[Biological effects of solar radiations]. Main biological effects of ultra-violet radiations are discussed. At a dermatological point of view, most of these effects are harmful: actinic erythema, cutaneous cancerogenesis (basal and spinous cell carcinomas, malignant cutaneous melanomas). Photo-immunosuppression and photoaging of the skin are discussed. Two benefic actions of sunlight are reviewed: synthesis of vitamin D3 and positive action of visible radiations on human psychism.


Islet amyloid polypeptide-producing pancreatic islet cell tumor. A clinical and biochemical characterization. Islet amyloid peptide (IAPP) or amylin is a recently discovered polypeptide without settled physiology in man. We present a patient with an endocrine pancreatic tumor secreting huge amounts of IAPP-like immunoreactivity (20,000 mol/l) and a concomitant development of diabetes mellitus. The release of insulin and pancreatic polypeptide (PP) was totally absent after an oral glucose load and a mixed meal, respectively. Tumor secretion of IAPP-like immunoreactivity seemed to be influenced by cholinergic mechanisms and by nutrients. The observed effects on insulin and PP secretion by high circulating levels of IAPP-like immunoreactivity may be of beneficial value for further studies of the physiology of IAPP in man.


[Long-term therapy of a metastasizing pancreatic vipoma using the somatostatin derivative octreotide]. A patient with metastatic VIP-producing pancreatic tumor was successfully treated with subcutaneous octreotide, an analogue of somatostatin, for more than 4 years. The profuse diarrhea was rapidly controlled and the plasma concentrations of the hormones (VIP, neurotensin, gastrin, pancreatic polypeptide) fell to nearly normal within 2 months. Because of asymptomatic increase in tumor size, we added chemotherapy 2 years later. Since the drug is rapidly effective and well tolerated, it will probably become the therapy of choice in this syndrome.


[Platelet receptors: nomenclature--structure--function. 1]. Current knowledge on platelet receptors, their structure, function and interaction with their ligands, and the various nomenclatures, is summarized. The role of platelet receptors in thrombosis and hemostasis, and in cell adhesion and invasion, is discussed. Many of the receptors are members of large families of cell-adhesion molecules (such as the integrin family of cell-adhesion receptors) and have common subunits. While some receptors are exclusive to platelets and megakaryocytes (e.g. GP IIb/IIIa), others may be found on several other cell types (such as the receptors for epinephrine or vitronectin). In addition, the receptors for the most important platelet agonists and the platelet binding sites of factors of plasmatic coagulation and fibrinolysis are reviewed.


Activation of mitogen-activated protein kinase kinase by v-Raf in NIH 3T3 cells and in vitro. Mitogen-activated protein (MAP) kinases are 42- and 44-kD serine-threonine protein kinases that are activated by tyrosine and threonine phosphorylation in cells stimulated with mitogens and growth factors. MAP kinase and the protein kinase that activates it (MAP kinase kinase) were constitutively activated in NIH 3T3 cells infected with viruses containing either of two oncogenic forms (p35EC12, p3722W) of the c-Raf-1 protein kinase. The v-Raf proteins purified from cells infected with EC12 or 22W viruses activated MAP kinase kinase from skeletal muscle in vitro. Furthermore, a bacterially expressed v-Raf fusion protein (glutathione S-transferase-p3722W) also activated MAP kinase kinase in vitro. These findings suggest that one function of c-Raf-1 in mitogenic signaling is to phosphorylate and activate MAP kinase kinase.


Development and characterization of a human monoclonal antibody probably detecting the leukocyte differentiation antigen CD39. A human monoclonal IgM antibody, referred to as TU223, has been produced. The reactivity of TU223 was tested in various cells and cell lines by complement-dependent microcytotoxicity test and fluorescence-activated cell sorter analysis. The antigen defined by TU223 was expressed on Epstein-Barr virus-transformed B-cell lines and on some Burkitt's lymphoma cell lines, but was not expressed on normal T cells, B cells or erythrocytes. In addition, expression of the antigen defined by TU223 was also induced on B cells activated by Epstein-Barr virus or pokeweed mitogen, and on T cells activated by phytohemagglutinin, concanavalin A, pokeweed mitogen or recombinant interleukin-2. However, no expression of the antigen detected by TU223 was induced at all on recombinant interleukin-4-treated B cells or macrophage-like cell line U937. When the ability of TU223 and various mouse monoclonal antibodies to bind to human differentiation antigens was compared, interestingly, the reactivity of TU223 was found to be very similar to that of mouse monoclonal antibody CD23 (H107), which reacts with Fc epsilon receptor II. Two-color analysis revealed that the antigen defined by TU223 is expressed on the cell surface of certain lymphoid cells expressing CD23 antigen. However, it can be concluded that the antigen defined by TU223 is clearly distinct from Fc epsilon receptor II, based on assay of cross-blocking between H107 and TU223. The surface antigen on B85 cells recognized by TU2232 had the molecular size of 80-82 kiloDaltons as determined by immunoblotting analysis.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of dithiocarbamates on testicular toxicity in rats caused by acute exposure to cadmium. N-Benzyl-D-glucamine dithiocarbamate (BGD), N-p-isopropylbenzyl-D-glucamine dithiocarbamate (PBGD), and diethyl-dithiocarbamate (DED) were compared for their protective effects against the testicular toxicity in rats induced by acute exposure to cadmium. Rats were injected subcutaneously with 109CdCl2 (3 mg Cd and 74 kBq of 109Cd/kg) and 30 min later, they were injected intraperitoneally with the chelating agents (0.4 or 3 mmol/kg). Cadmium injection increased lipid peroxidation and concentrations of hemoglobin and Ca in the testes, decreased the testicular weight, and caused sterility. The treatment with BGD (0.4 mmol/kg) did not satisfactorily protect against the testicular toxicity of cadmium. The administration of PBGD or DED at a dose of 3 mmol/kg significantly prevented the increase in the lipid peroxidation and hemoglobin concentration in the testes, the decrease in the testicular weight, and the sterility caused by cadmium. PBGD and DED significantly decreased the cadmium concentration in the testes, but DED increased the cadmium concentration in the kidney and brain. Only DED significantly prevented the increase in the testicular Ca concentration after cadmium. These results indicate that PBGD and DED protect against the sterility caused by cadmium in rats and that the effect of DED to increase the brain level of cadmium is more dangerous than the lack of effect of PBGD to prevent the increase in the testicular Ca level. The protective effects of PBGD and DED against the cadmium-induced testicular toxicity presumably result from a decrease in the cadmium concentration in the testes.


