Spironolactone antagonism of aldosterone action on Na+ transport and RNA metabolism in toad bladder epithelium. In earlier studies, aldosterone increased the incorporation of precursors into a class of cytoplasmic RNA with the characteristics of messenger RNA (mRNA), in toad bladder epithelium. In the present studies, this effect was analyzed further with a competitive antagonist, spironolactone (SC-9420). Paired hemibladders were labeled with 3H-uridine (30 min pulse - 140 min chase), with or without aldosterone (3.5 x 10(-8) M, 7 X 10(-8) M) in the presence or absence of SC-9420 (7 X 10(-6) M, 2.5 X 10(-5) M) at molar ratios of 200:1 to 280:1. Cytoplasmic RNA, either the total phenol-SDS extract or polyadenylated-RNA (poly(A)(+)-RNA) obtained by oligo-deoxythymidylate-cellulose (oligo(dT)-cellulose) chromatography was analyzed in linear 5 -- 20% sucrose gradients. Eight sets of experiments were completed in which the short-circuit current (scc) was monitored for 180 min and the incorporation of 3H-uridine (30 min pulse -- 150 min chase) was simultaneously determined on pools of epithelia from 5 to 10 hemibladders. The fractional change in scc correlated linearly with the fractional change in 3H-uridine of 12S cytoplasmic RNA (r=0.95, p less than 0.001). The poly(A)(+)-RNA fraction had no detectable rRNA or tRNA and gave a heterogeneous pattern, typical of mRNA, in the sucrose gradients. In the presence of exogenous aldosterone, SC-9420 inhibited the incorporation of 3H-uridine into poly(A)(+)-RNA (particularly 12S). These results support the inference that induction of mRNA mediates the action of aldosterone on Na+ transport.


Epstein-Barr virus genome studies in Burkitt's and non-Burkitt's lymphomas in Uganda. Burkitt's lymphoma (BL) has been widely investigated and has attracted attention because of the possible etiologic role of the Epstein-Barr virus (EBV). To further determine the role of EBV in the causation of this tumor, we measured EBV-specific nuclear antigen (EBNA) and EBV DNA using immunofluorescence and nucleic acid hybridization techniques, respectively. Of 34 BL biopsies, 27 tissues (79%) were EBNA-positive, whereas none of the 25 non-BL biopsy tissues were EBNA-positive. Of 15 BL tumors tested, 14 (93%) were EBV DNA-positive with a mean of 39 (range, 8-86) EBV genome equivalents per cell. Each of the 15 non-BL biopsy specimens subjected to nucleic acid hybridization had less than two virus genome equivalents per cell, although all had serologic evidence of past EBV infection. The findings further supported the possible etiologic role of EBV in African BL and negated the passenger hypothesis. The EBV genome could, therefore, be used as a separating marker between African BL and non-BL lymphomas.


Hepatitis B virus infection and primary hepatocellular carcinoma. Ninety-three patients with biopsy-proven primary hepatocellular carcinoma (PHC) from Uganda, Zambia, and the United States were examined for serologic evidence of hepatitis B virus (HBV) infection. Patients were tested for hepatitis B surface antigen (HBsAg) and its antibody (anti-HBs), antibody to the hepatitis B core antigen (anti-HBc), hepatitis B e antigen (HBeAg), and its antibody (anti-HBe). Active HBV infection, as indicated by positive tests for HBsAg (with or without anti-HBs) and anti-HBc (without anti-HBs), was present in 62% of PHC patients (58 of 93), in contrast with 10% of African controls (9 of 90), and less than 1% of most United States adult populations reported in the literature. The presence of HBeAg or anti-HBe was rare among PHC patients and controls.


A solid-phase radioimmunoassay for Epstein-Barr virus-associated membrane antigen prepared from B95-8 cell culture supernatants. Epstein-Barr virus(EBV)-associated membrane antigen (MA) was concentrated from B95-8 cell culture media by precipitation with polyethylene glycol followed by chromatography on Bio-Gel A-50m. In a RAJI cell-binding assay, MA-positive material could only be found in the void volume of the column. After ultracentrifugation all antigenic activity appeared in the pellet, which suggested that MA was present in aggregates, presumably fragments of cellular membranes and/or virus envelopes. The MA-containing preparation was photopolymerized in polyacrylamide gel. The homogenized gel was used in a solidphase radioimmunoassay with 125l-labeled IgG from an Anti-MA positive reference serum and an anti-MA negative control serum. The specificity of the reaction was confirmed in blocking tests with anti-EBV positive and negative sera. A good correlation was found between the results obtained in the radioimmunoassay and the results obtained in direct immunofluorescence tests for the detection of MA. The existence of at least two subspecificities of the MA complex could be confirmed by this radioimmunoassay.


Epidemiology of cancer of the testis in upstate New York. We collected data on the 434 individuals reported with cancer of the testis to the New York State Tumor Registry, 1960-64, from upstate New York. We compared these with the 410 members of a random sample of the upstate population interviewed from 1959 to 1962. A high risk of developing cancer of the testis was associated with professional occupations, native-born parentage, rural residence, and having been married, especially while young. These findings paralleled some other studies, as well as our earlier inquiry. Each of these factors carried a higher risk even when considered in the context of the other traits, and risk increased with an increase in the number of characteristics possessed.


Activation of Epstein-Barr virus in hybrid cells. Human-primate hybrid cell lines were established by fusion of African green monkey kidney cells (VERO) with lymphoblastoid cells from patients with infectious mononucleosis (IM)(IMK101) and from Burkitt's lymphoma culture (HR1K). Both Epstein-Barr virus (EBV)-specific antigens and EBV particle-containing cells increased in the hybrid lines (IMK1-1/VERO,HR1K/VERO). Treatment of the hybrids with 5-bromodeoxyuridine induced more antigen-positive and more virus-containing cells. EBV could be activated from IM lymphoblastoid cells by fusion of the lymphoblastoid cells with the VERO cells. The increase of viral antigens and virus particles may have been due to the cellular interaction between VERO cells and the lymphoblastoid cells or to a possible derepressor supplied by the VERO component of the hybrid. Virus derived from the HR1K cell line was replicated in the human-primate hybrid, but further investigation may be necessary to determine if it was identical to the EBV derived from the human cell line.


Ultracytochemical localization of glucose-6-phosphatase in Chang rat hepatoma in vivo and in vitro. The standard lead precipitation method was used for ultracytochemical localization of glucose-6-phosphatase (G-6-Pase) in the in vivo and in vitro forms of the Chang rat hepatoma and in the normal adult rat liver. Reaction product was visualized as very fine particulate within the cisternae of the nuclear envelope and endoplasmic reticulum. Cytochemically, the amount of the G-6-Pase reaction product in both forms of the tumor cells was obviously less than that in the normal hepatocytes. Apparently, the enzyme was not completely deleted from the hepatoma cells. The results supported some biochemical data of certain other hepatomas. The successful ultracytochemical localization of G-6-Pase in cultured hepatoma cells has not been reported previously.


Lamella-particle complexes in nuclei of owl monkey kidney cells infected with herpesvirus saimiri. A complex consisting of ribosome-like particles and a striated lamella occurring in stacked, tubular, and twisted ribbon-like configurations was observed in the nuclei of confluent monolayer cultures of primary owl monkey kidney cells infected with Herpesvirus saimiri. They were similar to the cytoplasmic cylindroid structures seen in some human leukemias and in the lymphoma of the northern pike.


Degradation of basement membrane by murine tumor cells. Tumor cells from the murine T241 fibrosarcoma, which rapidly and reproducibility produces pulmonary metastases, were tested in vitro for their ability to degrade isolated pulmonary basement membrane. Degradation of basement membrane substrate was quantified by the culture of the substrate with tumor cells and measurement of the solubilized hydroxyproline and hexose glycoprotein at neutral pH. It was found that tumor cells collected in the tumor venous drainage were associated with a significantly greater solubilization of basement membrane than were tumor cells obtained from the primary tumor mass. Tumor cells were also assayed for their ability to solubilize type I collagen purified from human dura. Venous effluent tumor cells solubilized collagen to a significantly greater level than primary tumor cells, spleen cells, or liver cells. These findings raised the possibility that metastasizing tumor cells may be a distinct tumor subpopulation with regard to invasive potential.


Heterogeneity of karyotype and growth potential in simian virus 40-transformed Chinese hamster cell clones. Five clones of Chinese hamster cells transformed with simian virus 40 (SV40) were isolated from methylcellulose and characterized as to Giemsa-banded karyotype, DNA content, saturation density, agglutination with concanavalin A, and tumorigenicity. Chromosome analysis and DNA content studies at early passage revealed that the genetic complement for all clones was predominantly near tetraploid. All cultures examined contained a proportion of hypertetraploid cells. Nonrandom chromosome changes included at least one broken No 1 chromosone in 80% or more of the cells in each clone, and fewer sex chromosomes than anticipated from the ploidy of the cells. Several abnormal marker chromosomes tended to recur. These changes were more pronounced in the cells cultured from tumors formed by three of the clones. A karyotypically stable stem line was not noted for any of the clones or tumors. The functional significance of the karyotypic heterogeneity was assessed by means of cloning efficiencies both on plastic and in methylcellulose.


Nucleotide-induced inhibition of surface sialyl transferase activity on cultured Burkitt's lymphoma cells. Sialyl transferase activity was demonstrated on the surfaces of intact, cultured lymphoblastoid cells (RAJI) derived from a Burkitt's lymphoma. Pretreatment of the cells with neuraminidase increased the labeled sialoprotein by severalfold. A nunber of nucleotides were effective in decreasing the amount of sialoprotein assembly. CMP was the most effective inhibitor. UMP, AMP, and GMP were also inhibitory, but to a lesser degree. The diphosphate derivatives were similarly inhibitory, but generally less active than their monophosphate counterparts. The cyclic nucleotides were the least effective of all nucleotides tested; cCMP and cAMP showed a small degree of activity, whereas cUMP and cGMP were without effect. These studies indicated that a number of noncyclic and cyclic nucleotides can influence the activity of the sialyl transferase system.


Concanavalin A and the production of bovine leukemia virus antigen in short-term lymphocyte cultures. The influence of the mitogen concanavalin A (Con A) on the production of bovine leukemia virus (BLV) antigen in short-term lymphocyte cultures was determined by means of a single radial immunodiffusion test. Con A did not affect viral antigen production in peripheral blood lymphocytes from 60% of both experimentally and naturally infected cattle. Antigen production was stimulated by Con A in lymphocytes from 28% of the cattle, but it was inhibited in lymphocytes from 12%. Similar results were also obtained with lymphocytes from both blood and lymph nodes from 10 cattle with lymphosarcoma and from 10 clinically normal cattle with histologically normal lymph nodes. In sheep and goats, Con A had no effect on lymphocytes from 50%, stimulated BLV production in 43%, and inhibited BLV production in 7%. These results indicated that lymphocytes should be cultured with and without Con A to identify every BLV-infected animal.


