pulmonary hyperperfusion. However, preferential nuclide accumulation may occur in either lung, regardless of shunt function. Consequently, the degree of right-to-left shunting must be determined to fully assess a surgical anastomosis. This technique also allows for assessment of the reduction of right-to-left shunting after intracardiac repairs of congenital cardiac abnormalities.


Role of chronic low-level lead exposure in the aetiology of mental retardation. Water-lead levels were measured in the homes occupied during the first year of life by 77 mentally retarded children aged two to six years and 77 non-retarded matched controls, and in the homes occupied by their mothers during pregnancy. The water-lead content was significantly higher in the retarded group, and the probability of mental retardation was significantly increased when water lead exceeded 800 mug. per litre. Blood-lead levels were also significantly higher in the retarded group. It is concluded that lead contamination of water may be one factor in the multifactorial aetiology of mental retardation and that every effort should be made to reduce the lead content of drinking-water.


Regression of warts. An immunological study. Altogether 173 patients with warts were under observation for at least 3 and in most cases 6 months. In 80% of the patients wart-virus antibodies were present and could be measured by immunodiffusion (I.D.) and in 20% also by complement-fixation (C.F.) techniques. The mean duration of the warts in patients with C.F. antibodies was 0-6 years and in the others 1-9 years. The occurence of C.F. antibodies (IgG) was associated with rapid healing; 75% of these patients were cured during the first 2 months of the observation period. In contrast, of the patients with antibodies measurable only by the I.D. technique (IgM and/or low titres of IgG), only 16% were cured during the first 2 months and they had a fairly constant cure-rate (approximately 9% per month) during the 6 months' observation period. The results indicate that the cure of warts is partly connected with immunological phenomena, especially with the presence of C.F. antibodies. In other cases wart regression may be mainly a non-immune process, perhaps due to a limited lifespan of wart cells.


Isolation of genital mycoplasmas from blood obtained shortly after vaginal delivery. Blood obtained from three hundred and twenty-seven women within a few minutes of vaginal delivery was cultured for genital mycoplasmas (Mycoplasma hominis and T mycoplasmas). Twenty-six (8%) of the women had genital mycoplasmas isolated from their blood. Ten women had blood-cultures which contained M. hominis. Fifteen women had blood-cultures which contained T mycoplasmas. Both M. hominis and T mycoplasmas were isolated from the blood of one woman. Only one of two hundred and seventy-three blood-cultures obtained 1 or more days after delivery contained mycoplasmas. The isolation of M. hominis, but not of T mycoplasmas, from the blood was associated with vaginal colonisation and a serological response to the homologous isolate. Two of ten women whose blood contained M. hominis gave birth to stillborn infants. In contrast, there were only four (1.3%) stillbirths among the infants born to the three hundred and one women whose blood did not contain mycoplasmas. This difference is statistically significant.


Normal serum-lipids in renal-transplant patients. In contrast to previous reports, most patients (78%) with a successful renal transplant being followed up at the University of Kentucky Medical Center had a normal serum-lipid profile. The patients with hyperlipidaema (22%) had normal fasting insulin levels; they received similar immunosuppressive therapy but were significantly older and more obese than those with normal lipids. This lower prevalence of hyperlipidaemia and the absence of fasting hyperinsulinaemia are tentatively attributed to the use of alternate-day corticosteroid therapy in stable renal-transplant patients. If confirmed, the relative infrequency of hyperlipidaemia in patients of alternate-day corticosteroid therapy would be an additional advantage of that therapy over a daily regimen.


Effects of intravenous hyperalimentation of plasma-lipoproteins in severe familial hypercholesterolaemia. The plasma-lipoprotein response to intravenous hyperalimentation was studied in three patients with severe familial hypercholesterolaemia. Hyperalimentation substantially lowered plasma concentrations of cholesterol, low-density lipoproteins (L.D.L.P.), and high-density lipoproteins. The chemical composition of L.D.L.P. did not change. Triglyceride levels increased slightly in two patients and decreased in the third. Turnover of radiolabelled L.D.L.P. was disturbed as L.D.L.P. concentration fell but then returned to normal. The mechanism by which intravenous hyperalimentation rapidly lowers plasma-cholesterol in severe hypercholesterolaemia is unknown.


Treatment of acute haemorrhage of pituitary tumours. Sudden headache, visual loss, and mental change developed in three patients with unsuspected pituitary tumours. In each case stereotaxic transnasal cannulation of the intrasellar mass demonstrated a liquid haematoma under moderate pressure which could be easily aspirated. Restoration of vision and relief from neurological disturbances was prompt. Radiation therapy was then administered to prevent recurrence.


Doxycycline treatment and human infertility. The role of mycoplasmas in infertility was studied in 120 couples. During the twelve months of the study 27 couples (22-5%) conceived. T mycoplasmas were isolated from 63% of these couples, and Mycoplasma hominis from 18%, compared with 56% and 13%, respectively, in those who did not conceive. 88, with primary infertility of unascertained cause, took part in a controlled trial with doxycycline. The couples in the trial were allocated randomly to three groups: 30 received doxycycline, 28 received a placebo, and 30 couples were untreated. Although a twenty-eight-day course of doxycycline eradicated M. hominis and T-strain mycoplasmas from 27 (96%) of the 28 couples harbouring them, the rate of conception was no higher in those treated with the drug than in control groups. It is  concluded that mycoplasmas are not associated with primary infertility and that, although doxycycline eradicates them, this drug is of no benefit in the treatment of primary infertility of unascertained cause.


Bone-marrow-profileration patterns in acute myeloblastic leukaemia determined by pulse cytophotometry. The proliferation patterns of normal and leukaemic bone-marrow were studied by measuring the D.N.A. content of large numbers of cells by pulse cytophotometry (P.C.P.). In nineteen normal bone-marrow samples an average of 66-3% of the bone-marrow cells were in the G1 phase (2n D.N.A.), 26-1%  in the S phase (2n smaller than D.N.A. smaller than 4n), and 7-5% in G2+M phase (4n D.N.A.). The percentages of S-phase cells determined by autoradiography and P.C.P. correlated well, both in normal and in leukaemic bone-marrow. In 25 patients with untreated acute myeloblastic leukaemia (A.M.L.) lower percentages of cells were found in S and G2+M phases, indicating a smaller proliferating pool compared with normal bone-marrow. The likelihood of a complete remission being attained in A.M.L. with the first treatment course was correlated with the percentage of S-phase cells present before treatment. At remission in A.M.L. the proliferation pattern was restored to normal.


Antiglobulin production to altered IgG in rheumatoid arthritis. Conformationally altered IgG molecules have been detected in the serum of patients with rheumatoid arthritis. A hypothesis is presented that specific T-cell unresponsiveness to autologous IgG can be bypassed through the recognition of the altered IgG by competent B lymphocytes. The recognition of altered IgG is mediated through membrane Fc receptors (which may themselves be different in rheumatoid arthritis) and this favours stimulation of those cells carrying a specific receptor for an antigenic part of the molecule. A particular cellular arrangement may be required for complete antigenic stimulation and antiglobulin production. The resultant antiglobulin can have the same binding affinity for autologous and homologous IgG since the antigenic part of the molecule need not be a structurally altered site.


Early events in atherogenesis.

Early events in atherogenesis. Most of the factors that are thought to be involved in atherogenesis can act by producing increased endothelial permeability. They may cause damage in their own separate ways or through their effects on blood-platelets. Several constituents of platelets could produce vascular damage that may result in intimal oedema and subsequent atherosclerosis.


Antivenom in sea-snake bit poisoning.

