Chemotherapy of disseminated breast cancer. Current status and prospects. Chemotherapy once relegated to end stage patients has markedly improved with the use of combinations. Response rates with single agents have improved from 15 to 35%, to 50 to 70%, using combinations with an increase in complete response rates to about 25%. A series of four studies completed by the Eastern Cooperative Oncology Group over the past eight years typifies the improvement in response rates achieved by combinations as compared to single agents. Survival gain can be demonstrated for responders vs non-responders; however with current combinations, there is an apparent plateau in response rates (55 to 60%), durations of response (eight months) and survival for responders (18-22 months) as compared to survival of non-responders (six to eight months). Further improvement in response rates may occur by searching for new agents, combining hormonal and immunostimulation with chemotherapy or by sequencing non-crossresistant combinations. However, since most patients with breast cancer present with local or regional disease but go on to die of disseminated cancer, major improvements in survival are most likely to occur by treating this neoplasm as a systemic disease through cobmining effective local therapy with systemic treatments.


Kinetic, hormonal and clinical studies with aminoglutethimide in breast cancer. Approximately one-third of patients with metastatic breast carcinoma respond to surgical ablative therapy but the morbidity associated with these procedures has limited their use to highly selected patients. Consequently, a chemical method of adrenal suppression was developed using a potent inhibitor of adrenal steroid synthesis, aminoglutethimide, in combination with a synthetic glucocorticoid, dexamethasone. While this regimen effectively blocked adrenal function, it was complicated by a drug interaction in which aminoglutethimide accelerated the metabolism and reduced the bioavailability of dexamethasone. To overcome this problem, a new regime using aminoglutethimide and hydrocortisone, a glucocorticoid less susceptible to altered metabolism, was developed. Kinetic studies confirmed that aminoglutethimide does not interact with hydrocortisone to alter its rate of metabolism. Hormone measurements established that 1000 mg of aminoglutethimide and 40 mg of hydrocortisone daily suppressed DHA-sulfate, androstenedione, estrone, estradiol and aldosterone to a greater extent than the prior protocol using aminoglutethimide and 2-3 mg of dexamethasone. Patients experienced objective tumor regression with equal frequency while receiving the aminoglutethimide-hydrocortisone regimen or aminoglutethimide and dexamethasone and the overall rate of response in 50 evaluable patients was 38%. Side effects occurred frequently in the first few weeks of treatment but disappeared nearly uniformly thereafter. The present aminoglutethimide-hydrocortisone regimen is simple, non-toxic, effective in inhibiting estradiol synthesis and capable of inducing tumor regression as frequently as previously reported with adrenalectomy.


Gonadotropin receptors in experimentally induced ovarian tumors in mice. Gonadotropic hormones are required for the induction and maintenance of tumors arising in ovaries that have been transplanted to the spleens of gonadectomized mice. The characteristics of gonadotropin receptors for human chorionic gonadotropin (HCG)-luteinizing hormone on cells from these tumors of varying size, age, and morphology have been determined. The specific binding of 125I-labeled HCG to cells obtained by collagenase digestion, 15 to 65 weeks postimplantation from granulosa cell or luteinized cell, or mixed granulosa-luteal tumors was analyzed by Scatchard plot. Neither the size, weight, duration of implantation, nor histological morphology affected the receptor-binding affinity [equilibrium dissociation constant (Kd), 6 X 10(-10) M], and, presumably, the receptor is qualitatively similar. In contrast, the number of HCG receptors per cell increased 17-fold and was related to the degree of morphological luteinization of the tumor. HCG-sensitive adenyl cyclase was also demonstrated and compared to HCG binding in a highly luteinized tumor.


Effect of somatostatin on growth hormone release by MtT-F4 rat pituitary tumor in vitro. Fluoride-stimulated adenylate cyclase is demonstrated inisolated tumor cells of transplantable rat pituitary tumor MtT-F4 in vitro. The intracellular cyclic adenosine 3':5'-monophosphate is lowered in the cells incubated in the presence of synthetic somatostatin. Contrary to the findings reported for normal pituitary, however, the immunoreactive growth hormone release does not change when either somatostatin or phosphodiesterase inhibitors are present in the incubation medium. The presence of dibutyryl cyclic adenosine 3':5'-monophosphate (5 mM) in the incubation medium does not change the rate of growth hormone release by isolated tumor cells.


Epidemiologic study of prostatic cancer: preliminary report. In a preliminary report on a small fraction of the projected subject population, some evidence in support of both sexual activity and venereal transmission hypotheses is accumulating. Prostatic cancer cases, in contrast to hospitalized and/or neighborhood controls, are beginning to show greater proportions of selected sexual activities compatible with venereal transmission of an infectious agent, such as number of sexual partners, use of prostitutes, prior venereal disease, and genital infections in the spouse. Patients with prostatic cancer also appear to have had higher fertility and more prostatic cancer in blood relatives than controls. Age at first intercourse and at first marriage are lower among the cancer patients than among the controls. Antibody titrations for herpesvirus and cytomegalic virus, although currently not revealing striking disparities in positivity, tend to show higher titers among the cancer cases.


Intensive chemotherapy of small cell bronchogenic carcinoma. Thirty-two patients (27, extensive disease; five, regional disease) with histologically documented small cell carcinoma entered a randomized study to determine the efficacy of intensive induction chemotherapy. The necessity of a protected environment (laminar air flow room) during this treatment was also evaluated. Patients received high-dose or standard-dose cyclophosphamide, methotrexate, and CCNU (CMC) during the first 6 weeks of treatment. Subsequent maintenance therapy consisted of standard-dose CMC until disease progression. In 23 patients treated with high-dose chemotherapy there were responses in 96% (30% complete). Standard-dose chemotherapy gave responses in 45%, none of which were complete. Median survival correlated with completeness of response and was 16+ months for the seven complete responders. Patients receiving high-dose CMC spent an average of 10 days with neutrophil counts less than 1000/mm3. There was only one documented, non-fatal infection. High-dose chemotherapy gives better responses and longer survival than previously utilized standard doses of the same drugs. Patients could safely be treated in the hospital ward.


Properties of the H-4-II-E tumor cell system. I. Growth and cell proliferation kinetics of an experimental hepatoma. Growth and cell proliferation kinetics of hepatoma H-4-II-E and its tissue culture derivative have been studied to establish the characteristics of an in vivo--in vitro solid tumor model. The H-4-II-E line, originating from the Reuber H-35 hepatoma, can be maintained and studied either in cell culture or as a transplantable solid tumor in ACI male rats. In addition it allows for the in vitro assay of cell survival following treatment of animal tumors in situ. In vivo, hepatoma H-4-II-E is rapidly growing tumor with a mean doubling time of 49-2 hr. The cell cyle time is 39-1 hr with a cell loss factor of 0-32. Retrospective examination of tumor specimens obtained during the establishment of the H-4-II-E tumor system demonstrates that both structural as well as cell population changes have occurred. The biological characteristics of the primary tumor (H-35) and an early intermediate stage (H-35tc2) are compared with H-4-II-E and the histopathological, growth and cell kinetic changes are discussed.


Properties of the H-4-II-E tumor cell system. II. In vitro characteristics of an experimental tumor cell line. The growth and cell proliferation characteristics of the H-4-II-E cell line, giving rise to hepatoma H-4-II-E when inoculated into male ACI rats, were studied in vitro. Following seedling of 2 x 10(5) cells into culture dishes, exponential cell growth occurs in cultures fed both at 24 hr and 48 hr intervals with a population doubling time of 18-4 hr. Plateau phase growth conditions are established on day 7 and day 5 for cultures fed at 24 hr and 48 hr intervals respectively. Both the plateau phase cell density and the maintenance of plateau phase appear dependent on the frequency of feeding. For cultures fed daily, the transition from exponetial growth to plateau phase results from both a reduction in the number of proliferating cells (99% v. 35%) as well as an elongation of the cell cycle (17-7 hr v. 128-4 hr). The cell proliferation characteristics of the culture are further discussed in reference to both cell growth and feeding schedules of other cell lines.


Modulation of in vitro erythropoiesis: enhancement of erythroid colony growth by cyclic nucleotides. Mammalian erythropoiesis, as assayed by erythroid colony formation in vitro, is enhanced by cyclic adenosine nucleotides and agents which are capable of raising intracellular cyclic AMP (cAMP) levels. With canine marrow cells as target, this enhancement was shown to be specific for cAMP and its mono- and dibutyryl derivatives. Adenosine and its derivatives, such as AMP, ADP and ATP, and other cyclic nucleotides, such as cGMP, dibutyryl-cGMP, cCMP and cIMP and sodium butyrate were inactive. The phosphodiesterase inhibitor, RO-20-1724, and the adenyl cyclase stimulator, cholera enterotoxin, both markedly increased colony numbers. Studies with tritiated thymidine showed that about 50% of the cells responding to either erythropoietin (ESF) or dibutyryl cAMP (db-cAMP) were in DNA synthesis. However, by unit gravity sedimentation velocity analysis, the peak of ESF-responsive colony forming cells sedimented more rapidly (8-7 +/- 0-2 mm/hr) than the peak of db-cAMP-responsive cells (7-5 +/- 0 mm/hr). These results demonstrate that adenyl cyclase-linked mechanisms influence in vitro erythropoietic proliferation and suggest that other hormones and simple molecules might interact with surface receptors and thus modulate the action of ESF at the cellular level.


Relationships between membranous organelles in amoebae studied by electron microscopic cytochemical staining. The intracellular location of a variety of enzymes was studied in Amoeba proteus with the use of electron microscopic cytochemical methods, in an attempt to assess the relationships between different membranous organelles. One group of enzymes, including nucleoside diphosphatases (IDPase, UDPase, GDPase, ADPase), carbamoyl phosphatase, alkaline phosphatase, and BAXD oxidase was localized mainly in the rough endoplasmic reticulum, nuclear envelope, and convex side of the Golgi apparatus. Esterase activity had a similar localization except that the Golgi apparatus was "stained" throughout most of its extent. A second group of enzymes was found in Golgi cisternae and vesicles, and in come vacuoles. This group included acid phosphatase, thiamine pyrophosphatase, and aryl sulfatase. Some enzymes previously detected in cytoplasmic membranes of other cells, including glucose-6-phosphatase, showed little or no activity in amoebae. The results suggest that there are chemical similarities and probable functional relationships between the rough endoplasmic reticulum, the nuclear envelope, and the convex side of the Golgi apparatus. On the other hand, the concave pole of the Golgi apparatus, aggregates of smooth tubules and vesicles, and the cell surface appear more closely related to one another than to the endoplasmic reticulum and the convex side of the Golgi apparatus. The cytochemical similarity between the Golgi apparatus and certain vacuoles such as food vacuoles may reflect the role of the Golgi apparatus in the formation of lysosomes. The locations of reaction products of the various enzymes in amoebae are compared with observations reported for other cell types.