Macrophage-mediated in vitro sensitization of T-lymphocytes. I; Detection of murine leukemia virus-associated antigens. Cytotoxic effector T-lymphocytes were produced in vitro by sensitization of spleen cells on monolayers of syngeneic macrophages that had been fed with radiation leukemia virus-containing cell extracts or with supernatants of virus-producing cell cultures. The sensitized lymphocytes were cytotoxic to cell lines that expressed viral antigens. Secondary mouse embryo fibroblasts were little affected. Sensitization via macrophages appeared to be a useful system for identification of viral antigens on surfaces of various target cells, as well as for tests of the protective effect of such lymphocytes against tumor growth in vivo.


Liver and skeletal muscle mitochondrial function following burn injury. The possibility of altered mitochondrial function consequent to burn injury was investigated. Mitochondria isolated from liver or skeletal muscle of burn-injured rats (20% tbs) were compared at 3 days postburn to shams and normal controls. Mitochondrial yields were the same for all groups. ADP;O ratios were in the theoretical ranges expected and did not differ among burn, sham, and normal animals. Respiratory control ratios (RCR's) were decreased in liver mitochondria, averaging 71.7% of normal for burned animals compared to 95.8% for the sham group. The loss of respiratory control in liver mitochondria implies inefficient use of substrate chemical energy and could contribute to postburn hypermetabolism. The different response of muscle mitochondria as compared to liver suggests that alterations may be organ specific.


Solubilization of the Epstein-Barr virus-determined nuclear antigen and its characterization as a DNA-binding protein. The Eptstein-Barr virus (EBV)-determined nuclear antigen (EBNA) was solubilized from isolate nuclei of two EBV-transformed cell lines- Raji and AW-Ramos, by high-salt treatment. Its DNA-binding properties were studied by DNA-cellulose chromatography and a 51Cr release complement fixation assay. EBNA binds to both double-stranded and single-stranded calf thymus DNA, showing a higher affinity to double-stranded DNA. There was no detectable difference in the DNA binding of EBNA prepared from Raji and AW-Ramos cells.


Polyacrylamide gel electrophoretic analysis of herpes simplex virus type 1 immunoprecipitates obtained by quantitative immunoelectrophoresis in antibody-containing agarose gel. Crossed immunoelectrophoresis was used to characterize herpes simplex virus type 1 (HSV-1) antigens produced by infected HEp-2 cells. We report on a method for analyzing the polypeptide content in individual antigen-antibody precipitates eluted from the second-dimensional agarose gel. Four glycoprotein antigens of HSV-1, Ag-8, Ag-11, Ag-6, and Ag-3, were isolated and analyzed for polypeptide content. The molecular weights of the polypeptides are presented.


Inhibition of viral RNA methylation in herpes simplex virus type 1-infected cells by 5' S-isobutyl-adenosine. 5' S-isobutyl-adenosine (SIBA), a structural analogue of S-adenosylhomocysteine, reversibly blocks the multiplication of herpes simplex type 1 virus. In the presence of SIBA, viral protein synthesis is inhibited. After removing SIBA the synthesis of proteins starts rapidly again. The new polypeptides are mainly alpha proteins (Honess and Roizman, J. Virol. 14:8-19, 1974,), normally the first to be synthesized after infection. The rapid synthesis of proteins after release of inhibition seems to be directed by mRNA formed in the presence of SIBA as indicated by experiments using actinomycin D but which was undermethylated as shown by analysis of methyl groups on RNA. SIBA inhibits the methylation of mRNA and especially that of the 5' cap. Capping of mRNA thus seems to be essential for efficient translation. The analogue affected various methylations to different extents.


Isolation and comparison of murine leukemia virus-related glycoproteins from AKR and New Zealand mice. The major glycoprotein (gp70) of murine leukemia virus occurs free of virus in the serum and body fluids of certain strains of mice. These glycoproteins were isolated from New Zealand Black mouse (NZB) ascites fluid and from AKR and New Zealand White mouse (NZW) serum by immunoaffinity chromatography and were compared by immunological tests and peptide mapping. Glycoproteins gp70-NZB and gp70-NZW were indistinguishable by all criteria tested and were more closely related to gp70 from Moloney leukemia virus than was gp70-AKR.


Biological and biochemical studies of cells transformed by simian virus 40 temperature-sensitive gene A mutants and A mutant revertants. The growth properties of hamster cells transformed by wild-type Simian virus 40 (SV40), by early SV40 temperature-sensitive mutants of the A complementation group, and by spontaneous revertants of these mutants were studied. All of the tsA mutant-transformed cells were temperature sensitive in their ability to form clones in soft agar and on monolayers of normal cells except for CHLA-30L1, which was not temperature sensitive in the latter property. All cells transformed by stable revertants of well-characterized tsA mutants possessed certain growth properties in common with wild-type-transformed cells at both temperatures. Virus rescued from tsA transformants including CHLA30L1 was temperature sensitive for viral DNA replication, whereas that rescued from revertant and wild-type transformants was not thermolabile in this regard. T antigen present in crude extracts of tsA-transformed cells including CHLA30L1, grown at 33 degreeC, was temperature sensitive by in vitro immunoassay, whereas that from wild-type-transformed cells was relatively stable. T antigen from revertant transformants was more stable than the tsA protein. Partially purified T antigen from revertant-transformed cells was nearly as stable as wild-type antigen in its ability to bind DNA after heating at 44 degrees C, whereas T antigen from tsA30 mutant-transformed cells was relatively thermolabile. These results further indicate that T antigen is a product of the SV40 A gene. Significantly more T antigen was found in extracts of CHLA30L1 grown to high density at the nonpermissive temperature than in any other tsA-transformed cell similarly grown. This is consistent with the suggestion that the amount of T antigen synthesized in CHLA30L1 is large enoughto allow partial expression of the transformed phenotype at the restrictive temperature. Alternatively, the increase in T antigen concentration may be secondary to one or more genetic alterations that independently affect the transformed phenotype of these cells.


Purification of hepatitis A virus from chimpanzee stools. Hepatitis A virus antigen was purified from early acute-phase chimpanzee stools by a rapid three-step procedure using 7% polyethylene glycol precipitation, CsCl banding, and Sepharose 2B column chromatography. Electron microscopic examination of the hepatitis A virus entigen preparation revealed highly purified hepatitis A virus particles.


Inverted repetition in the chromosome of pseudorabies virus. An electron microscope examination of pseudorabies virus DNA single strands after self-annealing shows a loop of single-stranded DNA at one end of the molecule contiguous to a double-strand region. The molecule then terminates in a further single-stranded region that does not form a loop. It is suggested that the DNA contains a sequence of 13.3 x 106 daltons at one end, which is repeated internally with opposite polarity. The segment of the genome separating the repeats has a double-strand molecular weight of 5.4 x 106. The whole native DNA has a molecular weight of 90 x 106 to 95 x 106.


Strandedness of Pichinde virus RNA.

Strandedness of Pichinde virus RNA. The Pichinde virus RNA did not possess the following characteristics of eucaryotic mRNA: polyadenylic acid sequence, capped methylated structure, and ability to direct protein synthesis in vitro. Polysomal RNA extracted from cells infected with Pichinde virus reannealed with 32P-labeled virus RNA, protecting about 60% of the latter against RNase degestion. The polyadenylic acid-containing polysomal RNA also reannealed to the 32P-labeled virus RNA to approximately the same extent. These indicate that the major part of the genomic RNA of Pichinde virus is negative stranded.


Virus-secific transcription in 3T3 cells transformed by the ts-a mutant of polyoma virus. Virus-specific RNA transcription has been measured in 3T3 cells transformed by the ts-a mutant of polyoma virus by RNA-excess hybridization to the separated strands of polyoma DNA. In two cloned sublines maintained at 39 degrees C, the nonpermissive temperature for the A gene function, RNA transcripts of a large fraction of the "early" strand are detected in both nuclear and cytoplasmic RNA fractions, but no "late" strand transcription is detected. Temperature shift to 31.5 degrees C, the permissive temperature, induces viral DNA replication and virus production accompanied by late strand transcription. In two independently derived noninducible cell lines, L strand transcription is never observed, even after cultivation at the permissive temperature. A smaller fraction of the E strand is transcribed in each noninducible cell than in its inducible parent, and this difference is further characterized as a lack of transcripts of portions of HpaII restriction endonuclease fragments 2 and 6.


Analysis of intracellular feline leukemia virus proteins II. Generation of feline leukemia virus structural proteins from precursor polypeptides. The synthesis and processing of feline leukemia virus (FeLV) polypeptides were studied in a chronically infected feline thymus tumor cell line, F-422, which produces the Rickard strain of FeLV. Immune precipitation with antiserum to FeLV p30 and subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to isolate intracellular FeLV p30 and possible precursor polypeptides. SDS-PAGE of immune precipitates from cells pulse-labeled for 2.5 min with [35S]methionin revealed the presence of a 60,000-dalton precursor polypeptide (Pp60) as well as a 30,000-dalton polypeptide. When cells were grown in the presence of the proline analogue L-azetidine-2-carboxylic acid, a 70,000-dalton precursor polypeptide (Pp70) was found in addition to Pp60 after a 2.5-min pulse. The cleavage of Pp60 could be partially inhibited by the general protease inhibitor phenyl methyl sulfonyl fluoride (PMSF). This partial inhibition was found to occur only if PMSF was present during pulse-labeling. Intracellular Pp70 and Pp60 and FeLV virion p70, p30, p15, p11, and p10 were subjected to tryptic peptide analysis. The results of this tryptic peptide analysis demonstrated that intracellular Pp70 and virion p70 were identical and that both contained the tryptic peptides of FeLV p30, p15, p11, and p10. Pp60 contained the tryptic peptides of FeLV P30, P15, and P10, but lacked the tryptic peptides of P11. The results of pactamycin gene ordering experiments indicated that the small structural proteins of FeLV are ordered p11-p15-p10-p30. The data indicate that the small structural proteins of FeLV are synthesized as part of a 70,000-dalton precursor. A cleavage scheme for the generation of FeLV p70, p30, p15, p11, and p10 from precursor polypeptides is proposed.