Antivenom in sea-snake bit poisoning. Among a series of 101 patients bitten by sea-snakes in Malaya in the years 1957-64, 80% were fishermen. Bathers and divers are occasionally bitten. Before sea-snake antivenom became available the mortality-rate (despite the high toxicity of sea-snake venom) was only 10%; however, of 11 with serious poisoning, 6 died. Subsequently 10 patients with serious poisoning received specific sea-snake antivenom; 2 patients, admitted moribund, temporarily improved but died, and 8 patients recovered dramatically. In serious poisoning the suitable dosage of intravenous sea-snake antivenom is 3000-10,000 units; in mild poisoning 1000-2000 units should suffice.


Total body and serum potassium during prolonged thiazide therapy for essential hypertension. Serum-potassium and total body potassium (T.B.K.) were measured serially for 1 year in a group of eighteen patients with mild essential hypertension. The patients were receiving a single daily dose of 10 mg. bendrofluazide without potassium supplements. At 12 months the mean value for serum-potassium (3-86 mmol per litre) was significantly lower than the mean pre-treatment value (4-26 mmol per litre). There was no significant decrease in T.B.K. in the same period. Diastolic blood-pressure fell significantly, and there were no apparent side-effects from the medication. In the group as a whole, a reduction of about 1 mmol per litre in serum-potassium was associated with an average reduction of 10% in T.B.K., but there was a large individual variation. The amount of potassium loss during the period of study did not seem to be clinically significant. It is suggested that routine potassium supplements are not essential in the treatment of uncomplicated essential hypertension with thiazide diuretics.


Detection of circulating immune complexes in pathological human sera. The results of attempts to detect circulating immune complexes in sera by two methods are described. Both rely on the binding of the first complement component (C1) to the complex for its detection. The results, using these methods, of attempts to show which patients have complexes which contain nucleic acid and hence possibly viral antigens in the complexes are discussed. The anti-complementary complexes found in some patients seem to be associated with immune-complex vascular disease.


Faecal shedding of hepatitis-A antigen.

Faecal shedding of hepatitis-A antigen. Serial stool specimens from two individuals with experimental hepatitis-A infection were examined and the shedding pattern of hepatitis-A antigen (HAAg) was determined by immune electron microscopy. HAAg particles were detected at least 5 days before the development of abnormal transaminase levels and jaundice, but not later than the day of peak transaminase levels. The pattern of early faecal shedding of HAAg particles correlated well with the early infectivity of faeces and accorded with the suggestion that HAAg is involved in the aetiology of hepatitis-A infection.


Elective induction of labour. A randomised prospective trial. In a prospective, randomised trial, 111 obstetrically normal pregnant women, who had elective induction of labour performed between 39 and 40 weeks, were compared with 117 controls who were managed expectantly until 41 weeks. Compared with the controls, the patients who had elective induction of labour had significantly less meconium staining in labour and a smaller blood-loss after delivery. The mean length of labour, the amount of pethidine used, and the Apgar scores at 1 minute were similar in the two groups. In the electively induced group, the caesarean-section rate was lower and the use of epidural analgesia more common than in the controls, but the differences were mot statistically signficant. The hour of delivery was similar in the two groups, suggesting that convenience to medical and nursing staff would not be greatly changed by elective induction of labour. There was no evidence that the hazards to mother and child were increased by elective induction, and its use might improve perinatal mortality by reducing the number of unexplained mature stillbirths.


Complement changes and disseminated intravascular coagulation in Plasmodium falciparum malaria. Complement activation as shown by increased clearance of radioactive C1q and reduction in serum-C3 level was found in fifteen (83%) of eighteen patients studied during Plasmodium falciparum infection. Six patients had haemostatic defects suggesting disseminated intravascular coagulation (D.I.C.), and five others had other clinical complications. A correlation was found between reduction of C3 and clinical complications as well as with the degree of anaemia and with thrombocytopenia. Moreover, the most severe thrombocytopenia and the most severe reduction of C3 levels were found in those patients with D.I.C. An immune reaction associated with complement activation in P. falciparum infection is believed to contribute to injury of red blood-cells and platelets and to promote the development of D.I.C. and other serious clinical complications.


Resistance to slimming: adaptation or illusion? Twenty-nine women who claimed that they could not lose weight on prescribed diets were isolated in a country house and fed 1500 kcal. per day for 3 weeks. Nine women maintained weight within plus or minus 1 kg. and were characterised by low basal (B.M.R.) and daily metabolic rates and by a long previous history of dieting. The remaining women did lose weight, but there was much individual variation. B.M.R. is a good indicator of probable weight loss as a result of dieting. In the group as a whole it was more closely related to body fat than to lean body mass. It was also related to fat-cell number as determined by biopsy, but not to fat-cell size. These findings suggest that among a group of would-be slimmers who claim to be unable to lose weight there will be some who have become metabolically adapted to a low-energy diet and others whose inability to lose weight is illusory.


Metronidazole for Crohn's disease.

Metronidazole for Crohn's disease. Five patients with Crohn's disease for 4 months to 12 years were treated with metronidazole. Four of them improved after 2-4 weeks' treatment. In three of them corticosteroids and sulphasalazine could be withdrawn. One patient did not improve until 4 months of metronidazole treatment had passed and corticosteroids were reintroduced. In three patients metronidazole was discontinued after 4-6 months of treatment. Two of them had no signs or symptoms of relapse 3 and 6 months later. The third patient is free from symptoms but laboratory data indicate disease activity.


Correlation of the blood-level of a pregnancy-associated alpha-macroglobulin with the clinical course of cancer patients. The variations in serum level of a pregnancy-associated alpha-macroglobulin (P.A.M.) were studied during the treatment of breast-cancer patients. Good correlation was found between P.A.M. concentrations and the course of the disease--levels rose prior to the clinical recognition of metastatic disease and decreased significantly on successful treatment. Periodic P.A.M. determinations may allow detection of tumour recurrence while it is still at a treatable stage.


Changing composition of human milk and early development of an appetite control. It is suggested that the changes in composition of human milk during the feed are associated with development of an appetite-control mechanism in breast-fed babies. If such a mechanism exists it would be destroyed by artificial feeds of constant concentration and flavour.


International outbreak of Salmonella Eastbourne infection traced to contaminated chocolate. Between Dec. 4, 1973, and Feb. 15, 1974, 80 cases of infection due to Salmonella eastbourne, previously a rare isolated serotype in the United States, were reported from twenty-three States. An additional 39 cases were reported from seven Provinces in Canada during a similar period. A telephone case-control study implicated Christmas-wrapped chocolate balls manufactured by a Canadian company as the vehicle of transmission. S. eastbourne was subsequently isolated from several samples of leftover chocolate balls obtained from homes where cases occurred. Investigation of the factory revealed that the contaminated Christmas and Easter chocolates, and a few chocolate items for year-round sale, had been produced between May and October, 1973. Bacteriological testing of samples taken at the plant implicated cocoa beans as the probable source of the salmonella organisms which, in the low-moisture chocolate, were able to survive heating during production. This outbreak and the finding of salmonella of other serotypes in chocolates produced by another manufacturer suggest that chocolate-related salmonellosis may be a significant public-health problem.


Controlled prospective study of the effect on liver function of multiple exposures to halothane. Patients who had received halothane within a periof of one year and who required another anaesthetic were allocated at random to be given halothane or a control anaesthetic, the control being fiben using halothane-free apparatus. There were 76 patients entries in each group. Serum-glutamic-oxaloacetic-transaminase (S.G.O.T.) levels were measured before the anaesthetics and serially postoperatively for two to three weeks. The S.G.O.T. levels in the halothane group were significantly higher than in the controls. High levels were confined to patients who had had less than four previous halothane anaesthetics, increases above normal in the remainder and in the controls being rate. 1 patient in the halothane group had an S.G.O.T. of 440 I.U. per litre and hepatocellular necrosis on liver biopsy. 2 patients in the halothane group whose S.G.O.T.s rose to more than twice normal showed a similar reaction to re-exposure to halothane, although they had not shown a reaction to an intervening control anaesthetic.