[Effects of saline infusion on the activity of erythrocyte sodium-potassium pump in normal subjects and in patients with essential hypertension]. The effects of a 2 litre isotonic saline infusion on erythrocyte Na,K pump activity and urinary sodium excretion (UNaV) were evaluated in 20 patients with essential hypertension and 15 normotensive subjects. The effect of preincubation of normal erythrocytes in plasma from hypertensive patients on erythrocyte Na,K pump was also studied. Before saline infusion no significant differences were found between normotensive and hypertensive subjects in the mean values of intracellular sodium and potassium concentration, ouabain-sensitive Na efflux in erythrocytes and urinary sodium excretion. Erythrocyte Na,K pump activity decreased significantly (p less than 0.01) after saline infusion in both groups of subjects. The reduction was significantly lower in hypertensives than in normotensive. delta UNaV was significantly higher in hypertensive patients than in normotensive subjects (25 +/- 4 vs 14 +/- 2 mmol/h; p = 0.04). Only in normal subjects a significant correlation was found between the difference in Na,K pump activity pre and post saline infusion and delta UNaV (r = 0.52; p less than 0.05). Plasma from hypertensive patients obtained before saline infusion significantly (p less than 0.01) inhibited Na,K pump of erythrocytes from normal subjects; plasma taken after saline infusion produced a marked increase of this inhibition, significantly (p less than 0.01) higher than serum taken before the infusion.


Stable isotopic tracers of lead mobilized by DMSA chelation in low lead-exposed rats. The ability of DMSA to mobilize skeletal lead or effect a redistribution of endogenous lead to other target organs in low lead-exposed organisms is unclear. Discrepant results of past studies of DMSA and other lead chelators (e.g., CaNa2EDTA) may be due, in part, to experimental differences and difficulties in distinguishing mobilized skeletal lead from other endogenous or exogenous lead sources. Therefore, the influence of DMSA on the mobilization and redistribution of lead in skeletal and soft tissue compartments of low lead-exposed female Wistar (115-125 g) rats was investigated using ultraclean stable lead isotope tracer techniques. Rats that had been reared on a low lead-level diet (lead intake approximately 80 ng Pb/g body/day) were fed 206Pb-enriched drinking water (210 ng Pb/ml) for 1.5 days and then were chelated with a single ip injection of a 0.11 mmol/kg dose of DMSA. Blood, kidney, brain, tibia, urine and feces were collected 24 hr after chelation and analyzed for lead concentrations by graphite furnace atomic absorption spectrometry and for lead isotopic compositions by thermal ionization mass spectrometry. These analyses demonstrated that DMSA chelation significantly increased (15-fold) the diuresis of labile soft tissue lead, but not skeletal lead. DMSA also appeared to effect a redistribution and input of a comparable amount of lead to the skeleton and smaller relative amounts of lead to the soft tissues (blood, kidney) of the chelated animals. The clinical significance of these latter observations beyond the context of this preliminary study is not clear.


Arthropod toxins as leads for novel insecticides: an assessment of polyamine amides as glutamate antagonists. In the search for new toxins, preferably with new sites of action, the polyamine amides represent a new class of compounds with potential as insecticides and as pharmaceutical agents due to their antagonism of ligand-gated cation channels. In particular, they are potent antagonists of the L-glutamate receptors of insect skeletal muscle. In this paper, we report on synthetic studies to produce hybrid analogues based upon the argiotoxin spider toxins and philanthotoxin-433 which is obtained from a solitary, parasitic wasp. We speculate upon possible modes and sites of action for these antagonists and we discuss their potential as insecticides and in the possible treatment of ischaemic damage. The synthesis and characterization of 4-hydroxyphenylpropanoylspermine is reported and the locust muscle biological assay is described. Using this pharmacological screen, structure-activity relationships have been determined in our laboratories. These are reviewed in the light of the current literature. Voltage clamp studies of the synthetic analogue philanthotoxin-343 and the effects of this polyamine amide on glutamate receptors expressed in Xenopus oocytes are outlined. In conclusion, a description of our current ideas and understanding of the many sites and modes of action of the polyamine amides, based both upon our own studies and also upon those recently reported, is presented.


Dermonecrotic and lethal components of Loxosceles gaucho spider venom. Loxosceles gaucho spider venom causes a typical dermonecrotic lesion in bitten patients and rarely causes lethal systemic effects. Gel filtration on Sephadex G 100 of L. gaucho spider venom resulted in three fractions: fraction A, containing the higher mol. wt components (approximately 35,000); fraction B, containing lower mol. wt components (approximately 15,000); and fraction C, containing very low mol. wt components (probably small peptides). The dermonecrotic and lethal activities were detected exclusively in fraction A. The venom and fraction A produced large dermonecrotic lesions in rabbits with necrosis spreading by gravity to the skin of the lateral body wall. Analysis by SDS-PAGE showed that the proteins contained in fraction A are approximately 35,000 and 33,000 mol. wt. Immunoblotting analysis showed that the proteins responsible for the dermonecrotic and lethal activity are very immunogenic and the first to be detected by antibodies during the course of immunization.


Effects of toxin Ts-gamma, purified from Tityus serrulatus scorpion venom, on the isolated rat atria. By using a pair of silver/silver-chloride electrodes it was possible to record, simultaneously, the atrial electrogram and the atrial contractile force of rat atria, in an organ bath, containing Krebs-Ringer solution (30 degrees C, pH 7.4, bubbled with 95% O2 and 5% CO2). Addition of toxin Ts-gamma, purified from Tityus serrulatus scorpion venom, into the bath (1 microgram/ml), evoked complex effects characterized by an initial reduction of both rate and contractile force, followed by increase of force and reduction of rate and finally by reduction of both rate and force. The increase of contractile force was prevented by metoprolol and is, therefore, adrenergic in nature. The reduction of rate was concomitant with changes in the atrial electrogram in which a positive P wave was replaced by a diphasic P wave, while the positive Ta wave was depressed. Experiments with tetrodotoxin, atropine and physostigmine indicate that these effects are due to the release of acetylcholine from vagal endings.