Genetic studies of the ploidy of Moloney murine leukemia virus. An assay for Moloney murine leukemia virus was developed that made use of the production of morphologically altered foci in nonproducer mouse cells (15F) carrying murine sarcoma virus. Wild-type (wt) virus gave a ratio of titers at 39 degrees C/34degrees C = 1.05 +/- 0.45 (standard deviation;n = 20). A spontaneous, thermosensitive (ts) mutant of Moloney murine leukemia virus, ts3, defective in a late viral function, gave 39 degrees C/34degrees C = 0. A murine cell line (TB) was mixedly infected with ts3 and wt (multiplicities of infection, 7.8:4.3), cloned after infection, and shown to be infected by both viruses. At 34 degrees C it produced wt, ts, and particles of mixed parentage. The heterozygotes (hz) had ratios of assays 39 degrees C/34 degrees C = 0.06 to 0.84 (mean, 0.36). To eliminate possible interference by multiploid particles with determination of the proportions of the three types of particles, the virus produced by the mixedly infected, cloned cell line at 34 degrees C was distributed by velocity sedimentation in a sucrose gradient, and virus was picked from the lightest part of the gradient. The proportions of ts, wt, and hz were 0.27, 0.26, and 0.47. Those particles identified as hz segreated ts, wt, and hz in the proportions 0.24, 0.27, and 0.49, respectively. These values were not significantly different from those predicted from a diploid model of the genome.


Isolated trigeminal neuropathy. An unusual complication of carcinoma of the lung. The occurrence of isolated metastasis to the trigeminal nerve or ganglion form a distant, solid tumor is rare. This communication describes a patient with carcinoma of the lung; an isolated trigeminal neuropathy developed and at autopsy proved to be secondary to metastasis to the trigeminal ganglion. The appearance of a trigeminal neuropathy in a patient with an active solid tumor elsewhere in the body is most likely due to metastasis.


Control of renin secretion.

Control of renin secretion. The present study was designed to examine the interrelationship between the intrarenal vascular receptor and the sympathetic nerve, beta-adrenergic system, for renin secretion in the anesthetized dog. 1) A reduction in renal arterial pressure from a control pressure to 100 mmHg changed neither ther flow rates of all cortex zones nor renin secretion. Further reduction of renal arterial pressure to 75 mmHg resulted in a significant increase of renin secretion and a decrease of blood flow in the outer cortex. Intrarenal arterial infusion of norepinephrine at a control pressure increased a renin secretion. However, norepinephrine infusion at a reduced pressure suppressed the renin release with a recovery of the vascular resistance to the control level. These results suggest that the changes in the degree of blood flow and pressure in the renal afferent arterioles are not essential for the renin secretion,but renin secretion by the pressure reduction might be related to the autoregulatory capacity of afferent arterioles in the outer cortex. 2) At 5 min of hemorrhagic period (75 mmHg) arterial PRA elevated in control, and phenoxybenzamine and propranolol treated groups and any significant difference in responses was not observed among groups. However, at 60 min of hemorrhagic hypotensive period PRA in control and phenoxybenzamine treated groups further increased, but PRA in propranolol treated group was not alter from its 15 min value. These results indicated that the roles of vascular receptor and renal sympathetic nervous sytem in receptor and renal sympathetic nervous system in renin secretion might be separated, and that the renal sympathetic nervous system did not relate to the early response of renin release, but related to the late response. 3) Intrarenal arterial infusion of cAMP and DbcAMP resulted in a significant increase of renin release. In addition, CaC12 solution was infuesed into the renal artery and a significant rise in renal venous PRA was observed within 5 min of infusion. These data suggested that a beta-adrenergic receptor-adenyl cyclase-cAMP system was involved in the control of renin secretion, and that since the intracellular effect of cAMP was partly related to the change of intracellular Ca distribution, its change resulted in an increase in renin secretion.


Viral growth in splenic megakaryocytes of mice experimentally infected with mouse hepatitis virus, MHV-2. Electron microscopic and immunofluorescence studies were made on the spleen of mice infected intraperitoneally with mouse hepatitis virus, MHV-2. Virions and their budding process were demonstrable in reticulum cells as well as megakaryocytes at early stage of infection with a marked decrease in number of peripheral platelets, while histiocytes and macrophages were also found to have a large number of virions at later stage.


Isolation and some properties of temperature-sensitive mutants of mouse fibroblasts with respect to morphological transformation. We have isolated from a mouse fibroblast strain C3H2K several temperature-sensitive mutants that express their transformed phenotype at low (33 degress C) but not at high (38 degrees C) temperature. The mutant clones exhibited a piled up and criss-cross pattern of growth and a high saturation density at 33 degrees C, but flat density-inhibited monolayer and a low saturation density at 38 degrees C. The temperature-dependent exprestion of transformed phenotype was fully reversible upon temperature shift, and seemed to require cell division. Plating efficiencies of the clones in liquid medium were slightly higher at 33 degrees C than at 38 degrees C, but the cells were unable to form colonies in soft  agar medium.


Nasoencephalopathy of mice infected intrananasally with a mouse hepatitis virus, JHM strain. A mouse hepatitis virus, strain JHM, grown on DBT cell culture was inoculated intranasally into ICR-SLC weanling mice, and histopathological lesions were studied in relation to viral growth. In the spleen virus titer reached a peak of 10(3) PFU/0.2G 48 H after inoculation, and later it decreased gradually. No virus was detected from the liver throughout the experiment, while some early inflammatory reactions appeared in the spleen and liver without any further development. At 48 h postinoculation there existed degeneration and necrosis in the nasal mucosa and submocosa. In the brain and spinal cord active viral growth was seen at 48 h postinfection or later. In the olfactory bulb mitral cells were also affected with accumulation of glial cells and some meningitis. At 72 to 96 h postinoculation, degeneration of neurons and glial cells were remarkable in the tructus olfactorius, cortex of lobus piriformis, septa pellucidum and commissura anterior accompanying meningitis. At 120 h postinfection, pyramidal cells in the hippocumpus were also degenerated and necrotized, and nodular proliferation and collapse of glial cells, small foci of demyelination and perivascular cuffing were seen in the interbrain. At 144 h postinoculation or later, the lesions developed through the whole brain including the pons and medulla oblongata as well as spinal cord. Brain virus titers showed 10(5) PFU/0.2g at 120 h and 10(4) PFU/0.2g at 144 h postinfection. In mice surviving at 168 hr after inoculation severe demyelinating lesions were observed despite of a decreased virus titer. These findings suggest that intranasally inoculated virus might invade the olfactory bulb through the tractus olfactorius and then produce necrotizing lesions, extending later towards the posterior parts of the central nervous system.


Experimental double infection of Japanese encephalitis virus and herpes simplex virus in mouse brain. In our laboratory, 64 autopsy cases of Japanese encephalitis patients were examined by immunofluorescence. Three patients showed some evidences of double infection of Japanese encephalitis virus and herpes simplex virus. Experiments were done to see the mechanisms of double infection. In doubly infected mice, Japanese encephalitis virus antigen was localized in the herpes simplex virus infected areas in the brain. These results suggested that "blood brain barrier" was broken by herpes simplex virus infection and Japanese encephalitis virus gained access to the susceptible cells.


Production and properties of theta-toxin of Clostridium perfringens with special reference to lethal activity. theta-Toxin of Clostridium perfringens produced in a synthetic medium showed high toxicity. The mouse was killed with 5-10 hemolytic units of the toxin. Neutralization experiments showed that lethal and hemolytic activities were due to the same toxic entity. The amount of theta-toxin expressed in lethal activity reached more than 30% that of alpha-toxin in the synthetic medium SM67. Although the activities of alpha-and theta-toxins were not additive in terms of LD50, increase in the ratio of theta-toxin to alpha-toxin resulted in reduction of the survival time of the mouse injected with a lethal dosis of the mixture of the two toxins when compared with alpha-toxin alone. Unlike those produced in other media, the hemolytic and lethal activities of theta-toxin produced in the synthetic medium was not activated by reduction with thioglycollate, even after partial purification. In other respects, the toxin was not different significantly from those reported in the past. The toxic form was detected also in complex medium. It was suggested that theta-toxin may be produced as a nascent entity.


Ascending spinal tracts of the spino-bulbo-spinal reflex in cats. Twenty-six chloralosed cats were employed in order to determine spinal ascending pathways of the spino-bulbo-spinal (SBS) reflex evoked by stimulation of the sural nerve. 1. Partial spinal transection of the dorsal part of the lateral funiculus abolished the SBS reflex ipsilateral to sural nerve stimulation. 2. By recording spinal cord potentials in response to sural nerve stimulation two pathways were established in the dorsolateral funiculus as the spinal ascending tracts of the SBS reflex; one is the direct pathway to the bulbar reticular-formation (direct spino-reticular tract) and the other one (indirect spino-reticular tract) is the relayed by the lateral cervical nucleus. Direct stimulation of the dorsolateral funiculus at the lumbar level elicited the SBS reflex. 3. Short-latency unit discharges were recorded from axons of the direct spino-reticular tract by sural nerve stimulation. These axons were discharged antidromically by stimulation of the bulbar reticular formation. 4. Intracellular recordings from the neurons of the lateral cervical nucleus revealed that spike potentials, riding on EPSPs, were induced by sural nerve stimulation and antidromic firings were obtained by stimulation of the bulbar reticular formation. 5. Neurons originating the spino-reticular tract, direct and indirect, were located in the Rexed V-VII laminae in the lower lumbar segments. They were fired monosynaptically by sural nerve stimulation and antidromically by stimulating the dorsolateral funiculus of the lumbar segments. Among them, some were activated antidromically by stimulating the bulbar reticular formation.


[Relationship between the kininase- and angiotensin- converting activity under normal conditions and in experimental myocardial infarct]. In rat experiments the depressor and pressor responses to kinins and angiotensins recorded in the carotid artery were compared as to the different methods of their administration. With intravenous injection, the responses to bradykinin and kallidin were lower, and to angiotensin I--higher, than with their intra-aortic administration. The responses to angiotensin II remain identical under these conditions. The administration of all the preparations against the background of 2,3-dimercaptopropanol (unithiol) resulted in levelling of the responses irrespective of the mode of administration. In experimental myocardial infarction induced in rats by ligation of the coronary artery a reduction of the kinin-destructing and an increase of the angiotensin-converting activity was noted in the early postinfarction period. The response of the peripheral vessels to angiotensin, bradykinin and noradrenaline is decreased, while that to isoproterenol remain unchanged. The obtained results support the concept of the existence of the kininase and angiotensin-converting activity connected with a common biochemical factor of pulmonary circulation. Changes in the proportion of these functions under normal conditions and in cases of pathological states are interpreted as a particular regulation mechanism of the vascular tone and arterial pressure.