Developmental immunohistology of melanotrophs in Xenopus laevis tadpoles. The adenohypophysial primordium of Xenopus laevis tadpoles at stages 33/34 to 46 (Nieuwkoop and Faber, 1956) were examined immunohistologically for alpha-MSH, beta-MSH and ACTH. beta-MSH was demonstrated from stage 37/38 onwards, and alpha-MSH from stage 39. No signs of ACTH production were detected. alpha-MSH and beta-MSH occurred in the same cells. No differences were found in the intensity of immunofluorescence between tadpoles which were kept on a black and a white background. The present study lends no support to the hypothesis concerning the derivation of alpha-MSH from ACTH. The observations made suggest that the morphological formation of the pars intermedia is accomplished during stages 37/38 to 39.


Morphological aspects of the hypothalamic-hypophyseal system. VII. The tanycytes: Their relation to the hypophyseal adrenocorticotrophic function. An ultrastructural study. The ultrastruct of the tanycyte ependyma in male 160-180 g Wistar albino rats was studied under normal conditions and in experiments involving long-term suppression of ACTH secretion and its long-term stimulation. The former was accomplished by daily (for 8 days) intraperitoneal administration of dexamethasone phosphate at low (5 microgram/100 g) and high (100 microgram/100 g) concentrations. The effectiveness of suppression of the hypothalamic-hypophyseal-adrenal system in the experimental animals was judged by their reaction to two-minute ether stress (determination of plasma corticosterone) and by the results of measurement of the adrenal weights. Stimulation of ACTH secretion was achieved by bilateral adrenalectomy; the animals were examined on days 8, 10, 14, and 22 following the operation. The results obtained were in agreement with the previously established fact that there is a negative correlation between tanycyte activity and hypophyseal adrenocorticotropic function (Akmayed and Fidelina, 1974). They also testified to the predominant involvement of the median eminence tanycyte ependyma (beta-tanycytes according to the author's nomenclature) in these relationships. It is supposed that correlations are regulated by a feedback mechanism and attest to the involvement of beta-tanycytes in the inhibiting control of hypophyseal adrenocorticotrophic function. The mechanism of this control may be explained alternatively: either the tanycytes transport ACTH-suppression substances (catecholamines, corticosteroids, ACTH) from the CSF to the hypophyseal portal system or they themselves secrete substances possessing ACTH-suppressive activity. The authors distinguish several types of vesicles in the beta-tanycytes, the number of which changed with experimentally induced shifts in hypophyseal adrenocorticotrophic function. These vesicles are discussed in connection with the transport and secretory activity of the tanycytes and are considered to be a possible substrate of the hypothalamic inhibiting effect on ACTH secretion.


Cyclic AMP-induced cytolysis in S49 cells: selection of an unresponsive "deathless" mutant. Wild-type S49 lymphoma cells respond to cyclic adenosine 3', 5'-monophosphate (cAMP) by inducing cAMP phosphodiesterase, halting growth in the G1 phase of the cell cycle and subsequently dying. By using a counter selection procedure, we have isolated a new class of mutants of S49 cells termed "deathless" that are resistant to cytolysis, but otherwise respond like the wild-type cells to cAMP. Upon removal of the cyclic nucleotide, D-cells resume their normal growth. Unlike all other cAMP-resistant mutants of S49 cells isolated until now, the D- mutant has a functionally normal cAMP-dependent protein kinase and retains normal ability to induce phosphodiesterase and arrest cell growth in G1. It is probable that the altered gene product of the D- mutant is distal to protein kinase and in a biochemical pathway separate from that of cAMP induction of phosphodiesterase or growth arrest. The D- mutant may facilitate studies of the mechanism of cAMP-induced cytolysis and growth regulation in S49 cells.


Developmentally regulated cyclic AMP-dependent protein kinases in Dictyostelium discoideum. Two apparently distinct species of cyclic AMP-dependent protein kinase appear during the first 1-2 hr of development in Dictyostelium discoideum; no such activity can be detected in vegetative cell extracts. These two kinases are similar in properties to the type I and II cyclic AMP-dependent protein kinases found in a number of mammalian tissues. Their time of appearance supports the idea that one or both mediate the effects of cyclic AMP on gene expression early in Dictyostelium development.


Biochemical transformation of mouse cells by fragments of herpes simplex virus DNA. Mouse L cells lacking the enzyme thymidine kinase (LMTK-) have been converted to a TK+ phenotype by infection with fragmented HSV2 strain 333 DNA. The DNA fragments used were either unique, produced by cleavage with the restriction endonucleases Eco RI and Hild III, or randomly produced by mechanical shearing. Survival in HAT medium was used initially to establish the TK+ phenotype; clones possessing the ability to grow in selective medium were picked on the basis of differing morphology and growth rates. Cytosol extracts of these clones possessed virus-specified TK activity identical to that present in cells lytically infected with HSV2, as indicated by thermolability and mobility on polyacrylamide gel electrophoresis. The transformed cells also exhibit HSV-specific immunofluorescence. Based on these transformation studies, it is possible to assign a map location to the TK gene on the HSV genome.


Suppression of replication of SV40 and polyoma virus in mouse-human hybrids. Mouse-human heterokaryons are permissive for the replication of both SV40 virus and polyoma virus. If the hybrids which develop from these heterokaryons segregate human chromosomes (mouse greater than human hybrids), the hybrids are permissive for replication of polyoma virus but not for replication of SV40 virus. If the subsequent hybrids segregate mouse chromosomes (human greater than mouse hybrids), such hybrids support the replication of SV40 virus but not the replication of polyoma virus, even when the hybrids contain at least one copy of each mouse chromosome. This indicates that during the transition from heterokaryon to hybrid cell, suppression of expression of species-specific function(s) required for the replication of these species-specific viruses occurs in parallel with the direction of chromosome loss and suppression of nucleolus organizer activity.


Continuous tissue culture cell lines derived from chemically induced tumors of Japanese quail. Several continuous tissue culture cell lines were established from methylcholanthrene-induced fibrosarcomas of Japanese quail. The lines consist either of fibroblastic elements, round refractile cells or polygonal cells. They show transformed characteristics in agar colony formation and hexose uptake, and most are tumorigenic. Their cloning efficiency in plastic dishes is not increased over that of normal quail embryo fibroblasts. The quail tumor cell lines do not produce endogenous avian oncoviruses and fail to complement the Bryan high titer strain of Rous sarcoma virus; those tested lack the p27 protein of avian oncoviruses. Most of the cell lines are susceptible to subgroup A avian sarcoma viruses, but are relatively resistant to viruses of subgroups C, E and F as compared to normal quail embryo fibroblasts.


Similarities of two hepatitis A virus strains. In outbreaks of type A hepatitis in Los Angeles, USA, and Rosario, Argentina, virus particles were isolated from faeces. The geographically diverse strains were identical in appearance and serological reactivity. They differed only in buoyant density, but other workers have also obtained inconsistent results in estimating this. We conclude that the virus strains in the two epidemics were identical.


[Degradation by phytohemagglutinin of the antiviral state induced by interferon]. Phytohaemagglutinin (PHA M and P forms), when added to L 929 cells previously treated by murine interferon, degrades the antiviral state and restores virus sensitivity in the cells. This degradation depends on the amount of the lectin, the contact period with the cell and the presence of PHA membrane receptors. The importance of membrane configuration not only in the induction but also in the maintenance of the antiviral state is discussed.


[Ontogenetic analysis of the theta rhythm during paradoxical sleep in rats]. In the rat, an ontogenetic study of theta rhythm during paradoxical sleep shows a clear evolution between the 14th and the 30th postnatal day. During this period, its mean frequency increases from 5 to 7 cycles per second, and its spectrum shows significant changes. These parameters (mean frequency and spectral components) of the theta rhythm could be a useful index of brain maturation in the rat.


[Transformation, in vitro, of cerebral hamster cells by polyoma virus]. A cell line called HCxPy was obtained in vitro by transformation of dissociated hamster brain cell cultures by polyoma virus. The first foci of transformed cells became evident 90 to 120 days after viral infection. This cell line is now at the 46th passage. The cells appear tumorigenic for hamsters after subcutaneous and intracerebral injection. They carry the polyoma virus T and cell surface antigens. Good evidence for astrocytic differentiation can be found by morphological examination of the tumours and of the cultured cells.


[Does the diabetes caused by viral infection in mice depend on immune reactions?]. Normal DBA2 mice become highly diabetic after infection with the EMC virus. But 500 R X-irradiation before infection inhibits the increase of mean blood glucose levels thus indicating the participation of immune reactions in the appearance of diabetes.


[Desensitization by human chorionic gonadotropin (hCG): demonstration of a negative control of gonadotropin receptors by hCG at the level of testicular Leydig cells]. A single injection of hCG (500 IU) to intact and hypophysectomized prepubertal rat increases plasma testosterone levels and decreases hCG receptors in the testicular Leydig cells for 4 to 5 days. In intact animals testosterone exhibit a rapid increase within hours and a delayed paradoxical increase between 60 and 96 hours. In hypophysectomized animals the first peak is not present. In both intact and hypophysectomized rats hCG receptors are almost undetectable between 10 and 96 hours following hCG injection. Since receptor occupancy cannot account for this phenomenon it is concluded that hCG is exerting a negative control on its own receptors.


[Detection, by direct lymphocyte immunocytotoxicity, of antigens in cells transformed by Rous virus]. The hamster cells transformed by the Rous Sarcoma Virus (V.S.R.) evidenced surface antigenic alterations that were detected by a method of cellular mediated immunocytotoxicity. Immune hamster lymphocytes were added to tritiated proline prelabeled target cells. These lymphocytes were able to lyse specifically the V.S.R. transformed cells.


[Demonstration of an alpha adrenergic effect on the adipocytes of dog adipose tissue]. Phentolamine, an alpha blocking agent, potentializes the adrenergical lipolytic effect of isolated fat cells of Canine omental adipose tissue when their average diameter is greater than 80 micrometers. This effect can be found also in isolated fat cells of mesenteric adipose tissue, spontaneously less sensitive with catecholamines. Phenylephrine, alpha sympathomimetic, partially inhibits the lipolytic effect stimulated by isoprenaline.