Foot-and-mouth disease: the risk for Great Britain after 1992. Mass annual vaccination against foot-and-mouth disease, previously applied by eight member states in the European Community (EC), was progressively phased out during 1990-91. The other four member states (the United Kingdom, Denmark, the Republic of Ireland and Greece) either never have vaccinated or ceased to do so several years ago. The EC should increase its international competitiveness if it maintains its present foot-and-mouth disease-free, non-vaccinating status. Freedom from disease and a harmonised disease control policy will also permit unrestricted movement of livestock and animal products throughout the EC when the single market is completed in 1992. Vaccination against foot-and-mouth disease on continental Europe has greatly reduced the number of outbreaks during the last 30 years and this reduction has been of indirect benefit to Great Britain. However, the cessation of vaccination will result in a higher proportion of fully susceptible cattle and in the event of outbreaks will increase the likelihood of the rapid dissemination of virus and increase the risk that the infection will enter Great Britain. The main risks of entry are likely to be associated with live animals in which the disease can be mild or inapparent, ie, sheep and goats, and with airborne virus originating from pigs on the nearby continent especially in Brittany and the Benelux countries where they are present in very high densities.


Expression of the Epstein-Barr virus latent membrane protein (LMP) in insect cells and detection of antibodies in human sera against this protein. Recombinant baculoviruses containing the complete LMP and truncated LMP genes were generated and high levels of the LMP proteins were expressed in Spadoptera Frugiperda insect cells. A specific rabbit antiserum directed against the N-terminal part of LMP was obtained by immunizing the rabbits with Escherichia coli-expressed trpE-N-terminal part of LMP fusion protein. A total of 127 human sera were studied for their immune response to the recombinant full-length LMP. In immunofluorescence analysis, all sera tested showed no detectable reaction with the recombinant full-length LMP. In immunoprecipitation-immunoblotting analysis, however, sera from patients with nasopharyngeal carcinoma (5/22), patients with Hodgkin's disease (16/27), patients with other diseases exhibiting high EA-IgG titers (3/52), and VCA-IgG-positive healthy individuals (2/26) were shown to contain antibodies against this recombinant LMP. The expressed LMP proteins provided a sufficient and economic source of the proteins for further serological and biological studies.


Herpes simplex virus type 1 gene UL13 encodes a phosphoprotein that is a component of the virion. The UL13 open reading frame of herpes simplex virus type 1 (HSV-1) has been expressed in insect cells by a recombinant baculovirus and in Escherichia coli. In the latter case, the UL13 gene was fused to the gene for glutathione S-transferase (GST) to allow high-level expression of an 80-kDa GST-UL13 fusion protein. Antibody raised against the fusion protein reacted specifically with the 55-kDa UL13 gene product expressed by the recombinant baculovirus. This antibody also recognized a late phosphoprotein in HSV-1-infected cell lysates and a component of purified HSV-1 virions, both with the same electrophoretic mobility as the baculovirus-expressed protein. The virion component was efficiently phosphorylated in vitro by a virion-associated protein kinase. Using the same antibody, the probable homolog of the UL13 gene product was identified in HSV-2-infected cells and purified virions.


Characterization of the large tegument protein (ICP1/2) of herpes simplex virus type 1. ICP1/2 (also designated VP1/2) is a 270-kDa structural protein of herpes simplex virus type 1 (HSV-1) which is located in the tegument region of the virion. In this report we describe the production of a polyclonal antiserum specific for ICP1/2 and the use of this antiserum to examine the synthesis, processing, and intracellular localization of the viral polypeptide. Pulse-labeling studies indicated that ICP1/2 is synthesized late during infection, being initially detectable between 8 and 9 hr postinfection with the rate of synthesis continuing to increase until 11 to 12 hr postinfection. Further studies on the expression of ICP1/2 in the presence or absence of viral DNA replication indicated that the synthesis of the polypeptide is absolutely dependent on viral DNA replication. These results suggest that ICP1/2 represents a gamma 2 (true late) gene product. Additionally, we have performed experiments to determine if ICP1/2 is post-translationally modified in HSV-infected cells. These studies indicated that ICP1/2 is phosphorylated on serine residues; however, we found no evidence to suggest that the protein is glycosylated. Using subcellular fractionation and indirect immunofluorescence techniques, we have determined that ICP1/2 is diffusely distributed throughout the nucleus and cytoplasm of HSV-infected cells with no specific compartmentalization of the polypeptide.


Construction of a US3 lacZ insertion mutant of herpes simplex virus type 2 and characterization of its phenotype in vitro and in vivo. We have constructed and characterized a mutant of herpes simplex virus type 2 (HSV-2) which was inserted a modified lacZ gene, placed under the control of HSV-1 beta 8 promotor, into the US3 protein kinase gene. The mutant, L1BR1, could not induce the virus-encoded protein kinase activity, but could replicate in Vero cells as efficiently as the parental virus. When the biological properties of L1BR1 were examined in mice by using four routes (footpad, intraperitoneal, corneal, and intracerebral) of infection, the mutant displayed the route-dependent reduction of virulence; after inoculation by footpad and intraperitoneal routes, the mutant was more than 10,000-fold less virulent than the parental virus, but it exhibited only about a 10-fold decrease in virulence following the corneal and intracerebral infection. In the intraperitoneal inoculation into adult mice, the replication of L1BR1 in the liver and spleen was severely restricted, but in newborn mice the mutant could grow as well as the parental virus in these organs. The adoptive transfer of peritoneal macrophages from adult mice resulted in a marked inhibition in the replication of L1BR1 in the liver and spleen of newborn mice, while the transfer exhibited little or no effect on the production of the wild-type virus in these organs. We also found that the mutant, unlike the parental virus, could not replicate in precultured peritoneal macrophages from adult mice. Taking these observations together, it seems likely that L1BR1 lost the ability to overcome the mononuclear-phagocytic defense system and thereby lost its pathogenicity by intraperitoneal and footpad routes. Furthermore, the mutant was shown to be rescued by a 4.8-kb HindIII/Xbal fragment containing the entire US3 open reading frame. However, we could not rule out the possibility that some of the phenotypes of L1BR1 are due to mutations in the US3-neighboring genes, US2 and US4.