[Chiasmatic syndromes-diagnostic possibilities with computerized tomography (CT)]. The normal suprasellar subarachnoid space (so-called "suprasellar hexagon") and its content are fairly well demonstrated by CT. Abnormal appearance (obstruction, stenosis, asymmetry) of this space is always suspect of a suprasellar space occupying lesion. Enhancing (intravenous application of iodine containing contrast medium) helps to delineate the exact tumor extension. The point of origin--important for deciding on the nature of a lesion--is often difficult to determine. The CT-pattern itself is not specific for a certain kind of tumor. We have observed a typical chiasmatic syndrome with all meningiomas of the tuberculum sellae, with 75% of the chromophobe adenomas and with 50% of the craniopharyngiomas. Absence of this syndrome was due to high position of the tumor (craniopharyngiomas) or to parasellar extension of the lesion (chromophobe adenomas). Diagnosis of recurrent tumor is difficult; it must be based on the post-operative CT-examination. Chiasmatic syndromes not due to the pressure effect of a space occupying lesion (i.e. chiasmatic syndromes of vascular, toxic or degenerative origin) can not be assessed directly by CT.


[Endocrinology of the chiasma syndromes (author's transl)]. A wealth of new knowledge about the function of the hypothalamo-hypophyseal system has been accumulated over the past ten years and progress keeps going at a fast pace. Several factors have contributed to this explosive growth: Pituitary hormones can now be measured by radio-immunoassay in body fluids, hypothalamic stimulating and inhibiting factors have been identified and are available for clinical studies and dynamic tests have been developed for detailed investigations of minor disturbances in the pituitary control system. A tremendous gain in accuracy of clinical diagnosis and endocrine follow-up is one of the consequences. Furthermore, new clinical pictures have emerged such as several variants of the hyperprolactinemia-hypogonadism syndrome or the concept of pituitary microadenoma. At the same time, neuropharmacology is also developing at a tremendous pace and exceedingly interesting perspectives linking neurotransmission with endocrinology and with various diseases of the brain are emerging. One highlight is the discovery of bromoergocryptine as a dopaminergic agonist, active in suppressing prolactin secretion as well as in treating Parkinsonism. In a second part of this presentation 5 cases with pituitary disease are presented. They were selected to cover part of the wide range of clinical signs and symptoms produced by tumours of the pituitary region. There is indeed no good relationship between ophthalmo-neurologic symptoms, destruction of the bony sella and endocrine disturbances.


[Functional histopathology of pituitary neoplasms (author's transl)]. The generally accepted classification of pituitary adenomas into eosinophilic, basophilic, mixed, and chromophobe types has not been very useful since no clear correlation exists between the staining character of the tumor cells and the clinical syndrome produced. The ultrastructural examination of different adenoma types with clinically manifest endocrine activity (acromegaly, amenorrhea-galactorrhea-syndrome, Cushing's disease) shows that the type of hormone secreted can only be determined in few cases. The ultrastructure is more representative of the activity of the secretory process than of the type of the product. Histoimmunological methods achieve the specific identification of the various types of adenomas with endocrine symptomatology. The so-called "chromophobe" adenomas, which only manifest signs of a space occupying lesion with pituitary insufficiency of varying degree and compression of the visual pathways, represent a mixed group. The majority of the cases (about 60%) shows increased prolactin secretion with amenorrhea or loss of potency, but without galactorrhea or gynecomastia. A small group of cases may either produce normal hormones (growth hormone or prolactin) at a very low rate, or secrete hormone fragments or abnormal, as yet undeterminable substances. The oncocytomas, which stain with eosin in light microscopy, seem to suffer from a defective metabolism and therefore may have lost the ability of hormone production.


Adverse effects of mouse hepatitis virus on ascites myeloma passage in the BALB/eJ mouse. During experimental serial passage of ascites myelomas through BALB/cJ mice, unexpected illness and premature deaths occurred. Postmortem examination of affected mice revealed focal or diffuse discolored depressed areas in the liver and, in some cases, splenomegaly. Histopathologic findings consisted of focal to diffuse areas of necrosis with minimal leukocytic infiltration. Aerobic and anaerobic bacterial cultures of livers and spleens from affected mice were negative. Mouse hepatitis virus (MHV) was isolated from livers of clinically ill mice and from the ascites myeloma lines. An MHV contaminated ascites myeloma line, when passed into nude (nu/nu) mice, killed the animals in 6 days; the virus was isolated from livers of inoculated mice. Attempts to determine the source of the infection were unsuccessful. Serologic survey of newly acquired mice indicated no evidence of antibodies to MHV while mice in holding rooms had titers that ranged from 1:10 to 1:40. Two solid myeloma lines (being maintained by subcutaneous passage) were negative for MHV when tested by virus isolation techniques, and nine lines were negative to 11 murine viruses when tested by mouse antibody production assay. Attempts to demonstrate Eperythrozoon coccoides in control BALB/cJ mice were unsuccessful. Because of the outbreak, changes were made in animal handling procedures. A colony of BALB/cAn mice negative to MHV antibodies was established to provide animals for experimental passage of tumors, and animals in both the breeding and transfer room were placed under filter tops. The results were encouraging. In the four newly established tumor lines, one having been passed 46 times, no illness or unexplained deaths were observed.


Radiological evidence of lymphatic drainage of bone marrow cavity in long bones. Injection of Lipiodol UF into the distal epiphyses of tibia and femur in the dog was followed within several minutes by visualization of lymphatics draining the bone. Some of the radioopaque material was retained in the bone marrow cavity and tiny lymph vessels originating in that area could be seen.  Contrast medium filled also the popliteal lymph node. The problem of migration of bone marrow lymphocytes through the lymphatics to the regional lymph nodes is discussed.


The role of the adrenal cortex in human essential hypertension: keynote address. The data presented establish that in early or mild essential hypertension there is a state of inappropriate hypermineralocorticoid activity represented by the sum of aldosterone and 18-hydroxy-11-deoxycorticosterone concentrations in the plasma. This disturbance, associated with a "normal" or excessive salt intake, would produce the arteriolar cationic changes in sodium, potassium, calcium, or magnesium leading to hypersensitivity or hyperresponsiveness of the arteriolar actomyosin to normal levels of circulating norepinephrine or angiotensin. The nature of the cationic changes in the arteriolar cells responsible for the increased tonicity of the arteriolar actomyosin, which is the fundamental cause of essential hypertension, remains to be elucidated.


Hypothesis about the mechanism of protein folding. A three-step mechanism of protein folding, proposed in our previous paper, is applied here to postulate the nature of the intermediates in the folding of rubredoxin, ferricytochrome c, and lysozyme. Contact maps are calculated for these three proteins, and it is shown that they contain much information (such as the polarity of residues in contact regions) about the structure of the native protein. Elementary processes are described for the formation of contact regions. Based on these concepts, details of the pathways of folding these three proteins from the unfolded to the native structure are postulated, focusing on the formation of ordered backbone structures (such as alpha-helical, extended, and chain-reversal conformations) in step A of the three-step mechanism and on the formation of contact regions in response to medium-and long-range interactions in steps B and C. It was found that chain reversals can often play an important role in forming contact regions in step A (short-range interactions) and in step B (medium-range interactions) but not in step C.


Use of the telephone for short-term follow-up. We have instituted a telephone appointment system (TAS) to follow patients with well-defined relatively minor problems. The patient is asked to call a special phone number at a specified date and time to obtain test results or information. A senior physician reviews each chart beforehand and indicates in lay terms, the information to be given to the patient by the clerk. Initial contact was made with 235 (58%) of the 404 scheduled telephone appointments reviewed during a four-week period. Physician data review time averaged one minute per chart and two full-time clerks were required. Although no improvement in patient compliance was found, significant savings of patient, physician, and nursing time are obvious. Additional advantages include decreased total patient load and the opportunity for senior physicians to assess the quality of care and provide feedback to the emergency department house officers.


Metabolism of cholesteryl esters of very low density lipoproteins in the guinea pig. Very low density lipoproteins from guinea pig plasma, endogenously labeled with 3H in both the esterified and free cholesterol moieties, were obtained from serum collected 20 hr after the intravenous injection of 3H-cholesterol into donor animals. When these lipoproteins were injected into recipient guinea pigs, the esterified 3H-cholesterol was rapidly cleared from the plasma; 24% was in the liver in 5 min and 54% in 15 min. A smaller fraction of the esterified cholesterol appeared in other plasma lipoprotein fractions, with 3H in the low density lipoproteins reaching a peak of 9%-18% of the injected esterified 3H-cholesterol between 30 and 60 min after the injection. The results indicate that most of the esterified cholesterol in very low density lipoproteins of guinea pig plasma is removed directly by the liver and a minor fraction is transferred to low density lipoproteins. The pattern of labeling of cholesteryl esters of high density lipoproteins in these experiments suggests that their low concentration in the guinea pig is accompanied by a rapid turnover rate.


Studies on the minimum reproducible unit of staphylococcal L-forms. The minimum size of a reproducible unit of staphylococcal L-forms was determined by filtration and electron microscopic methods. Ultrathin sections of an induced strain of Staphylococcal L-forms (STA-EMT-1) in liquid medium revealed several types of structures, all of which were bound by a single membrane and most of which possessed ribosome-like granules. Many of the small granules were less than 0.3 micron and were attached to the membrane of the large bodies. Using a serial filtration method, it was observed that viable L-forms were still detected in 0.22 micron filtrate, but the viable cell count of L-forms decreased in number with the decrease in pore size of membrane filters. A fractionation technique, using L-forms filtered through a membrane filter with a 0.45 micron pore size, revealed that there were three classes of small bodies but only the first class with ribosome-like granules over approximately 0.2 micron in diameter seems to be able to reproduce.


Screening for breast cancer by mammography: an Australian experience. Four years' experience of screening well women for breast cancer by mammography is described. Twenty-six cancers were detected in 11,927 women (2-2 per 1,000), of which 20 were unsuspected by the patient and 12 were impalpable. Of the 26 cancers detected, 18 had no histological evidence of tumour in axillary lymph nodes, and 10 were impalpable and had unaffected axillary nodes. Four patients whose mammograms did not suggest malignant disease were subsequently found to have cancer, within 12 months.