Controlled trial of repeated halothane anaesthetics in patients with carcinoma of the uterine cervix treated with radium. 39 patients with carcinoma of the uterine cervix who were treated with radium and required repeated general anaesthetics were randomised to halothane and control groups. Their serum-alanine-aminotransferase (S.G.P.T.) levels were measured before each general anaesthetic, and those patients whose S.G.P.T. levels rose above 100 I.U. per litre were freed from the restriction determined by the initial allocation and treated as indicated clinically. None of the 21 patients in the control group had S.G.P.T. levels rising above 100 I.U. per litre. 4 out of 18 patients in the halothane group developed S.G.P.T. levels above 100 i.u. per litre before their third radium treatment. None of these had any symptoms or alteration in other liver-function tests, but liver biopsies in 2 of these patients showed changes characteristic of Hepatitis. Arbitrary selection of 18 out of the 39 patients would only give rise to the degree of abnormality observed in the halothane-treated group with a probability of about 0-02. In the patients studied who required repeated general anaesthetics at short time intervals, the monitoring of S.G.P.T. levels before each operation was useful screen for liver damage and may have reduced postoperative hepatic necrosis by preventing further anaesthetics with halothane when the liver was already damaged.


Enhanced bactericidal activity of phagocytes from patients with chronic granulomatous disease in the presence of sulphisoxazole. The decrease in the frequency and severity of bacterial infections in four of five children with chronic granulomatous disease (C.G.D.) on long-term sulphonamide therapy was out of proportion to demonstrable direct antibacterial effects of the drug. In an attempt to determine the mechanism for this apparent protective effect, the killing of sulphisoxazole-resistant Escherichia coli and Staphylococcus aureus by leucocytes from five patients with C.G.D. was studied in the presence or absence of 5-50 mug. per ml. sulphisoxazole. With leucocytes from all five patients there was a modest but reproducible enhancement of bactericidal activity in the presence of the sulphonamide. Studies of phagocytosis-associated oxidative metabolism in patients' cell have not revealed a metabolic basis for improved killing.


Asthma mortality associated with pneumothorax and intermittent positive-pressure breathing. Many patients with acute asthma are treated with intermittent positive-pressure breathing (I.P.P.B.) devices. The development of pneumothorax in two fatal cases and its association with I.P.P.B suggests that this complication may be an important hazard in the treatment of acute asthma.


Thioguanine as primary treatment for chronic granulocytic leukaemia. In an attempt to convert the bone-marrow population from Philadelphia-chromosome (Ph-1) positivt to partially or wholly Ph-1 negative, thioguanine was used as primary therapy in seven patients with chronic granulocytic leukaemia (C.G.L.). Although eight episodes of neutropenia were induced, prolonged remission or conversion to Ph-1-negativity was not achieved in any patient. However, thioguanine was at least as effective as busulphan for initial therapy in C.G.L., and had the advantage of rapid reversibility of haemopoietic depression when discontinued. Thioguanine merits further evaluation as an agent for the management of C.G.L. in its chronic phase.


Portacaval shunt in four patients with homozygous hypercholesterolaemia. Four patients with homozygous hyperbetalipoproteinaemia who had proved resistant to intensive medical therapy have undergone portacaval shunt. During 3 weeks of total parenteral alimentation before the operation, the serum-cholesterol decreased significantly in each of the four patients. During the fat-emulsion phase of hyperalimentation, one patient experienced a rise in cholesterol. Postoperatively, the size of xanthomas decreased in all cases; serum-cholesterol levels rose above those achieved during hyperalimentation, and this rise continued for as long as 3 months before a further reduction was recorded. In patients with angina, this improved considerably; and in most cases bruits decreased in itensity. The results at this stage do not, however, suggest portacaval shunt for all patients with homozygous hypercholesterolaemia.


Survival of the genetically incompatible fetal allograft. A hypothesis is put forward to account for the non-rejection of the fetal allograft, based on shared surface-repellent molecules (S.R.M.s) which prevent close apposition of maternal and fetal immunocompetent cells. This hypothesis is shown to be compatible with the major theoretical requirements of the immune system. Thus it provides possible explanations for the self/not-self recognition process, tolerance to organ transplants, the development of autoimmune disease, and high and low zone tolerance. It is also argued that the recognition of self is important with reference to cellular antigens, but may not be essential for humoral antigens.


Infectious mononucleosis: model for self-limiting lymphoproliferation. Evidence for the extensive lymphoproliferation which characterises infectious mononucleosis (I.M.) is summarised, and some of the mechanisms which are though to control lymphoproliferative activity in this disease are discussed. The main host responses which appear to limit lymphoproliferation in I.M. include the development of humoral antibodies against the Epstein-Barr (E.B.) virus-associated membrane antigen (present on the E.B. viral envelope and on E.B.-virus-infected cells) and cellular immune responses directed against E.B.-virus-containing lymphocytes. Recent evidence is reviewed which indicates that E.B. virus preferentially infects B lymphocytes and that these E.B.-virus-containing cells, which are altered antigenically, evoke a massive response in the host T cells which do not carry E.B. virus; in the presence of E.B.-virus-infected B cells, T cells are transformed and become cytotoxic for B cells. Some of the general implications of the predominant T-cell response in I.M. are discussed with particular relation to autoantibody formation and antigenic competition. The possible role of the distinctive heterophil antigens and antibodies in determining the self-limiting course of I.M. is briefly considered. The general status of I.M. as a self-limiting, albeit intense, lymphoproliferative disease is appraised and the tenuous relationship between I.M. and irreversible lymphoproliferative states is discussed.


X-linked recessive progressive combined variable immunodeficiency (Duncan's disease). Of 18 boys in Duncan kindred, 6 died of a lymphoproliferative disease. They exhibited a subtle, progressive combined variable immunodeficiency disease characterised by benign or malignant proliferation of lymphocytes, histiocytosis, and alterations in concentrations of serum-immunoglobulins. Infectious mononucleosis occurred during or preceding terminal events in at least 3 of the cousins. Fever, pharyngitis, lymphadenomegaly, hepatosplenomegaly, atypical lymphocytosis, and a spectrum ranging from agammaglobulinaemia to polyclonal hyper-gammaglobulinaemia occurred. At necropsy, the thymus gland and thymic-dependent areas in the lymph-nodes and spleen were depleted of lymphocytes. Diffuse infiltrates composed of lymphocytes, plasma cells, and histiocytes, some containing erythrocytes, invaded the haematopoietic organs, viscera, and central nervous system. In addition, 2 half-brothers had lymphomas of the ileum and central nervous system. Approximately half the boys, including the half-brothers, were affected, and girls were spared, implying sex-linked recessive inheritance. Various lymphohistiocytoses resemble Duncan's disease, but it is distinctive from them in the mode of inheritance or by histiological characteristics. This study suggests that the Epstein-Barr virus or other viruses triggered the fatal proliferation of lymphocytes and that progressive attrition of T-cell functions allowed uncontrolled lymphoproliferation.


Oral contraceptives and breast cancer. Progress report of an epidemiological study. 322 women aged 16-45 years undergoing treatment for newly diagnosed breast cancer, together with 502 matched control patients, have been interviewed at six hospitals. The data suggest that the use of oral contraceptives is unrelated to the risk of breast cancer. While this result is reassuring, it is important to remember that it mst be many years yet before the question of the possible relation between oral contraceptives and breast cancer is finally settled.