[Induction of endogenous murine C-type viruses. Attempt at discrimination by the pI of polypeptide p30 and the aspect of cellular transformation]. Endogenous ecotropic and xenotropic murine C-type viruses induced in K-Balb-3T3 cells treated with iododeoxyuridine (IdU) were selected by infection of appropriate indicator cells. The isoelectric point (p1) of the major viral polypeptide (p30) was found to be 6.1 for the ecotropic virus (class I), and 5.7 for the xenotropic virus (class II). An isoelectric form (iso p30) of pl 6.5 was observed in the initial induction peak. In addition, the pattern of cellular alteration in NRK cells at its onset varied according to the pseudotype, the class I pseudotype inducing round cell foci while the foci associated with the class II pseudotype consisted of fusiform cells.


Angiotensin (A I, A II, A III) receptor characterization. Correlation of prostaglandin release with peptide degradation. We examined the ability of the angiotensins (A I, A II, A III) to release a prostaglandin E (PGE)-like substance in the isolated Krebs' perfused kidney and mesenteric vasculature of the rabbit by parallel bioassay. In the kidney, the order of potency for PGE release was A II greater than A III greater than A I with ED50's of 36, 100, and 500 pmol, respectively. In the mesenteric preparation, on the other hand, the order of potency was A III greater than A II greater than A I with ED50's of 75, 125, and 500 pmol, respectively. During one transit through the kidney 72-76% of bioassayable A I and A II was degraded. A III was 89% metabolized. In contrast, the mesenteric vasculature inactivated only 27% of A II and 23% of A III. This data suggests an inverse relationship between renal peptide degradation and PGE release. For characterization of the renal angiotensin receptor-mediating PGE release, dissociation constants (KB) of the competitive angiotensin antagonists [IIe7]-A III and [Sar1, IIe3]-A II were determined with each angiotensin. KB values of the individual antaganists were not significantly different with A I, A II, or A III; this finding suggests that one renal angiotensin receptor is involved with PGE release.


Evaluation of the dual-precipitation method by comparison with the ultracentrifugation method for measurement of lipoproteins in serum. We evaluated the dual-precipitation method for quantitative measurement of lipoproteins as described by Wilson and Spiger [J. Lab. Clin. Med. 82, 473 (1973)] for normo- and hyperlipemic sera, by comparison with the results obtained with ultracentrifugation. If serum with an above-normal triglyceride concentration is analyzed, the very-low-density lipoprotein cholesterol value obtained with the precipitation method is usually too low. For measurement of high-density lipoprotein cholesterol the ultracentrifugation and precipitation procedures give comparable results, but the latter method is preferred because sinking pre-beta-lipoproteins present in the high-density lipoprotein fraction isolated by means of the ultracentrifuge may result in falsely high values for cholesterol in that fraction. Therefore, at least for the determination of very-low-density lipoprotein cholesterol in hyperlipemic serum, the use of an ultracentrifuge remains necessary. Because few laboratories have an ultracentrifuge at their disposal, it seemed important to look at the stability of sera in view of the forwarding of samples. Also, a way of increasing the efficiency of the ultracentrifuge was studied. Sera can be stored for a week at 4 degrees C or for 54 h at room temperature without noticeable effect on lipoprotein values. Moreover, reliable values can be obtained with an ultracentrifugation time of 8 h (0.8 X 10(8) g-min).


Optimal conditions for assaying human lactate dehydrogenase by the lactate-to-pyruvate reaction: Arrhenium relationships for lactate dehydrogenase isoenzymes 1 and 5. Optimal reaction conditions to sassay human lactate dehydrogenase (lactate-to-pyruvate) were established for isoenzymes 1 and 5 at 25, 30, and 37 degrees C in diethanolamine and 2-amino-2-methyl-1,3-propanediol. Different substrate concentrations are required at each temperature. The conditions permit measurement of lactate dehydrogenase 1 and 5 with the lowest substrate concentrations that allow for the highest equal sustainable efficiency in measuring both isoenzymes. About 95% of each isoenzyme activity is measured if the assay is performed within the first minute after the reaction is initiated even for activities as high as triple the upper limit of normal. The Arrhenius relationship is different for each isoenzyme, but results obtained for each at one temperature can be compared with results at another temperature by use of simple conversion equations. Assays at 25 and 30 degrees C are more economical and less variable than assays at 37 degrees C.


Separation of hydroxyproline-containing protein from C1q (a subcomponent of complement) in serum. Precipitation of the euglobulin fraction from serum separates C1q from another hydroxyproline-containing protein(s), which remains in the serum-minus-euglobulin fraction. The presence of C1q in the euglobulin fraction has been verified by radial immunodiffusion assay, by sensitivity to collagenase, by hydroxyproline content, and by electrofocusing. The presence of another hydroxyproline-containing protein(s) in the serum-minus-euglobulin fraction has been verified by the same criteria. The isoelectric pH range of C1q is 6.1-7.0; the isoelectric pH range of the other hydroxyproline-containing protein(s) is 4.2-5.5.


Enzyme kinetics in single cells: concept and model. We describe a technique whereby it is possible to measure enzyme activity in a singel cell. The model chosen involved the measurement of myeloperoxidase activity in a polymorphonuclear neutrophil leukocyte. Details of the apparatus are described. The experiments necessary to optimize the reaction conditions are summarized. The nature of the phases of the reaction are described. The technique appears to have further application in measurement of the activities of other enzymes in leukocytes or other cells.


Studies on the pre-alpha-lipoprotein in patients with familial lecithin: cholesterol acyltransferase deficiency. The present study shows that regular alpha1- and pre-alpha-lipoproteins cannot be detected in serum of patients with familial lecithin: cholesterol acyltransferase (LCAT) deficiency. After electrophoresis on agarose gel only one single band of albumin mobility was observed in the alpha1-pre-alpha-region. In contrast to sera of normal subjects neither the anodic front nor the cathodic part of this region revealed any lipoprotein bands in the patients studied. The lack of the cathodic part might be related to a low amount of alpha1-lipoprotein. The apparent lack of the anodic front could be related to a low amount of "albumin-Apo-A-I-containing lipoprotein" (AAL). AAL was not detected with conventional methods in LCAT deficient sera. The alpha1-lipoprotein was made up of two immunologically identical peaks, both of which had a Sudanophilic character. After incubation of lysolecithin with albumin and AAL and subsequent thin layer chromatography, a significant lysolecithin-binding capacity of AAL was demonstrated, superior to that possessed by albumin.


Radiotheraphy in chemodectoma of the glomus jugulare. Twenty-five patients treated for glomus jugulare tumours over a 13-year period are reviewed, the majority (20) receiving radiotherapy only. Only two patients developed tumour recurrence, one following inadequate-dose radiotherapy, the other after surgery. Both are doing well following radiotherapy for their recurrent disease. Two patients were lost to follow-up and five have died of unrelated disease, four with no tumour reactivation over a prolonged period. The remaining 16 patients have been maintained in continuous tumour control for periods ranging from 4 to 15 years. While these tumours are by nature slow growing and usually not malignant, they demand radical treatment due to the symptomatology they create. Radiotherapy would seem to offer excellent prospect of satisfactory tumour control.


Endodermal sinus tumor of the mediastinum. Ultrastructural study. This is the eighth case report of an endodermal sinus tumor of the mediastinum (first case studied by electron microscopy). The ultrastructure of the tumor was found to mimic that of the normal yolk sac and was also similar to that of three previously reported cases of endodermal sinus tumor of the ovary, thus confirming the correctness of Teilum's original interpretation of the tumor as arising in germ cells and differentiating towards extraembryonic structures.


[Tryptic pancreatitis in small-cell bronchial carcinoma (author's transl)]. In two cases of typical small-cell bronchial carcinoma (one of them peripheral, small and clinically not diagnosed) destructive parenchylmal metastases in the pancreas, carcinomatous lymphoangiosis and infiltration of the larger excretory ducts by tumour cells were demonstrated. In both the clinical course was dominated by tryptic pancreatitis with treatment-refractory hypokalaemia. Since pancreatic metastases occur in about 10% of bronchial carcinomas, tryptic pancreatitis is, however, only rarely observed. It is, therefore, likely that autodigestion occurs only if in addition to parenchymal destruction by the metastases there are other factors. These may be blockage of lymphatic flow by carcinomatous lymphangiosis and duct stenosis by tumour infiltration of the walls. Hypercorticism due to ectopic ACTH production by the small-cell carcinoma may also be factor, but is without proof.


Extramammary perianal Paget's disease: report of a case. The case report of a patient with extramammary perianal Paget's disease who developed three histologic types of adenocarcinoma of the rectum over a period of six years is described.


A radioreceptor assay for insulin: direct measurement of dog pancreatic vein serum insulin. A simple radioreceptor assay for insulin rat liver membranes as receptor sites, with sufficient specificity precision, and sensitivity to detect 10 ng or 276 muU/ml of serum insulin, has been developed. In the presence of standard porcine insulin at the concentration of 1.0 ng/tube, approximately 8% of 125I-porcine insulin was bound to the plasma membranes and ninety-five per cent of this binding was inhibited by 1.0 microgram of standard insulin per tube. Four animal insulins inhibited the binding of 125I-insulin while ACTH, glucagon, human growth hormone, and oxytocin were inert. Insulin values in dog pancreatic vein sera obtained during and after glucose loading and measured by the present radioreceptor assay agreed well with immunoreactive insulin. The ratio of IRI to the measurement by radioreceptor assay was 1.09 +/- 0.18 for the same sera.


Effect of 1alphaOH-vitamin D3 in a case of pseudohypoparathyroidism. A 37-year-old woman with typical features of pseudohypoparathyroidism (chronic tetany, paresthesia, persistent hypocalcemia, round face, short stature, short metacarpals and metatarsals, cucutaneous calcification and lack of response to exogenous and endogenous parathyroid hormone as regards urinary phosphate and cyclic AMP excretion) was treated with oral administration of 2 microgram/day of 1alphaOH-vitamin D3. Serum calcium started to rise within 3 days returning to the normal level with disappearance of symptoms referrable to hypocalcemia. Such a favorable effect of a small dose of 1alphaOH-vitamin D3 in a patient with typical hypoparathyroidism suggests an important role of disturbance of 1alpha-hydroxylation of vitamin D3 by the kidney in the pathogenesis of calcium and phosphorus abnormality in this disease. Forty-four cases of pseudohypoparathyroidism in the Japanese literature were briefly reviewed.


Cyclic AMP accumulation in cerebral cortical slices: effect of carbamazepine, phenobarbital, and phenytoin. Recent investigations have suggested that abnormal increases in brain cyclic 3',5'-adenosine monophosphate (cAMP) may play a role in epileptogenesis. Therefore, the effect of three commonly used antiepileptic drugs on cAMP accumulation in rat cortex slices was investigated. Ouabain, a depolarizing agent which produces seizures when applied to rat cortex, produced a five- to sevenfold increase in cAMP accumulation, and both carbamazepine and and phenytoin inhibited this increase. Ouabain stimulation may be mediated by the release of endogenous adenosine, and carbamazepine antagonized adenosine stimulation of cAMP accumulation whereas phenytoin did not. Carbamazepine had no effect on adenosine efflux. The augmentation of cAMP accumulation by norepinephrine was inhibited by carbamazepine and phenobarbital but slightly increased by phenytoin. If increases in brain cAMP are involved in epileptogenesis, the antagonism of cAMP accumulation by antiepileptic drugs may play a role in their anticonvulsant action.


