Cytoplasmic oestradiol-binding sites and their relationship to oestradiol content in the endometrium of cattle. Twenty-three cows and heifers were killed at known times during the oestrous cycle or during the first 35 days of pregnancy. Duplicate cytosol preparations were made from the endometrium of each uterine horn and both the binding-site concentration and the oestradiol level were determined for each sample. During the cycle, the oestradiol concentration was only 0-2 to 1-7% of the concentration of binding sites which varied considerably between Days 19 and 5 (47,665 +/- 7538 sites/cell, mean +/- S.E.M.) and Days 6 to 18 (7060 +/- 444 sites/cell). The concentration of binding sites remained low in pregnant animals (6689 +/- 492), although the oestradiol concentration was high about 20 days after insemination, resulting in almost 14% of the sites being occupied. Five inseminated animals in which no conceptus was found when they were slaughtered 19 to 22 days later had low concentrations of binding sites but two animals had high levels of oestradiol with 13% and 15%, respectively, of their cytoplasmic sites being occupied. It is suggested that these animals had recently lost their conceptuses. Two ovariectomized cows and one non-cyclic animal contained high concentrations of oestradiol-binding sites in the uterine cytoplasm. No significant difference was found between the uterine horn adjacent to the ovary with the CL and the contralateral horn in early pregnancy or during the luteal phase of the oestrous cycle. An animal killed 1 week after parturition contained fourfold more sites in the involuting horn than in the opposite horn. It is suggested that progesterone plays a major role in regulating oestrogen-induced replacement of cytoplasmic binding sites.


Never too old to climb.

Never too old to climb. This nurse-administrated program has added another dimension to the hospital environment. It provides a real challenge and opportunity for reaching out in planning and implementing a program for the elderly, long-term psychiatric hospitalized resident. It has been successful because of a sincere and dedicated staff who consistently communicated, verbally and non-verbally, that "we cared"--by listening, keeping open minds, establishing and maintaining an environment conducive to growth, acceptance, encouragement, social contacts, and creating a genuine relationship based on trust. It was through these media that we achieved a milestone with the new program. Demonstration of concern, patience and interest seemed to have been a motivating force which stimulated a desire to achieve optimum growth for a majority of the residents, which has affected every service in this hospital. The freedom to plan and execute, with administrative support, challenged staff, who reached out and grasped the opportunity to turn away from the traditional ways of providing care to new ways of improving and delivering quality care to the elderly, long-term psychiatric residents. This program has been in not, however, been a panacea for all residents. Assuming responsibility, freedom to make decisions and thoughts of leaving the hospital have been somewhat difficult adjustments for a few residents. As of December 1974, approximately 5 per cent of those placed in the community were returned to the structured environment of the hospital. The successes, however, have far outnumbered the adversities in that 34 have been discharged and three are ready to be discharged in the near future.


Termination of pregnancy after accelerated lactation in the rat. III. Effects of hormonal treatments. The relationship was investigated of the hormones associated with pregnancy and lactation and the termination of pregnancy that occurs when rats mated at the post-partum oestrus are allowed to suckle a large litter after implantation. The primary cause for pregnancy termination was found to be an insufficient level of progesterone with the possible need for a primary or synergistic dose of oestrogen. The progesterone deficiency was not due to the high levels of prolactin present in nursing rats, since pregnancy termination could not be produced by prolactin administration in the absence of accelerated lactation, and could not be prevented by blocking prolactin secretion after accelerated lactation. Administration of LH to rats after accelerated lactation did prevent termination of pregnancy.


Rupture of ovarian follicles at ovulation. The appearance of mammalian follicles at the time of ovulation has been described by numerous observers. Importance features to which attention has been directed are: changes in the vascularization of the follicle, the appearance of an avascular area known as the macula pellucida or stigma, the formation of small blood clots near the stigma and some extravasation of blood, and the rupture of the follicle at the stigma and exist of the oocyte and follicular fluid. The first major advance beyond this descriptive stage came with the demonstration that antral pressure does not increase prior to follicle rupture. This observation eliminated a number of hypotheses concerning ovulation and focused attention on the deterioration and weakening of the follicle wall at the stigma. That the follicle wall weakness prior to rupture is established beyond doubt by gross observations of its increased fragility during various manipulative procedures, by stress-strain measurements in vivo and in vitro, and by histological observations. Progressive weakening of the stigma region by thinning and degeneration is a fundamental aspect of the preovulatory maturation of follicles, and it would appear to be a necessary prerequisite to rupture.


Interrelationships between prostaglandins, cyclic AMP and steroids in ovulation. Ovulation is a complex phenomenon, involving a series of biochemical events within the ovary, leading to the rupture of the follicle. This paper summarizes recent studies in our laboratory of some of these biochemical changes using the rabbit as an experimental model. It has been shown in our laboratory that isolated Graafian follicles obtained from oestrous rabbits synthesize steroids and cyclic AMP when incubated in vitro. Luteinizing hormone added to the incubation medium increased steroidogenesis and cyclic AMP synthesis many fold. When follicles were isolated from rabbits at different times following the ovulatory stimulus (mating or HCG injection) it was found that the in vitro response to LH in terms of steroidogenesis and cylcic AMP synthesis was lost as ovulation approached. In contrast, when prostaglandins (PGF and PGE) were measured in rabbit Graafian follicles it was found that the PGF and PGE levels increased as ovulation approached. From these data and from reports in the literature, we have developed a hypothetical model for ovulation in the rabbit which may help in a better understanding of the ovulatory process.


A phlogistic function of PGE1 in calcium pyrophosphate dihydrate crystal-induced inflammation. Promotion of calcium pyrophosphate dihydrate (CaPPD) crystal-induced inflammation by prostaglandin (PG) E1 has been demonstrated by two new techniques: (1) Rats deficient in essential fatty acids, biological precursors of PG, developed less footpad swelling than did normal rats following injections of unheated CaPPD. Addition of one ng of PGE1 to these crystals resulted in normal swelling. (2) This phlogistic effect of PGE1 was also demonstrated in normal rats fed a normal dietwho received injections of CaPPD crystals heated to 200 degrees C for three hours. The heated crystals induced less footpad swelling than did unheated crystals. When one ng of PGE1 was added to the suspensions of heated crystals the resultant swelling approximated that obtained by unheated crystals. The possible role of PGE1 in mediating CaPPD crystal-induced inflammation is suggested. An analysis of urate and CaPPD crytal-induced inflammation is presented with comments setting forth the concept that these metabolic crystal disorders are membrane diseases.


Calcium pyrophosphate dihydrate deposition disease without chondrocalcinosis. The pseudogout syndrome is usually associated with radiographic evidence of articular cartilage calcification. Eight patients who had joints containing calcium pyrophosphate dihydrate crystals were studied. Extensive radiographic evaluation was obtained in seven patients and a limited evaluation in the other. None had evidence of chondrocalcinosis. Six had distinctive radiographic abnormalities of the wrists consisting of radiocarpal joint space narrowing and sclerosis, and subchondral cystic degeneration of the carpal bones. We conclude that calcium pyrophosphate dihydrate deposition disease and pseudogout can occur without radiographic evidence of chondrocalcinosis and that the diagnosis can be suggested by characteristic radiographic abnormalities of the wrists.


Predominance of synovial fluid lymphocytes in early rheumatoid arthritis. Evaluation of 158 synovial fluids, mainly from patients with rheumatoid arthritis (RA) and the crystal deposition fiseases (CDD), revealed differences in the differential white blood count and numbers of cytoplasmic inclusion bodies (CIB) present. Lymphocytes and CIB were increased in RA, suggesting a different pathologic mechanism from that of CDD. Lymphocyte-like cells contained CIB implying phagocytic capability. Lymphocytes were the initial predominant cell type in RA of less than six weeks disease duration. Later polymorphonuclear leukocytes increased, possibily representing a secondary non-specific inflammatory response similar to that seen in CDD.


Isolation of foamy virus from rhesus, African green and cynomolgus monkey Leukocytes. Foamy virus (FV) was recovered regularly from the leukocyte of rhesus and cynomolgus monkeys and somewhat less often from African green monkey leukocytes. Virus was found in virtually all organs of experimentally infected rhesus monkeys. No illness or pathologic abnormalities were noted in these animals or in any of the naturally infected animals in spite of the prolonged period of viral persistence in various organs and tissues.


Herpesvirus antibodies and antigens in patients with cervical anaplasia and in controls. Antibody activity to herpesvirus hominis type 2 (HVH-2) in 151 women cured of cervical carcinoma (60 in situ, 91 invasive) and in 106 controls differed significantly, especially between the in situ (73%) and control (17%) groups. Sera of 57 patients with cervical atypia showed an increased antibody activity to HVH-2 (58%), compared with that of the 57 matched controls (23%). Antibodies to cytomegalovirus (CMV) were detected more frequently in sera of women with atypia (61%) than in sera of women with cervical disorders other than atypia (42%) or in sera of healthy controls (33%). HVH antigens were present in cervical cells from patients with atypia and from matched controls. Not only exfoliated (imprints) but also cultured and cocultured cervical cells contained HVH antigens; there was no correlation between antigen positivity and antibody activity to HVH-2. Our data further supported the association between HVH-2 and cervical anaplasia and indicated that CMV may also be implicated in its etiology.


Suppression of spontaneous in vitro transformation of autologous leukocytes by plasma from convalescent and postconvalescent infectious mononucleosis patients. All 14 plasma samples from 13 convalescent and postconvalescent infectious mononucleosis (IM) patients suppressed the spontaneous in vitro transformation of autologous leukocytes. In contrast, only 2 of 8 plasma samples from patients with acute IM suppressed this transformation. All 7 patients whose blood was tested both in the acute and convalescent or postconvalescent phases of IM showed either a conversion in transformation suppression status from negative to positive or an increase in the strength of transformation suppression. Thus recovery from IM appeared to be associated with the ability of plasma to suppress the in vitro spontaneous transformation of autologous leukocytes.


Loss of ability to synthesize collagen in fibroblasts transformed by rous sarcoma virus. Transformation of chick embryo fibroblasts by Rous sarcoma virus inhibited their ability to synthesize collagen. Kinetic experiments showed that 72 hours after infection, collagen synthesis was reduced by 90%. Nontransforming Rous-associated viruses did not inhibit collagen synthesis. The inhibition resulted from the failure of the cells to synthesize collagen polypeptides rather than from a decrease in the activity of prolyl hydroxylase; the levels of prolyl hydroxylase were fourfold those in uninfected cells. The addition of dibutyryl cyclic AMP and theophylline, alone or together, did not restore collagen synthesis.