Hormonal pattern of relapse in hyperthyroidism. 22 patients with Grave's disease were followed up for up to a year after antithyroid drug therapy was discontinued. Clinical assessment and serum T3, T4, and thyroid-stimulating-hormone (T.S.H.) estimations were done serially and simultaneously. Serum T3 or T4 concentrations may be elevated briefly in the first few weeks after antithyroid drugs are stopped, as a rebound effect not necessarily indicative of subsequent relapsf. Clinical relapse of hyperthyroidism with subsequent improvement on antithyroid drugs occurred in 13 patients. Of these 13, serum T3 concentrations became elevated before serum T4 concentrations in 5, thus predicting the subsequent development of clinical hyperthyroidism. In the remaining 8 patients who relapsed, serum T4 was elevated a month before the serum T3. Hyperthyroidism was diagnosed clinically after elevated serum T3 concentrations in 11 patients and at the same time in 2 patients. The mean period of "biochemical hyperthyroidism" in these 11 patients was 12 weeks, with a range of 1 to 56 weeks. During this period 9 of the 11 had minor clinical changes attributable to hyperthyroidism. It is concluded that serial estimations of serum T3 provide the most reliable method of monitoring relapse in hyperthyroidism.


Second malignancies complicating Hodgkin's disease in remission. The incidence of second tumours occurring in the course of Hodgkin's disease has been investigated in a series of 452 patients treated with standard chemotherapy or radiotherapy, combination chemotherapy alone, intensive radiotherapy alone, or both intensive radiotherapy and combination chemotherapy administered in sequence. 16 tumours were noted. When analysed according to mode of treatment, 6 cases occurred in a group of 62 patients who received both modalities. When analysed for age, sex, and man-years of follow-up, this group appears to have 14-5 times the risk of developing a second tumour. However, that subgroup which had a complete remission after intensive radiotherapy followed by a relapse of disease, prior to receiving combination chemotherapy, had the highest risk with 18-5 times greater incidence of second tumour than expected. It is noteworthy that, of the 16 second tumours, 2 were acute myeloid leukaemia; in both cases a similar chromosomal abnormality (45 chromosomes, C-group deletion) was noted. The mechanism of oncogenesis may represent a combination of the immunosuppressive effects and cellular effects of those forms of treatment.


Fitzgerald factor: a hitherto unrecognised coagulation factor. A newly recognised asymptomatic coagulation-factor deficiency is reported in a 71-year-old man. This factor seems to operate at an early stage in the intrinsic coagulation pathway and participates in other Hageman-factor-mediated biological reactions. The patient has an atypical cellular inflammatory response as observed by the skin-window technique.


Cell-mediated immunity to liver in patients with alcoholic hepatitis. Mononuclear-cell responses to liver extracts were studied by a migration-inhibition assay in patients with alcoholic liver disease, viral hepatitis, chronic alcoholism without evidence of liver disease, and in healthy individuals. Patients with acute alcoholic hepatitis demonstrated liver-antigen-induced inhibition of migration (migration index [M.I]equal 0-58 plus  or minus 0-08, mean plus or minus S.D.), while patients with cirrhosis, alcoholism, and acute viral hepatitis, as well as healthy volunteers, did not demonstrate such a response (M.I. 0-92 plus or minus 0-13, 0-90 lus or minus 0-10, 0-86 plus or minus 0-18, 0-99 plus or minus 0-04, respectively). It is concluded that cell-mediated immunity to normal or damaged liver tissue may act to perpetuate alcoholic hepatitis and thereby contribute to the development of cirrhosis.


Lymphocyte-dependent antibody and renal graft rejection. A patient with preformed cytotoxic lymphocyte-dependent antibody (L.D.A.) against the specific renal transplant donor rapidly rejected the grafted kidney. The characteristics of rejection were not those of hyperacute rejection due to cytotoxic complement-dependent antibody or of acute rejection is attributed to the cytotoxic activity of L.D.A. The avoidance of transplants based on positive crossmatches by the L.D.A. test or at least the carrying out of the L.D.A. crossmatch is advised.


Etomidate, a new water-soluble non-barbiturate intravenous induction agent. This paper describes clinical experimence, in 100 patients, with the new non-barbiturate, water soluble induction agent, etomidate. This drug produces sleep in one arm/brain circulation-time. There was no effect on pulse-rate, a slight fall in blood-pressure, and a low frequency of apnoea. Involuntary movements were noted, and in some patients these were severe. Etomidate warrants further evaluation as an intravenous induction agent.


Atypical spherocytosis, a disease of spleen as well as of red blood-cells. It is proposed that atypical spherocytosis is a result of severe spherocytosis combined with splenic dysfunction. The spleen exercises a "conditioning" effect on the red blood-cell but is insufficient in the "culling" and "pitting" functions. As a result, a large population of severely damaged erythrocytes is  circulating in the peripheral blood giving rise to atypical autohaemolysis.


Day-transfusion centre for patients with thalassaemia major. In August, 1973, the Royal Children's Hospital in Melbourne, Australia, established a day-transfusion centre for treatment of children with thalassemia major. The centre is described, and the results of the first 4 months of operation were compared with a similar period before the centre opened, for the same fifty children. A significant decrease in total number of nights spent in hospital, hours spent at hospital for each blood-transfusion, days lost from school or unusual activity, and travel cost was noted. The transfusion-rate rose, but this probably reflected a change of policy rather than an effect of the centre.


[Diagnosis and control of therapy of thyroid disorders by TRH-test (author's transl)]. TSH-measurement before and after stimulation with TRH proved to be of good value in the diagnosis of thyroid dysfunctions, better than other parameters including J-131-test. During treatment of hypothyroid disorder TRH-test, often repeatable, allows estimation of T4 substitution dosis. In Graves disease therapy, furthermore, should be judged by in-vitro or clinical parameters.


[Basophil leucocytes and hyperlipidemia (author's transl)]. In explorations with 42 patients, who have got 2 g of fat per kg body weight, Heidsick, a member of our hospital, discussed quantitative and qualitative changing in basophil leucocytes in relation to the cholesterol and triglyceride content of the blood and the body weight. He showed, that there was an increase of basophil leucocytes under the load of fat and a changing in their cellular structure. There was no significant relation to the cholesterol level in blood, but there was such one in a group of healthy persons to the triglyceride level. Important was also a significant correlation between the body weight and the number of basophilic leucocytes.


[Propafenon, a new antiarrhythmic agent for the treatment of ectopic and rapid cardiac arrhythmias (author's transl)]. Propafenon (SA 79, Fenoprain, Baxarytmon) is a new antiarrhythmic agent with "quinidine-like" localanaesthetic direct membrane and beta adrenergic blocking actions. The drug was given to 50 patients with various types of cardiac arrhythmias. The effective and compatible dose was 70-140 mg administered intravenously in 1 to 3 minutes. Therapeutic success was observed in ectopic beats and paroxysmal tachycardias of both atrial and ventricular origin. Propafenon is effective on the sinus node, the atrial wall and the atrioventricular muscle; the rate of sinus node and ectopic pacemakers is lowered. The atrioventricular and intraventricular conduction time may be increased. Within the therapeutic dose Propafenon has only minimal cardiodepressive activity. Propafenon should not be given in patients with severe heart failure, hypotension and shock and atrioventricular block.