The equine herpesvirus type 1 (EHV-1) homolog of herpes simplex virus type 1 US9 and the nature of a major deletion within the unique short segment of the EHV-1 KyA strain genome. The DNA sequence of the short (S) genomic component of the equine herpesvirus type 1 (EHV-1)KyA strain has been determined recently in our laboratory. Analysis of a 1353-bp BamHI/PvuII clone mapping at the unique short/terminal inverted repeat (Us/TR) junction revealed 507 bp of Us and 846 bp of TR sequences as well as an open reading frame (ORF) that is contained entirely within the Us. This ORF encodes a potential polypeptide of 219 amino acids that shows significant homology to the US9 proteins of herpes simplex virus type 1 (HSV-1), EHV-4, pseudorabies virus (PRV), and varicella zoster virus (VZV). The US9 polypeptides of the two equine herpesviruses exhibit 50% identity but are twice as large as their counterparts in HSV-1, PRV, and VZV. All five US9 proteins are enriched for serine and threonine residues and share a conserved domain of highly basic residues followed by a region of nonpolar amino acids. DNA sequence and Southern blot hybridization analyses revealed that the Us of EHV-1 KyA differs from the Us of EHV-1 KyD and AB1 in that the ORFs encoding glycoproteins I and E and a unique 10-kDa polypeptide are deleted from the KyA genome. These data demonstrate that the predicted 10-kDa protein unique to EHV-1 is nonessential for replication in vitro and that EHV-1 glycoproteins I and E, like their equivalents in HSV-1 and PRV, are also nonessential. These findings and those reported previously by this laboratory and others reveal that the Us segment of EHV-1 comprises nine ORFs, two of which, US4 and 10-kDa ORF, are unique to EHV-1. The gene order of the Us is US2, protein kinase, gG, US4, gD, gI, gE, 10 kDa, and US9.


Modulation of the cellular phenotype by integrated adeno-associated virus. The adeno-associated virus (AAV) rep gene encodes a series of overlapping, multifunctional, nonstructural proteins (Rep proteins) which regulate the viral life cycle and which are also capable of trans-regulating nonviral gene expressions (reviewed in Berns, 1990, Microbiol. Rev. 54, 316-329). To investigate the expression of the AAV rep gene in a cellular chromosomal context, SV40-transformed Chinese hamster embryo (OD4) cells were infected with an AAV/neo hybrid virus and progeny resistant to the antibiotic G418 were selected and amplified. Chromosomal integration and RNA transcription of the AAV and neo DNA inserts were confirmed by Southern and Northern blotting procedures. One of the G418R cell lines stably expressed a protein which reacted specifically with AAV anti-Rep antiserum in Western immunoblots. The stable integration of AAV rep DNA, which did not interfere with cell proliferation under normal growth conditions, was associated with two changes in cellular phenotype: eight of nine lines were markedly more sensitive to UV light (254 nm) than were the parental OD4 cells; and seven of the nine lines had lost the capacity to promote SV40 origin DNA amplification in vitro, in contrast to the parental OD4 cells. OD4 cells transformed to G418R by AAV/neo DNA constructs with a deleted rep gene, or by a neo DNA construct lacking AAV DNA, did not display these phenotypic changes. It is suggested that stable integration of the AAV rep gene interferes with cellular processes connected with DNA repair and gene amplification.


Molecular and structural basis of hemagglutination in mengovirus. The molecular and structural basis of mengovirus hemagglutination (HA) was investigated by the comparison of nucleotide sequences of the entire capsid coding regions of an HA+ variant, two HA- mutants, 205 and 280, and two HA+ revertants of 205. The mutants were selected after acridine mutagenesis of mengovirus-37A, a heat-stable and HA+ variant that is neurotropic in mice. HA+ revertants of mutant 205 were isolated from brain tissue of mice inoculated with mutant 205. The nucleotide sequences were determined by consensus RNA sequencing using genomic RNA templates from purified virions. Two nucleotide differences were observed in the VP1 coding region of the RNA genomes of mutants 205 and 280 in comparison to the RNA sequences of 37A and the revertants. Interpretation of these data predict substitutions of two consecutive amino acids at residues 1231 (K to R) and 1232 (P to S) of VP1 which form part of the H-I loop of VP1 found at the icosahedral fivefold axis. Analysis of the amino acid substitutions in the context of the three-dimensional structure of the mengovirus-M capsid indicated that hemagglutination most likely involves residues found at the icosahedral fivefold axis and probably does not involve the residues that form the putative cellular receptor binding site (the "pit"). Eleven amino acid differences were observed between the structural proteins of mengovirus-M and 37A, five in VP1, three in VP2, and three in VP3.


Mutation of the HANA protein of Sendai virus by passage in eggs. A study was made to elucidate the effect of host cells on the HANA protein of Sendai virus. Two strains of Sendai virus were isolated from an epidemic in an animal laboratory by inoculating the lung homogenate of a moribund mouse either into LLC-MK2 cells (Oh-L) or into the allantoic cavity of embryonated eggs (Oh-E). Oh-E agglutinated chicken red blood cells at 37 degrees (HA37+), while Oh-L did not (HA37-). When Oh-L was passaged in eggs, conversion of the HA37- virus to the HA37+ virus readily occurred. A single point mutation was recognized on the HANA protein of the HA37+ virus either at position 525 (Gln----Arg) or at position 198 (Leu----Phe). Hl test with monoclonal antibody revealed conformational changes around the receptor binding site. Neuraminidase activity was also affected by these mutations. The changes in these biological activities of the HANA protein seemed to allow the HA37+ virus to replicate in eggs. On the contrary, the HA37+ virus replicates as efficiently as the HA37- virus in LLC-MK2 cells and no reversion to the HA37- virus was observed. The overall results indicate that the passage of Sendai virus in eggs resulted in selection of viruses possessing a specific mutation on the HANA protein. The pneumopathogenicity in mice was not significantly different between the HA37- virus and the HA37+ virus, suggesting the existence of genes other than the HANA gene that determine mouse pathogenicity.