Neoplasms in strains of splenectomized mice after a single 7,12-dimethylbenz[alpha]anthracene treatment. The effects of splenectomy on carcinogenesis by a single 10-mg dose of 7,12-dimethylbenz[alpha]anthracene (DMBA) given in olive oil by gavage was tested on BTOs, C57BL/60s, C3H/HeOs, and BALB/cOs mice. The splenectomy, performed a week before the DMBA was given, did not affect physical status or the incidence of acute toxic death of animals. DMBA-treated animals developed neoplasms at a significantly higher rate than did untreated mice. Splenectomy did not influence the overall incidence of neoplasms. Observed tumors in DMBA-treated groups were those of skin, forestomach, colon, liver, lung, adrenal, ovary, breast, hematopoietic-lymphoreticular system, and vascular system, depending on the strain. Types of DMBA-treated neoplasms were affected by prior splenectomy, depending on the strain: Splenectomy inhibited lung adenomas in BALB/cOs females and hepatomas in C57BL/60s females; splenectomy enhanced skin neoplasms in C57BL/60s and squamous cell carcinoma of the forestomach in BTOs males. The most significant change was in the incidence of the group of lymphomas. Myelogenous leukemia was increased in DMBA-treated groups of all strains, but splenectomy inhibited the development of this type of lymphoma.


Simian virus tumor antigen: a virus-specific and preexisting cell protein. The tumor (T) antigen of simian virus 40 (SV40)-transformed SV3T3 cells was immune precipitated with T antiserum obtained from hamsters bearing SV40-induced tumors and with rabbit antiserum to hamster cells. The resulting antigen-double antibody complexes were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Host components of SV40 T antigen with molecular weights of 70,000 and 30,000 were detected.


Histologic and electron microscopy observations on diethylnitrosamine-induced hepatomas in small aquarium fish (Oryzias latipes). Liver neoplasms were induced in medakas (Oryzias latipes) by the addition of diethylnitrosamine (DENA) to their aquarium water at levels of 15-135 ppm for 8 weeks. After 13 weeks, 21 to 32 fish had developed hepatomas. Medakas are useful for further studies because they are highly susceptible to the carcinogenic effect of DENA, and the time for tumor induction is relatively short. Histologic type differed in the lesions of different fish and also within individual tumors. Some were typical trabecular hepatomas, others were anaplastic hepatomas or cholangiomas, or mixtures of these. Electron microscopy revealed an extensive rough-surfaced endoplasmic reticulum in a lamellar pattern, many mitochondria, and several round lysosomes in tumor cells. A few fat droplets with occasional crystalline ghosts were sometimes in the cytoplasm. The Golgi apparatus was not conspicuous. Some cells had highly developed microvilli that showed differentiation toward structures resembling bile capillaries.


Murine oncornavirus activation in the pancreas during infection with Venezuelan equine encephalitis virus. Electron microscopic studies of the pancreases of 3-week-old ICR Swiss mice infected with Venezuelan equine encephalitis virus and killed 6 days post inoculation (at which time they were moribund) indicated significantly more type-C particles than were found in uninfected controls. This phenomenon was apparently possible because of the microanatomy of the pancreas, in which interstital spaces allowed accumulation of virus particles.


Differential response to cytochalasin B among cells transformed by DNA and RNA tumor viruses. Mouse, hamster, rat, human, and chick cells were transformed by RNA and DNA tumor viruses: simian virus 40, adenovirus type 7, Kirsten mouse sarcoma virus (Ki-MuSV), Moloney mouse sarcoma virus, and Rous sarcoma virus. All cultures of transformed cells grew to high concentration densities. Normal and transformed cells were treated with cytochalasin B (CB) at concentrations preventing cytoplasmic cleavage. Cells altered by DNA tumor viruses responded to CB with numerous nuclear divisions resulting in highly multinucleated cells. All but one line of cells transformed by RNA tumor viruses responded to CB with usually only one and occasionally two nuclear divisions. Only binucleated cells were formed. One clone of CB-treated BALB/c mouse embryo fibroblasts transformed by Ki-MuSV showed numerous cells with four and five nuclei. HOWEVER, IN CONTRASt to cells transformed by DNA viruses, few cells had seven or more nuclei. These results suggest that, in the presence of CB, cells transformed by DNA tumor viruses show uncontrolled nuclear division, whereas cells tranformed by RNA tumor viruses show controlled nuclear division.


Effect of adeno-associated virus on cancer expression by herpesvirus-transformed hamster cells. Infection of herpes simplex virus type-2-transformed hamster tumor cells with adeno-associated virus type 1 before inoculation into hamsters specifically delayed the appearance of palpable tumors and increased the survival time of the animals. The data indicated that a defective virus of humans can influence cancer expression by a virus-transformed cell.


Inhibition of in vitro growth of lymphoma cells by macrophages from tumor-bearing mice. Spleen cells from C57BL/6 mice bearing primary tumors induced by the Moloney strain of murine sarcoma virus (MuSV) strongly inhibited the uptake of tritiated thymidine (3H-TDR) by RBL-5 lymphoma cells in a 48-hour growth-inhibition assay (GIA). This activity was first detected 7 days after MuSV was injected; it peaked at 14 days, and was usually no longer detectable after 18-21 days. It could be detected at effector cell/target cell ratios between 20:1 and 5:1, at which normal spleen cells had a growth-promoting effect. The effector cells in the GIA were not T cells, and various depletion experiments suggested that they were macrophages. Macrophages of a purity of over 95% were obtained in the glass-adherent fraction of thioglycollate-induced peritoneal exudate cells (PEC). PEC were growth inhibitory when obtained from either normal or MuSV tumor-bearing mice. However, at effector cell/target ratios of 2.5:1, only PEC from MuSV tumor-bearing mice had an effect; PEC from normal mice were inactive. Activity of spleen cells in the GIA appeared distinct from T-cell-dependent specific cytotoxicity, which was not affected by removal of macrophages. Activity in the GIA was nonspecific, and target cells which do not cross-react with RBL-5 cells were equally inhibited. Furthermore, spleen cells from mice bearing primary tumors induced by 3-methylcholanthrene were also fully active against RBL-5 cells. Supernatants from spleen cell cultures obtained from mice 14 days post injection with MuSV also inhibited the incorporation of 3H-TDR by RBL-5 cells in vitro. However, this effect seemed to be an artifact, since the tumor cells proliferated equally well in the presence or absence of the supernatants. In contrast, the direct effect of spleen cells from MuSV tumor-bearing mice was reflected both by an inhibition of cell proliferation and by inhibition of 3H-TDR incorporation.


Local antitumor activity of a primary and an anamnestic response to a syngeneic guinea pig hepatoma. After intradermal (id) injection, the line-10 hepatoma grew progressively in nonimmune guinea pigs, whereas the line-1 hepatoma grew for approximately 2 weeks, developed central necrosis, ulcerated, and regressed. Growth of the line-10 hepatoma was suppressed when line-10 hepatoma cells were mixed with antigenically distinct line-1 hepatoma cells before id injection into syngeneic strain-2 guinea pigs. Mixture of line-10 with irradiated line-1 or viable strain-2 embryo cells did not inhibit tumor growth. Preimmunization of recipients to line-1 cells abrogated the suppression of tumor growth from mixtures of line-1 and line-10.


DNA polymerases in polyoma virus-infected mouse kidney cells. Infection of arrested mouse kidney cells by polyoma virus results in the induction of the cellular 6-8S DNA polymerase activity. Levels of this enzyme increase two- to threefold in the cytoplasm but seven- to tenfold in nuclei and nuclear extract, suggesting an accumulation of the enzyme in the nucleus. Experiments using the inhibitor of DNA synthesis, fluordeoxyuridine, indicate that this accumulation is linked to active DNA synthesis. The activity and cellular distribution of the small 3.4S DNA polymerase remains unchanged.


Effect of hypertonic conditions on protein synthesis in cells productively infected with simian virus 40. Hypertonic medium selectively suppressed the synthesis of most host cell polypeptides relative to the synthesis of simian virus 40 capsid polypeptides and a minority of cellular polypeptides, notably histones. Under optimal hypertonic conditions, the synthesis of the major capsid polypeptide (VP1) is enhanced about sevenfold relative to host polypeptide synthesis. Because of the small amounts of the other nonhistone capsid polypeptides (VP2) and VP3) present in cell lysates, it was difficult to quantitate the extent, if any, of their enhancement. The maintenance of the restricted pattern of protein synthesis caused by hypertonic medium was dependent on continual peptide chain initiations. The resistance of viral protein synthesis to hypertonic conditions provides a means of detecting relatively low levels of intracellular viral protein synthesis. Analysis of the specific activity of the acid-soluble [3H]lysine pool indicated that the rate of incorporation of [3H]lysine into protein was an overestimation of the actual rate of overall protein synthesis occurring in cells exposed to hypertonic as compared to isotonic conditions. Since it is likely that both cellular and viral protein synthesis draw lysine from a single pool, this change in pool specific activity does not affect the analysis of relative rates of protein synthesis at a given level of tonicity.


Differential expression of helper viral structural polypeptides in cells transformed by clonal isolates of woolly monkey sarcoma virus. Cell lines transformed by woolly monkey sarcoma virus (WSV) in the absence of infectious virus production were analyzed for the expression of woolly monkey helper viral p30, p12, and gp70 antigens. Several lines produced high levels of both p30 and p12, whereas gp70 was not detectable. One transformed clone expressed only p12, and in another cell line, none of the helper viral antigens were detected. The properties of each sarcoma virus bred true upon transmission, indicating that each variant represents a distinct genotype. The different cell lines were examined with respect to properties characteristic of the transformed state. The in vitro growth properties and oncogenicity of each WSV-transformed clone were indistinguishable, indicating that transformation by WSV occurs independently of the expression of at least three helper viral polypeptides.


Qualitative complementation test for temperature-sensitive mutants of herpes simplex virus. A simple, rapid, qualitative method for classifying temperature-sensitive mutants of herpes simplex virus into functional complementation groups has been developed. The positive reaction observed in this test reflects the ability of mutant pairs to interact by both complementation and recombination.


Inhibition by aurintricarboxylic acid and polyethylene sulfonate of RNA transcription of vesicular stomatitis virus. The in vitro activity of the ribonucleoprotein-dependent RNA transcriptase of vesicular stomatitis virions was found to be completely inhibited by low concentrations of aurintricarboxylic acid (ATA) and polyethylene sulfonic acid (PES) when these inhibitors were added before the start of the RNA polymerase reaction. However, if RNA synthesis was allowed to occur before ATA or PES was added, RNA synthesis continued for a short time (10 min or less) in the presence of either inhibitor at a concentration which completely inhibited uninitiated enzyme. The ability to continue to synthesize RNA in the presence of ATA or PES only developed if all four nucleoside triphosphates were present during the preincubation period prior to the addition of the inhibitors. The protection was apparently not due to the released products of RNA polymerization. The results are interpreted as indicating that ATA and PES probably  inhibit some reaction other than elongation of RNA chains, and this reaction might be one involved at or near initiation sites.