Effect of carbamazepine on the in vitro uptake and release of norepinephrine in adrenergic nerves of rabbit aorta and in whole brain synaptosomes. The effects of carbamazepine, in vitro, on adrenergic neuronal and whole brain synaptosomal uptake and release of tritiated norepinephrine (3H-NE) were assessed. At 10(-4) M, carbamazepine inhibited 3H-NE uptake by 22% in rabbit thoracic aorta and in brain synaptosomes. At the same concentration, carbamazepine inhibited stimulation-induced release of 3H-NE by 42.6% and inhibited isometric contraction in rabbit ear artery helical strips by 31.6%. At 10(-5) M, carbamazepine exhibited a 17.6% inhibition of 3H-NE uptake in brain synaptosomes in the absence of effects on transmitter release. Cocaine, 10(-4) M, and imipramine, 10(-4) M, inhibited uptake by 88% and 85%, respectively, in aorta, and cocaine, 10(-4) M, inhibited synaptosomal uptake by 67.7%. Since antiepileptic blood levels of carbamazepine range between 1.3 and 3.0 X 10(-5) M, it was concluded that the observed effects of carbamazepine are insufficient to account for the anticonvulsant action of the drug. However, the blockade of 3H-NE uptake by brain synaptosomes at 10(-5) M serves to explain the recently described analeptic activity of this agent.


Acrodermatitis enteropathica. II. Zinc deficiency and ultrastructural findings. The basic defect in acrodermatitis enteropathica (A.E.) is zinc deficiency caused by zinc malabsorption. The clinical symptoms disappear and serum zinc levels normalize after oral treatment with zinc. A report is given on two siblings suffering from A.E., both treated with oxyquinolines for a long period with changing clinical success. A permanent clinical remission could be achieved by treatment with zinc-sulphate at doses of 110-220 mg daily. The serum zinc levels normalized. The correlation between the zinc concentration of the hair and the kind of therapy was not very close. As we have shown in our first communication, the Paneth cells of the intestinal mucosa display ultrastructural changes in form of an unhomogeneous structure of the cytoplasm, formation of giant granules, and inclusion bodies. The zinc-therapy led to a complete normalization of the pathological changes in the Paneth cells. Thus, the changes in the Paneth cells in A.E. are the result and not the cause of zinc deficiency.


Prognosis in carcinoma of the urachus.

Prognosis in carcinoma of the urachus. The literature regarding the prognosis of carcinoma of the urachus is reviewed. Factors affecting the prognosis are discussed. En bloc partial cystectomy, if feasible, is the form of treatment which gives the best long-term survival rate. A series of 5 cases is presented with a 13-year survival rate of 40%.


Age changes in the neuromuscular system of rats. Data relating to the effects of aging on the neuromuscular system of rats have been reviewed and summarized. Age effects of muscle, nerve and neuromuscular junction are presented only in a descriptive, qualitative manner, primarily due to paucity of well controlled experimentation in this area. Certain morphological and biochemical changes taking place with aging of the neuromuscular system are discussed, but no conclusive hypotheses can be presented at this time explaining diminution of motor activity in old animals.


Differentiation of opiate and neuroleptic receptor binding in rat brain. For 6 large series of compounds derived from the piperidine moieties of spiperone, pimozide, haloperidol, pethidine, fentanyl and 4-methocarboxy-fentanyl, IC50 values were determined in the opiate and neuroleptic binding assay using [3H]-fentanyl and [3H]-haloperidol as ligands, respectively. The specificity and difference between both receptors were demonstrated on the basis of several criteria (1) the sterospecificity of the binding (2) the significant correlation between the in vitro activity of the drugs and their pharmacological potency in vivo, not withstanding some discrepancies which are probably of pharmacokinetic and/or metabolic origin (3) the ability to discriminate between morphinomimetic and neuroleptic drugs by the differential affinity for their specific receptro (4) the structure--activity relationships derived from the in vitro data indicating that the structural requirements for high affinity binding in vitro parallel those for high in vivo potency (5) a demonstration, on the basis of physicochemical principles, of the difference in binding mechanism between morphinomimetics and neuroleptics to their respective receptor.


[Effect of insulin, adrenaline and cyclic adenosine monophosphate on the activity of transport adenosine triphosphatases and pyruvate dehydrogenase of the rat myocardium under normal conditions and in insular insufficiency]. Experiments with rats were set up to study the interaction of hormones with Na+, K+-ATP-ase and Ca2+-ATP-ase of the sarcoplasmatic reticulum fragments and with pyruvate-dehydrogenase of the myocardial mitochondria in healthy rats and the ones with alloxan diabetes. Insulin was found to activate, while epinephrine and c-AMP to inhibit Na+, K+-ATP-ase. The Ca2+-ATP-ase is activated with epinephrine and c-AMP. In alloxan diabetes Na+, K+-ATP-ase is inhibited and Ca2+-ATP-ase and pyruvate-dehydrogenase--activated.


[Effect of diethylene-triaminopentamethylphosphonic acid complex on various hematologic and biochemical indicators in experimental animals]. A successive triple introduction of organophosphorus complexon DTPP (sodium-dicalcium salt of diethylene-triaminopentamethylphosphonic acid) in a dose of 500 mg/kg to albino male rats did not produce any marked effect on the composition of the peripheral blood. Some shortening of the initial period in blood coagulation process was noted. DTPP produced a moderate stimulation of the iron-containing enzymes (catalase, peroxidase, cytochroxidase) which led to an increased oxygen consumption. The blood serum and urine potassium, sodium and calcium content did not undergo any significant changes, while the magnesium concentration in the blood serum dropped by 37 pc and in the urine rose by as much as twice as compared to controls. This bears evidence to an elevated discharge of magnesium from the organism following the action of the DTPP complexon.


[Effectiveness of combination of various antihypoxic drugs in acute hypoxic hypoxia]. In experiments with mice evidence was obtained that a combination of apressin with obsidan and diprazine; of adenosine with nicotinamide-adenine-dinucleotide (NAD) and also a mixture of both combinations produce a marked protective effect in hypo and normobaric hypoxic hypoxia.


[Effect of cytostatic drugs, administered against the background of the action of insulin and thyroxine, on the processes of glycolysis and biosynthetic processes of neoplastic cells]. Experiments conducted on albino rats with a grafted tumour of transplantable alveolar mucinous carcinoma PC evidence that the cytostatic effect of thiophosphamide and sarcolysin equally grows in strenght with simultaneous introduction of thyroxin or insulin into the organism. In the same conditions the antineoplastic action of phthorafur is in a greater measure determined by the level of insulin and to a lesser degree by that of thyroxin. An analysis of the results subsequent to a biochemical examination (determination of the pyruvate, lactate, thiamine concentration in the blood and in the tumour, the rate of the 14C-glyxine incorporation in the protein of cancer cells) bears proof to the fact the neoplasm continues to definetly react in response to exogenously introduced hormones, this bringing changes into the kinetic and dynamic properties of the cytostatics.


General thermodynamic considerations of receptor interactions. Assuming that the biological response is directly proportional to the fractional degree of receptor occupancy, the mathematical relationships between free ligand concentration, receptor occupation and biological activity are developed for a number of equilibrium models. The models considered include simple 1:1 binding with and without conformational changes in the receptor, the coupled binding of two distinct effectors to a single macromolecule, and a system involving indirect coupling between two effectors that bind to two distinct components of the receptor system. This latter model is elaborated into the concept of a domain of receptor-enzyme pairs such that occupation of a single receptor may activate the entire domain of enzymes. This model can explain discrepancies between activation and binding isotherms as has been found with some beta-adrenergic agonist-sensitive adenylate cyclase systems.


Mechanisms and sites of action of newer angiotensin agonists and antagonists in terms of activity and receptor. From the myotropic and vasopressor activities of the numerous analogs of angiotensin II, it has been determined that the phenyl group of position 8 possesses the information for biologic response while the aromatic side groups in positions 4 and 6, the guanido group in position 2 and the C-terminal carboxyl are involved in binding to the receptor site. Removal of a side group of the C-terminal phenyalanine yields peptides that bind to the receptor. While many of these have low agonist properties, all have antagonist properties. Modifications in the aromatic side groups affect conformation of the octapeptide. This change may relate to receptor binding but sufficient data are not yet available to determine a correlation pattern. A proposed conformation for angiotensin is given as well as an artist's concept of angiotensin II binding to its membrane receptor utilizing the groups known to be involved in binding. Both angiotensin II and III [des-Asp] angiotensin II stimulate the biosynthesis and release of aldosterone from adrenal glomerulosa cells. Sufficient data are not yet available to determine whether the conversion of angiotensin II to angiotensin III is neccessary for the steroidogenesis activity.


[Effect of natural exogenous estrogens on the response to LH-RH (author's transl)]. LH-RH in doses of 25 and 50 microgram,, was injected intravenously into normal women pre-treated or not with conjugated natural estrogen. An inhibition of the  release of LH in response to 25 as well as 50 microgram of LH-RH, was observed in women receiving either dose of estrogens (0.628 and 1.25 mg).


Nesidioblastosis: the pathologic basis of persistent hyperinsulinemic hypoglycemia in infants. Morphologic and quantitative analysis of seven cases based on specific immunostaining and electron microscopy. Seven surgical specimens of pancreas, obtained at laparotomy from infants suffering from persistent hyperinsulinemic hypoglycemia, were analyzed by qualitative and quantitative immunocytochemistry and by electron microscopy. In five cases a multifocal ductuloinsular proliferation, in one a focal adenomatosis, and in one a solitary encapsulated nodule (adenoma) were observed. Combinations of the different patterns of proliferation were seen in six cases. Budding off from the ductular epithelium and interposition of endocrine cells between ductular epithelial cells were prominent features common to all cases. An almost fivefold increase of the mean total area occupied by endocrine tissue was found over that of age-matched controls. Four cell types were seen to participate regularly in the proliferation, and their ratios were remarkably constant in all cases, mean figures being 62:21:9:8% for B:A:D:D1 cells, respectively. The ratio of B cells per total endocrine area in nesidioblastosis was very close to that per islet of the controls (62:59%). Since common features were found in all or in the majority of cases, it is suggested that the various patterns of proliferation are merely morphologic variations of the same basic defect. Nesidioblastosis may result from inappropriately controlled development of the endocrine pancreas that is not arrested but carries on beyond birth and during infancy. The application of specific immunocytochemistry as a necessity for full appreciation of the extent of endocrine proliferation is stressed.