Viral infections of the autonomic nervous system and its target organs: pathogenetic mechanisms. The possibility that viruses can infect the peripheral autonomic nervous system (ANS) and cause a variety of acute, chronic or recurrent diseases, including such disorders as peptic ulcer, is proposed. By altering the functional activity of autonomic neurons or by seeding the target organs of these neurons with virus, infection of the ANS may produce a broad spectrum of clinical manifestations. Whether these manifestations are anatomically circumscribed or widespread will depend upon the route by which autonomic infection is acquired as well as the character of interaction between the virus and the infected autonomic neuron. In addition, it is proposed that autonomic ganglia may serve as reservoirs of latent viruses, insuring their preservation and intermittent transmission in the human community.


Detection of [3H]cyclic AMP in the transcription complex of rat liver mitochondria. By equilibrium ultracentrifugation and electrophoresis in agarose gel data were obtained on the binding of [3H]cyclic adenosine monophosphate ([3H]cAMP) to the transcriptional complex of mitochondria with the density 1.825 g/ml in CsCl ethydium bromide. Treatment of the complex with RNase, DNase and pronase change the density of [3H]cAMP bound material; rifampicin prevents detection of [3H]cAMP. Study of inner membrane lysates showed that [3H]cAMP is bound to structures active in RNA and protein synthesis in mitochondria. The conclusion is made that cAMP is involved in formation of the transcriptional complex much as it is involved in initiation of transcription in bacterial operons.


Studies on the synthesis and structure of mitochondrial DNA in cells infected by Rous sarcoma viruses and on the occurrence of intramitochondrial virus-like particles in certain RSV-induced tumor cells. 1. The synthesis of mitochondrial DNA in CEF in vivo at 3,4 and 6 days after infection with RSV (Schmidt-Ruppin, subgroup A) was progressively stimulated 2 to 4-fold as compared with that in uninfected CEF cells grown in parallel. 2. The stimulation of mtDNA synthesis in vivo upon transformation was found to be temperature dependent when the thermosensitive mutant of RSV, T5, was used to infect the cells. 3. In contrast to mtDNA synthesis, nuclear DNA synthesis did not differ in transformed and uninfected cells, nor did it change significantly upon temperature shifts. 4. MtDNA (monomeric and catenated dimeric forms) in transformed and uninfected CEF replicate by displacement synthesis. Various replication intermediates are described. 5. The restriction endonuclease EcoRI cleaves closed circular mtDNA from CEF at one specific site. 6. Heteroduplex molecules formed between nicked circular and/or EcoRI cleaved mt DNA molecules from uninfected and transformed CEF revealed, with a few exceptions, no detectable base sequence heterogeneity in at least 98% of cases. 7. Intramitochondrial virus like particles (IMV) are described in hamster tumor cells. The evidence suggests both engulfment of cytoplasmic particles by mitochondria and the presence of intramitochondrial incomplete forms of particles. Bromodeoxyuridine was found to enhance the frequency of IMV.


Biosynthesis of subviral oncogenic particles (virosomes) in mitochondria of Rous sarcoma and Rauscher murine leukemia cells. Experimental evidence for the presence and biosynthesis of subviral, leukemogenic particles in the isolated mitochondria of spleen cells of mice infected with Rauscher murine leukemia (RML) virus is presented. These subviral particles sediment at a density of 1.27-1.29 g/ml and induce splenomegaly and RML three weeks after i.v. or i.p. administration to white mice. Virosomes have been labelled with [32P]phosphate in the isolated mitochondria from RML spleen cells and high molecular weight (70S) [32P]RNA has been isolated from these subviral, leukemogenic particles. Rauscher virus group specific antigens were detected by immunodiffusion in the inner membrane and matrix fraction of the mitochondria of RML spleen cells. These results together with our earlier findings strongly suggest that mitochondria of the transformed cells participate in the biosynthesis of RNA tumor viruses. Possible mechanism of the penetration of viral genetic information of RNA tumor viruses into mitochondria of tumor cells in vivo is discussed.


Integration and regulation of mitochondrial assembly in yeast. The interactions between the mitochondrial and nucleocytoplasmic systems required for mitochondriogenesis have been investigated at several different levels. Those involved in the formation of functional enzyme complexes have been studied using cytochrome oxidase: this multimeric (2 X 7 and 2 X 6 subunits for enzymes from yeast and beef heart respectively) has been resolved, and the mitochondrial contribution has been shown to be dispensible for catalytic function proper. Using novel mutants, with a mitochondrial mode of inheritance, a mitochondrial gene product localized in the oligomycin-sensitive ATPase has been implicated in the assembly not only of this complex, but of cytochrome oxidase as well. Interactions required for the genetic competence of the mitochondrial system have become apparent as a result of studies in the mechanism of action of the highly effective mitochondrial mutagen ethidium bromide. This agent first becomes covalently inserted into mitochondrial DNA and, after its excision, eventually results in extensive degradation of the macromolecule. The excision reaction has now been shown to be performed by a complex between the oligomycin-sensitive ATPase and a DNA-binding protein presumably involved in recognizing the damage. On the level of replication and expression of the mitochondrial genome studies using thermolabile mutants have demonstrated that these processes appear independent of the replication of nuclear DNA but not of its expression.


Structure of cytochrome c oxidase from baker's yeast - a progress report. Preparation of four subunits for amino acid sequence determination and attempts to localize the cytochrome c binding site. Cytochrome c oxidase from the inner membrane of yeast mitochondria consists of seven nonidentical protein subunits, three being synthesized on mitochondrial ribosomes (molecular weights I: 43 K, II: 34 K, and III: 24 K) and four being made on cytoplasmic ribosomes (molecular weights IV: 14 K, V: 12 K, VI: 12 K, and VII: 4.5 K). In the present study all four cytoplasmically synthesized subunits of the enzyme were isolated on a large scale using ion exchange chromatography and gel filtration. Their amino acid composition as well as their amino- and carbosy-terminal amino acid residues have been determined. Sequence determinations of subunits IV and VI are already in an advanced state. The sequence of subunit VI is characterized by a large amino-terminal stretch dominated by charged amino acid residues followed by a cluster of hydrophobic amino acids. The binding site of yeast cytochrome oxidase for cytochrome c was studied by chemical crosslinking experiments. The formation of a disulfide bridge between the two proteins was observed by using cytochrome c from yeast modified with 5-thionitrobenzoate at the cysteinyl residue in position 107. Alternatively, a disulfide between yeast cytochrome c and the oxidase could be formed directly by oxidation with copper phenanthroline. Gel electrophoresis of the crosslinked complexes in sodium dodecyl sulfate revealed a new protein band with an apparent molecular weight of 38 K. This new band appears to be derived from cytochrome c and from subunit III of cytochrome oxidase.


Studies of the UV-sensitivity of virus-specific RNA synthesis in cells infected or transformed by adenovirus 5 and SV 40. The effect of UV-irradiation on host and virus-specific RNA synthesis in cells infected or transformed by tumor viruses (Ad 5 and SV 40) was studied. It was found that the synthesis of host and Ad 5 RNA in infected KB cells was almost euqally inhibited upon UV-irradiation; the transcription of the EcoRI produced fragements--fragment B was inhibited to a greater extent than fragment A, suggesting that the transcription of the whole Ad 5 genome starts from the left side. The transcription of viral sequences in transformed cells, on the other hand, was more resistant to UV-irradiation than the transcription of the host ones. The results obtained suggest that the small size of the virus transcriptions, or their location at the beginning of large host transcriptions, may be responsible for the observed data.


[A new diet for inducing early experimental atherosclerosis in rabbits]. Authors by a new combined diet consisting of cholesterol vitamin D3 and sour cream produced clinico-biochemical changes characteristic for atherosclerosis in a very short period in rabbits. This diet caused severe lipoproteinaemia, hypercholesterinaemia and microscopis and macroscopic changes in the aortic wall. Authors assume, that this diet is suitable for producin atherosclerosis in a short period.


Immunoblastic lymphadenopathy with centrofollicular PAS-positive inclusions. The authors describe the clinical and histopathologic picture of immunoblastic lymphadenomegaly in a 66-year-old patient with allergic antecedents. The condition of the patient ameliorated following cortisone and antibiotic therapy, but adenomegalia relapsed after several months. PAS-positive granular inclusions were found in the macrophage cells in the germinative centres of the follicles, showing the aspect of a blockade with nuclear significance.


Congenital cytomegalovirus infection.

Congenital cytomegalovirus infection. The overall prevalence of congenital cytomegalovirus infection among the offspring of a highly immune young female population was 2.4 per cent (23 of 939). To ascertain whether the presence of anticytomegalovirus antibodies protects the developing fetus, we examined the offspring of 239 prospectively studied women. Despite substantial levels of preconceptional antibodies, intrauterine cytomegalovirus infection occured in seven of 208 (3.4 per cent) seroimmune women. Three neonates with congenital infection were born to 31 initially seronegative women. All the congenitally infected infants had subclinical involvement. Maternal humoral immunity may not protect the fetus against congenital cytomegalovirus infection. Neutralization kinetics and restriction enzyme analysis with endonucleases (EcoR-1 and HinD 111) demonstrated antigenic and genetic homology between viral strains isolated from two siblings consecutively infected in utero, indicating that repeat maternal infection with the same virus is transmissible to sequential products of conception.


Hypoglycemia of infancy and nesidioblastosis. Studies with somatostatin. We treated a two-month-old infant with servere intractable hypoglycemia and nesidioblastosis with continuous glucose infusions (0.75 g per kilogram per hour) via a central venous catheter. Preprandial glucose levels on this regimen were 37+/-2 mg per deciliter (+/-S.E.M.). Basal serum insulin levels were within normal fasting levels for this age group but inappropriately elevated for the blood glucose levels. The beta cells were exquisitely sensitive to infusions of synthetic cyclic somatostatin, with a dose-dependent rise in blood glucose and concomitant suppression of serum insulin levels. There was only minimal suppression of plasma glucagon levels. Single subcutaneous injections of 50 microng of protamine zinc somatostatin raised preprandial blood glucose levels to 83+/-3 mg per deciliter for four to five days although preprandial hormone levels were unchanged. These findings indicate that hypoglycemia of infancy is a hyperinsulin state with abnormal basal regulation of insulin secretion.