Altered DNA synthesis in a mutant of Salmonella typhimurium that channels bacteriophage P22 toward lysogeny. Pox-1, a mutant of Salmonella typhimurium, strongly channels P22 toward lysogeny. Viral DNA synthesis in this slow-growing mutant is delayed to a greater extent than viral protein synthesis. The relative enhancement of c2 repressor synthesis results in much higher repressor/DNA synthesis ratios in Pox-1 than in wild-type cells. This probably accounts for the high frequency of lysogenization.


Structure of B77 sarcoma virus RNA: stabilization of RNA after packaging. Extracellular maturation of Bratislava 77(B77) sarcoma virus RNA involves a stabilization of linkage between 35S subunits since the Tm of 60 to 70S RNA in the presence of 0.5 M NaCl increases from 56 to 67.5 C as the age of the virus increases. This stabilization process is strongly temperature dependent; the rate at 45 C is increased fourfold over the rate at 37 C. As a result of the instability of the immature RNA, denaturation to subunits occurs at room temperature during phenol extraction. This dissociation can be prevented by increasing the NaCl concentration during the extraction. The data support a model which proposes that RNA subunits of oncornaviruses exist as assembled 60 to 70S RNA even in immature virions but that the linkages between subunits are stabilized as a function of time. These linkages appear to be maintained by nucleotide base pairing rather than by protein. However, isolated RNA does not undergo stabilization, suggesting that some other component of the virion is necessary for the process to occur.


Transcription of simian virus 40. V. Regulattion of simian virus 40 gene expression. RNA "exhaustion type" hybridization was used to measure the complementarity of nuclear and cytoplasmic viral RNA to the early (E) and late (L) simian virus 40 (SV40) DNA strands. This type of hybridization measures the amount of labeled RNA complementary to each of the two DNA strands, rather than the fraction of each SV40 DNA strand that is homologous to SV40 RNA. At 48 h after infection, about 5% of the nuclear newly synthesized viral RNA was complementary to the E-strand (- strand) and 95% was complementary to the L-strand (+ strand). This proportion was independent of the labeling time, indicating similar accumulation of the E- and L-RNA transcripts in the nucleus. The nuclear E- and L-viral RNA transcripts sedimented in a similar manner on sucrose gradients. Of the cytoplasmic viral RNA only about 1% was complementary to the E-strand, these molecules sedimenting at 19S, whereas 99% were complementary to the L-strand and sedimented at 19S and 16S. The abundance of E-RNA transcripts in nuclei of cells infected with serially passaged virus was about four times higher than that in nuclei of cells infected with plaque-purified virus; however, the size and proportion of the corresponding cytoplasmic E- and L-RNA transcripts was independent of the type of virus used to infect the cells. According to these results at least two control mechanisms regulate viral gene expression in productively infected cells, one operates at the trnascriptional level and the second at the post-transcriptional level.


Host influence on the activity of genes c1 and c3 in regulating the decision between lysis and lysogency in bacteriophage P22. A Polymyxin B-sensitive mutant of Salmonella typhimurium (Pox-1) channels all infecting wild-type P22 toward lysogenization. The efficiency of this channeling is sufficiently high that P22c+ (wild type) cannot form plaques on Pox-1; phage mutants defective in repressor synthesis (P22c1, c2, c3) or refractory toward repressor (P22vir B) can form plaques. The lytic growth of all phages which have a functional c1 gene is retarded in Pox-1; this retardation is seen even in phages which cannot make repressor. We present experiments which are consistent with the explanation that the retardation is an exaggeration of a normal regulatory event. In a wild-type host, P22 genes c1 and c3 products, host RNA polymerase, and other host factors (?) interact at a promotor site (c27) IN THE PHAGE DNA. This interaction promotes repressor synthesis and represses transcription of lytic genes. In the mutant Pox-1, a host product involved in viral DNA synthesis and transcription is altered. The altered host product results in stronger retardation of lytic gene transcription. The importance of this interaction in the decision between lysis and lysogeny is discussed. The mutant Pox-1 alters the expression or activity of another phage gene. Gene c3 product is absolutely required for lysogenization in this host, although it is not so required in wild-type S. typhimurium.


Biochemical and biophysical characteristics of diarrhea viruses of human and calf origin. Polyacrylamide gel electrophoretic analysis of purified preparations of human and calf diarrhea viruses indicated eight polypeptide components, or possibly nine in the case of the calf diarrhea virus. Thermal denaturation and analytical studies of the calf diarrhea virus genome showed it to consist of 11 double-stranded segments of RNA. The placing of the human and calf diarrhea viruses together with other similar viruses into a genus separate from reovirus and orbivirus, but within the family Reoviridae, is discussed.


Simian virus 40-specific proteins in HeLa cells infected with nondefective adenovirus 2-simian virus 40 hybrid viruses. The synthesis of simian virus 40 (SV40)-specific proteins in HeLa cells infected with the nondefective adenovirus 2 (Ad2)-SV40 hybrid viruses, Ad2+ND2, Ad2+ND3, Ad2+ND4, and Ad2+ND5, was investigated. Infected-cell proteins were labeled with radioactive amino acids late after infection, when host protein synthesis was shut off, and analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. All polypeptides normally seen in Ad2-infected cells were found in cells infected by the hybrid viruses. In addition to the Ad2-specific proteins, cells infected with Ad2+ND2 contain two SV40-specific proteins with apparent molecular weights of 42,000 and 56,000, cells infected with Ad2+ND4 contain one protein with an apparent molecular weight of 56,000, and cells infected with Ad2+ND5 contain one protein with an apparent molecular weight of 42,000. Cells infected with Ad2+ND3 do not contain detectable amounts of proteins not seen during Ad2 infection. Pulse-chase experiments demonstrate that the SV40-specific proteins induced by Ad2+ND2, Ad2+ND4, and Ad2+ND5 are metabolically unstable. These proteins are not present in purified virions. Two nonstructural Ad2-specific proteins have been demonstrated in Ad2 and hybrid virus-infected cells which have a smaller apparent molecular weight after a short pulse than after a pulse followed by a chase. The molecular weight increase during the chase may be caused by the addition of carbohydrate to a polypeptide backbone.


N-terminal amino acid sequences in the major capsid proteins of foot-and-mouth disease virus types A, O, and C. Sequences of amino acids at the N-termini of virus proteins VP1, VP2, and VP3 were determined for foot-and-mouth disease virus types A12 strain 119, O1Brugge and C3Resende. In the polyacrylamide gel electrophoresis system used to purify the proteins, VP3 migrated faster than VP1 or VP2; and in the virion, VP3 could be cleaved by trypsin into VP3a and VP3b. The N-terminal amino acids for each of the virus types were glycine in VP1, aspartic acid in VP2, and threonine in VP3. No divergences in sequence across the virus types were indicated until at least the fourth position in VP1, and the third in VP3. For virus types A12, O1 and C3, the sequences were, respectively: for VP1 (Gly-ile-phe,pro,val---), (Gly,ile,phe---) and Gly-ile-phe,ala---); for VP2 (Asp,X,met---), (Asp---) and Asp-leu---); and for VP3 (Thr-thr-ala-thr---), (Thr-thr-ser---) and (Thr-thr---). Unresolved mixtures of VP3a and VP3b, from either A12 or O1 viruses, appeared to have the N-terminal amino acids threonine, which is presumed to be the same threonine as in uncleaved VP3 and serine, which is generated by the tryptic cleavage.


Properties and origins of infectious rhinovirus type 14 particles of different buoyant densities. Isopycnic centrifugation of rhinovirus type 14 (RV14), purified from infected HeLa or KB cell cultures, into CsCl gradients resolved two bands of infectious virus particles with buoyant density values of 1.409 +/- 0.007 (H virus) and 1.386 +/- 0.004 (L virus) g/ml. Only H virus was detected by incorporation of radiolabeled uridine into viral RNA, and H virus accounted for the majority of infectivity in gradients. H and L virus could not be differentiated by plaque morphology, extent of neutralization by RV14-specific antiserum, or particle size. Electron microscope studies showed that most L-virus particles were associated with an amorphous material. Treatment of L virus with proteolytic enzymes or rebanding L virus in CsCl gradients resulted in recovery of the majority of infectivity as H virus. Virus purified from cell-free fluids from infected HeLa or KB cell cultures banded only as H virus. HeLa cell cultures challenged with purified H virus and harvested at 3 h postinoculation for virus purification yielded only infectious H virus. Both H and L viruses were detected in cell cultures that had been challenged with purified H virus and harvested at 12 h postinoculation. The data suggest that H virus represents progeny virus, whereas L virus represents sequestered infectious virus particles which become associated with an amorphous material and do not enter into viral replicative processes.


Proteins specified by herpes simplex virus. XIII. Glycosylation of viral polypeptides. In the course of herpes simplex virus 1 (HSV-1) replication in human epidermoid carcinoma no. 2 cells, the synthesis and glycosylation of host cell proteins ceases and is replaced by the synthesis and glycosylation of virus-specified polypeptides. Analyses of the synthesis of viral glycoproteins show that the glycosylation of viral polypeptides occurs late in the virus growth cycle and that certain of the precursors to major vital glycoproteins are members of the gamma group of polypeptides, i.e., polypeptides synthesized at increasing rates until 12 to 15 h postinfection. Viral glycoproteins are formed by stepwise additions of heterosaccharide chains to completed precursor polypeptides. The precursor and the highly glycosylated product are separable by gel electrophoresis and are localized in different fractions of infected cells. Within 15 min of their synthesis, precursor polypeptides acquire heterosaccharide chains of about 2,000 molecular weight, which contain glucosamine but little or nor fucose or sialic acid. Both precursor and product of this first stage of glycosylation are absent or present in low concentrations in the surface membranes of the infected cell and in the virion. The partially glycosylated product is then conjugated further in a slow, discontinuous process to form the mature glycoprotein of the virion and plasma membrane. These mature products bear large heterosaccharide units with molecular weights greater than 4,000 to 5,000; these contain fucose and sialic acid as well as glucosamine. Heterosaccharide chains from infected and uninfected cells are distributed among discrete size classes and the smallest chains consist of multiple saccharide residues.


Comparison of the virion proteins specified by herpes simplex virus types 1 and 2. Purified herpes simplex virus type 2 (HSV-2) virions were found to contain approximately the same number of polypeptides as HSV type 1 (HSV-1) virions. Comparisons of the structural proteins specified by five independent HSV-2 isolates revealed some minor differences in their electrophoretic profiles on sodium dodecyl sulfate-acrylamide gels; certain invariant features of the electrophoretic profiles, however, allowed clear differentiation between all the HSV-2 isolates and HSV-1.