In-vitro observations on isolated adipose tissue cells from hyperobese subjects. Isolated adipose tissue cells were prepared from subcutaneous samples obtained from nine morbidly obese subjects weighing from 187 to 306% of ideal body weight. The responsiveness of these adipocytes to a number of test substances was determined by measuring cellular cyclic AMP concentration at one-half hour and glycerol release at four hours. Theophylline (10(-3) M) and epinephrine (10(-5) M) stimulated lipolysis; theophylline stimulated an increase in cyclic AMP, while epinephrine failed to prompt a significant change in the nucleotide. Neither the alpha blocker, phentolamine (10(-5) M), nor the beta blocker, propranolol (3 X 10(-5) M), affected lipolysis or cyclic AMP; when these agents were incubated in combination with epinephrine, changes occurred indicative of the presence of both alpha and beta adrenergic receptor sites. Insulin significantly reduced both basal and stimulated lipolysis but failed to affect cyclic AMP. With minor exceptions, adipocytes from hyperobese subjects behaved similarly to cells from unselected donors; at the concentration used, there was no evidence of resistance to insulin.


[The Landry-Guillain-Barré-Strohl-syndrome. Prognosis in adults (author's transl)]. The clinical aspect of acute polyradiculitis (Landry-Guillain-Barré-Strohl syndrome) of 85 patients is resumed. 52 of these 85 patients were seen after an average of 5, 1 years, 8 were dead at the time of control and 25 could not be traced. Two patients died during the acute phase of polyradiculitis corresponding to a letality of 2%. 24 of the subjects (46%) had recovered completely, 28 (54%) showed some residual symptoms. Eleven patients (21%) had some residual weakness of hands and/or feet. Out of these eleven six (11% of total) were so much disabled that they received a disablement pension. 17 patients with slight sensory disturbances and/or some loss of reflexes were not handicapped in their every-day life. 20 patients were examined electrophysiologically during the follow-up. All the five patients with a pathological EMG also showed some clinical residual signs. A positive correlation between pathological conduction velocities in the median and/or deep peroneal nerve and clinical residua was only inconstantly found. Patients with severe tetraparesis during the acute phase had more often residual symptoms. The time from the maximum motor deficiency to the beginning of recovery was longer in the latter group than in patients without residual symptoms (23 respectively 8 days) while there was no difference of the time between the beginning of motor weakness and the maximum motor deficiency in the two groups. A treatment with ACTH and/or corticosteroids seemed not to influence the long term prognosis of the illness.


Establishment and characterization of a cell line derived from a spontaneous murine lung carcinoma (M109). The Madison lung (M109) tumor cell line, initiated from a "spontaneous", anaplastic murine lung carcinoma, has been propagated continuously in vitro for more than 300 cell generations. Cytogenetic analysis revealed a mouse karyotype with a mode of 78 chromosomes (2n = 40). Three distinct marker chromosomes were identified by trypsin-giemsa banding. The cells piled up in culture and had a short generation time and high plating efficiency. Electron microscopy revealed highly undifferentiated cells with little rough endoplasmic reticulum, an abundance of free polysomes, the presence of few and often odd-shaped mitochondria, lipid bodies and phagocytic vacuoles. Virus particles of the C-type were found frequently. The subcutaneous transplantation of M109 cultured cells at a relatively low cell inoculum produced highly metastatic tumors in syngeneic BALG/c mice.


Induction of melanogenesis in vitro in the epidermal melanoblasts of newborn mouse skin by MSH. The number of epidermal melanocytes positive to the dopa reaction increased when skin explants from newborn mice were cultured with MSH or dbc-AMP. These agents seem to induce melanogenesis in the pre-existing melanoblasts. This hormone-induced melanogenesis is suppressed by actinomycin D or cycloheximide, suggesting that the initiation of melanogenesis in the epidermal melanoblasts requires de novo transcription and translation.


Intercellular immunological controls and modulation of cyclic AMP levels. Some doubts. We have re-examined two sets of observations put forward to support the hypothesis that rises in cAMP levels induced by vasoactive amines and prostaglandins are involved in the intercellular control of immunological and inflammatory processes. (1) This hypothesis is said to be supported by the fact that cholera toxin, which raises cAMP levels in lymphoid tissue in vivo, is immunosuppressive. However, we found that cholera toxin inhibited antibody production only if given in doses causing gross destruction of lymphoid tissue. This sort of evidence, therefore, cannot be used to support a hypothesis about homoeostasis under physiological conditions. (2) The hypothesis is also said to be supported by the claim that vasoactive amines, prostaglandins, cholera toxin and methyl xanthines, which raise cAMP levels in cells in vitro, also inhibit the formation of haemolytic plaques by spleen cells from mice immunized with sheep red cells. However, we were unable to confirm this claim, except when the experimental conditions were such that cells were killed or other artefacts operated.


The action of thiols on lymphocyte membranes. Short-term incubation with cysteine in concentrations not altering cell viability increased the amount of -SH groups on the surface of rat lymphocytes. Maximal effect was achieved in 10 min using 2 mM cysteine, which concentration also induced immediate total loss of response to adrenaline, as measured by intracellular cyclic AMP level. Neither of these effects was observed after incubation of lymphocytes with glutathione or dithiothreitol.


Specific immune lysis of paramyxovirus-infected cells by H-2-compatible thymus-derived lymphocytes. Mice exposed to paramyxovirus (Sendai) generate specifically sensitized thymus-derived lymphocytes (T cells) which, in an in vitro 51Cr release assay, interact only with virus-infected target cells sharing strong transplantation antigens. Reciprocal exclusion of cytotoxic T-cell activity is found for Sendai virus, lymphocytic choriomeningitis virus and ectromelia virus. Immune T cells are detected as early as 3 days after intraperitoneal inoculation with a large dose of Sendai virus, and cytotoxicity is generally maximal on days 5-7. Lysis is restricted to interactions where sensitized lymphocytes and virus-infected target cells (fibroblasts, tumour cells or macrophages) are compatible at the K or the D locus of one H-2 haplotype. Identity of immune response (Ir) genes is neither sufficient nor necessary. Levels of T-cell responsiveness show some variation with H-2 type. Cytotoxic T-cell activity associated with H-2b is less than that recognized for H-2k or H-2d. These differences are, however, not obviously related to Ir gene control.


Serological relatedness of herpes simplex viruses. Type-specificity of antibody response. The serological relatedness of forty-seven strains of type 1 and type 2 herpes simplex virus was investigated by reciprocal and non-reciprocal neutralization kinetics. Early rabbit antisera divided the virus strains into two distinct groups where confident indentification of virus type was possible. Hyperimmune mouse and rabbit antisera did not divide the two virus types into two distinct non-over-lapping groups. The extent of overlap varied with the particular attribute of the virus being studied. The virus types were best discriminated by their neutralizability by type 1 antisera and least well by their neutralizability by type 2 antisera. The results of reciprocal kinetic neutralization test with hyperimmune mouse antisera were analysed by multi-dimensional cluster analysis. Hyperimmune mouse or rabbit antisera could not be discriminated with respect to their immunogenic type by their absolute neutralization rate constants against either type 1 or type 2 virus, but could be distinguished on a group basis by their relative neutralizability against both virus types (antiserum specificity attribute); however, using this latter criterion, the type of immunogen could only be predicted in seven of the forty antisera under test. 'Early' mouse antisera could also be distinguished as groups by their absolute k-values against type 1 herpes virus. Thus, immunogenic identification, on other than a group basis, was unreliable. The specificity of a given serum was inversely related to its titre. There was a positive correlation between the specificity of a given virus strain and of its corresponding antiserum.


Immunoglobulin synthesis by an SV-40-induced hamster lymphoma. A transplantable lymphoid tumour induced in Syrian hamsters by SV-40 was shown to synthesize a 7Sgamma2 monoclonal immunoglobulin of the same idiotype was also found in the membrane of the tumour cell. The induction of immunoglobulin synthesizing tumours by SV-40 should be useful for studies on immunoglobulin synthesis, lymphoid cell receptors and tumour immunity.


The antigenicity of chondrocytes.

The antigenicity of chondrocytes. Stimulation in vitro of allogeneic and syngeneic mixed lymphocyte cultures suggests the presence of differentiation antigens on the surface of the chondrocyte. This may provide part of the answer to the nature of the chronicity of certain arthritic processes.


Peripheral blood lymphocyte response to acute infections in humans. The response of subpopulations of peripheral venous blood lymphocytes to systemic bacterial or viral infections was studied. B-lymphocytes were defined by the presence of surface binding sites for mu chains, which were determined by immunofluorescent staining. T-lymphocytes were defined by the ability to form active sheep cell rosettes. Virus-infectible lymphocytes, which may represent activated T-lymphocytes, were defined by the ability to support virus replication. Patients with bacterial infections had an increase in the B-lymphocytes of peripheral venous blood, whereas patients with viral infections had an increase in T-lymphocytes as compared to controls. The number of virus-infectible lymphocytes was increased in patients with bacterial infections but not in patients with viral infections. These studies suggest that subpopulations of human peripheral blood lymphocytes vary in response to different types of infectious agents.


Comparative effects of host and viral factors on early pathogenesis of Marek's disease. A series of experiments on the early pathogenesis of Marek's disease was conducted according to a uniform scheme. In each experiment, there was a single variable-age, genetic strain, or virus strain. Virus assays from spleen, buffy coat, and bone marrow, and fluorescent antibody tests on spleen, bursa of Fabricius, and thymus were conducted on five birds per group daily from the 3rd through the 10th day postinoculation. From these data, it was apparent that the response could be divided into two periods: 4 to 6 days = early; 8 to 10 days = late. Serological tests showed all groups except the 1-day-old group to have neutralizing antibody by the end of the 10-day period. With few exceptions, none of the variables tested exerted any appreciable influence on the level of virus growth in spleen, bursa, or thymus during the early period. High levels of infection occurred in all birds during that period. Changes in infection pattern which occurred during the late period were significant and could be correlated with occurrence of Marek's disease in test samples of birds held until 7 weeks after infection. Infectivity levels dropped appreciably in the case of resistant N-line birds given JM virus, and, during the late period, infection levels were significantly higher in GA-infected birds than in those given viruses of lower virulence. Whereas the virus titers during the 8- to 10-day period usually reflected the eventual clinical pattern of Marek's disease, the levels of viral antigen (fluorescent antibody tests) were much less consistent. One further experiment conducted by the same uniform scheme demonstrated no significant effects on early pathogenesis or course of Marek's disease in birds given continuous oral medication with amino-ureido-sulfone.