Premature chromosme condensation in the bone marrow of Chinese hamster after application of bleomycin in vivo. The Chinese hamster bone marrow was used as a test system in vivo to analyse the chromosome-danaging effect of bleomycin. Both chromosome and chromatid aberrations were found. Mitoses with aberrations (Ma) show a linear dose-effect relationship after a recovery time of 24 h, the same hold true for cells with micronuclei (Cm) and for mitoses with premature chromosome condensation (PCC). The dose-effect relationships for Ma, Cm and PCC run parallel to each other with Ma at the highest and PCC at the lowest level (Ma greater than Cm greater than PCC). The time-effect relationships for Ma, Cm and PCC show that after 12 h recovery time there are no PCCs but the highest frequencies of Ma and Cm indicating that most cells are in their first post-treatment mitoses or Gi-phases at this fixation time. In addition to the frequency determinations autoradiographic analysis were performed to clarigy the nature of the PCCs. The results are interpreted as follows: bleomycin induces chromosomal aberrations that in turn give rise to micronuclei by means of lagging chromatin, main and micronuclei eventually become asynchronous in their cell cycles and mitosing main nuclei induce PCC in the micronuclei.


Induction of serum Haemophilus influenzae type B capsular antibodies in adult volunteers fed cross-reacting Escherichia coli 075:K100:H5. Serum Haemophilus influenzae Type b (HITB) anticapsular antibodies were induced in adult volunteers by feeding of either of two strains of a cross-reacting Escherichia coli 075:K100:H5. In all the fed volunteers colonization for a finite period lasting up to eight weeks occurred, without adventitious reactions. Nine of 14 normal volunteers and one of two volunteers previously immunized with the Type 6 H.influenzae polysaccharide responded with a greater than twofold increase in serum Type 6H. influenzae antibodies. These antibodies induced by Esch. coli were specific for the capsular polysaccharide and had bactericidal activity. The safety of this procedure, the comparable results in laboratory animals and the identification of other cross-reacting strains of Esch. coli with meningococcal and pneumococcal capsular polysaccharides suggest that colonization with these nonpathogenic organisms at birth may provide a general method of preventive immunization to diseases caused by encapsulated bacteria.


Formalin infiltration of the ductus arteriosus. A method for palliation of infants with selected congenital cardiac lesions. To circumvent the decreased pulmonary blood flow associated with closure of the ductus arteriosus in newborns with heart defects, we infiltrated buffered formalin solution into the wall of that structure to delay its closure. In four infants with pulmonary atresia in whom shunts had been unsuccessful or were technically not feasible, this procedure produced rapid improvement of arterial oxygen tension that was maintained in three infants for one to nine months. The other died of complications of attempted shunt procedures. In an infant with interrupted aortic arch and a large ventricular septal defect, formalin infiltration of the ductus and pulmonary arterial banding alleviated cardiac failure and improved lower-body perfusion. Formalin infiltration of the ductus is an effective palliative technic for treating certain congenital cardiac defects. No adverse effects have been noted.


Metabolic parameters in women with metastatic breast cancer. The metabolic mechanism for increased circulating free fatty acids in post-menopausal women with metastatic breast cancer was investigated. Hormone and metabolic response to glucose and growth hormone were compared to cancer patients and control subjects; thyroid, adrenal and pituitary function were evaluated. The results of these studies indicated that breast cancer patients had glucose intolerance and delayed and prolonged insulin secretion, increased basal growth hormone levels and insensitivity of adipose tissue to growth hormone. Cortisol and protein-bound iodine levels were normal and there was no lipolytic factor in the sera of breast cancer patients. The changes observed in breast cancer patients were not attributable to age, obesity, inanition or stress. These metabolic abnormalities may characterize host susceptibility to breast cancer or be effects of tumor.


Unique and repetitive sequences in multiple genes for feather keratin. Embryonic chick feather keratins are a family of homologous polypeptide chains. The mRNA coding for these has been obtained in a pure state and transcribed into complementary DNA (cDNA) using the reverse transcriptase from avian myeloblastosis virus. Studies on the kinetics of hybridisation and reannealing of cDNA indicate that there are 25-35 different keratin mRNA species in the embryonic chick feather, and a total of 100-240 keratin genes in the chick genome. Each keratin gene contains both a unique and a repetitive sequence. It is proposed that the repetitive sequences are the keratin coding sequences and that the unique sequences correspond to untranslated regions.


Benign prostatic hyperplasia. Early recognition and management. At the present time the medical management of prostatic hyperplasia remains primarily in the investigative stage. Various prostatic inhibitory substances, primarily hormonal in nature, have been studied in animals and humans. The work of Scott with cyproterone acetate in a group of 13 patients with nodular hyperplasia demonstrated improvement of the obstructive symptoms in 11 patients, an increased flow rate in 9, reduction of residual urine in 8, decrease in prostatic size in 7, and epithelial histologic changes in 8. Nevertheless in spite of these and other encouraging preliminry results, the principal therapy for genign prostatic hyperplasia is surgical relief of bladder neck obstruction by prostatectomy which has a mortality rate of less than one per cent. The effects of medical therapy remain uncertain and are employed primarily in patients who are poor surgical risks.


Suprapubic and retropubic prostatectomy for prostatic hyperlasia. The historical aspects, preoperative evaluation including preoperative note and postoperative flow sheet examples, and the surgical techniques of suprapubic and retropbuic prostatectomies are described in detail. These procedures allow relief from symptomatic prostatic hyperplasia with minimal morbidity and mortality.


Cerebrospinal fluid proteins in multiple sclerosis. Various CSF proteins were studied in 255 definite multiple sclerosis patients at various disease stages and compared with corresponding values obtained from 174 controls. The CSF changes in acute multiple sclerosis patients included a significant increase of total proteins and of gamma globulin, IgG, IgA, IgM, alpha-2 ceruloplasmin, 7S-gamma-1, and cytotoxic index for nerve cells in tissue culture, and significant decreases of pre-albumin, alpha-1, and alpha-2 and of the beta/gamma globulin ratio. The CSF levels of IgG, IgA, and IgM remained significantly higher in steroid-treated multiple sclerosis patients than in controls, but the levels often were significantly reduced while patients were on treatment or in remission. During remission or treatment with ACTH and/or steroids, the alpha-2 ceruloplasmin, 7S-gamma-1, and cytotoxic index were significantly reduced and the pre-albumin, alpha-1, and alpha-2 globulin classes and the beta/gamma ratio showed a tendency to return to normal.


Unusual demyelinating disease. A form of diffuse-disseminated sclerosis. A young man, in good health until the age of 22, developed progressive personality changes, lethargy, motor diffuculities, urinary bladder dysfunction, and convulsions. Spinal fluid abnormalities included monocytic pleocytosis and selective increase of gamma globulins. The clinical features and the structural lesions in the central nervous system are reminiscent of disseminated-diffuse sclerosis. Oligodendrocytes appeared normal in number and showed nonspecific abnormalities. Comparison of the structural lesions found in this case with those described in seven reports of demyelination indicates that the separation of this syndrome, as a distinct entity, may not be justified. We suggest that such cases be classified as myelinoclastic disorders, in the same category with multiple sclerosis and its variants.


[Tuberculosis of the tracheobronchial lymph nodes in ambulatory practice]. While endemic T.B. is undoubtedly receding, sources of infection are still widespread and knowledge of the pathology of the bacillus is still required. A short account of tracheobroncheal lymphadenopathy is accompanied by radiological illustration of personal cases treated without hospitalisation.


[Therapeutic effect of a pharmacologic combination of choleretics and digestive enzymes in exocrine pancreatic insufficiency]. Phosphorylcholine and homocysteine have an important choleretic action and also potentiate exocrine pancreatic secretion by way of stimulation and a more effective preparation of the substrate against attack by lipolytic enzymes. The protection offered by pancreatic enzymes in the correction of the digestive insufficiency in the endoluminal stage is also known. An analysis was therefore made of the action of an association of phosphorylcholine homocysteine and digestive enzymes in cases of exocrine pancreatic insufficiency and primary or secondary malabsorption. This action was evaluated on the basis of the reduction of elimination of steatorrhoea. For this purpose, faecal lipids were determined with the method proposed by van de Kramer et al. The most significant results were obtained in cases where digestive insufficiency was most marked, where correction of the pancreatic enzyme deficiency and improved biliary function were particularly required.