Method of examining viral RNA metabolism in cells in culture: metabolism of vesicular stomatitis virus RNA. A method is described for radioactively labeling viral RNA and then quickly halting further incorporation of radioactive precursor into RNA so that the fate of the labeled RNA can be followed. Small complementary RNAs synthesized during vesicular stomatitis virus infection in the presence of cycloheximide do not metabolize to virion length molecules when protein synthesis inhibition is reversed.


Amounts of Epstein-Barr virus DNA in somatic cell hybrids between burkitt lymphoma-derived cell lines. The amounts of Epstein-Barr virus DNA in somatic cell hybrids between human lymphoid cell lines were found to be higher than in the parental cell lines, although the hybrids were not induced to spontaneous virus production.


Should patients with zoster receive zoster immune globulin? Zoster developed in 55 patients, and 15, generalized disease occurred. Twelve of 14 patients with dissemination had prompt appearance of both complement-fixing (CF) antibody and fluorescent antibody against varicella-zoster (VS) membrane antigen (FAMA). The geometric mean titers of VZ antibody from the first to the fifth and the sixth through the tenth days following onset of zoster were similar in patients with localized and generalized involvement. Two patients with disseminated zoster had undetectable CF antibody, but VZ FAMA was demonstrable. Because the clinical course of zoster appears to be unrelated to the serologic response to VZ virus, it is unlikely that administration of antibody with zoster would be benefit in this disease.


The respiratory effects of chloromethyl methyl ether. A prospective study of 125 chemical workers was carried out for ten years to investigate the incidence of lung cancer. Some of the men were exposed to chloromethyl methyl ether containing bis(chloromethyl) ether as an impurity. Bronchogenic carcinoma was markedly increased among them, with a strong dose-response relationship. An unexpected inverse relationship was noted between smoking and the incidence of lung cancer. The neoplasms (all small-cell carcinomas) occurred in relatively young men. Symptoms of chronic bronchitis were reported more often among men exposed to chloromethyl ether, and a dose-response relationship was apparent, with smoking a cofactor. Ventilatory function was not significantly affected by chemical exposure. Periodic screening over the first five years of the study showed a decrease in chronic coughing and an increase in dyspnea while chemical exposure was diminishing.


Adenoid cystic carcinoma (cylindroma) of the trachea masquerading as asthma. A 40-year-old woman was admitted with upper airway obstruction; chest roentgenogram with tomograms, laryngoscopy, and bronchoscopy verified the presence of a subglottic tumor. Spirometry disclosed a maximum inspiratory flow rate more reduced than in the maximum expiratory flow rate, suggestive of upper airway obstruction. The pathologic conditions were consistent with cystic adenoid carcinoma (cylindroma) of the trachea. The patient underwent tracheostomy and total laryngectomy with an uneventful recovery. No evidence of local extension or distant metastases were evident at time of surgery.


Cortisol secretion in Nelson syndrome: Persistence after "total" adrenalectomy for Cushing syndrome. In two patients who were severely pigmented (Nelson syndrome) following bilateral adrenalectomy for Cushing syndrome, symptoms of hyper-cortisolism developed while they were receiving only physiologic steroid replacement. Cortical assays proved that endogenous cortisol production had not been obliterated. Even after total adrenalectomy, steroid measurement should be performed to guard against adrenocortical excess.


Interaction in culture between mouse ascites hepatoma (MH-134) cells and lymphoid cells of isologous mice. Thymocytes or lymphocytes of mesenteric lymph nodes were obtained from mice bearing subcutaneously mouse ascites hepatoma MH-134 for 5 to 40 days. These lymphoid cells were added into the cultures of MH-134 cells. Morphological changes of cells in the mixed cultures were observed by time-lapse cinemicrography for the period of 4 weeks. Lymphoid cells were phagocytosed by MH-134 cells, and, in most cases, the tumor cells did not undergo any damage due to the phagocytosis. The exceptional cases were as follows: When MH-134 cells were mixed with thymocytes from mice bearing MH-134 tumor for 5 days, MH-134 cells phagocytosed thymocytes but some of them died later. In the mixed cultures of MH-134 cells and thymocytes or mesenteric lymph node lymphocytes from mice bearing MH-134 for 14 or 15 days, MH-134 cells phagocytosed lymphoid cells but died later by the burst of cytoplasm. By the burst many lymphoid cells phagocytosed appeared from the broken cytoplasm of MH-134 cells and, in some cases, the lymphoid cells looked to be alive. These findings suggest the possibility that lymphoid cells attack tumor cells not only from the cell surface but also from the inside being phagocytosed by tumor cells.


Pathogenicity of mouse hepatitis virus for mice depending upon host age and route of infection. Day-old to 4-week-old mice from a breeder colony which had been seromonitored to be free from mouse hepatitis virus infection, were tested for susceptibility to the virus by different routes of inoculation. After intraperitoneal, intravenous and intracerebral inoculation of 10(2) or more plaque-forming units of the virus, mice of all ages died of acute hepatitis. While day-old mice died also after subcutaneous, intranasal and peroral routes of inoculation, those 3 weeks or more of age resisted to infection by these routes. To intranasal inoculation mice 1 and 2 weeks of age were fully susceptible but some of the resisted to peroral inoculation. In the course of non-fatal infection in 4-week-old mice after intranasal inoculation, viremia and production of some hepatic lesions were recognized and infection became fatal in association with cortisone treatment. The results suggested that the intranasal route of infection may be of importance for spreading of infection in mouse breeding colonies in which inapparent infection is prevailing.


Carcinogenesis in tissue culture 24: tumorigenicity and aggregate-forming capacity of mammalian cells in culture. Twenty two kinds of strains and substrains of mammalian cells were cultivated on a gyratory shaker to obtain cell aggregates. The relationship between the morphology of aggregates and tumorigenicity of the cells was investigated. In some groups of cells, the size of aggregates corresponded with their tumorigenicity, e.g. among rat liver cells untreated, hepatoma cells produced by back-transplantation of rat liver cells after the treatment with a chemical carcinogen in tissue culture, and the hepatoma cells passaged serially through rats. The correspondence, however, was generalized to not all the cell strains. It was impossible to identify the tumorigenicity of cells by their aggregate-forming capacity.


Sex difference in the development of fatty liver by orotic acid. Effects of orotic acid on liver lipid accumulation and incorporation of methionine [methyl-14C] into liver phosphatidylcholine and protein, and into serum beta-lipoprotein were studied. Male and female rats of Wistar strain were fed a semisynthetic diet supplemented with 1 per cent orotic acid for 7 days. Feeding of orotic acid induced a marked fatty liver in female rats, but not in males. In female rats, radioactivity in liver phosphatidylcholine was significantly decreased by orotic acid, and that in liver protein was slightly decreased. In male rats, incorporation of methionine [methyl-14C] into liver phosphatidylcholine and protein was unchanged between the control and the rats fed orotic acid. Radioactivity in serum beta-lipoprotein was decreased to a greater extent in female rats than in males. These results suggest that sex difference in the development of fatty liver may be due to the difference in the effect of orotic acid on liver phosphatidylcholine biosynthesis.


[Familial glucocorticoid insufficiency (author's transl)]. Familial glucocortivoid insufficiency--also called hereditary lack of response of the adrenals to ACTH-has not yet been described in Europe. Isolated glucocorticoid insufficiency combined with intact secretion of aldosterone and high plasma level of ACTH are characteristic. The diagnostic difficulties are demonstrated by the deaths before diagnosis of 13 siblings of 21 patients from 9 families. One such family is described. The first of 3 children died at the age of 3-1/2 years after a two day illness interpreted as encephalitis. Another boy, aged 4-1/2 years, had a hypoglycemic attack. 3 months later he died in status epilepticus after a short feverish illness. At autopsy the adrenals were very small. Histologically only the glomerulous zone was developed. An the hypophysis there was hyperplasia of the R cells producing ACTH. The 6 year old sister had been pronouncedly pigmented from the age of a few months onwards. Age in terms of height and bone development corresponded to 8-1/2 and 7-1/2 years. After intravenous insulin and synacthen plasmacorticoids could not be found. With normal and low supply of salt, the following data were found: Renin-activity 2.9 and 5.6 ng/ml p.h. respectively; rate of aldosterone secretion 62.6 and 151.5 mug/24 hrs.; average aldosterone plasma concentration 9.39 and 27.7 ng/100 ml respectively; MCR 666 and 5471 p/24 hrs. Plasma ACTH in this patient (5086--7200 pg/ml) and, post mortem, in her brother (8100 pg/ml), were extremely reaised.


[Results of catamnestic, neurological and psychological investigations in children suffering from cytomegaly virus infection, diagnosed in neonates (author's transl)]. Report on results of follow-up in 10 children, in 9 of whom clinical findings at the age between 6 and 11 months suggested the diagnosis of connatal cytomegaly. All 10 children secreted cytomegaly virus in the urine. In 3 of them post-natal infection appears probable. Follow-up was carried out at the ages between 3 and 5.9 years. Physical development was delayed in 4, severe neurologic damage occurred in 2, medium severe or minimal, in part reversible, neurologic signs in 3 childrens. The 2 most severely neurologically damaged children could not be examined psychologically. The IQ of the other 8 was between 92 and 105. 3 children showed in individual other tests only minor, insignificant loss of performance, 2 more pronounced defects which suggested cerebral damage.


[Investigation of cyclic renal adenosinmonophosphate (yamp)-excretion in pseudohypoparathyreoidism (author's transl)]. Report on a boy aged 6-1/2 years with pseudohypoparathyroidism. This is due to a genetic lack of PTH inducible adenocyclase. Even during vitamin D treatment renal camp formation by giving PTH extract cannot be stimulated, while in this situation. PTH shows a clearly phosphaturic effect. Whether this is due to a PTH potentiated vitamin D effect is discussed. The assay of camp in urine before and after stimulation with PTH extract has proved the most certain diagnostic criterion.


[Ultrasound in the differential diagnosis of Wilms tumors (author's transl)]. The usefulness of ultrasound for distinguishing between Wilms tumor and hydronephrosis is shown in a case where radiographic signs were not clear. Ultrasound always permits differentiation between cystic and solid tumors.


[Plasma ACTH and Nelson tumors in bilaterally adrenalectomized Cushing patients]. Bilaterally adrenalectomized patients (18) because of Cushing's syndrome were studied. 16 of these 18 persons exhibited fairly high plasma ACTH levels. Distinctly elevated plasma ACTH levels were observed in 6 of them. 5 patients of this group showed an enlarged sella and deep pigmentation of the skin. 4 of the latter group had a mucoid cell adenoma which was removed selectively. After the surgery the ACTH values were found to be within normal range without any significant endocrine dysfunction of the pituitary. A long-term ACTH survey revealed that the ACTH levels remained within the ranges of bilaterally adrenalectomized Cushing patients without concomitant enlargement of the sella or pigmentation.