Human rotavirus in lambs: infection and passive protection. A human stool filtrate containing rotavirus which was administered orally to gnotobiotic lambs caused diarrhea, virus excretion, development of antibodies to rotavirus, and pathological changes in the villi of the small intestine. Thus, lambs may serve as experimental animals for the study of human rotavirus infections. This model system was used to study passive protection. Human immunoglobulin G (IgG) containing antibody to rotavirus was fed to lambs 24 to 78 h after birth, and the lambs were infected with lamb-passaged human rotavirus when 30 h old. The lambs treated with IgG did not develop diarrhea, and virus excretion was delayed in onset and shortened in duration. It may be possible to make similar use of IgG to protect children at risk in a rotavirus outbreak. The treatment did not prevent the lambs developing antibody to rotavirus.


The effect of imipramine on isolated skeletal muscle preparations. Imipramine (2-10 microng/ml) noncompetitively inhibited acetylcholine responses of the frog rectus abdominis muscle, and markedly inhibited the contracture produced by carbachol and succinylcholine without affecting the contracture produced by KCl, caffeine, and chlorpromazine. The twitch responses to indirect and direct stimulation of the rat phrenic nerve-diaphragm and the frog sciatic nerve-gastrocnemius were first augmented and then depressed markedly and irreversibly by imipramine (5-20 microng/ml). The indirect stimulation was inhibited earlier and to a greater degree than the direct stimulation. The blockade in the nerve-sartorius developed and progressed quickly without prior augmented responses, and with a parallel time course for indirect and direct stimulation. On the frog rectus, physostigmine antagonised whereas d-tubocurarine and CaCl2 increased the imipramine-induced inhibition. In the nerve muscle preparations, physostigmine, CaCl2 and KCl did not affect the neuromuscular blockade produced by imipramine; tubocurarine (0.05 microng/ml) markedly increased the blocking effect of imipramine (20 microng/ml) on the rat phrenic nerve-diaphragm. The results have been discussed in relation to the memberane stabilizing and the calcium releasing actions of imipramine.


Cell-mediated cytotoxicity to herpes-infected cells in humans: dependence on antibodies. Herpes simplex virus type 1 (HSV-1)-infected human skin fibroblasts were used as target cells in a 51Cr release assay, using peripheral blood mononuclear cells from HSV-seropositive and -seronegative donors as effector cells. Cytotoxicity was exerted by ordinarily prepared lymphoid cells but could be reduced by extensive washing of the effector cells. The antibody dependence of the system was shown by the recovery of activity through addition of positive serum or medium used for the early washes of effector cells. Three donors found to be seronegative in the usual serological tests were shown to be seropositive in this test. It is proposed that the assay can be used as a very sensitive serological test.


Effector cell involved in cell-mediated cytotoxicity to cells infected with herpes simplex virus type 1. Experiments were performed to analyze the characteristics of the effector cells involved in cell-mediated killing of herpes-infected fibroblasts. Effector cells were fractionated according to adherence and phagocytic properties, and lymphocytes were depleted of T-cells, and EA- and EAC- (antibody-coated bovine erythrocytes and bovine erythrocytes coated with rabbit anti-bovine antibodies and C5-deficient mouse serum, respectively) rosette-forming cells by differential centrifugation. Column passage was used as an alternative to depletion of B-cells and F(c)-receptor-positive cells. Antibody-dependent, cell-mediated cytotoxicity was found to be associated with nonphagocytic cells carrying low-avidity F(c) receptors. Some of these had surface immunoglobulin; others lacked both B- and T-cell markers. Low levels of killing were found without addition of anti-herpes antibody to the test system, and this killing seemed to depend on cells belonging to the same subpopulation as the antibody-dependent effector cells.


Passive immunity to feline leukemia: evaluation of immunity from dams naturally infected and experimentally vaccinated. Antibodies against feline leukemia virus (FeLV) and the feline oncornavirus-associated cell membrane antigen (FOCMA) were transferred from pregnant cats to their suckling kittens. All of these kittens were protected against infection and oncogenesis by virulent FeLV when challenged at 2 weeks of age. Suckling kittens acquired 25 to 100% of maternal virus-neutralizing and FOCMA titers by 3 days of age, and titers underwent linear decay to undetectable levels by 2 to 3 months of age. FOCMA antibody in dams and kittens was identified as immunoglobulin G (IgG) by use of goat anti-human IgG serum, which cross-reacts with feline IgG in the indirect membrane immunofluorescence test for FOCMA antibody. In an attempt to induce protective maternal antibody by vaccination, 10 pregnant cats were immunized by three to five weekly intramuscular injections with purified FeLV inactivated by ultraviolet irradiation. After the course of immunization, neither virus-neutralizing nor FOCMA antibody was detectable in the dams or in 19 kittens born to these cats. When these kittens were challenged with FeLV at 2 weeks of age, 18 of 19 developed persistent viremia and FeLV-related disease.


Protection against herpes simplex virus infection in mice by Corynebacterium parvum. Corynebacterium parvum administered in mice prior to herpes simplex virus (HSV) infection significantly protected them against lethal encephalitis. This was seen both with a mouse strain highly susceptible to HSV and with one relatively resistant to HSV. Mice immunosuppressed by cyclophosphamide and showing an increased mortality after HSV infection were also protected by C. parvum pretreatment. However, C. parvum given simultaneously with or after HSV infection did not exert a therapeutic effect.


Increase in serum IgE levels of ovalbumin-sensitized cats and the detection of elastase and collagenase activities in secretions of sensitized feline alveolar macrophages challenged in vitro. The feline model of respiratory hypersensitivity induced by intraperitoneal injection of ovalbumin has been studied. IgE serum antibodies were present for 3-10 weeks following sensitization, with maximum titers occurring between 50 and 70 days. Similarly, peak passive cutaneous anaphylaxis reactions occurred between 50 and 70 days. Alveolar macrophages, obtained by tracheal lavage at 65 days after sensitization, produced elastase like and collagenase-like secretions 48 h after challenge with ovalbumin in culture. Macrophages from nonsensitized cats did not produce these secretions. It is hypothesized that reaginic antibodies and sensitized alveolar macrophages, such as those found in the cat model, may be responsible in part for the destruction of lung tissue found in long-term respiratory diseases, similar to fibrosing alveolitis and pulmonary emphysema in man.


Preferential enhancement of IgE antibody formation by Bordetella pertussis. Preferential enhancement of IgE antibody response was observed in BALF/c mice by the administration of Bordetella pertussis with antigen (DNP-Salmonella). Correlation between B cell mitogenic activity and adjuvant action among B. pertussis, Salmonella, lipopolysaccharide of Escherichia coli and Ficoll was examined but was not found. Thymus-derived cells seemed necessary to develop adjuvant action of B. pertussis since antibody response in athymic nude mice was not influenced by B. pertussis. Helper function of adoptively transfered spleen cells was enhanced by immunization of the donor mice with carrier antigen in the presence of B. pertussis. The magnitude of enhancement was greatest in IgE class. The results indicated that preferential enhancement of IgE antibody formation by B. pertussis is mediated by the augmentation of carrier-specific helper function.


Differential induction of Epstein-Barr virus-related antigens in heterokaryon cultures. Different paterns of induction of Epstein-Barr virus (EBV)-related antigens were observed in heterokaryons produced by Sendai virus-mediated fusion of producer and non-producer human lymphoblastoid cells with various other cell types. EBV-related early antigens (EA) and viral capsid antigen (VCA) could obviously be induced in heterokaryons between producer cells (P3HR-1 and QIMR-WIL), normally expressing these natigens at very low frequency, and human FL or HeLa cells. Positive cells were detected as early as 3 h after fusion and there often followed a rapid increase in positive cells. In contrast, in heterokaryons between non-producer cells (Raji and NC-37) and FL or HeLa cells, only EA but not VCA was induced. EA induction was also evident in fusion of human lymphoblastoid cells with monkey cells (Vero) but with mouse cells (L-M(TK-) C11D and MCB-2) no EBV induction occurred. The EBV induction in heterokaryons was significantly enhanced by 5-iododeoxyuridine treatment.


Comparative studies on adult donor lymphocytes infected by EB virus in vivo or in vitro: origin of transformed cells arising in co-cultures with foetal lymphocytes. Co-cultures were set up between equal numbers of mononuclear cells from the blood of EB virus-infected individuals, either acute IM patients or healthy seropositive adult donors, and foetal cord blood mononuclear cells of the opposite sex. The cell lines arising in the co-cultures were of mixed origin, with foetal cells predominating in many cases. In contrast, when mononuclear cells from seronegative adult donors were first infected with EB virus in vitro and then 5 to 12 days later co-cultured with a large excess of foetal cells of the opposite sex, the cell lines which arose were almost exclusively derived from the adult donor despite the fact that a small minority of the virus-infected adult cells released infectious virus capable of transforming the co-cultivated foetal cells. The experiments suggest that EB virus-infected cells present in the blood of IM patients and seropositive donors do not possess the capacity for unlimited in vitro growth shown by seronegative adult donor lymphocytes experimentally infected with the virus.


Bovine leukemia virus specific antibodies among French cattle. I. Comparison of complement fixation and hematological tests. Complement fixation (CF) and hematological studies were performed on 517 cows living in normal conditions in different geographical areas of France. The animals belonged to three different categories: (1) multiple or single case herds, in which lymphosarcomas had been detected in the past five years. (2) Leukemia free herds of high risk regions. (3) Leukemia free and apparently unexposed herds. Positive animals were found with both methods in the first two categories but not in the third. Persistent lymphocytosis and CF antibodies were more frequent in leukemia than in exposed but leukemia free herds. Approximately 2 times more animals were found positive by serologic than by hematologic tests. The mean geometrical titer of CF antibodies was higher in lymphocytotic than in normal animals and highest in lymphosarcomatous cows. Persistent lymphocytosis was first detected in 3-years-old animals whereas 22% of younger cows were positive in the CF test.


Immunologic studies of the low molecular weight DNA binding protein of murine oncornaviruses. A low molecular weight, highly basic DNA-binding protein was purified from several oncornaviruses by the sequential procedures of gel filtration in guanidine-hydrochloride, DEAE-cellulose chromatography and affinity chromatography on single-stranded DNA sepharose. The binding protein from Rauscher and woolly monkey type-C viruses was the fastest migrating of the virion proteins in SDS-polyacrylamide gels and thus is designated p10 according to previous convention although our estimates of molecular weight were 8-9,000 daltons. The binding protein from these two viruses was resolved into two bands by acid-urea electrophoresis although only a single NH2 terminal amino acid was detected (S. Oroszlan, personal communication), thus indicating charge heterogeneity. Antibody to Rauscher virus p10 species-specific in gel diffusion and complement-fixation tests and did not exhibit cross-reactivity with other virion proteins. A DNA-binding protein was also detected in preparations of mouse mammary tumor virus. This purified protein had an apparent molecular weight of 12,500, was the second fastest migrating component in the virus preparations, and was antigenically unrelated to the mouse type-C virus p10.