Expression of SV40 receptors on apical surfaces of polarized epithelial cells. We have investigated the interaction of SV40 virions with polarized monkey kidney epithelial cells. Virions were tagged with biotin to facilitate their detection and were found to retain full infectivity. When polarized Vero C1008 cells were incubated with biotinylated virions followed by a strepavidin-rhodamine conjugate, distinct cell populations were identified which expressed very different levels of SV40 receptors. The parental Vero C1008 cells yielded three types of cell clones which exhibited low, moderate, or predominantly high levels of SV40 binding. Virus-binding assays to each of these clones as well as to parental Vero C1008 cells indicated that the level of SV40 receptor expression is cell-cycle-dependent. The cellular receptors for influenza A virus (WSN strain) were also found to be distributed heterogeneously on polarized epithelial cells. In contrast, in several types of nonpolarized cells, SV40 receptors were found to be uniformly distributed over the monolayer. SV40 binding was not found to correlate with HLA expression on Vero C1008 cells or other cell types. Also in contrast to SV40 receptor expression, which is restricted to the apical domain, HLA was found to be distributed on both apical and basolateral domains of Vero C1008 cells.


Structural, functional, and immunological characterization of bovine herpesvirus-1 glycoprotein gl expressed by recombinant baculovirus. The major glycoprotein complex gl of bovine herpesvirus-1 was expressed at high levels (36 micrograms per 1 x 10(6) cells) in insect cells using a recombinant baculovirus. The recombinant gl had an apparent molecular weight of 116 kDa and was partially cleaved to yield 63-kDa (glb) and 52-kDa (glc) subunits. This processing step was significantly less efficient in insect cells than the analogous step in mammalian cells, even though the cleavage sites of authentic and recombinant gl were shown to be identical. The oligosaccharide linkages were mostly endoglycosidase-H-sensitive, in contrast to those of authentic gl, which has mostly endoglycosidase-H-resistant linkages and an apparent molecular weight of 130/74/55 kDa. Despite the reduced cleavage and altered glycosylation, the recombinant glycoprotein was transported and expressed on the surface of infected insect cells. These surface molecules were biologically active as demonstrated by their ability to induce cell-cell fusion. Fusion was inhibited by three monoclonal antibodies specific for antigenic domains I and IV on gl. Domain I maps to the extracellular region of the carboxy terminal fragment glc and domain IV to the very amino terminus of the glb fragment, indicating that domains mapping in two distinct regions of gl function in cell fusion. Monoclonal antibodies specific for eight different epitopes recognized recombinant gl, indicating that the antigenic characteristics of the recombinant and authentic glycoproteins are similar. In addition, the recombinant gl was as immunogenic as the authentic gl, resulting in the induction of gl-specific antibodies in cattle.


Modulation of the human papillomavirus type 16 induced transformation and transcription by deletion of loci on the short arm of human chromosome 11 can be mimicked by SV40 small t. The human papillomavirus (HPV) type 16 enhancer-promoter has been shown to be active in human fibroblasts with a deletion on the short arm of one chromosome 11 (karyotype 46,del(11)(p11.11p15.1)) but is virtually inactive in diploid human fibroblasts (Smits, Smits, Jebbink, and ter Schegget, 1990b, Virology, 176, 158-165). In diploid human embryonic fibroblasts, activation of the HPV16 enhancer-promoter could be achieved by expression of the SV40 small t. By cotransfecting SV40 small t cDNA together with HPV16 DNA into diploid cells, it was possible to increase the transforming activity of HPV16 by 10- 15-fold. Furthermore, SV40 small t was essential for the SV40 large T-induced morphological transformation of human diploid fibroblasts, whereas SV40 small t was dispensable for transformation of del-11 cells. We propose that, as a result of the deletion of loci on the short arm of chromosome 11 in del-11 cells, functions are expressed that mimic those of SV40 small t in transformation and trans-activation.


On the association of human beta 2 microglobulin with cell culture-grown human cytomegalovirus (HCMV). We studied the production of human beta 2 microglobulin (beta 2m) in mock-infected or human cytomegalovirus (HCMV) infected human embryonic lung fibroblasts (HEL) and the association of human beta 2m with HCMV virions. Titration of beta 2m by two-step sandwich enzyme immunoassay revealed that HEL released considerable amounts of human beta 2m into the culture medium and that the production of beta 2m was significantly enhanced by HCMV infection. The concentration of human beta 2m in the culture medium of HCMV-infected HEL reached 500 to 600 ng/ml, which corresponded to 7- to 12-fold of levels found in healthy adult urine. Immunoprecipitation assays showed that HEL-grown HCMV bound a significant amount of endogenous beta 2m, but the viruses were efficiently neutralized by either human hyperimmune anti-HCMV globulin or anti-HCMV monoclonal antibody even when treated with a large amount of human beta 2m or with dialysed urine. Thus it seems unlikely that the binding of beta 2m by HCMV is involved in masking the viral antigenic site necessary for neutralization.


A highly attenuated host range-restricted vaccinia virus strain, NYVAC, encoding the prM, E, and NS1 genes of Japanese encephalitis virus prevents JEV viremia in swine. A highly attenuated strain of vaccinia virus (NYVAC) was engineered to express the Japanese encephalitis virus (JEV) prM, E, and NS1 genes or the prM and E genes. The recombinant viruses were tested as vaccine candidates in pigs, a natural host of JEV. JEV-neutralizing and hemagglutination-inhibiting antibodies appeared in swine sera 7 days after immunization with 10(8) PFU of the recombinant viruses and increased after a second dose at 28 days. The JEV levels detected in the serum after JEV challenge (d56) of the swine with 2 x 10(5) PFU of JEV were significantly reduced in animals inoculated with the recombinant viruses. These results demonstrate the ability of these NYVAC-vectored recombinants to protect pigs from JEV viremia.


Sequences within the feline immunodeficiency virus long terminal repeat that regulate gene expression and respond to activation by feline herpesvirus type 1. We constructed a series of deletion mutants of feline immunodeficiency virus (FIV) long terminal repeat (LTR) to identify the regions that regulate gene expression and are responsive to trans-activation of FIV LTR by feline herpes-virus type 1 (FHV-1). We demonstrated that sequences between -124 and -79, and between -21 and -32 (relative to the cap site) are essential for gene expression of FIV in Felis catus whole fetus 4 (fcwf-4) cells. Further, we demonstrated that the sequence between -63 and -23 responds to trans-activation of FIV LTR by FHV-1 in fcwf-4 cells.