[Criteria for choice of therapeutic procedures in lymphomas]. Surgery, high-energy radiotherapy and cyclic polychemotherapy are best indicated for the treatment of Hodgkin's disease and other forms of lymphoma. Overall evaluation will determine the most suitable management in each case. Clinical andpathological stage, the histological picture, the degree of vascular invasion and the immunological reactivity of the patient are the most important factors in such an evaluation. Close cooperation between the haematologist, histologist, radiologist, surgeon, radiotherapist, pharmacologist and immunologist will obviously be required.


[Current problems in therapy of evolutive malignant granuloma. Our experience with a new scheme of polychemotherapy]. Treatment of advanced Hodgkin's disease by associating vincristine, adriblastine, prednisone and procarbazine is compared with other widely employed protocols. Most subjects in the series had already received one or more drugs and had relapsed. In spite of this, 60% were still in complete remission one year after the administration of 6 courses of the new association. An account is given of maintenance therapy in Hodgkin's disease once remission is obtained, and the problems thus raised are illustrated.


[Palliative demolitive surgery].

[Palliative demolitive surgery]. Nine cases of untreatable tumour in which radical surgery was employed palliatively are presented.  Three hemipelvectomies for recurrent rhabdomyosarcoma were performed.  In one case, death occurred postoperatively, probably as a result of pulmonary embolism.  One patient survived for 8 months, while the other is still alive after three years.  Of two cases in which interscapulothoracic disarticulation was performed, survivals of 9 and 5 months were observed in subjects with fibrosarcoma in a mastectomy site and recurrent sarcoma of the humerus with ling metastases.  Survival to 7 months was obtained in a case of sarcoma of the maxilla, while three patients with squama cell cancer of the mouth floor, chondrosarcoma of the mandible and botryoid sarcoma of the tonsillar fossa are still living after periods of 10 months to 2 yr.  Though devoid of schematic indications, palliative demolition surgery can be considered in borderline cases where the operative risk is not high.  Irrespective of "quantity", the "quality" of life remaining to the patients can be made compatible with the psychophysiological limits of the human personality.


[Four years of clinical experience with the cardio-eurhythmic action of verapamil in intravenous administration]. Personal clinical experience using Verapamil intravenously in 200 patients with a number of arrhythmia conditions is reported. The effectiveness of the drug in controlling supraventricular paroxysmal tachycardia and high ventricular frequency atrial fibrillation in particular is pointed out. Good results were also obtained in atrial flutter, in sinus tachycardia, in extrasystole conditions and, at times, in ventricular paroxysmal tachycardia. Clinical experience limits contraindications to atrioventricular block, severe shock, excessive digitalization, and the associated use of beta block substances.


[Origin of macular edema caused by pars planitis]. In four cases with clinical diagnosis of "pars planitis" fluorescein angiography was performed and macular area observed. Permeability disturbances were noted in retinal capillary area. This points to the vitreous changes being secondary to an inflammation of superficial retinal layers. For this reason in those conditions the name "hyalo-retinitis" used by Gärtner seems to be more correct than the limiting diagnosis of "pars planitis".


Subacute thyroiditis. A diagnosis in otolaryngology. Subacute thyroiditis is often unrecognized and patients may be treated for pharyngitis, laryngitis, otitis media or similar disorders connected with otolaryngology. It is conceivable that these patients may seek treatment in ENT departments. The difficulties encountered in making a correct diagnosis are illustrated here on the basis of 12 consecutive cases. Eight of these patients were initially misdiagnosed resulting in useless therapy. Subacute thyroiditis is a self-limiting illness, and permanent myxoedema is rare. Nevertheless, one case with this sequela is reported.


[Specific desensitization of respiratory pollen allergies]. Under the influences of the pollen season and of specific desensitization, the serum IgE concentrations undergo modifications. During the pollen season, the antigenic stimulation induces a rapid synthesis of IgE antibodies which manifest themselves by a post-seasonal pic. The immunotherapy causes an increase of the total and specific IgE during the first few months of treatment. Afterwards the serum levels decrease progressively and after two years of treatment are inferior to the initial values. The same evolution is found in all the subjects, but with variable intensities. Forming parallely are specific blocking antibodies belonging to the IgG class which play, without a doubt, a role in the clinical amelioration obtained during the firsttwo years of treatment.


[Value of calcium gluconate labelled with 99mTc in the detection of intracranial tumors]. In a preliminary communication, the authors compare the success coefficients of pertechnate 99m-Tc and of a new labelled molecule: calcium gluconate 99m-Tc, in a group of 32 patients. The results obtained showed the undoubted superiority of the gluconate in comparison with the pertechnate in the detection of intracranial metestases, and in particular those of the posterior fossa. It proved equally effective as bleomycin labeled with 57-Co. Amongst the other types of tumour studies, the use of the gluconate seems to be particularly indicated for neoplasms of the posterior fossa and in those close to the base of the skull and venous sinuses, given the marked reduction in "background noise" obtained when this method is used.


[Role of (serum and secretory) IgA deficiency in chronic respiratory disorders. A proposal 21 cases]. The current possibility of measuring at one and the same time serum IgA and sputum SIgA, has led to precision in knowledge of IgA deficits in respiratory pathology. The authors report 21 cases detected in a group of 1000 patients (adults, adolescents and children), suffering from various chronic respiratory disorders and who had either total deficits in serum and sputum IgA (6 cases) or partial deficits (15 cases - mixed [5], serum IgA [5i1, sputum IgA [5]. In 9 cases the assoicated respiratory disorder was bronchiectasis, in 7 recurrent rhino-tracheo-bronchitis and in 5 asthma. In no cases were any extra-respiratory manifestations noted, in particular digestive disturbance or auto-immune disease. In some cases there was an associated deficit in IgE and, much less commonly, in IgG or M. Cellular immunity was not altered. The authors then discuss the place of IgA and SIgA deficitis in the pathogenesis of chronic respiratory pathology as well as their substitutive treatment using natural human immunoglobulins and the results thereof.


[Alpha fetoprotein in umbilical cord blood]. Quantitative measurement of alpha-foetoprotein was made on the cord blood of 158 new-born of gestational age ranging between 15 and 43 weeks. The technique used was that of simple radial immunodiffusion of Mancini which made possible the exact measurement of A.F.P. in all the specimens apart from 3. The correlation coefficient between the A.F.P. level in cord blood and gestational age is significant (r equals 0,85 for a risk p equals 0,001). However, the degree of difference at a given point in pregnancy and the difference in levels found in certain cases of twin pregnancies, suggest that factors other than gestational age (certain complications of pregnancy, for example) may influence foetal A,F.P.


[Is the prostatic adenoma of hormonal origin]. Prostatic adenoma develops at the site of the periurethral glands. The concentration of 5alpha-dihydrotestosterone seems greater at the center of the adenoma than in the overlying prostate. This suggests that medical treatment of prostatic adenoma might be feasible using substances inhibiting the 5alpha-reduction of testosterone.