Incomplete viral genome in a non-virogenic mouse tumour cell line (RVP3) transformed by Prague strain of avian sarcoma virus. Two cell lines, RVP3 and RVA4, derived originally from mouse tumors induced by the Prague and Schmidt-Ruppin strain of RSV, respectively, were studied. tall attempts failed to induce infectious virus production in RVP3 cells by fusion with chicken fibroblasts even if the cells were infected with avian leukosis viruses. Also, attempts to rescue the viral genome by transfection were unsuccessful. RVP3 cells harboured 31-45% of the viral genome sequences, as was shown by molecular hybridization, and therefore they were designated cryptovirogenic. The tumour cell line RVA4 did not contain any detectable viral sequences. The significance of the detection of the incomplete Rous virus genome sequences in mammalian cells is discussed.


Circadian variation in plasma melanocyte stimulating hormone activity in the rat. Non--stress levels of plasma melanocyte stimulating hormone (MSH) and corticosterone were determined at 3-h intervals during controlled light--dark cycles in adult, male rats. Significant 24-h periodicity was demonstrated for both plasma MSH and corticosterone. Whereas plasma MSH values were not as high during the early evening hours as during the early morning hours, plasma corticosterone levels showed a marked rise during the early afternoon and crest values occurred just before the lights off phase of the 24-h light--dark cycle. These results indicate that in the rat, the level of MSH in plasma fluctuates with a low amplitude circadian frequency which is not in phase with that observed for plasma corticosterone.


E.s.r. of spin-trapped radicals in aqueous solutions of amino acids. Reactions of the hydroxyl radical. The radicals produced by reactions of hydroxyl radicals with amino acids in aqueous solutions have been investigated. Hydroxyl radicals were formed by U.V.-photolysis of hydrogen peroxide and the short-lived amino acid radicals were spin-trapped by tert-nitrosobutane and identified by electron spin resonance spectroscopy. Nineteen amino acids were studied, and several radicals were identified which have not been observed previously by other methods. Only side-chain radicals were identified for alanine, threonine, aspartic acid, asparagine, lysine, phenylalanine, tyrosine, proline and hydroxyproline; whereas for glycine the C(2) carbon radical was spin-trapped. Both C(2) carbon radicals and side-chain radicals were assigned to valine, leucine, isoleucine, serine, glutamic acid, glutamine, arginine and methionine.


Cytochemical evidence for aerobic pathways in Mycobacterium lepraemurium. Three enzymes of aerobic pathways (cytochrome c oxidase, peroxidase and catalase) and one key enzyme of the tricarboxylic acid cycle (succinate dehydrogenase) were investigated for their ultrastructural localization in M. lepraemurium in infected mouse liver and in cultures of M. fortuitum as a control. All four enzymes were localized in M. fortuitum. To M. lepraemurium only cytochrome c oxidase and peroxidatic activity were detected. The localization of the latter enzyme activity was different compared with M. fortuitum. Succinate dehydrogenase was not detected in M. lepraemurium but rather surprisingly was found in the membrane of the phagosomes containing the bacteria. It is concluded that M. lepraemurium can function aerobically and has either a glyoxalate pathway or is an obligate autotroph.


The effect of theophylline on the breakdown of the blood-aqueous barrier in the rabbit eye. A disruption of the blood-aqueous barrier in rabbit eyes was elicited by topical prostaglandin E2, infrared irradiation of the iris, or subcutaneous alpha-melanocyte stimulating hormone (alpha-MSH). The course of the inflammatory reaction was followed by photoelectrical measurements of the aqueous flare in the anterior chamber. Pretreatment with intravenous theophylline, a phosphodiesterase inhibitor, significantly increased the protein leakage caused by prostaglandin E2 and alpha-MSH, but the response to infrared irradiation was slightly but not significantly enhanced. Intravenous theophylline given in higher doses caused per se an aqueous flare increase, which could not be inhibited by pretreatment with topical indomethacin. Our results indirectly indicate that accumulation of intraocular cAMP promotes a barrier damage and that cAMP might be the common effector of the barrier breakdown caused by prostaglandin as well as by nonprostaglandin agents.


Adenylate cyclase and phosphodiesterase activity in rabbit ureter. Adenylate cyclase and phosphodiesterase enzyme activities were demonstrated in rabbit ureter. NaF, 10 mM, caused a 60.9 per cent increase in adenylate cyclase activity. Isoproterenol, 5 X 10-7 to 10-5 M induced a statistically significant dose-dependent increase in adenylate cyclase activity which was suppressed by propranolol, 10-7 M. Theophylline, 5 X 10-5 to 10-2 M, significantly inhibited phosphodiesterase activity. Thus, isoproterenol and theophylline, two agents that can relax ureteral segments previously contracted by a depolarizing concentration of potassium, could presumably increase cyclic AMP levels, isoproterenol by increasing synthesis and theophylline by decreasing degradation.


Activities of the androgen-responsive receptor of the prostatic plasma membrane. Evidence of the presence of an androgen-activated "gatekeeper" receptor in the human prostate plasma membrane is presented. Not only does testosterone in vitro very rapidly accelerate uptake of alpha-aminoisobutyric acid into prostate slices but also its metabolite, dihydrotestosterone, greatly increases the rates of phosphorylation and dephosphorylation of the cation-dependent ATPase of isolated membranes. The relationship of this regulatory unit to prostatic physiologic processes is discussed.


The direct effect of ultrasound upon Wilms' tumor in the rat. The possible activation of immunologic defenses against tumor by local ultrasonic irradiation was investigated. Sonication of one tumor of a dual implant destroyed that tumor only with no contralateral effect. Injection of tumor cells previously sonicated in vivo did not induced immunity to subsequent challenge with live cells. Immunofluorescent staining of host kidneys for antibody complexes was negative. It was concluded that sonication was only effective when directly applied to the tumor.


Further biological properties of the human syncytial virus. Some biological properties of the human syncytial virus have been examined. A 13-day plaque assay in whole human embryo fibroblasts (HEF) has been developed using a liquid (growth medium) overlay. The plaques were 0.7-2 mm in diameter and often showed a clear central zone with irregular edges. Pretreatment of HEF monolayers with the polycation DEAE-dextran for either 30 min or 1 h was found to enhance plaque formation by a factor of from 2- to 7-fold. The plaque assay procedure required cell cultures undergoing active cell division. Adsorption kinetics and growth cycle studies in HEF indicated a relatively long adsorption period (3 h) and a relatively prolonged latent period of 24 h. Even under optimal conditions, virus yields were low and did not exceed 1 PFU per infected cell. Like other animal syncytium-forming 'foamy' viruses, the human virus induced both intranuclear and cytoplasmic antigens detectable by immmunofluorescence and was also markedly labile to freezing and thawing.


Detection localization of Epstein-Barr virus-associated early antigens in single cells by autoradiography using 125I-labeled antibodies. (With 1 color plate). A characteristic feature of virus-transformed cells is the expression of virus-specific antigens usually detectable by immunological or radioimmune procedures. We report here an autoradiographic method for the detection and localization of such antigens in individual cells which combines the cellular specificity and radiolabeling sensitivity of the above procedures. As a test system we have studied the reaction between virus antigen in some Epstein-Barr virus (EBV) DNA-containing cell lines and anti-virus antibody specificities in certain human sera. EBV DNA-containing cell lines express an EBV-determined nuclear antigen (EBNA) and some lines also express the EBV-associated antigens, early antigen (EA) and virus capsid antigen (VCA), in a minority of cells in the population. By employing appropriate 125I-IgG-labeled human sera, we clearly show that EA can be detected without difficulty in cell lines known to spontaneously express this antigen. Moreover, the specificity and sensitivity of the present method is such that low levels of EA cand be detected at cellular concentrations which remain undetectable by conventional immunofluorescence.


Transforming activity and antigenicity of an Epstein-Barr-like virus from lymphoblastoid cell lines of baboons with lymphoid disease. Two lymphoblastoid cell lines were established from baboons with lymphoid disease. Cells of these lines were positive for complement and Fc receptors but lacked sheep cell receptors, theraby indicating B-cell origin. The cells contained antigens which cross-reacted with Epstein-Barr virus (EBV), viral capsid antigen (VCA), early antigen (EA) and membrane antigen (MA). Both lines released virus with in vitro transforming activity for lymphocytes of several primate species including humans. Cells of the original lines and transformed cells showed no staining for EB nuclear antigen (EBNA). The virus was neutralized by anti-MA positive baboon and human sera. Baboon virus and EBV had different but overlapping in vitro host-cell ranges.


Bovine type II interferon: activity in heterologous cells. The antiviral and cell growth inhibition spectra of bovine and human interferons are described. The antiviral activity of type II interferon in heterologous cells far exceeded that of the corresponding type I interferon. Antiviral activity of bovine type II interferon parallels cell growth inhibition activity both in thermal inactivation kinetics as well as in the rate of synthesis. Furthermore, the spectrum of antiviral activity against heterologous cells was the same as that seen for cell growth inhibition suggesting that one molecule may mediate both effects. The therapeutic implications of type II interferon prepared in animals for use in human medicine are discussed briefly.


Immune deficiency of T system with possible T cell regulatory activity defect. The occurrence of T system immunodeficiency in an infant together with excessive production of IgM and, to a lesser degree, of IgG and IgA, is an unusual combination. A case is reported in which an unremitting lung infection with lymphadenopathy and hepatosplenomegaly developed in a previously healthy two-month-old infant. Leukocytosis with lymphocytosis, monocytosis and eosinophilia was rapidly followed by leukopenia and lymphocytepenia after a blood transfusion for anemia. There was a transient clinical remission, but on relapse 10 days later, quantitative and functional T cell deficiency was found together with increased IgG and IgA and with IgM values reaching 50 times greater than normal. Thymic humoral factor was successful in vitro in increasing the number of identifiable T cells (E rosetts) as well as T cell function (leukocyte migration inhibition factor production). However, the infant died suddenly, and at autopsy evidence of a generalized inflammatory reaction compatible with a viral infection was found. The thymus was small, hypoplastic and hypocellular. It is speculated that the T system deficiency may have been acquired following Epstein-Barr virus infection, and that T cell regulatory activity of immunoglobulin production was defective.