Cellular immunity and herpesvirus infections in cardiac-transplant patients. We observed severe infection with herpes simplex virus in cardiac-transplant patients despite their high serum antibody levels to this virus. Therefore, we sought to correlate clinical susceptibility to two herpesvirus (simplex and zoster) infections with specific cellular immunity, assessed by the transformation and interferon responses of peripheral blood mononuclear cells to heat-inactivated antigens. Transformation and interferon response to herps simplex virus was maximally depressed immediately after transplantation, the time when severe and prolonged infection with herps simplex virus occurred. Six months to six years after transplantation, both clinical susceptibility and cellular immunity to herpes simplex virus were normal. Herpes zoster infections were more frequent than normal at all times after cardiac transplantation; depressed or absent cellular responses to the varicella zoster virus paralleled that susceptibility. In these patients the risk of severe herpesvirus infections correlated with depressed cellular immune responses to the specific viral agent involved.


[Determination of vitamin D3 in premixes and mixed feeds]. The authors developed a two-dimensional thin-layer chromatographic method for the qualitative and quantitative determination of vitamin D3 in premixes and mixed feeds. The procedure permits to determine I.U. = 0.128 micrograms of vitamin D3. The method is illustrated by the description of the analyses of 3 premixes and 5 mixed feeds.


[Antistatic finishing of plastics from food hygiene and toxicological viewpoints]. The authors investigated experimentally the migration behaviour of formed plastics with antistatic finish. The degree of molecular coating on the surface and the rate of diffusion of the antistatics used (ethoxylated amines and alkyl sulphonate) were calculated from the results of washing-off and continuous extraction experiments. The findings are indicative of a good depot effect, i.e. the antistatic action is steadily regenerated by secondary migration. Certain agents such as  polyethylene glycol enhance diffusion whereby the values for the migration in normal polystyrene are increased; but this increase is acceptable from the viewpoint of safety. In impact polystyrene, the limiting value for total migration is largely exceeded so that considerable contamination may be expected when it comes in contact with foods. From the viewpoint of food-toxicology, there are concerns about the high specific migration values of alkyl sulphonate. For this reason, its use in the food sector should be limited to such cases where direct contact with foods is excluded.


Immune surveillance--a powerful mechanism with a limited range. There is excellent evidence for the belief that immune surveillance mechanisms prevent the outgrowth of potentially neoplastic cells induced by horizontally transmitted, ubiquitous, potentially oncogenic viruses, indigenous to natural populations. Polyoma virus in mice, Marek's disease in the chicken, Herpesvirus saimiri in the squirrel monkey, and the Epstein-Barr virus in man are examples. There is much less evidence for immune surveillance against chemically induced tumors. It is argued that surveillance against the virus-induced tumors may have evolved by the selective fixation of appropriate immune responsiveness (IR) genes. It is important to distinguish between the breakdown of an existing surveillance mechanism, e.g., by immunosuppresion, and the lack of immune recognition, due to the deficiency of the IR gene equipment. Presently available in vitro lymphocytotoxicity tests are not yet developed to the point where they can reliably distinguish between these alternatives.


Spontaneous tumor regression: possible relationship to in vitro parameters of tumor immunity. Evidence is discussed from three animal systems (Shope papillomas in rabbits, Moloney sarcomas in mice, Schmidt-Ruppin-Rous sarcomas in Japanese quail) that immune reactions can be important in spontaneous tumor regression. In vitro studies performed in these systems indicate that blocking serum factors can thwart cell-mediated immune responses and that "unblocking" antibodies are often found in animals whose tumors have regressed. To what extent spontaneous regressions of human neoplasms are due to immunologic mechanisms is unknown; in 2 patients who had undergone spontaneous tumor regression, tumor cell cytotoxic lymphocytes and unblocking serum factors were detected. Better animal models and better knowledge about the mechanisms of tumor immune reactions are needed before tumor regression can be more successfully induced by immunologic manipulation (to what extent that will even be feasible in man is unknown).


Mechanisms of escape from immune surveillance. The escape from immune rejection is, like the development of drug resistance in microorganisms, a multi-pathway process. Selection favors the phenotype, i.e., the cell that can grow in spite of the immune rejection process, not the underlying mechanism. Experimental evidence documents the roles of host immunosuppression (genetic or environmental), immunoresistance at the cellular level, "sneaking through" (i.e., growth of a tumor to irreversible size prior to the mobilization of an appropriate immune response), lack of antigenic recognition, and blocking enhancement type reactions.


A pathologist looks at spontaneous regression of cancer. We  are all subjected to varying amounts of mutating, potential cancer-inducing events, which are cumulative. In most instances, the accumulations are repaired or are lethal. If a viable clone survives, it must proliferate in order to become manifest and eventually to overpower the host's normal regulatory and defense mechanisms. The proliferation may be incited and sustained by cocarcinogens, hormone excess, chronic infections such as schistosomiasis, and in the case of lymphoid cells, by immune incompetence. On the other side are the protective reparing enzymes, which serve to cut out the damaged segments of DNA and repair them as fast as they can. The normal homeostatic mechanisms tend to keep cell proliferation and activity within the beneficial bounds of need. The controlling forces that cause cells to stop reproducing themselves and differentiate to perform a useful function also tend to slow or even stop the useless proliferation if all the cells can undergo maturation to a postmitotic state. There are also the protective mechanisms of immunity: Sensitized B lymphocytes tag the antigenic sites of tumor cells and activate the T-cell macrophages to destroy the tagged cells.


Spontaneous regression of bronchogenic carcinoma. Two cases of spontaneous regression of bronchogenic carcinoma were discovered in the literature, and the pertinent findings are described. Six equivocal cases of spontaneous regression are also presented. The reasons for the scarcity of spontaneous regression of bronchogenic carcinoma are discussed; no apparent reasons for spontaneous regression in these two cases could be determined.


Spontaneous regression in choriocarcinoma and related gestational trophobalstic neoplasms. Gestational trophoblastic neoplasia represents a biologic spectrum of tumors progressing from the hydatid mole, to invasive mole, and to choriocarcinoma. This progression is reflected in increasing degrees of aneuploidy in the respective lesions. Just as there is a natural tendency for the rejection of the trophoblast of a normal pregnancy culminating either in parturition or in spontaneous abortion, rejection of tumors of trophoblast occurs at any point in the progression of the disease spectrum. The unusual effectiveness of chemotherapy in trophoblastic disease may be related to this natural tendency to rejection. This tendency, in turn, is thought to derive from the genetic disparity between the maternal host and the tumor tissue of fetal origin, since the fetus possesses both maternal and fetal antigens.


Photochemical attachment of cyclic AMP binding protein(s) to the nuclear genome. The nature of the cyclic AMP-receptor-nucleus interactions was examined by a novel combination of two photoreactions. A photosensitive derivative of cyclic AMP, N6-butyryl cyclic AMP, was covalently attached to its cytoplasmic receptod by photo-affinity labelling and this receptor complex was photo-crosslinked by the DNA in the rat liver nuclei. The photolytic reactions seemed to be specific since stable links were formed only when substantial noncovalent binding occured.


Sequence and structure of D-glyceraldehyde 3-phosphate dehydrogenase from Bacillus stearothermophilus. The glyceraldehyde 3-phosphate dehydogenase holoenzyme of Bacillus stearothermophilus possesses precise 222 symmetry: in this respect it differs from the reported structure of the lobster muscle enzyme. Pairs of active sites are linked through a flexible polypeptide loop which probably mediates the structural changes giving rise to cooperative effects. Three additional salt bridges made by each subunit to others would make a major contribution to thermostability of the tetramer.


Polymorphism of the major envelope glycoprotein (gp70) of murine C-type viruses: virion associated and differentiation antigens encoded by a multi-gene family. Structural comparison of the major envelope glycoproteins (gp70) from 35 different murine type C viruses and free gp70 expressed at various anatomical sites in the mouse showed that the gp70s are polymorphic products of a large multi-gene family encoding viral and differentiation antigens. Different proviruses are expressed in cells following distinct pathways of differentiation. When the various gp70s are grouped according to primary structure they fall naturally into viral host range classes, confirming the suspicion that C-type viral tropism is largely determined by the nature of the gp70 product expressed.


Dopamine"autoreceptors": pharmacological characterization by microiontophoretic single cell recording studies. The effects on the firing of single dopamine (DA) neurons in the substantia nigra (and adjacent ventral tegmental area) of a representative group of catecholamine agonists and antagonists were studied in rats using single cell recording and microiontophoretic techniques. Microiontophoretic application of DA or the DA agonist apomorphine depressed the firing of these cells; the DA antagonist trifluoperazine blocked this effect. However, the alpha-agonist clonidine had no depressant effect and the beta-agonist isoproteronol had only a weak depressant action on DA neurons. Furthermore, the alpha-antagonist piperoxane and the beta-antagonist sotolol were completely ineffective in blocking the depressant effects of DA. These results show that DA-sensitive receptors on the soma of DA neurons are pharmacologically distinct from alpha or beta adrenoreceptors. Because of their location and selective responsiveness to DA agonists, the catecholamine receptors on the soma of DA neurons appear best classified as DA "autoreceptors".


Possible feed-back inhibition of noradrenaline release by purine compounds. The contractile responses to transmural stimulation of, and the overflow of tritium from the rat portal vein prelabelled with 3H-noradrenaline were studied. The contractile responses of the rat portal vein were sustained throughout the period of stimulation. The tension developed did not decline when two consecutive periods of stimulation were compared. In contrast, the tritium overflow decreased during the second period of stimulation. Preincubation with 3 micronM phenoxybenzamine during 30 min increased 3-fold the tritium overflow during stimulation. Phentolamine and phenoxybenzamine were nearly equipotent in reducing the vascular response to stimulation. In contrast, phentolamine was less potent than phenoxybenzamine in increasing the 3H-noradrenaline overflow elicited by stimulation. The results obtained with phentolamine are interpreted in terms of a different potency of phentolamine to produce blockade of prejunctional and postjunctional alpha-adrenoceptors in the rat portal vein. ATP inhibited by 70% the tritium overflow induced by stimulation. The potency of ATP in inhibiting the overflow increased when the prejunction alpha-adrenoceptors were blocked. The purine compounds ATP, ADP, AMP and adenosine were roughly equipotent in inhibiting stimulation-induced tritium overflow. The tritium released by stimulation decreased when uptake and metabolism of adenosine were inhibited. Under physiological conditions, a prejunctional purinergic inhibition of noradrenaline release might be involved in an endogenously mediated negative feed-back regulatory mechanism. It is possible that the purinergic inhibition of the noradrenaline liberation elicited by stimulation plays a physiological role in tissues with both purinergic and adrenergic innervation.