Phenotypic and genetic polymorphisms among human herpesvirus-6 isolates from North American infants. Fifteen human herpesvirus-6 (HHV-6) isolates from North American infants with primary infection manifest as febrile or roseola (exanthem subitum) like illnesses were characterized phenotypically on the basis of their in vitro growth in continuous T-cell lines and primary human mononuclear cells and by their reactivity with monoclonal antibodies. All isolates replicated efficiently in primary human cord blood mononuclear cells, but five distinct patterns of viral replication in human cells lines were observed. Two of the HHV-6 isolates from infants were found to replicate in HSB-2 cells, a property associated with so-called group A viruses, which had previously been isolated only from adults. These same isolates also reacted with a panel of A-specific monoclonal antibodies. Genomic characterization of viral isolates using well-characterized restriction site polymorphisms indicated that these two isolates contained a mixture of both A- and B-type genomes, in different proportions. These data suggest that not all HHV-6 isolates can be categorized into one of two broad groups and that such segregation of HHV-6 isolates may in fact be misleading.


Nucleotide sequence of the envelope glycoprotein of Negishi virus shows very close homology to louping ill virus. Negishi virus, a member of the family Flaviviridae, was originally isolated in Japan, during an outbreak of Japanese encephalitis. Antigenically, however, Negishi virus resembles the tick-borne rather than the mosquito-borne flaviviruses. Monoclonal antibodies that bind louping ill virus showed a close antigenic relationship between louping ill and Negishi virus. The genes encoding the envelope glycoprotein of Negishi virus (strain 3248/49/P10) and louping ill virus (strain SB526) were cloned and sequenced. They showed a very close homology at both the nucleotide and deduced amino acid levels. Comparison with the known sequence of another strain of louping ill virus (strain 369/T2) and with other tick-borne flaviviruses showed that Negishi virus was more closely related to louping ill virus than to the other tick-borne viruses. The significance of this observation for virus evolution, virus distribution in the environment, and the potential use of nucleotide sequencing for rapid and precise identification of flaviviruses are discussed.


The correct sequence of the porcine group C/Cowden rotavirus major inner capsid protein shows close homology with human isolates from Brazil and the U.K. Amino acid sequence alignments between the human group C/Bristol and the published porcine group C/Cowden VP6 proteins have revealed a region of extreme sequence divergence. We have been unable to confirm the nucleotide sequence of the Cowden VP6 gene corresponding to this region of divergence. Direct sequencing of a PCR-amplified cDNA pool has revealed a frame shift, and three nucleotide changes, within the published sequence of the porcine (Cowden) VP6 gene. The corrected sequence of the porcine protein revealed a closer homology with VP6 from the Bristol strain and two new human group C rotavirus isolates. Atypical rotaviruses have been detected in the feces of children living in Belém, Brazil, and Preston, U.K. Direct sequencing of PCR-amplified cDNA corresponding to the VP6 gene of one isolate from each location confirmed the presence of a group C rotavirus. The complete nucleotide sequences of the VP6 genes from the group C/Belém and C/Preston rotaviruses contained an open reading frame of 1185 nucleotides (395 amino acids; deduced M(r) 44,669 Da). The Belém VP6 gene demonstrated 97.9% nucleotide homology with the human group C/Bristol VP6 gene and 83.4% nucleotide homology (91.6% deduced amino acid homology) with the corrected porcine group C/Cowden sequence. The Preston VP6 gene demonstrated 99.6% nucleotide homology with the human group C/Bristol VP6 gene and 84.0% nucleotide homology (91.6% deduced amino acid homology) with the corrected porcine group C/Cowden sequence. Remarkably, the deduced amino acid sequence of the Brazilian strain was identical to that of the U.K. isolates.


The herpes simplex virus type 1 immediate-early polypeptide ICP4 is required for expression of globin genes located in the viral genome. We infected Vero cells with ICP4-deficient herpes simplex virus recombinants bearing the rabbit beta-globin and human alpha 2-globin genes under the control of their own promoters and found that globin gene expression occurred only when ICP4 was provided in trans. These results demonstrate that ICP4 is required for the activity of globin promoters located in the viral genome and support the hypothesis that these cellular promoters are functionally equivalent to HSV early regulatory regions.


Nucleotide sequence of gene 5 encoding the inner capsid protein (VP6) of bovine group C rotavirus: comparison with corresponding genes of group C, A, and B rotaviruses. To further study the molecular characteristics of group (gp) C rotaviruses, we produced, cloned, and sequenced cDNA to gene 5 of the Shintoku strain of bovine gp C rotavirus. The resulting clone was specific for gene 5 and was genetically related to the human and porcine gp C rotaviruses, as demonstrated by Northern blot hybridization analysis. The Shintoku gene 5 is 1352 nucleotides in length and has one open reading frame encoding a polypeptide of 395 amino acids with a predicted molecular mass of 44.5 kDa. Comparative sequence analysis indicated that: (i) the Shintoku gene 5 protein shared 88.4 to 90.6% homology with the VP6 of the human (Bristol and 88-220) and porcine (Cowden) strains of gp C rotaviruses, but only low homology with the VP6 of bovine gp A (RF) and human gp B (ADRV) rotaviruses (41.3 and 16.3%, respectively); (ii) the predicted secondary structure was highly conserved among the gene 5 proteins of the bovine, porcine, and human gp C rotaviruses; and (iii) seven highly conserved regions were identified for the first time in the deduced primary amino acid sequences of gene 5 of gp C and gene 6 of gp A rotaviruses. However, only three of these highly conserved areas were present in the regions of VP6, where the secondary structure was predicted to be similar for the rotavirus strains examined. These three regions may contribute to common epitopes between the two groups of rotaviruses. Our results, in comparison with data for other rotaviruses, indicate that gene 5 of the bovine gp C rotavirus codes for the major inner capsid protein (VP6).


Molecular cloning of a novel human papillomavirus (type 60) from a plantar cyst with characteristic pathological changes. In a plantar cyst composed of the wall of the squamous cell layer and the horny inner substance in the lower dermis, we found characteristic pathological changes, such as cytoplasmic eosinophilic inclusions and vacuolated structure, and, immunohistochemically, the papillomavirus capsid antigen. The human papillomavirus (HPV) DNA cloned from the cyst showed no homology with other known prototypes of HPV (HPV 1 through HPV 59) by Southern blot analysis under stringent conditions and was named as HPV 60. HPV 60 DNA was found in three other cases of plantar cyst with the identical pathological changes, but not in a plantar cyst without such changes. The results suggest that HPV 60 has unique biological properties to induce a plantar cyst as a distinct type of cutaneous HPV.