Immunological escape mechanism in spontaneously metastasizing mammary tumors. Immunological and biochemical studies of spontaneously metastasizing and nonmetastasizing rat mammary carcinomas and their plasma membranes indicated that: (i) all spontaneously metastasizing tumors have little or no demonstrable glycocalyx, while all nonmetastasizing tumors have a thick glycocalyx; (ii) there is a direct relationship between the glycocalyx and immunogenicity, and an inverse relationship with the metastasizing capacity of tumor cells, properties which can be quantitated by levels of the plasma membrane marker enzyme 5'-nucleotidase (EC3.1.3.5;5'-ribonucleotide phosphohydrolase) activity; (iii) the absence of glycocalyx from the metastasizing tumor cell surface seems to result from its dissociation from plasma membranes, for solubilized cell surface antigen is readily found in the blood of metastasizing tumor bearing rats, while there was no detectable tumor cell surface antigen in the blood of the nonmetastasizing tumor hosts tested; (iv) both metastasizing and nonmetastasizing mammary tumors appear to have a common soluble cell surface antigen; (v) in addition to this common antigen, there is another membrane-bound antigen in the nonmetastasizing, immunogenic tumor cell surface which presumably is the tumor specific transplantation antigen; and (vi) this antigen is immunobiologically unique, but seems to be immunochemically related to the common soluble antigen. It is postulated that the lack of an immunogenic coat and/or the presence of solubilized tumor cell surface antigen in the blood may provide an immune escape mechanism for tumor cells by interfering with cell-mediated immune response of tumor hosts, leading to their dissemination.


Interspecies brain antigen detected by naturally occurring mouse anti-brain autoantibody. Normal mouse sera contain naturally occurring antibodies that are cytotoxic in the presence of rabbit complement for NB1, a cell line derived from a neuroblastoma adrenal metastasis of a spontaneous ovarian teratoma. The anti-NB1 antibodies can be specifically removed from normal mouse sera by absorption of the sera with homogenized brain tissue of mouse, rat, guinea pig, chicken, and man and by homogenized kidney tissue of mouse and man. The antigen recognized by anti-NB1 naturally occurring autoantibodies, designated mouse brain antigen-2 (MBA-2), is not present on other normal tissues or tumor cell lines tested. MBA-2 is distinct from previously described mouse brain antigens.


[The effect of emotional stressors on thyroid gland function]. Response of the normal and goiterous thyroid gland to the information on the forthcoming operation was studied in 12 patients with varicose veins of the lower limbs and in 67 patients with thyrotoxicosis; this was done by determination of protein-bound iodine (PBI). PBI proved to increase before the operation in both groups of the patients.  Due to undesirable hyperfunction of the thyroid gland prior to the operation in thyrotoxicosis, the psychodepressive premedication component should be increased.


Antibodies reactive with cell surface carbohydrates. Normal and immune sera from various animal species were fractionated on columns of Sepharose covalently coupled with the glycoprotein fetuin. Elution of the material bound to fetuin yielded low but reproducible amounts of protein, ranging from 0.02 to 0.2% of the protein mass of the input sera. This material has been identified by immunoelectrophoresis in agar and by zone electrophoresis on cellulose acetate as immunoglobulin. The Ig fractions bound and agglutinated erythrocytes of various species, and also bound to cells from various mouse tissues including heart, kidney, thymus, and spleen. In all cases, the binding was inhibited by glycoproteins such as fetuin and thyroglobulin, by a glycopeptide isolated from fetuin, and by some bacterial lipopolysaccharides. When the binding of these Ig fractions to mouse splenocytes was tested in the presence of 17 saccharides, no inhibition of binding was observed except by sialic acid, D-galactose, N-acetyl-D-glucosamine, and D-mannose, all of which showed partial inhibition. Inasmuch as these four saccharides are present on the carbohydrate moiety of fetuin, the results suggest that the isolated material is a carbohydrate-specific Ig (CS-Ig) fraction of serum capable of binding to the carbohydrate portion of cell surface receptors and glycoproteins. When bound to lymphocytes, these CS-Ig molecules induced redistribution (patching and capping) of cell surface receptors. Moreover, the CS-Ig fractions from chicken and rabbit sera were weakly mitogenic for mouse splenic lymphocytes. CS-Ig fractions are useful new reagents for studying glycoproteins and the interactions and activities of cell surface carbohydrates.


RNA-directed DNA polymerase from human leukemic blood cells and from primate type-C virus-producing cells: high- and low-molecular-weight forms with variant biochemical and immunological properties. RNA-directed DNA polymerase (reverse transcriptase) from leukocytes of individual leukemic patients can be grouped by velocity gradient analyses into two distinct classes, a low-molecular-weight (LMW) class of approximately 70,000 and a high-molecular-weight (HMW) class of 130,000 to 140,000. The reverse transcriptases from mammalian type-C viruses have with one exception (see text) been isolated as enzymes with molecular weights of 70,000. In this study, the reverse transcriptase from extracellular gibbon ape leukemia virus was also isolated only as the LMW class. However, the enzyme from gibbon virus-producing cells was isolated partially in the HMW form; this form was converted completely to the LMW form by treatment with 0.5 M KC1 and 0.5% Triton X-100 and could be re-converted to the HMW form by lowering the KC1 and Triton X-100 concentrations. A similar conversion from a HMW form to a LMW form was demonstrated with enzyme from human leukemic cells. The LMW form of the human and gibbon ape cellular enzymes utilized synthetic primer-templates in a similar fashion to viral enzyme, and this form was strongly inhibited by antisera (IgG) to reverse transcriptase from simian (woolly monkey) type-C virus. The HMW form of both enzymes utilized synthetic primer-templates less efficiently than the LMW form, and was resistant to inhibition by antipolymerase IgG of simian type-C virus. The HMW form of the cellular reverse transcriptases transcribed viral 70S RNA in the absence of synthetic primer relatively more efficiently than did the extracellular viral form. These data suggest that the HMW form is due in part to aggregation of the LMW form and in part to a cellular factor(s) which may affect both the form and function of intracellular reverse transciptase.


RNA-directed DNA synthesis by the DNA polymerase of Rous sarcoma virus: structural and functional identification of 4S primer RNA in uninfected cells. The RNA-directed DNA polymerase of Rous sarcoma virus requires a 4S RNA molecule as primer for the initiation of DNA synthesis on the viral 70S RNA genome. We have now functionally identified primer activity in uninfected cells on the basis of the capacity of cellular 4S RNA to actively participate in the initiation of DNA synthesis by the RNA-directed DNA polymerase of Rous sarcoma virus in vitro. This was accomplished by reconstitution experiments in which 4S RNA from uninfected avian cells was tested for its ability to restore template activity to the viral RNA genome from which all primer had been removed. Similar reconstitution experiments were employed to demonstrate a primer activity in the 4S RNA population of duck, mouse, and human cells. Primer activity appears to be absent in lower eukaryotic or prokaryotic cells. Unambiguous identification of the Rous sarcoma virus primer molecule in uninfected cells was accomplished by directly purifying a 4S RNA molecule from the bulk of host cell transfer RNA and establishing structural similarities between this cellular 4S RNA species and the Rous sarcoma virus primer by two-dimensional paper electrophoresis of oligonucleotides obtained from a T1 ribonuclease digest of the RNA species. We conclude that the Rous sarcoma virus DNA polymerase can utilize a host cell molecule as primer for the initiation of RNA-directed DNA synthesis in vitro.


Cell-surface-associated nucleic acid in tumorigenic cells made visible with platinum-pyrimidine complexes by electron microscopy. Platinum-pyrimidine complexes have been used as the sole electron-dense stains for electron microscopy on 18 types of mammalian cells. These recently discovered complexes react avidly with nucleic acids in vitro, are antitumor agents, and are highly soluble in water. In cells, they are selective for regions rich in nucleic acid and make visible the cellular chromatin, nucleolus, and ribosomes. In addition, cells that are tumorigenic exhibit electron-dense patches at the plasma membrane; normal cells do not. The results of treatment of ascites sarcoma-180 cells with concanavalin A, nucleases, glycosidases, or a protease suggest that the electron-dense patches are DNA, with neuroaminic acid involved in the surface binding.