The effect of metacaine (MS-222) on the activity of the efferent and afferent nerves in the teleost lateral-line system. 1. Free-swimming fish (Tilapia leucosticta and Rutilus rutilus) were used to determine threshold concentrations required for general anesthesia with metacaine (MS-222). The criterion for anesthesia was reached at a concentration of 1:1500 (w/w) with both fish, although the symptoms were somewhat more pronounced in Rutilus. 2. Recordings of efferent spontaneous activity in the lateral-line nerve were used as a measure of the central effect of metacaine during administration in the respiratory stream, as compared with respiration of fresh water. The reduction of activity at the end of a 3-min exposure to the drug was 70%. 3. Maximum recovery (90% of the initial activity) was reached after 30 min application of fresh water. 4. The various efferent impulse types were differentially affected by the anesthetic. 5. Direct recordings from sound-sensitive neurons in the medulla confirm the strong central effect of metacaine. 6. Irrigation of the lateral-line system with anesthetic solution also produced a reduction in afferent spontaneous activity; there was a distinct peripheral effect even at the threshold concentration. 7. Reactions of trigeminal nerve fibers to mechanical stimulation of the skin were reduced after application of a metacaine solution to the skin. This finding confirms the local anesthetic effect of the drug. 8. The anesthetic solution at a given concentration had a greater effect on the CNS than on the peripheral receptors. 9. The implications of the results with respect to the risk of misjudging the depth of general anesthesia are discussed. 10. Because of the complexity of its anesthetic efficacy, it is recommended that metacaine not be used for neurophysiological investigations, but rather that the animals be immobilized by means of muscle relaxants.


The actions of aminopyridines on avian muscle. The effects of 2-, 3-, and 4-aminopyridines were investigated on the isolated chick biventer cervicis nerve-muscle preparation and on nerve-free cell cultures of embryonic chick skeletal muscle.  All 3 compounds reversed tubocurarine blockade and augmented twitch height in indirectly stimulated biventer cervicis preparations. 4-Aminopyridine was approximately 10 times more potent than 2-, or 3-aminopyridine. Twitch augmentation was also seen in directly stimulated preparations but to a much lesser extent. The compounds did not have significant anticholinesterase activity, nor did they have any depolarizing activity when tested on nerve-free cultured muscle fibres. At high concentrations the aminopyridines produced a maintained contracture in the biventer preparations which was enhanced by neostigmine and inhibited by tubocurarine. It is suggested that the aminopyridines facilitate neuromuscular transmission by increasing acetylcholine release in response to nerve stimulation, and that the compounds can also increase spontaneous transmitter release.


Catecholamine synthesis in rat brain after axotomy: interaction between apomorphine and haloperidol. Axotomy of the ascending monoaminergic fibers by means of a complete cerebral hemitransection stimulated the formation of dopa during 30 min after inhibition of the aromatic amino acid decarboxylase with 3-hydroxybenzylhydrazine HCl, 100 mg/kg i.p., in c. striatum and the dopamine-rich part of the limbic system. Apomorphine, 0.5 mg/kg i.p., antagonized the accumulation of dopa not only on the intact but also on the lesioned side. Haloperidol, 2 mg/kg i.p., stimulated dopa accumulation on the intact side but could not further stimulate the increase in dopa caused by transection. When both drugs were given together, the inhibitory effect of apomorphine was fully counteracted by haloperidol on both sides. In the predominantly noradrenaline-innervated occipito-temporal cortex dopa formation was slightly higher on the lesioned than on the intact side and was not markedly influenced by apomorphine. In the rest of the hemispheres the apomorphine-induced decrease in  dopa formation was more pronounced on the intact than on the lesioned side and was fully antagonized by haloperidol. The dopamine concentration was slightly higher in the lesioned c. striatum as compared to the intact side irrespective of the drugs administered. In c. striatum and the limbic system haloperidol caused a decrease in dopamine on the intact side which was not antagonized by additional treatment with apomorphine. Hemitransection caused a decrease in noradrenaline especially in the hemisphere portion. Neither apomorphine nor haloperidol or both drugs in combination changed the latter effect. In general, the tyrosine concentration tended to be higher on the lesioned than on the  intact side in all brain structures investigated. The data support the view that a local receptor-mediated feedback mechanism exists which is controlling dopamine synthesis even in the absence of impulse flow.


Opiate receptor identification in vitro and in vivo. Using tritiated opiates with a high specific radioactivity, a high affinity binding to brain homogenates has been observed in vitro. Strong evidence was given that this type of binding is due to the interaction of labelled opiates with their specific receptors in the brain, since the ability of a wide variety of opiates to inhibit this binding corresponds very well with the potency of the drugs to produce their pharmacological effects in vivo. We have found that the simultaneous injection of labelled opiates into animals together with high doses of nonradioactive opiates decreases the brain concentration of the labelled drug. This displacement from brain satisfies criteria similar to those used for the identification of the opiate receptor in vitro. Furthermore, opiates produce their pharmacological effects in the same dose range in which they are able to displace a radiolabelled opiate from the brain. This strongly suggests that the receptor occupation by opiates can be identified by concentration changes in the brain of living animals.


Endogenous ligands of opiate receptors.

Endogenous ligands of opiate receptors. From brain and from pituitary tissue of vertebrates several peptides - with molecular weights between 500 and 3500 daltons - have been isolated, which behave like opiates in opiate receptor binding assays, in isolated tissue preparations and in the intact animal.


[Transmission of reticulofugal activity through the ventromedial group of propriospinal neurons in cats]. Responses of propriospinal interneurons localized in the ventromedial regions of the cat lumbar cord to the reticulospinal tract stimulation were studied. Monosynaptic excitatory action usually accompanied by di- and polysynaptic components was found in most units. Strong single stimuli were able to evoke discharges only in some cells, while temporal summation was necessary in many others. Excitation of neurons was followed by a short postactivation inhibition limiting the maximal discharge frequency.


[Extracranial metastasis of primary brain tumours]. This study deals with the visceral metastasis of glioblastomas, ependymomas, oligodendrogliomas and medulloblastomas. Taking a sery fo four cases in the medical literature out of one hundred and fifteen cases published since 1961 and selected according the WEISS criterias, allows to indicate the delay in apparition of these metastasis as well as their elective localisation depending on the anatomopathological type of the original cerebral tumor. It does not give any precise indication to why these visceral metastasis appear. It seems to be very likely that the apparition of these secondary tumors is due to the failure of the shole defence systems of the body, favoured by the relative long survical of these patients and the surgical opening of the brain envelops.


Cyclic 3'5'-adenosine monophosphate in the hypothalamo-neurohypophysial system of normal, NaCl-treated and lactating rats. The presence and production of cyclic 3', 5'-adenosine monophosphate (cAMP) were investigated in the hypothalamus and neural lobe of the rat. Theophylline (concentrations from 10(-3) to 8 X 10(-3) M) increased the in vitro content of cAMP in the isolated neural lobe and in hypothalamic tissue samples containing supraoptic (SO) or paraventricular (PV) nuclei. Acetylcholine (ACH; 10(-2) and 10(-4) M) or carbachol (10(-4) M) did not increase cAMP content in the isolated neural lobe. Small increases were apparent (p less than 0.05, t-test for paired samples) in the hypothalamus. The amounts of cAMP were significantly higher in isolated neural lobes but not in hypothalami of NaCl-treated or lactating as compared to control rats. Presence of cAMP in the neural lobe and activation of adenylate cyclase under stimulated hormone release conditions indicate a possible involvement of cAMP in the process of neurohypophysial hormone secretion.


Rat hypothalamic corticotropin-releasing factor (CRF) content remains constant despite marked acute or chronic changes in ACTH secretion. Hypothalamic corticotropin-releasing factor (CRF) activity was only slightly increased by 1 min ether stress and was unaltered by 2.5-10 min ether stress, 15-day adrenalectomy, 11-day hypophysectomy or 3-day dexamethasone treatment. There was a slight tendency for the hypothalamic CRF activity to be higher in the p.m. than in the a.m. Basal hypothalamic deafferentation did not significantly affect hypothalamic CRF content in the a.m. or p.m. compared to intact controls. We conclude that hypothalamic CRF content maintains relative constancy under conditions of marked acute or chronic stimulation or suppression of ACTH secretion.


[Sleep characteristics during 1st night in laboratory]. Three goups, each composed of five individuals were investigated: students, patients and controls aged 20--45 years. The characteristic features of the first sleep in the laboratory were compared with the sleep during experimental pre-examination stress in students and preoperative stress in patients. It was found that the first sleep in the laboratory had the same features as the sleep in experimental stress. The process of adaptation to the conditions of sleep in the laboratory during the 2nd and 3rd night was studied. In the case of patients no adaptation was observed during three successive nights.


[Cytochemical studies of peripheral white blood cells in Wilson's disease]. Using cytochemical methods the authors studied the activity of certain lysosomal enzymes and cytochrome oxidase in peripheral blood leucoytes in 22 patients with Wilson's disease. The control group comprised 50 healthy blood donors. It was found that the activity of acid phosphatase in the lymphocytes of patients was higher than in controls, the mean indices being respectively 90.50 +/- 8.95 and 60.38 +/- 3.95. The activity of beta-glucuronidase was found to be lower in the lymphocytes of patients, the mean value was 25.10 +/- 8.59 in patients and 64.91 +/- 5.78 in controls. The activity of cytochrome oxidase was lower in the granulocytes of patients with Wilson's disease than in controls, the mean values being 54.5 +/- 12.14 and 156 +/- 15.41 respectively. The activity of acid phosphatase in granulocytes as well as that of non-specific esterase in lymphocytes was similar in both groups. Decreased antigen degradation in Wilson's disease may be due not only to liver cirrhosis but also to disturbances in the metabolism of white blood cells, including, among others, decreased activity of cytochrome oxidase. The rise of the activity of acid phosphatase and reduced activity of beta-glucuronidase indicate chronic antigenic stimulation of lymphoid system.


[Case of spinal cord injury at the site of its junction with the medulla]. The authors describe a case of spinal cord injury close to the site of its junction with the medulla. The injury was caused by the sharp end of joiner file. Following the injury a neurological syndrome developed indicating selective direct injury to the pyramidal tract in the left lateral funiculi of the cord and to the pathways of deep sensation on the left side, as well a transient injury to the pathways of superficial sensation on the side.