Characterization of an oligomerization domain and RNA-binding properties on rotavirus nonstructural protein NS34. Intermolecular interactions between polypeptide chains often play essential roles in such biological phenomena as replication, transcription, translation, transport, ligand binding, and assembly. We have initiated studies of the functions of the rotavirus SA114F gene 7 product by sequence analysis and expression in insect cells. This nonstructural protein, NS34, is a slightly acidic protein, and its secondary structure is predicted to be 78% alpha-helix, with several heptad repeats of hydrophobic amino acids being present in its carboxy half. NS34 was found in oligomers when analyzed in insect cells, in SA11-infected MA104 cells, and in cell-free translation reactions. Investigation of the multiple electrophoretically distinct forms of NS34 showed they were all composed of homooligomers. Deletion mutants constructed and tested for oligomerization showed that the carboxy terminus of the protein, containing the predicted heptad repeats, was responsible for oligomerization. A basic region present in NS34 of group A rotaviruses, found to be 40% conserved in NS34 of group C rotavirus, is a candidate for a functional domain of this protein. NS34, which was found to be associated with the cytoskeleton fraction of cells, also interacts with viral RNA. These results make it likely that NS34 plays a central role in the replication and assembly of genomic RNA structures.


Involvement of FOS and JUN in the activation of visna virus gene expression in macrophages through an AP-1 site in the viral LTR. Gene expression of visna virus is highly restricted in monocytes, but is induced when monocytes differentiate into macrophages. A previous study on differential regulation of visna virus gene expression revealed that a specific AP-1 site in the long terminal repeat of the viral DNA is required for phorbol-ester-induced gene expression in macrophages (Gabuzda, Hess, Small, and Clements, Mol. Cell. Biol., 9, 2728-2733). In the present investigation, we examined the association of two DNA binding proteins, the proto-oncogene proteins FOS and JUN, with this AP-1 site in the visna virus LTR. We demonstrated that the concentrations of these two proteins and their mRNAs increased in U937 cells after phorbol ester induction. Furthermore, the binding of cellular proteins from the U937 nuclear extracts to this AP-1 site was significantly decreased in the presence of antibodies to JUN and FOS. In vitro-translated JUN protein also binds to this AP-1 sequence, and this binding is enhanced by the FOS protein. These results indicate that JUN and FOS are directly involved in the differential regulation of visna virus gene expression.


Genetic evolution and tropism of transmissible gastroenteritis coronaviruses. Transmissible gastroenteritis virus (TGEV) is an enteropathogenic coronavirus isolated for the first time in 1946. Nonenteropathogenic porcine respiratory coronaviruses (PRCVs) have been derived from TGEV. The genetic relationship among six European PRCVs and five coronaviruses of the TGEV antigenic cluster has been determined based on their RNA sequences. The S protein of six PRCVs have an identical deletion of 224 amino acids starting at position 21. The deleted area includes the antigenic sites C and B of TGEV S glycoprotein. Interestingly, two viruses (NEB72 and TOY56) with respiratory tropism have S proteins with a size similar to the enteric viruses. NEB72 and TOY56 viruses have in the S protein 2 and 15 specific amino acid differences with the enteric viruses. Four of the residues changed (aa 219 of NEB72 isolate and aa 92, 94, and 218 of TOY56) are located within the deletion present in the PRCVs and may be involved in the receptor binding site (RBS) conferring enteric tropism to TGEVs. A second RBS used by the virus to infect ST cells might be located in a conserved area between sites A and D of the S glycoprotein, since monoclonal antibodies specific for these sites inhibit the binding of the virus to ST cells. An evolutionary tree relating 13 enteric and respiratory isolates has been proposed. According to this tree, a main virus lineage evolved from a recent progenitor virus which was circulating around 1941. From this, secondary lineages originated PUR46, NEB72, TOY56, MIL65, BR170, and the PRCVs, in this order. Least squares estimation of the origin of TGEV-related coronaviruses showed a significant constancy in the fixation of mutations with time, that is, the existence of a well-defined molecular clock. A mutation fixation rate of 7 +/- 2 x 10(-4) nucleotide substitutions per site and per year was calculated for TGEV-related viruses. This rate falls in the range reported for other RNA viruses. Point mutations and probably recombination events have occurred during TGEV evolution.


[The etiological structure and epidemiological characteristics of viral hepatitis among servicemen]. Selected studies on etiological structure of hepatitis A were conducted among servicemen, using modern high-sensitive orrhological methods of diagnosis. 325 serums obtained from infectious patients were examined in the laboratory. On the basis of this analysis the authors make a conclusion concerning a rather high level of fecal-oral viral hepatitis among servicemen of the 40th Army, which differed from hepatitis A.


Loss of V protein expression in human parainfluenza virus type 1 is not a recent event. The P gene of paramyxoviruses usually contains an alternate overlapping ORF coding for a short cysteine-rich domain called V. This domain was thought to be a common feature of these genes as it is present in ten paramyxoviruses. However, the V ORF region of the P gene of one strain of human parainfluenza-virus type 1 (isolated in 1957) was recently found to be closed by no less than 9 stop codons. To determine whether the absence of the V ORF here might be due to the long adaptation of this strain in culture, 3 other more recent isolates were examined. Small differences in the V region were found, but all had conserved the vast majority of the stop codons. Moreover, examination of most of the intercistronic regions of the PIV1 genome failed to uncover a cryptic V gene. The V ORF is then unlikely to be a common feature of paramyxovirus P genes.


Gene translocations in poxviruses: the fowlpox virus thymidine kinase gene is flanked by 15 bp direct repeats and occupies the locus which in vaccinia virus is occupied by the ribonucleotide reductase large subunit gene. By sequencing a fragment of 7351 bp the fowlpox virus thymidine kinase gene has been found to map to a position within the equivalent of the vaccinia HindIII I fragment. The deduced gene arrangement in fowlpox virus is I3, X, TK, I5, I6, I7, I8, G1, indicating that the homologue of the vaccinia I4 gene has been replaced by two genes X and TK. The non-essential TK gene has therefore replaced another non-essential gene, I4 (the ribonucleotide reductase large subunit) in this region. The X/TK insertion in fowlpox virus is precisely flanked by direct repeats of 15 bp suggesting that the translocation event may have involved transposition. The % identities between the fowlpox virus and vaccinia virus proteins ranged between 58.5% and 31.3%.