[Psoriasis: A general disease? Can psoriasis be explained biochemically?]. The biochemical problems of psoriasis are discussed, especially two modern hypothesis about the pathogenesis of this skin disease. 1. DHEA-deficiency leading to an increase of G-6-PDH inaugurated by Holzmann et al. 2. cAMP deficiency in psoriatic lesions as a cause of psoriasis hypothized by Voorhees and Duell. A synopsis of the physiology of the steroid hormone DHEA (synthesis, regulation, conjugation and excretion) is given. There are doubts that this hypothesis is correct. Recent findings have indicated, that there is no decrease of cAMP in the psoriatic lesion--in contrast there is an increase of this cyclic nucleotid. It means that this theory is also doubtfull. Treatment of an hypothetical decrease of cAMP by AMP is not possible, because this AMP will not penetrate in the cell and cannot be metabolized to cAMP.


[Staphylodermia superficialis circinata. The 5th obligatory cutaneous paraneoplasia]. Clinical observations on two cases of staphylodermia superficialis circinata are reported. This rare variation of superificial staphylococcal skin infection is identical with the "erythema necroticans migrans". As this cutaneous manifestation is highly associated with malignant internal diseases it must be regarded as a "cutaneous paraneoplasia". Out of 14 cases of erythema necroticans migrans, so far published this dermatosis occured in 13 patients suffering from pancreatic cancer. Association with pancreatitis was demonstrated in one case. In both cases herein reported the cutaneous manifestations were associated with a carcinoma of the pancreas and with cervix cancer. Extreme loss of weight, atrophic glossitis, therapy-resistant anemia and a slight diabetes are extra-cutaneous symptoms of this paraneoplastic syndrom.


[X-ray irradiation of dermatoses in the genital area]. Report on empirical data of X-ray treatment of dermatoses in the genital area, concerning indications, technique and results. Irradiations are mainly successful in large patches of Bowen's disease and Bowen's carcinoma as well as in Queyrat's erythroplasia and Paget's disease. Carcinomata of the penis should not exceed the stages T1-T2. Regression of Peyronie's disease is accelerated. Psoriatic lesions disappear following Grenz-rays without further topical treatment. Grenz-rays can also prevent or delay local recurrences of herpes simplex eruptions in the genital area.


Evaluation by electron microscopy of the integrity of iodinated plasmalipoproteins. Iodination of proteins and lipoproteins is a widely used "in vitro" labelling procedure in metabolic, autoradiographic and various other studies. However, all available iodination techniques have involved the possible damage to the proteins by self-irradiation, oxidizing agents, the alkaline milieu or by the introduction of iodine into the molecular structure itself. To evaluate the integrity of iodinated lipoprotiens, we observed the electron microscopic appearance of normal and iodinated rabbit very low density lipoproteins (VLDL) by negative staining with phosphotungstic acid. Iodination up to a molar iodine/protein ratio of 2.89 did not results in any change of shape, size or aggregating tendency of the particles. No stacks or disk-like particles like those of various hyperlipoproteinemic states were found. We conclude that electron microscopy is a valuable tool in assessing the morphological appearance of lipoprotein iodination, but it should be complemented by other techniques.


Effects of picric acid-formaldehyde fixation on the LH(HCG) receptors' binding activity in the rat testis. A histochemichal model for qualitative and quantitative studies. PAF (picric acid-formaldehyde) fixation of rat testis for a short time at 0-4 degrees C was found to give satisfactory histological results and to preserve most of the specific binding activity of LH(HCG) receptors. Investigations of the characteristics of the hormone-receptor reaction after mild PAF fixation indicated that this reaction was not substantially affected in hormne receptor affinity and its own specificity; only the capacity of the receptors was lowered by about 20%. A histochemical model is presented whose main features are: fixation of testis tissue in PAF; freezing in liquid nitrogen and cutting in the cryostat; radiolabelled hormone-receptor binding reaction performed on the sections; autoradiography to reveal the binding reaction. The utility of the method for qualitative and quantitative receptor studies and its possible application to biopsy and surgical specimens, are discussed.


Incidence of minor physical anomaly in autism. Children diagnosed as autistic were matched by age and sex with 74 control subjects and examined for presence of minor physical anomalies. Of the 16 anomalies scored, autistic children demonstrated a significant accumulation greater than the number exhibited by normal children. Three of the stigmata--low seating of ears, hypertelorism, and syndactylia--were expressed differentially in the two groups, and high palate as well as unusual cranial circumference were significantly high in both groups. Clusters of stigmata that might be associated with known chromosomal disorders could not be identified. The increased number of anomalies suggests that among autistic children such congenital markers indicate a deviant intrauterine experience.


Phototaxis and membrane potential in the photosynthetic bacterium Rhodospirillum rubrum. Cells of the photosynthetic bacterium Rhodospirillum rubrum cultivated anaerobically in light show phototaxis. The behavior of individual cells in response to the phenomenon is reversal(s) of the swimming direction when the intensity of the light available to them abruptly decreases. The tactic response was inhibited by antimycin, an inhibitor of the photosynthetic electron transfer system. The inhibitory effect of antimycin was overcome by phenazine methosulfate. Motility of the cells was not impaired by antimycin under aerobic conditions. Valinomycin plus potassium also inhibited their phototactic response; however, valinomycin or potassium alone had no effect. A change in membrane potential of the cells was measured as an absorbance change of carotenoid. Changes in the membrane potential caused by "on-off" light were prevented by antimycin and by valinomycin plus potassium, but not by antimycin plus phenazine methosulfate nor valinomycin or potassium alone. The results indicated that the phototactic response of R. rubrum is mediated by a sudden change in electron flow in the photosynthetic electron transfer system, and that the membrane potential plays an important role in manifestation of the response.


On the role of the tubulin nonexchangeable GTP site in bovine neurotubule assembly. A procedure for radiolabeling the terminal phosphoryl group of the tubulin nonexchangeable GTP site using bacterial acetate kinase and acetyl-32P is described. Warming such samples to 37 degrees results in microtubule assembly and  hydrolysis of the nonexchangeable site GTP in a parallel fashion. Removal of the microtubule-associated protein fraction from lebeled tubulin prevents hydrolysis and assembly, and recombination of these components restores both processes again in a parallel fashion. These and other experiments indicate that the nonexchangeable site GTP hydrolysis and assembly are intimately linked. The experiments also demonstrate that GRP is not required at the exchangeable nucleotide site for assembly to occur.


Tuna cytochrome c at 2.0 A resolution. III. Coordinate optimization and comparison of structures. Optimum coordinate sets have been obtained for ferrocytochrome c and the two symmetry-independent molecules of ferricytochrome c from tuna at 2.0 A resolution by making the best fit of models with standard bond lengths and angles to the experimental electron density maps (1977) J. Biol. Chem. 252, 759-785, as a preliminary to full refinement with 1.5 A data. Both the Diamond model-building programs and locally developed minicomputer routines were tried, with the latter preferred for economy and ease of operation, although both gave satisfactory results. Atomic coordinates are available on microfiche or from the Brookhaven Protein Data Bank. Using the two ferricytochrome molecules as a control, no differences between oxidized and reduced cytochrome molecules can be seen that are outside the probable limits of accuracy of the 2.0 A analysis. Rotation and subtractive difference map comparisons also show no conformation changes. If believable differences do appear in the course of the 1.5 A refinement now underway, these should be no more than minor breathing of main chain or adjustment of side chains.


Glycosaminoglycan synthesis by cultured human skin fibroblasts after transformation with simian virus 40. The synthesis of glycosaminoglycans by human skin fibroblasts derived from normal subjects, Hurler and Marfan patients before and after transformation by SV40 virus has been studied. Virus transformation results in a marked increase in hyaluronic acid synthesis in normal and Hurler fibroblasts and, to a lesser extent, in Marfan fibroblasts which show augmented synthesis of this polysaccharide before transformation. There is also an increase in heparan sulfate synthesis but a moderate decrease in dermatan sulfate synthesis on transformation. Incubation of transformed fibroblasts with 4-methylumbelliferyl-beta-D-xyloside results in a marked increase in synthesis of free chondroitin sulfate chains. The synthesis of hyaluronic acid, but not of dermatan sulfate, is inversely proportional to cell density in normal fibroblasts but not in transformed fibroblasts.


Cyclic 3':5'-nucleotide phosphodiesterase. Stimulation of bovine brain cytoplasmic enzyme by lysophosphatidylcholine. Brain cytoplasmic cyclic 3':5'-nucleotide phosphodiesterase (EC 3.1.4.17) requires an endogenous Ca2+-binding protein for ful activity. We now show that lysophosphatidylcholine also effectively enhances activator-deficient phosphodiesterase activity. Stimulation by both ligands was immediate and reversible; both rendered the enzyme more thermally labile, decreased the energy of activation, and increased the Vmax of phosphodiesterase without affecting its apparent Km for adenosine 3'5'-monophosphate. However, the cofactor requirements of the two ligands were different. Although the protein activator gave a greater stimulation than lysophosphatidylcholine, the simultaneous presence of the two gave a stimulation comparable to lysophosphatidylcholine, suggesting that the effect of the latter was predominant. Phosphodiesterase was also stimulated by oleic acid, cardiolipin, and phosphatidylinositol, albeit to a less extent.


Use of N-chlorosuccinimide/urea for the selective cleavage of tryptophanyl peptide bonds in proteins. Cytochrome c. The conditions and utility of the N-chlorosuccinimide/urea (NCS/urea) reagent for the selective cleavage of tryptophanyl peptide bonds in proteins is demonstrated with cytochrome c. At low concentrations of NCS/urea the oxidation of thioether side chains in cytochrome c is the predominant reaction. Methionyl residues are oxidized to sulfoxide and the heme-thioether bridge is partially cleaved. At 10-fold excess of NCS/urea reagent, cleavage of the tryptophanyl peptide bond is optimal at approximately 50% yield in several species of cytochrome c studied. Analytical data on isolated horse cytochrome c peptide fragments demonstrate lack of modification and cleavage at tyrosyl and histidyl residues. However, at high concentrations of NCS/urea reagent (30-fold) unexpected conversions of methionine to sulfone and cysteine to cysteic acid in intact proteins are observed. This is in contradistinction to the absence of sulfone in NCS/urea-reacted amino acid mixtures. The mechanisms of halogenation and cleavage by N-bromosuccinimide, N-iodosuccinimide, and N-chlorosuccinimide are discussed. It is porposed that the selectivity with respect to halogenation by N-chlorosuccinimide is due to the insignificant participation of molecular chlorine in the NCS/urea reaction. A mechanism of halogenation and cleavage by NCS at tryptophan is also offered.