Paracrystalloids from mitochondria in thyroid follicular cells. Normal occurrence and experimental induction. The ultrastructure of the follicular cells of the thyroid gland of bats caught in early June, when the euthyroid state is being restored following the phase of great thyroid activity that occurs at arousal, was studied. Several ultrastructural features were found which had not been observed in animals caught at other periods of the annual life cycle. The most striking ultrastructural feature was the presence in many cells of prominent, large membrane-enclosed paracrystalloid bodies. The paracrystalloids exceeded 6.0 mum in length and were approximately 1.0 mum. in width. Examination of transition forms showed that paracrystalloids were derived from mitochondria and were formed by the reorganization of the cristae into parallel closely apposed membranes. The membranes run longitudinally in loops from one end of the mitochondrion to the other. Similar mitochondrial alterations were observed in rats by duplicating the thyroid conditions of the June bat. Rats were given thiouracil for 10 weeks followed by L-triiodothyronine injection. These animals all showed similar mitochondrial-paracrystalline alterations which were not seen in hypophysectomized animals or animals given L-triiodothyronine alone. Mitochondria from thyroids of rats given thiouracil alone showed an increase in size and number and changes in shape and internal structure. The results of the bat and rat studies suggest that rapid shutdown of active follicular cells appears to promote thyroid paracrystalloid formation from mitochondria.


Virus-induced abortion. Studies of equine herpesvirus 1 (abortion virus) in hamsters. A hamster-adapted strain of equine herpesvirus-1 (equine abortion virus) caused severe hepatic degeneration in both pregnant and nonpregnant hamsters and, in addition, regularly induced abortion in pregnant hamsters inoculated at midgestation. In nonpregnant hamsters, the only consistently affected organ was the liver despite a prolonged viremia. Newborn animals usually died 1 to 2 days after inoculation; adults died 5 to 9 days after inoculation. In pregnant hamsters, the virus had a tropism for the placenta as well as the liver. The placental infection was confined almost exclusively to one cell type in the fetal portion of the placenta: the trophoblast cells of the syncytiotrophoblast zone. Necrosis of this zone led to fetal death and abortion. Infection of the fetus did not occur.


Oligodendrocytes and their myelin-plasma membrane connections in JHM mouse hepatitis virus encephalomyelitis. Mice infected with JHM strain of mouse hepatitis virus develop a demyelinating encephalomyelitis. Myelin sheaths are stripped off axons by invading macrophages after degeneration of the infected oligodendrocytes. The derivation of the virus from granular cytoplasmic particles that condense around and bud into endoplasmic reticulum is demonstrated. The infected oligodendrocytes undergo hypertrophic changes prior to degeneration. Hypertrophic cells are characterized by abundant microtubules, filaments, mitochondria, aggregates of electron-dense particles, and numerous, unusual plasma membrane connections to myelin lamellae. Vacular and hydropic changes are prominent in degenerating cells. The significance of finding infected oligodendrocytes with altered myelin-plasma membrane connections is discussed with reference to the pathogenesis of recurrent, postinfectious demyelination known to develop subsequent to acute virus infections.


Studies of the oligomycin-sensitive ATPase from yeast mitochondria. Reconstitution of ATP-32Pi exchange in the presence of phospholipids. A purified preparation of the oligomycin-sensitive ATPase from yeast mitochondria has been shown to elicit an oligomycin- and uncoupler-senstitive ATP-32Pi exchange in the presence of phospholipids. Reconstitution was normally achieved by dialysis of an ATPase-phospholipid-cholate mixture. Following this procedure, vesicles with diameters between 200 and 1,500 A were seen by electron microscopy. As in mitochondria, ATPase activity in the reconstituted system was stimulated by a range of uncouplers which inhibited ATP-32Pi exchange. These and other findings suggest that the coupling mechanism may still be intact within the ATPase complex.


Magnetic resonance and kinetic studies of the mechanism of membrane-bound sodium and potassium ion- activated adenosine triphosphatase. EPR and water proton relaxation rate (1/T1) studies of partially (40%) and "fully" (90%) purified preparations of membrane-bound (Na+ + K+) activated ATPase from sheep kidney indicate one tight binding site for Mn2+ per enzyme dimer, with a dissociation constant (KD = 0.88 muM) in agreement with the kinetically determined activator constant, identifying this Mn2+-binding site as the active site of the ATPase. Competition studies indicate that Mg2+ binds at this site with a dissociation constant of 1 mM in agreement with its activator constant. Inorganic phosphate and methylphosphonate bind to the enzyme-Mn2+ complex with similar high affinities and decrease 1/T1 of water protons due to a decrease from four to three in the number of rapidly exchanging water protons in the coordination sphere of enzyme-bound Mn2+. The relative effectiveness of Na+ and K+ in facilitating ternary complex formation with HPO2-4 and CH3PO2-3 as a function of pH indicates that Na+ induces the phosphate monoanion to interact with enzyme-bound Mn2+. Thus protonation of an enzyme-bound phosphoryl group would convert a K+-binding site to a Na+-binding site. Dissociation constants for K+ and Na+, estimated from NMR titrations, agreed with kinetically determined activator constants of these ions consistent with binding to the active site. Parallel 32Pi-binding studies show negligible formation (less than 7%) of a covalent E-P complex under these conditions, indicating that the NMR method has detected an additional noncovalent intermediate in ion transport. Ouabain, which increases the extent of phosphorylation of the enzyme to 24% at pH 7.8 and to 106% at pH 6.1, produced further decreases in 1/T1 of water protons. Preliminary 31P- relaxation studies of CH3PO2-3 in the presence of ATPase and Mn2+ yield an Mn to P distance (6.9 +/- 0.5 A) suggesting a second sphere enzyme-Mn-ligand-CH3PO2-3 complex. Previous kinetic studies have shown that T1+ substitutes for K+ in the activation of the enzyme but competes with Na+ at higher levels. From the paramagnetic effect of Mn2+ at the active site on the enzyme on I/T1 of 205T1 bound at the Na+ site, a Mn2+ to T1+ distance of 4.0 +/- 0.1 A is calculated, suggesting the sharing of a common ligand atomy by Mn2+ and T1+ on the ATPase. Addition of Pi increases this distance to 5.4 A consistent with the insertion of P between Mn2+ and T1+. These results are consistent with a mechanism for the (Na+ + K+)-ATPase and for ion transport in which the ionization state of Pi at a single enzyme active site controls the binding and transport of Na+ and K+, and indicate that the transport site for monovalent cations is very near the catalytic site of the ATPase. Our mechanism also accounts for the order of magnitude weaker binding of Na+ compared to K+.


Estrogen and estrogen receptors of breast cancer. Human breast cancer can be divided into a group that contains specific receptor sites for estrogen and a group without such specific estrogen-binding sites. The presence of specific estrogen receptors in some tumors indicating hormonal dependency has been shown to be of predictive value for endocrine treatment. This would greatly improve therapeutic planning for patients with breast cancer. Tumor tissue from 52 patients was investigated for content of both cytosol estrogen and estrogen receptor. In addition, the total tumor estrogen was also determined in 14 of these tumors. The results of this investigation show two distinct groups: one group containing both estrogen receptor and estrogen and a second group with no receptor but with measurable amount of estrogen. Tumors with estrogen receptors have higher tissue levels of estrogen than tumors without specific estrogen receptor. Even in the absence of estrogen recptor, however, most tumor tissue examined contained a measurable amount of estrogen.


Protective effect of methylprednisolone on the heart during ischemic arrest. Although corticosteroids have been shown to stabilize lysosomal membranes and prevent release of hydrolytic enzymes, the mechanism of membrane stabilization remains obscure. The few reports regarding the use of steroids in myocardial ischemia have been conflicting. This study was undertaken to determine if a pharmacologic dose of the glucocorticoid methylprednisolone would protect the heart during ischemic cardiac arrest. A randomized double-blind study was performed in 25 dogs. Biochemical and hemodynamic parameters were assessed during and after cardiopulmonary bypass and after 30 minutes of ischemic cardiac arrest. Animals were divided into two groups. Group I served as controls and consisted of dogs injected intravenously with the vehicle of methylprednisolone 18 hours and 1 hour prior to experiment. Group II comprised dogs injected with methylprednisolone, 30 mg. per kilogram, IV, at the same time periods. Blood pH, gases, and electrolytes were measured; aortic, left atrial, and left ventricular pressures were monitored; the first derivative of the left ventricular pressure (dp/dt max.) was also determined. Arterial and coronary sinus blood samples were assayed for lactate levels and activity of the lysosomal enzyme, beta-glucuronidase. Left ventricular muscle was assayed for the nucleotides cyclic adenosine 3',5' monophosphate (AMP) and cyclic guanosine 3',5' monophosphate (GMP). Following restoration of coronary flow, mean aortic and left ventricular systolic pressures and left ventricular contractility as determined by dp/dt max. and dp/dt max./IP were depressed in both groups as expected but were significantly higher in Group II than in Group I (p less than 0.05). An increase in levels of both cyclic nucleotides occurred in each group during ischemia, but this increase in cyclic GMP was significantly greater in Group I (p less than 0.05). beta-glucuronidase activity and myocardial potassium loss as determined in coronary sinus blood were both significantly greater in Group I than in Group II (p less than 0.05). Results of this study demonstrate that pretreatment with a pharmacologic dose of methylprednisolone significantly enhances cardiac recovery after ischemia. Lysosomal membrane stability and modulation of cyclic GMP levels may be critical determinants in the mechanism of cardiac ischemia.


Uremic neuropathy. III. Controlled study of restricted protein and fluid diet and infrequent hemodialysis versus conventional hemodialysis treatment. The effect of a treatment schedule of restricted protein and fluid intake and of infrequent hemodialysis (schedule 1) as compared to a conventional hemodialysis treatment schedule (schedule 2) on the presence and severity of peripheral neuropathy has been studied in a small group of patients using a crossover design. Using the neurologic evaluation and quantitative assessment of cutaneous sensation and of nerve conduction as indices of peripheral nerve function, we have not demonstrated any worsening of peripheral nerve function from markedly curtailing the frequency of hemodialysis and modifying the diet. Because of the small size of the study, the preponderance of patients receiving the test treatment schedule first, and the possibility that nerve function slowly worsens with time, it is not possible to say with certainty that the test or the control treatment schedule might adversely affect peripheral nerve function. If such an effect is present it would appear to be small.