[Osteopetrosis (Albers-Schoenberg disease). III. Current nosographic aspects and relations to other condensing osteopathies]. A malignant paediatric variety and an adult variety of Albers-Schönberg disease are normally distinguished. On the basis of recent findings and personal observation it would appear advisable to accept two different courses of Albers-Schönberg disease in adults: one resembling the malignant infant form and the other with slow, practically asymptomatic (apart, obviously, from the skeletal lesions) course allowing for prolonged survival. This classification is of considerable practical importance for prognosis and therapeutic purposes. Other hereditary-familial and constitutional condensing osteopathy pictures exist that present radiological stigmata similar to those seen in Albers-Schönberg disease. The interest of the relations between A-S disease and certain of these condensing osteopathic conditions is obvious.


[Analysis of surgical treatment of 592 cases of cancer of the stomach]. A series of 592 cases of gastric carcinoma operated on between January 1960 and December 1974 at the S. Croce Hospital at Cuneo is reported. The following observations were made: greater frequency in males (68.25%). The following localizations: cardiac portion 31 cases; fundus 20; body 93 and antropyloric 249. Cancer ulcer in 5.06% of cases. Cancer on the gastric stump in ulcer resections in two cases. The following operations were performed: Gastric resection in 325 cases: operative mortality (O.M.) 5%; 5-year survival: 18%. Total gastrectomy in 38 cases: O.M. 13.1%; 2-year survival: 21%, 5-year survival 5.2%. E.A.G. in 61 cases; O.M. 17.8%; survival: 6 months; maximum 26 months. Gastrostomy in 16 cases; Expl. laparotomy in 150 cases. Survival was only a few months in these two latter groups. Out of all operations 5-year survival was 6.7% and 27.4% in so-called curative interventions. In principle, gastric resection is still considered to be the most valid approach.


Estrogens and endometrial carcinoma.

Estrogens and endometrial carcinoma. A group of 205 women with endometrial carcinoma was matched for age, parity, and year of operation with a group of 205 women who had had hysterectomies for benign disease. In the former group, 32 patients had used conjugated estrogens, while in the latter group 12 had used this hormone, yielding a relative risk of 3.1 (P = 0.0008). Users of other forms of systemic estrogens showed similar elevations in relative risk. Relative risk was related to duration of use, progressing from no evidence of risk among those using the hormone for less than 5 years to an 11.5-fold greater risk for those using it for 10 years or more. Risk was also related to the strength of the medication. The relative risk for users of the 1.25-mg tablets was 12.7 as compared to a two- to fourfold greater risk among users of lesser strength tablets.


Endometrial carcinoma and oral contraceptive agents. Six patients who took oral contraceptive agents for 5 to 18 years developed endometrial neoplasia. Endometrial adenocarcinoma occurred in 4 of these patients and severe adenomatous hyperplasia occured in 2. Five of the 6 patients took sequential agents; 1 patient used a combined agent. An additional patient who took Premarin and Provera sequentially developed adenocarcinoma of the endometrium. Eighteen cases of endometrial adenocarcinoma and 7 cases of adenomatous hyperplasia in patients with long-term sequential oral contraceptive use have previously been reported by others. Progestogens may not be completely protective against the endometrial cancer-causing potential of the estrogens, especially in the sequential regimens.


[Keractoconjunctivitis caused by type 10 adenovirus]. 4 cases in which type 10 adenovirus was isolated are described in connection with a new form of viral keratoconjunctivitis. The predominant symptoms, after disappearance of the benign-type conjunctivitis signs by the 5th day, are those of edema of the lids, chemosis, multiple corneal erosions and folds in Descemet's membrane, reaching their maxima by the 15th day. The course of the disease is of roughly 30 days.


[HLA-SD-antigens in cervix cancer and recurrent herpes genitalis patients (author's transl)]. The frequency of 36 HLA-SD antigens were investigated in 42 patients with cervix cancer, 36 female patients with recurrent Herpes genitalis and 450 healthy controls. The only significant difference between the 3 groups was an increase in the frequency of BW 15 in the cervix cancer patients. The patients with recurrent Herpes genitalis--who were sero-epidemiologically determined to have a higher risk of developing a cervix carcinoma-showed no significant difference in their HLA-SD antigen frequencies in comparison with the controls and the cervix carcinoma patients.


[On the histogenesis of uterine mixed tumors (author's transl)]. Heterologous tumors of the uterus are commonly designated as Müllerian mixed tumors. Their origin is attributed to dispersed embryogenic isles of Müllerian duct. The always high age of the afflicted woman plus the localization in post-climacteric polyps, speak against this assumption. Five personal observations have motivated the more intensive occupation with the histiogenesis of these cases. This has led to the conclusion that these mixed tissue formations do not come from dispersed cell isles, but that the multipotency of proliferated cells has only appeared in later age in connection with a metaplasia.


The tumor enhancing property of herpes simplex virus type-2 (HSV-2). The s.c. infection of DBF-1 mice with HSV-2 has a tumor enhancing effect on simultaneously i.m. implanted MCA-induced syngeneic spindle cell sarcoma. Thus with the dosage used here, the first palpable tumors appeared on day 6 in the virus treated group as compared to day 9 p.i. in the sham treated batch. Further, more tumors were formed: 75% among the infected and 40% among the sham injected. The tumor yield could be modulated: no difference between the 2 groups was found when the tumor cell dose was increased sufficiently to advance the first appearance of tumors to less than 6 days. The same result was obtained when the neoplastic cells were implanted 3 days ahead of injection of the virus. When the dose of the cells was decreased, no tumors were found in the sham treated batch, whereas there were some in the infected group. Previous in vitro mixing of the neoplastic cells with virus completely prevented tumor formation, probably due to their lysis by the virus. In fact, in tissue cultures, MCA cells become lytically infected with HSV-2. It is speculated that a similar situation occurs in acutely ill mice which die before the 13th day p.i., i.e., the tumor cells are destroyed by the virus. In fact, those animals, as a rule are free of neoplasmas. Thus it appears that an acute HSV-2 infection prevents MCA sarcoma whereas a latent infection promotes its growth. The possible reasons for this tumor enhancement caused by HSV-2 infection are discussed.


Neoplasms of the nasopharynx in children. Neoplasms of the nasopharynx are rare in children, but they threaten the child's life when they do occur. The nasopharynx tends to harbor dysontogenetic neoplasms. After classification into benign and malignant groups, nasopharyngeal neoplasms in children can be further characterized according to the age of the patients in which the clinical manifestations usually appear. Dermoids and teratomas are the most frequently encountered neoplasms of the nasopharynx in infants and may produce airway obstruction and dysphagia. Among the benign tumors of the nasopharynx in children, the juvenile angiofibroma deserves the most attention. With the onset in puberty, these neoplasms may cause recurrent massive bleeding and orbital and intracranial complications. Evaluation of the extent of the neoplasm and the source of the blood supply has been improved with bilateral selective internal and external carotid angiography. Intracranial and orbital invasion is regarded as an indication for radiotherapy. Surgery has been made somewhat safer by preoperative estrogen therapy and angiographic embolization of the major arterial supply. Patients with squamous cell carcinoma of the nasopharynx have immunologic similarities to patients with Burkitt's lymphomia and infectious mononucleosis; The etiologic role of the Epstein-Barr virus is considered. The parts played by radiation therapy, surgery, chemotherapy, and cryosurgery in the treatment of children with carcinoma of the nasopharynx are discussed. The value of radical neck dissection after radiation therapy is critically reviewed. The prognosis in patients with carcinoma of the nasopharynx is better in females than in males and better in children than in adults.


[Tubular and intestinal phosphate transport in vitamin D-resistant (hypophosphatemic) rickets (author's transl)]. Renal clearance and intestinal absorption studies were performed to investigate the renal and intestinal handling of phosphate under various conditions in two female patients with VDRR. The two types of transepithelial phosphate transport in the renal proximal tubule and intestinal mucosa are discussed. The defective parathormone sensible transport mechanism is slightly depressed under the influence of parathormone and activated after treatment of vitamin D in one patient, in the other no further inhibition by PTH could be observed. Electrolyte excretion in urine remains stable during treatment with vitamin D 3. Therapy consists of administration of oral phosphate and of pharmacologic doses of vitamin D.


Cyclic nucleotide phosphodiesterase activities of the fetal and mature human cerebral cortex. Phosphodiesterase activities were examined in the supernatant and pellet fractions of a 30,000 X g preparation of brain tissues from human fetuses and young adults. Differences in total activity and distribution of the high and low Km activity enzymes for adenosine and guanosine 3',5'-monophosphate (cyclic AMP and cyclic GMP, respectively) were found. The mature cortex had 10 times more activity than the fetal brain for cyclic AMP hydrolysis and 15-20 times more activity for cyclic GMP hydrolysis. In the fetus, more activity for both nucleotides at both high and low concentrations is associated with the supernatant fraction. With maturity, a shift in localization of high Km activity for cyclic GMP and low Km activities for both nucleotides to the particulate fraction is observed. Michaelis constants for both mature and immature brains are similar. The Km values for cyclic AMP are 10(-4) and 10(-5) M and 10(-4) and 10(-6) M for cyclic GMP. The Vmax values differed by a factor of 10 between the high Km and the low Km forms for each substrate. Differences were observed between the fetus and the adult when the hydrolysis of one nucleotide was measured in the presence of varying amounts of the other nucleotide. Low concentrations of either nucleotide stimulated the hydrolysis of low concentrations of the other in the adult whereas, in the fetus, low concentrations of cyclic AMP inhibited cyclic GMP hydrolysis. At higher concentrations of either nucleotide, the addition of the other over a wide range of concentrations resulted in inhibition which was exaggerated in the fetus.


Auditory and visual defects resulting from symptomatic and subclinical congenital cytomegaloviral and toxoplasma infections. Sensorineural hearing loss was present in ten of 59 (17%) patients with congenital cytomegalovirus (CMV) infection (three of eight born with symptomatic and seven of 51 born with subclinical infection). The defect was bilateral in eight, moderate to profound in eight, and of progressive nature in two. Hearing loss did not occur in 21 patients with natal CMV infection nor in seven of 12 patients with congenital toxoplasmosis. Histopathologic and immunofluorescent studies of the inner ear in two of three neonates who died with severe infection revealed that viral antigens were widely distributed in cochlear structures. Eye pathology was associated only with congenital Toxoplasma (nine of 12) and CMV (seven of 43) infections. Visual impairments were more prominent and severe in those born with symptomatic infections, exclusively so with CMV. However, ocular defects, in particular chorioretinitis, developed after birth in five of eight patients born with asymptomatic congenital toxoplasmosis. These data firmly establish clinically inapparent congenital CMV infection as a major public health problem and confirm the fact that congenital toxoplasmosis may be associated with late-appearing, debilitating chorioretinitis.