Thermochemotherapy: synergism between hyperthermia (42-43 degrees) and adriamycin (of bleomycin) in mammalian cell inactivation. The sensitivity of cells exposed in vitro to the antibiotics bleomycin or adriamycin is only mildly increased at 41 degrees over that seen at 37 degrees. However, at 43 degrees a marked synergism between the effects of hyperthermia and drug is observed. This synergism can also be demonstrated to occur in solid tumors in vivo. Cells after bleomycin exposure at 37 degrees repair potentially lethal damage, and 43 degrees inhibits this repair. This inhibition may in part account also for the observed sensitization of the cells to bleomycin, but not to adriamycin, since for the latter no repair can be demonstrated. However, fluorescence measurements show that at 43 degrees much more adriamycin is able to enter the cells than at 37 degrees. The possible implications of the results for cancer treatment are discussed.


Synthesis of DNA complementary to separated human alpha and beta globin messenger RNAs. Human globulin messenger RNA, purified by oligo(dT)-cellulose column chromatography, is reproducibly separated into two bands by polyacrylamide gel electrophoresis in the presence of 99% formamide. The more rapidly migrating (fast) band is somewhat more abundant than the slow band in normal (nonthalassemic) total reticulocyte globin messenger RNA. In alpha-thalassemic (Hb H disease) messenger RNA, the slow band is 6.5 times more abundant than the fast band, whereas in beta-thalassemic messenger RNA the fast band is three times more abundant than a second band, which has a slightly greater mobility than the slow band of normal and alpha-thalassemic RNA. The RNA bands of nonthalassemic globin messenger RNA were eluted from the gel and efficiently transcribed into DNA copies by use of the RNA-dependent DAN polymerase of avian myeloblastosis virus. Hybridization of these copy DNAs to fast and slow band RANs and to nonfractionated normal, alpha-thalassemic, and geta-thalassemic messenger RNAs revealed that the eluted fast band RNA contains predominantly alpha-chain specific sequences, whereas the eluted slow band RNA contains predominantly beta-chain specific sequences. Nucleotide sequence analysis of 32-P-labeled RNA transcribed from the slow band copy DNA also indicated that the slow band RNA is beta messenger RNA.


Evolution of biosynthetic pathways: immunological approach. Through the use of specific immunoadsorbent columns, it is shown that Escherichia coli aspartokinase I-homoserine dehydrogenase I, aspartokinase II-homoserine dehydrogenase II, aspartokinase III, and homoserine kinase, enzymes involved in the same complex biosynthetic pathway, share antigenic determinants. This raises the question of a common origin for the four cibtenoirart kinases. (Aspartate kinase or ATP:L aspartate 4-phosphotransferase, EC 2.7.2.4; homoserine dehydrogenase or Lhomoserine:NADP oxidoreductase, EC 1.1.1.3; homoserine kinase or ATP:L-homoserine O-phosphotransferase, EC 2.7.1.39.)


Nature of the tumor-associated determinant(s) of carcinoembryonic antigen. The carbohydrate moiety of carcinoembryonic antigen could be sequentially degraded by repeated cycles of periodate oxidation, reduction, and mild acid hydrolysis (Smith degradation). After three complete degradations, all fucose and sialic acid, 80% of the galactose, 65% of the mannose, and about 40% of the N-acetylglucosamine were eliminated without impairing the ability of degraded carcinoembryonic antigen to react with specific antisera against the antigen. Inhibition studies in a carcinoembryonic antigen/rabbit anti-carcinoembryonic antigen precipitating system with oligosaccharides covering previously known internal structures of glycoproteins and presumably corresponding to the internal carbohydrate region of the antigen, demonstrated that none of the compounds tested was inhibitory. Nor could any inhibitory effect on the binding of carcinoembryonic antigen to antibody against the antigen in a radioimmunoassay system be domonstrated for the carbohydrate moiety prepared by hydrazinolysis or the glyco peptide fraction isolated after papain degradation of the antigen. However, if carcinoembryonic antigen is completely reduced and alkylated, with three intrachain disulfide bonds cleaved per 10-5 g, the immunological activity is reduced to 3-5% of untreated antigen. Furthermore, treatment of the antigen with 0.5 NaOH at 20 degrees for 2 hr completely abolished its ability to react with antiserum, whereas its ability to precipitate with a series of lectins was unchanged. No release of low-molecular-weight carbohydrate orchange in sugar composition of alkali-treated antigen was observed. Our tentative conclusion is that the carbohydrate moiety of carcinoembryonic antigen does not contain the tumor-associated determinant(s).


Oncornaviral protein modulation in mouse uterine tissue by estrogen (38467). Treatment of ovariectomized NIH Swiss mice with estrogens elevated the level of the murine leukemia virus group specific protein and the activity of an RNA-directed DNA polymerase in the uterus. The extent that these markers were raised was dependent on the relative biological potency of the estrogen and on the time interval following treatment. Increases in the levels of both viral marker proteins were evident within 24 hr of treatment and were highest at 48 hr. Subsequently, viral protein levels declined to pretreatment levels.


Selective immunosuppressive activity of steroids in mice inoculated with the Moloney Sarcoma virus (38503). A steroid, 6-chloro-17-hydroxypregna-1,4,6-triene-3,20-doine (CHP) that exhibits selective activity in several models of cellular immunity including an apparent inhibitory action on the elicitation of delayed hypersensitivity, was examined in a new, simple experimental model for assessing aspects of host-cell-mediated immunological competence. This model is based upon the capacity of the adult mouse to prevent the progressive growth of tumors induced by the Moloney sarcoma virus. Two steroids reported to have immunosuppressive activity in other assay systems, namely, cortisol and progesterone, were also studied. Control mice and those injected with CHP maintained their capacity to reject the tumor. In contrast, significant numbers of mice receiving a single large injection of cortisol or progesterone succumbed to progressive tumor growth under the experimental conditions used. The data indicate that CHP, while influencing selected parameters of cellular immunity, e.g., the elicitation of delayed hypersensitivity, does not decrease the capacity of the host to mount a defense against the progressive growth of the Moloney virus-induced sarcoma. The results indicate that CHP may be useful in modulating specific aspects of cellular immunity without altering others. In addition, the experimental model described provides a simple method of assessing the possible immunosuppressive effects of naturally occurring and synthetic agents on viral-induced tumor growth.


Effect of aza-substituted nucleotides on the starve-refeed response of rats. Rat liver glucose 6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) activities were increased by starvation-refeeding to levels above those found in rats fed ad libitum. The increases in enzyme activities above ad libitum-fed levels were prevented by 8-azaguanine and 6-azauridine, but not by 2-azauridine. Blood insulin levels were not affected at the time studied. Two aza analogs, 8-azaadenine and 5-azacytidine, proved to be too toxic in this type of studies. Since 8-azahypoxanthine, 8-azaxanthine and 5-azauracil were neither effective in preventing the enzyme overshoot, nor toxic to the animals, it was concluded that the toxiciyty to the animals of 8-azaadenine and 5-azacytidine is due to the compounds themselves rather than to the breakdown products.


Replication of IPN virus: a cytochemical and biochemical study in SWT cells. Although IPN virus failed to multiply at 30 degrees, it replicated at 16 degrees and 22 degrees in SWT cells. At 22 degrees the viral eclipse period lasted nearly 6 hr with maximal virion titers attained by 24 hr, whereas replication at 16 degrees was much slower. The replication of the virion was inhibited by 0.05 mug/ml of AD which did not interfere with the production of reovirus. Biochemical studies revealed that cellular DNA synthesis was markedly reduced (greater than 50%) soon after infection whereas total RNA synthesis was enhanced. The period of rapid increase in RNA synthesis paralleled the exponential production of infectious virus. Viral inclusion bodies, revealed by acridine orange-staining of virus-infected cells (SWT and RGG-2) late in the infectious cycle, were found to contain single-stranded RNA on the basis of their staining characteristics and sensitivity to RNase.