Steroid binding properties of estradiol receptors in human breast cancer. The incidence of a specific estradiol receptor among the Finnish breast cancer patients was investigated using methods involving dextran-coated charcoal or sucrose density gradient centrifugation techniques. An estradiol receptor was detected in 20 (71%) out of the 28 tumor specimens studied with the following binding site concentrations: 100-1000 fmoles/mg cytosol protein in 12 patients; 10-99 fmoles/mg cytosol protein in 6 patients and below 10 fmoles/mg cytosol protein in 2 patients the lowest detectable level being about 5 fmoles/mg cytosol protein. The apparent intrinsic association constant of the receptor for estradiol-17 beta ranged from 0.4-32 X 10(10) liters/mole in different breast cancer specimens. Estradiol receptor concentration did not seem to correlate well with the age of the patients or the microscopic structure of the tumor. The ligand-binding specificity of the receptor was studied with 25 different estrogen derivatives in 6 separate tumor specimens. The binding proteins in all these tumors showed very similar ligand specificities, despite differences in their histological types and estradiol-binding site concentrations. The phenolic hydroxyl group at C--3 was essential for an effective binding by the receptor, whereas certain modifications in the D-ring structure were well tolerated. As is the case with other steroid receptors, certain hydrophobic substituents seemed to increase the binding of the ligand by the breast cancer estradiol receptor. The in vitro binding affinity and the in vivo biological (estrogenic) potency of some of the steroids investigated did not correlate very well.


Changes in the composition and physico-chemical characteristics of serum lipoproteins during ethanol-induced lipaemia in alcoholic subjects. The effect upon serum lipids and lipoproteins of a large intake of ethanol (180 g in 6 hr) has been studied during the ensuing 24 hr in 21 subjects. These were all men who were habituated to heavy drinking but who were shown to be normolipaemic when maintained on a normal diet with a restricted intake of alcohol. In all the subjects studied, ingestion of this amount of ethanol induced an acute hypertriglyceridaemia which varied in intensity between individuals, with increases ranging from 26% to 377% above the basal value. On the other hand, no significant change occurred in plasma cholesterol. In addition to measurements of triglycerides and cholesterol, the plasma from all the subjects were examined by electrophoresis in agarose and polyacrylamide gels with photometric scanning of the gels. In a subgroup of ten of the subjects, the plasma lipoproteins were separated into very low density (VLDL), low-density (LDL), and high-density (HDL) fractions, which were also analyzed biochemically and by electrophoresis. Following ethanol ingestion, a mean increase of about twofold in triglyceride content, and a decrease in the cholesterol/triglyceride ratio was observed in VLDL, LDL, and HDL. The triglyceride/protein ratio decreased in VLDL and increased in LDL. On electrophoresis of the intact sera following ethanol ingestion visible particulate fat and a band which failed to permeate the gel ('chylomicron-like band') was observed in five out of ten subjects and eight out of these ten subjects showed a prominent prebetalipoprotein (pre-beta) band. This component was found to be present in the lipoprotein fraction of density greater than 1.006 Kg/1. Examination of this fraction from basal specimens revealed a pre-beta band in only two of the ten subjects and the intensity of this band increased in the corresponding fractions from the same subjects after ethanol ingestion. In an additional four subjects no pre-beta was detected in this fraction before ethanol but appeared after ethanol ingestion. No attempt has been made to characterize further the component with these characters but it is suggested that it is probably an intermediate lipoprotein (ILDL), that is, a lipoprotein with characteristics intermediate between VLDL and LDL.


Familial hyper-alpha-lipoproteinemia: studies in eighteen kindreds. A newly recognized familial hyperlipoproteinemia, familial hyper-alpha-lipoproteinemia, is described in 18 kindreds. Affected probands and relatives had distinctive elevations of alpha-lipoprotein cholesterol (C-HDL), slight elevations of total cholesterol, no elevation of LDL and VLDL cholesterol, and normal triglyceride levels. The proband and at least one additional first degree relative had distinctive elevations of C-HDL in 16 of 18 kindreds. Simple segregation analysis involving 84 offspring of 22 hyper-alpha X normal-alpha matings from these 16 kindreds revealed a ratio of hyper-alpha to normal of 37:47, a ratio not significantly different from 1:1 (chi 2(1) = 1.2), the ratio consistent with autosomal dominant transmission. Despite the suggestion of a major gene effect by this analysis, evaluation of the C-HDL distribution in kindred members failed to reveal bimodality, and familial correlation analysis revealed no parent-offspring correlation. The present data suggest that an environmental cause common to sibships is possibly important in causing the disorder. Longevity analysis demonstrated elongation of life expectancy for kindred members, and there was an apparent rarity of premature cardiac events.


Uptake of radioactive DOPA by Mycobacterium leprae in vitro. Our previous studies demonstrated that Mycobacterium leprae contains a characteristic o-diphenoloxidase which converts a variety of phenolic compounds to quinones in vitro. This enzyme was not present in any other mycobacteria tested. The results reported here deal with the uptake and binding of radioactive DOPA by M. leprae. The leprosy bacilli incubated with tritium-labelled DOPA, readily took up the substrate. The binding of DOPA by the bacilli was markedly inhibited by diethyldithiocarbamate. The organisms also bound tritiated norepinephrine. Mycobacterium phlei which does not oxidize phenolic substrates failed to bind DOPA. Cultures of melanocytes which contain o-diphenoloxidase took up tritiated DOPA. Catecholamine metabolism is known to be important in myocardial cells. Cultures of turtle-heart cells did not oxidize DOPA to quinone; however, these cells bound the labelled substrate. A cell line of fibroblasts derived from armadillo skin neither oxidized nor took up DOPA. The results indicate that, like melanocytes and turtle-heart cells, M. leprae probably possesses specific receptor sites for the binding and subsequent metabolism of phenolic substrates.


[Standardized fight against cancer in childhood (author's transl)]. The 5-year experiences of a center for pediatric oncology in a children's university hospital are reported. Currently (1-1-1975) 60 of 135 treated leukemic and 88 of 152 treated tumor patients are living. 1974, there were in the average 34 ambulatory complete blood counts per week, and in addition 263 bone marrow and 196 ambulatory lumbar punctures per year. The standardized routine of an outpatient visit is described. Advice for parents and family physicians are given. A standardized diagnostic approach for M. Hodgkin as a prototype is proposed, and a standardized therapy for Wilms' tumor. Of 24 thus treated children with Wilms' tumor 16 ("67 p.c.") survive, included 7 of 15 ("47 p.c.") with metastasized disease (observation time 21-71 months). Current treatment of 49 children with acute lymphoblastic leukemia resulted in continuous complete remission of 22 patients ("45 p.c.") (observation time 1-4 years), but 6 dead in complete remission due to infection. The need for specialized treatment centers with skilled persons is emphasized.


[About the plurality of the manifestations of cytomegalovirus infection in adult (author's transl)]. Three various cases of cytomegalovirus infection will be described. At first will be represented a double infection of cytomegalovirus and toxoplasmosis. In the second case it's about the organmanifestation of cytomegalovirus of the liver as hepatitis. At last will be discussed a severe infection of the central nervous system on the one hand in form of Guillain-Barré-syndrome, on the other hand manifestated as encephalitis.


[Experiences in the management of patients with ileo- and colostomy (author's transl)]. In the presented paper the management of stoma-patients has been discussed. Two topics had to be seen, first the psychological guidance of the patient and his reentry into society and second the management of the stoma itself. Beside of the sometimes unavoidable complications the patients are often molested by preventable inconviences as skin-irritation, soiling, and odors. New progresses have been made by the development of scentless skin pelottes and adhesive folias. With a new adhesive tissue which avoids the contact between the skin and the adhesive folia we can prevent the skin irritations previously nearly unavoidable.


[Different forms of hyperlipoproteinemia in diabetics (author's transl)]. Plasma lipoprotein patterns currently employed in attempts to identify different forms of hyperlipoproteinemia have been investigated in 113 hospitalized diabetics. For classification two methods have been compared: The first is based on lipid electrophoresis pattern in agarose gel coupled with the measurement of triglycerides and cholesterol. The second is based on plasma lipoprotein pattern obtained by separation of lipoproteins on cellulose acetate and following densitometry combined with estimation of cholesterol and beta-cholesterol and triglycerides in plasma. It could be demonstrated, that the results obtained in agarose system are not convertible to data obtained with the method for quantifying lipoproteins. By quantitative analysis only 4 p.c. of diabetics had type IIa, 4 p.c. type V, the others type IIb or IV. Graphic plots and calculated concentrations of lipoproteins gave differences in lipoprotein profiles between compensated and acidotic diabetics. In diabetes stage 1 most values are in the normal range, in stage 2 prebetalipoproteins increase and betalipoproteins decrease. In some case betalipoproteins are elevated and prebetalipoproteins diminished. In stage 3 with metabolic acidosis we observed an altered lipoprotein profile with confluence of beta- and prebeta-peak. The calculated concentration profile was also different from the others and revealed no certain quantitative information described for other electropherograms containing alpha, beta- and prebeta-bands. This phenomenon was frequently observed in patients with acute viral hepatitis and severe chronic liver disease. The pattern in diabetics is representative for patients with an excess of plasma lipids (the 2.5 fold of normal values in the mean). It is characterised as a broad beta band on the electropherogram similar to type III pattern. Presence of beta migrating lipoproteins in the ultracentrifugal supernatand fraction of d = 1006 could not be demonstrated.


Envelope and nucleocapsid antigens of Cytomegalovirus (CMV). A method for the purification and concentration of Cytomegalovirus nucleocapsids and particles complete with envelope is described. Antibody to either nucleocapsid or envelope antigens can be detected in human sera. In patients with CMV disease the antibodies to the envelope antigen could not be detected before the 3rd week after the beginning of disease whereas antibodies to the nucleocapsid were detected earlier.


Multiple regulation of nucleoside catabolizing enzymes: regulation of the deo operon by the cytR and deoR gene products. The protein and repressor nature of two regulatory gene products in E. coli has been demonstrated, employing mutants with either amber or thermosensitive mutations. The regulatory genes are the cytR and the deoR genes, both of which contribute to the regulation of the synthesis of nucleoside catabolizing enzymes. Enzyme levels in strains with concurrent mutations in both regulatory genes are considerably higher than the sum of the levels in strains with a cytR or a deoR mutation alone, indicating a certain co-operativity between the two repressor proteins. The glucose repression of enzyme levels observed in the double regulatory mutant is similar to that found in a cytR mutant, and much more pronounced than the glucose effect in a deoR mutant. A model of the promoter-operator region in the deo operon is proposed.


Information contained in the amino acid sequence of the alpha1(I)-chain of collagen and its consequences upon the formation of the triple helix, of fibrils and crosslinks. The molecule of type I collagen from skin consists of two alpha1(I)-chains and one alpha2-chain. The sequence of the entire alpha1-chain comprising 1052 residues is summarily presented and discussed. Apart from the 279 residues of alpha1(I)-CB8 whose sequence has been established for rat skin collagen, all sequences have been determined for calf skin collagen. In order to facilitate sequence analysis, the alpha1-chain was cleaved into defined fragments by cyanogen bromide or hydroxylamine or limited collagenase digestion. Most of the sequence was established by automated stepwise Edman degradation. The alpha1-chain contains two basically different types of sequences: the triple helical region of 1011 amino acid residues in which every third position is occupied by glycine and the N- and C-terminal regions not displaying this type of regularity. Both of these non-triple helical regions carry oxidizable lysine or hydroxylysine residues as functional sites for the intermolecular crosslink formation. Implications of the amino acid sequence for the stability of the triple helix and the fibril as well as for formation of crosslinks are discussed. Evaluation of the sequence in connection with electron microscopical investigations yielded the parameters of the axial arrangement of the molecules within the fibrils. Axial stagger of the molecules by a distance D = 670 angstrom = 233 amino acid residues results in maximal interaction of polar sequence regions of adjacent molecules and similarly of regions of hydrophobic residues. Ordered aggregation of molecules into fibrils is, therefore, regulated by electrostatic and electrophobic forces. Possible loci of intermolecular crosslinks between the alpha1-chains of adjacent molecules may be deduced from the dimensions of the axial aggregation of molecules.


Effect of NAD on flounder muscle glyceraldehyde 3-phosphate dehydrogenase. Flounder muscle (Pseudopleuronectes americanus) glyceraldehyde-3-phosphate dehydrogenase was characterized as to its stability towards various inactivating treatments in the presence and absence of the enzyme cofactor, NAD. Incubation of a partially purified enzyme preparation at urea concentrations greater than 2 M produced a very rapid inactivation. NAD greatly reduced the rate of inactivation at all the urea concentrations tested. Incubation of each of the three major muscle enzyme forms in 0.1 percent trypsin or chymotrypsin for forty-five minutes decreased the activity of each form by 65 percent and 55 percent, respectively. NAD (5mM) afforded complete protection to each enzyme form from proteolytic digestion by these two enzymes. Exposure of each form to 50 degrees or 20 mM ATP also led to gross inactivation which could be greatly reduced if the respective incubations were performed in the presence of 5mM NAD. NAD was also found to be required for the renaturation of the unfolded urea-denatured subunits to form the active tetramer.


Regulation of mammalian pyruvate dehydrogenase. In mammalian tissues, two types of regulation of the pyruvate dehydrogenase complex have been described: end product inhibition by acetyl CoA and NADH: and the interconversion of an inactive phosphorylated form and an active nonphosphorylated form by an ATP requiring kinase and a specific phosphatase. This article is largely concerned with the latter type of regulation of the complex in adipose tissue by insulin (and other hormones) and in heart muscle by lipid fuels. Effectors of the two interconverting enzymes include pyruvate and ADP which inhibit the kinase, acetoin which activates the kinase and Ca2+ and Mg2+ which both activate the phosphatase and inhibit the kinase. Evidence is presented that all components of the pyruvate dehydrogenase complex including the phosphatase and kinase are located within the inner mitochondrial membrane. Direct measurements of the matrix concentration of substrates and effectors is not possible by techniques presently available. This is the key problem in the identification of the mechansims involved in the alterations in pyruvate dehydrogenase activity observed in adipose tissue and muscle. A number of indirect approaches have been used and these are reviewed. Most hopeful is the recent finding in this laboratory that in both adipose tissue and heart muscle, differences in activity of pyruvate dehydrogenase in the intact tissue persist during preparation and subsequent incubation of mitochondria.


[Importance of humoral mediators for the pathogenesis and treatment of migraine (author's transl)]. The vascular mechanism of migraine attacks -- intracranial vasoconstriction during the prodromal stage and passive vasodilatation of extracranial arteries during the painful phase -- has been confirmed by modern methods. In addition, number of humoral factors -- plasmakinin, serotonin, histamine -- intervene in the chain of events that culminates in migraine headache. In addition to these three humoral mediators, the part played by tyramine in migraine of alimentary origin, the precipitating effect of oestrogen decrease in menstrual migraine and the hypothetical role of prostaglandins and prolactin are discussed. The multifactorial mechanism of migraine accounts for the very variable profiles of effective anti-migraine drugs.


[Polyneuropathy in vitamin B 12 and folic acid deficiency. Clinical and histopathological study with electron-microscopy analysis of the sural nerve]. One case each of pernicious anemia and folic acid deficiency with chronic malabsorption with disease of the cord and histologically demonstrated concomitant disease of the peripheral nerve system in the sense of a polyneuropathy are described. The histological findings of nerve obtained by biopsy show, in both cases, the loss of individual nerve fibers as an expression of a chronic axonal degeneration. The pathogenetic basis to be considered in these cases is presented.


[Secretion of insulin. Hypotheses at present under discussion (author's transl)]. Proinsulin is dissociated into insulin and C-peptide in the Golgi zone and in the beta-granules which fulfil a storage and transport function at the same time. Through an active transport process involving the microtubular and microfilament cytoplasmic system, the granules arrive at the cell wall and there they are emiocytotically excreted. At a still completely unknown spot on this secretory route, control of the secretion rate by calcium ions and cAMP-dependent phosphorylation seems possible. Calcium and cAMP as signal transmitters are again subject to the positive and negative modulating receptor-transmitted effect of adrenergic and cholinergic transmitter substances and beta-cytotropic peptide hormones. The basis of every modulation of the secretory function, however, is a secretion signal dependent on the concentration of glucose itself and glucose metabolites (only one?) on the one hand, and on the other on the energy resulting from endoxidation of glucose in the form of high energy phosphates.


[Clofibrate treatment of diabetics with hyperproteinemias (author's transl)]. The effect of a six weeks clofibrate treatment on lipid metabolism was investigated in a collective of 38 constantly maladjusted elderly diabetics with an average of 30% overweight and various forms of hyperlipoproteinemias. Clofibrate produced a significant lowering of the mean triglyceride and cholesterol levels, whereas no such effect was observed in an untreated control group. Related to the various types of hyperlipoproteinemia (IIa, IIb and IV), the group treated with clofibrate showed a rate of normalization of nearly 50%, while the rate of improvement in the untreated control group was 10%.


The transplanted kidney as a source of cytomegalovirus infection. To determine the incidence of cytomegalovirus infection in renal-transplant recipients we followed 32 prospectively for six months after operation. As judged by serologic change and virus isolation the infection rate for the entire group was 66 per cent (21 of 32 patients) - 59 per cent (13 of 22) for seronegative patients and 80 per cent (eight of 10) for seropositive patients. Of 10 seronegative patients who received kidneys from seronegative donors, only three became infected. However, of 12 seronegative patients who  received kidneys from seropositive donors, 10 became infected. Thus, there was a significant correlation between development of infection and seropositivity of the donor (P = 0.03), particularly when the recipient was seronegative (P = 0.02). Five possible and four definite recognizable clinical illnesses were associated with cytomegalovirus infection; all except two were in initially seronegative subjects who received kidneys from seropositive donors. Primary infection and disease in nonimmune recipients may be caused by cytomegalovirus transmitted by the kidneys of latently infected seropositive donors.


Cell-mediated immunity to Epstein-Barr-virus-transformed lymphoblastoid cells in acute infectious mononucleosis. Mononuclear peripheral blood leukocytes from 21 patients with infectious mononucleosis and 16 healthy controls were tested in a 51Cr-release assay for cytotoxicity against two human lymphoblastoid cell lines derived from the same donor. One line contained the Epstein-Barr virus (EBV); the other did not. Acute-phase leukocytes were significantly more cytotoxic against the EBV-infected cell line than were control leukocytes. Mean (+/- S.E.) lysis at a leukocyte-target-cell ratio of 100:1 was 10.6 +/- 1.6 per cent for patients and 3.4 +/- 0.6 per cent for controls (P less than 0.0005). Cytotoxicity correlated with the percentage of atypical lymphocytes. Cells of three patients with acute mononucleosis-like illnesses failed to show killing activity above those of normal controls. Cytotoxicity against the EBV-negative line was not significantly different for each group. The finding of cytotoxic cells in infectious-mononucleosis patients with atypical lymphocytes suggests that these cells operate in vivo to limit the proliferation of altered EBV-transformed B lymphoblasts.


Increased risk of endometrial carcinoma among users of conjugated estrogens. The possibility that the use of conjugated estrogens increases the risk of endometrial carcinoma was investigated in patients and a twofold age-matched control series from the same population. Conjugated estrogens (principally sodium estrone sulfate) use was recorded for 57 per cent of 94 patients with endometrial carcinoma, and for 15 per cent of controls. The corresponding point estimate of the (instantaneous) risk ratio was 7.6 with a one-sided 95 per cent lower confidence limit of 4.7. The risk-ratio estimate increased with duration of exposure: from 5.6 for 1 to 4.9 years exposure to 13.9 for seven or more years. The estimated proportion of cases related to conjugated estrogens, the etiologic fraction, was 50 per cent with a one-sided 95 per cent lower confidence limit of 41 per cent. These data suggest that conjugated estrogens have an etiologic role in endometrial carcinoma.


HBs-Ag-negative hepatitis in a hemodialysis unit: relation to Epstein-Barr virus. Eleven of 40 patients in a hemodialysis unit had clinical or biochemical evidence of hepatitis during a five-week period. The clinical disease was mild, being limited solely to dialysis patients. Epidemiologic investigation indicated that the incubation period was between 17 and 35 days and that 10 of 11 patients had been exposed to a single venous-pressure monitor before onset. Dried blood and evidence of blood reflux up the venous-pressure gauge suggested that cross-contamination of the blood of successive patients probably resulted in transmission of disease. No association with the hepatitis B surface antigen or anti-hepatitis B antibody was demonstrated, but 10 of the 11 patients with elevated transaminase levels had evidence of recent exposure, to Epstein-Barr virus as manifested either by Ox-cell hemolysin titers or rises in titers to viral capsid antigen.


Potentiation of hypoglycemic effect of sulfonylureas by halofenate. We investigated the possibility of a drug interaction between the antilipemic agent halofenate and sulfonylureas. Twelve young, healthy men were given 1 g of tolbutamide by mouth before and after 12 days of double-blind treatment with 1 g per day of halofenate, or placebo. There was a significant increase in serum tolbutamide at eight, 10 and 12 hours (P less than 0.01) and a significant (P less than 0.01) decrease in serum glucose at one, four and six hours after halofenate treatment, but not after placebo. In a long-term, double-blind study of halofenate or clofibrate treatment of patients with Type IV hyperlipoproteinemia, diabetic patients receiving a sulfonylurea and halofenate either required a reduction in the dose of the sulfonylurea or demonstrated significantly improved control of hyperglycemia (P less than 0.05) or both. No appreciable decrease in serum glucose levels was noted in diabetic patients receiving sulfonylurea and clofibrate. This interaction between halofenate and sulfonylureas is clinically important, especially in view of the association of hyperlipemia and diabetes.


Elevated histaminase (diamine oxidase) activity in small-cell carcinoma of the lung. To investigate the possible embryologic relation between small-cell carcinoma of the lung and medullary thyroid carcinoma, we measured plasma histaminase (an enzyme found in medullary carcinoma tissue) in 25 patients with small-cell tumors. The assays used histamine and putrescine as substrates. Thirty-two per cent of the patients by the histamine assay, and 31 per cent by the putrescine, had values greater than +2 S.D. from the mean for 63 normal persons. In contrast, among 20 patients with squamous and large-cell lung tumors, one (by the histamine assay), and two (by the putrescine) had elevated values. In four of five autopsy cases, histaminase was high in small-cell carcinoma tissue. The enzyme in plasma and in tumor behaved as classic histaminase in substrate specificity, and in response to inhibitors. The data support the proposed embryologic relation between small-cell lung carcinoma and medullary thyroid carcinoma, and further associate histaminase with some neural crest tumors.


Presence in normal human testes of a chorionic-gonadotropin-like substance distinct from human luteinizing hormone. The high rate of human chorionic gonadotropin production by testicular tumors caused us to investigate the possibility that normal human testes contain small amounts of that substance. Extracts of human testes obtained at autopsy demonstrated parallel inhibition curves to the human chorionic gonadotropin standard in a radioimmunoassay specific for the hormone. The immunoreactive material was adsorbed onto concanavalin A, a reaction characteristic of glycoproteins, and was eluted within the chorionic gonadotropin range on Sephadex G-100 column chromatography. Solubilized receptor proteins for the hormone could not be identified in the extracts. The demonstration that the normal human testes contain a glycoprotein similar or identical to human chorionic gonadotropin suggests that the fetal genome responsible for production of the hormone during pregnancy is not completely suppressed in the adult. Excessive productton of this glycoprotein may account for the high levels of human chorionic gonadotropin reported in the serum of patients with germ-cell tumors of the testes.


Evidence for viral hepatitis other than type A or type B among persons in Costa Rica. In studies of hepatitis in an endemic zone in Costa Rica, 103 patients were examined for antibodies against hepatitts A by the immune-adherence assay, for hepatitis B antigen and its antibody by radioimmunoassay and passive hemagglutination, respectively, and for antibodies against cytomegalovirus by complement fixation. Twelve cases were encountered in which both Type A and Type B hepatitis could be excluded on the basis of serologic testing. In all but one of these 12 patients, cytomegalovirus infection was also excluded. The patients had not had blood transfusions and available evidence pointed to person-to-person transmission. The illness in these patients was evidently neither hepatitis A nor hepatitis B and qualifies for consideration as the still hypothetical third type of hepatitis ("C"?).


X-ray analysis of glucagon and its relationship to receptor binding. X-ray analysis of the pancreatic hormone glucagon shows that in crystals the polypeptide adopts a mainly helical conformation, which is stabilised by hydrophobic interactions between molecules related by threefold symmetry. A model is presented in which the glucagon molecule exists in dilute solutions as an equilibrium population of conformers with little retention of conformers with little retention of structure, and in which the helical conformation is stablised by hydrophobic interactions either as an oligomer or as a complex with the receptor.


Mode of action of antirheumatic drugs on the cyclic 3',5'-AMP regulated glycosaminoglycan secretion in fibroblasts. The effect non-steroidal anti-inflammatory drugs on formation and release of glycosaminoglycans (GAG) and cyclic 3',5'-AMP levels was studied in embryonic mouse fibroblasts. The results were compared and correlated with the action of these drugs on cyclic 3',5'-AMP-dependent as well as independent protein kinase obtained from bovine diaphragm. 1. Phenylbutazone dose-dependently decreased cyclic 3',5'-AMP levels and GAG secretion both in unstimulated and PGE1 stimulated cells. 2. Indometacin decreased cyclic 3',5'-AMP levels and GAG secretion only in cells with elevated cyclic 3',5'-AMP levels after stimulation by PGE1. 3. Sodium salicylate decreased cyclic 3',5'-AMP levels in the presence and absence of PGE1. However, GAG secretion was reduced only in cells with elevated cyclic 3',5'-AMP levels, since the drug activated cyclic 3',5'-AMP-independent protein kinase activity, thus presumably precluding changes in GAG formation at low levels of cyclic 3',5'-AMP. 4. Mefenamic acid decreased cyclic 3',5'-AMP levels in cells stimulated by PGE1, whereas GAG secretion was increased both in the absence and presence of PGE1. This increase in GAG secretion was closely correlated to an enhanced cyclic 3',5'-AMP-dependent and independent protein kinase activity. The results indicate that non-steroidal anti-inflammatory drugs may exert their effects on GAG formation by interfering with cyclic 3',-5'-AMP formation or function.


Bovine adrenal cortex adenylate cyclase: properties of the particulate enzyme and effects of guanyl nucleotides. The preparation of a partially purified plasma membrane fraction from bovine adrenal cortex is described. Adenylate cyclase in this particulate preparation retained high sensitivity to ACTH and is also stimulated by 5'-guanylyl-imidodiphosphate [Gpp(NH)p]. GTP, in contrast to Gpp(NH)p, had very little intrinsic activity to stimulate activity to stimulate adenylate cyclase. GTP could however, with high affinity, inhibit the Gpp(NH)p effects on adenylate cyclase. When the concentration of creatine phosphate, a component of the ATP-regenerating system in the adenylate cyclase assay mixture, was lowered from 20 to 2 mM (at 0.1 mM ATP, 5 MM MG2+) GTP, dGTP and other nucleotides like ITP and much less UTP or CTP gained considerable intrinsic activity in the presence of ACTH to stimulate adenylate cyclase. The apparent affinities of the nucleotides for ACTH-stimulated adenylate cyclase from bovine adrenal cortex (at 2 mM creatine phosphate) were, GTP = dGTP greater than Gpp(NH)p greater than Gpp(CH2)p (5'-guanylyl-beta, gamma-methylene-diphosphonate) greater than ITP greater than UTP greater than CTP. These findings indicate that regulatory nucleotide binding sites exist for bovine adrenal cortex adenylate cyclase. Their specificity is similar to the nucleotide sites modulating angiotensin binding in bovine adrenal cortex plasma membranes (Glossmann et al., 1974a). The regulatory nucleotide binding sites for the adrenal cortex adenylate cyclase complex can also be identified under conditions where only Gpp(NH)p has high intrinsic activity (e.g. at 20 mM creatine phosphate) but other nucleotides like GTP act as antagonists. Both stimulants, ACTH and Gpp(NH)p, appear to remain firmly bound to the particulate membrane preparation, as suggested by preincubation experiments.


Chloramphenicol/thiamphenicol and cycloheximide as tools for the measurement of mitochondrial protein synthesis in vitro during organogenesis of rat embryos. 1. A test system was developed to allow the measurement of protein synthesis in vitro in mitochondria from tissues which were accessible only in small quantities. The subcellular fractions which could be isolated are not purely mitochondrial but contain other particles as well, mainly microsomal, which are also active in protein synthesis. The following differences between mitochondrial and microsomal protein synthesis in vitro were used to measure selectively the mitochondrial portion in cell fractions sedimenting between 600 and 10000 X g: selective inhibition of mitochondrial protein synthesis by chloramphenicol/thiamphenicol selective inhibition of microsomal protein synthesis by cycloheximide kinetics of amino acid incorporation a medium favoring mitochondrial protein synthesis Activity of mitochondrial protein synthesis was based on measurements of cytochrome oxidase, a mitochondrial marker enzyme. 2. The technique developed was used for the evaluation of mitochondrial protein synthesis in mammalian embryonic tissues. It may equally well be applied to other tissues available in small amounts and in cases where the isolation of highly purified mitochondrial fractions is met with difficulty. 3. Comparing the rate of 14C-phenylalanine incorporation into mitochondrial protein from rat embryos at different stages of gestation, it was found that mitochondria from 11=day-old rat embryos exhibit an approximately 30-fold higher capacity for protein synthesis than those of day 13-16. On day 12 the capacity is 6 times higher than on the following days.


Embryonic development and mitochondrial function. 1. Effects of chloramphenicol infusion on the synthesis of cytochrome oxidase and DNA in rat embryos during late organogenesis. Cytochrome oxidase, which is partially synthesized by the mitochondrion, was used  as a measure for the development of mitochondrial function in rat embryos during the late stage of organogenesis. For this  purpose the specific inhibitor of mitochondrial protein synthesis, chloramphenicol (CAP), served as a tool. Due to the rapid elimination rate of CAP from rats, a method for continuous infusion which would not cause immobilization to the animals was devised. 1. Pharmacokinetic studies proved that CAP reaches the embryo before placentation. Concentrations of CAP in the embryo are as high as they are in the maternal serum (about 20 mug/ml serum or g embryo) and thuse are sufficiently in supply for the inhibition of mitochondrial proteins synthesis, if 1000 mg/kg CAP are infused intravenously per 24 hrs. CAP is partially excluded from the embryonic compartment after the placental barrier has fully developed: whereas CAP concentration in the maternal serum remains at about 20 mug/ml, the concentration in the embryonic compartment drops to about 10 mug/g embryonic tissue during day 13 of gestation. 2. The average cytochrome oxidase activity per cell is very low (about 1 nmole O2/min X mug DNA-1) in embryonic tissue as it is in many other rapidly proliferating tissues. It is 15-60 times higher in slowly proliferating tissues, as, for example, the adult rat liver or brain (greater than 14 nmoles O2/min X mug DNA-1). 3. When the infusion technique is applied on day 12 of gestation, a sufficiently high concentration of CAP in embryonic tissue can be obtained to inhibit the synthesis of cytochrome oxidase. In constrast to tissues of an adult organism-as in the case of liver after partial hepatectomy- in embryonic tissues this limitation in the availablity of cytochrome oxidase appearently results in a critical reduction of energy production, which subsequently affects the DNA synthesis and embryonic growth. 4. The possible relevance and applicability of these experimental findings to man is discussed.


Time-course of the effect of alpha-methyl-p-tyrosine on ACTH secretion. 250 mg/kg i.p. of the methyl ester of alpha-methyl-p-tyrosine (alpha-MpT) produced a rapid increase (by 1 h) in immunoreactive plasma ACTH, whereas the hypothalamic norepinephrine (NE) concentration and plasma corticosterone (B) changed significantly only 6 h after the administration of the inhibitor. These data suggest that the norepinephrinergic inhibitory tonus of CRF-ACTH secretion depends on the availability of a rapidly utilized pool of transmitter, affected in a very short time by alpha-MpT.