Mapping of the RD phenotype of the Nancy strain of coxsackievirus B3. The RD variants of group B coxsackieviruses differ from their parental strains in having the ability to replicate in a human rhabdomyosarcoma cell line, RD. The nucleotide sequence of the P1 region of the RD variant of coxsackievirus B3 strain Nancy (CB3NRD) was determined by sequencing cloned cDNAs, obtained by PCR amplification. A comparison between the established nucleotide sequence and that of the P1 region from the parental virus revealed 12 point mutations which corresponded to six amino acid replacements. To identify if the P1 region is responsible for the phenotype of CB3NRD, a chimeric virus was constructed, using an infectious cDNA clone of CB3. The P1 region of the infectious cDNA was replaced by cDNA fragments from CB3N (parental strain Nancy) or CB3NRD and the resulting recombinants were assayed for their ability to infect and replicate in RD cells. The results showed that the RD phenotype of CB3NRD maps in the P1 region. Furthermore, a chimera which only contained the 5' part of the P1 region derived from CB3NRD and the remaining P1 sequence from CB3N was able to replicate in RD cells, suggesting that the VP2 polypeptide contains at least one determinant for the RD phenotype.


[Electron spin resonance spectroscopy as probes for spices. Detection of treatment with ionizing radiation]. Detection of irradiated spices by electron spin resonance (ESR) measurements was not successful in the past because a central line of unknown origin was detected in the ESR-spectra of both irradiated and unirradiated samples. Identification of irradiated samples by measuring the increase of intensity of this signal after irradiation is limited because the signal intensity decreases over a period of some weeks of storage and reaches the range of unirradiated samples. By changing the measurement conditions (low microwave power) we could detect two additional lines on both sides of the main signal. This line pair appears only in the spectra of irradiated spices. A similar line pair was found in the spectra of irradiated nutshells and possibly derives from cellulose radicals in the sample. For some spices, especially paprika, the identification of irradiated samples by detecting these additional lines was possible even after relatively long periods of storage.


Direct effect of ACTH on renin release in isolated perfused guinea-pig kidneys with adrenal glands. To investigate the direct effect of corticotropin (ACTH) on the renin-angiotensin-aldosterone system, isolated guinea-pig kidneys with adrenal glands were perfused with various doses of ACTH (0.1-1000 micrograms/l) and 0.3 mmol/l of dibutyryl cyclic AMP (cAMP) through each cannula inserted into the abdominal aorta and the inferior caval vein. Perfusate renin activity was increased in a dose-dependent manner by the addition of ACTH in a range of 0.1-1000 micrograms/l, and reached a plateau at 20 min with each dose. The perfusate cAMP level was dose-dependently increased with 10-1000 micrograms/l of ACTH. Perfusate renin activity was also markedly increased by the addition of dibutyryl cAMP. The same effects of ACTH on renin and cAMP secretions were observed in the kidney perfusion model from which the adrenal glands were excluded. Aldosterone secretion failed to respond to 0.1 micrograms/l of ACTH, and was increased by higher concentrations (1-1000 micrograms/l) in the same experiments. These results demonstrate that ACTH has a direct effect on renal renin release in a physiological concentration (0.1 micrograms/l), and that the action of ACTH is probably mediated by cAMP. The sensitivity of renin release to ACTH stimulation is no less than that of aldosterone secretion during ACTH infusion, so it is possible that ACTH is an important stimulator of the renin-angiotensin system.


Increase in the expression of thyroid hormone receptors in porcine granulosa cells early in follicular maturation. Thyroid hormone has been demonstrated to synergize with FSH to exert stimulatory effects on the differentiation of porcine granulosa cells. In order to further characterize the nature of thyroid hormone action on granulosa cells, the presence of triiodothyronine (T3) receptors in the nuclei of porcine granulosa cells was examined, and qualitatively and quantitatively compared during follicular maturation. Then, comparative abilities of granulosa cells from varying follicle stages to respond to T3 were assessed in terms of FSH-induced LH/hCG receptor formation and progesterone secretion. Furthermore, the expression of erb-A was analyzed using Northern blot hybridization of porcine granulosa cell RNA with a v-erb-A probe. Binding experiments with [125I]T3 showed that granulosa cell nuclei obtained from small follicles had a greater ability to bind [125I]T3 compared to those from large follicles. Scatchard analysis revealed the presence of nuclear T3 receptors with a single class of binding sites. There was little difference in the affinity of the T3 receptors during follicular maturation. By contrast, the number of the T3 receptors was higher in small follicle granulosa cells compared to that in large follicle granulosa cells. Thus, the increased T3 binding to small follicle granulosa cells relative to large follicle granulosa cells appears to be attributable to the increased number of the nuclear T3 receptors rather than to a change in the affinity. The magnitude of the stimulatory effects of T3 on granulosa cell functions was maximal in small follicle granulosa cells, but negligible in large follicle granulosa cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Arginine vasotocin binding component in the uterus (shell gland) of the chicken. The plasma membrane fraction of the uterus of the chicken was found to contain a component that shows specific binding to arginine vasotocin (AVT), arginine vasopressin (AVP) and oxytocin (OT). This binding component possesses a higher affinity to AVT than to AVP or OT, and the affinity to AVT was higher in laying hens than in non-laying hens and immature pullets, while the maximum number of binding sites per mg protein was less in the laying hens. Intramuscular injections of either estradiol-17 beta, progesterone or testosterone into the immature pullets for six consecutive days caused an increase in the affinity and number of binding sites. The results suggest that AVT receptors are present in the chicken uterus and that their binding properties are affected by ovarian steroid hormones.


Immunolocalization of cystatin A in neoplastic, virus and inflammatory lesions of the uterine cervix. Cystatin A was immunohistochemically demonstrated in the normal squamous epithelium of the uterine cervix, particularly in the parabasal and superficial cell layers whereas it was absent or scanty in the basal cells and in areas with parakeratosis. Cystatin A was also found in neoplastic lesions (dysplasia, carcinoma in situ and squamous cell carcinoma), but less abundant than in normal squamous epithelium. The immunoreaction in intraepithelial neoplasia was closely related to the degree of morphological maturation of the squamous cells with more abundant cystatin A in low grade dysplasia and less in high grade dysplasia and carcinoma in situ. In squamous cell carcinoma, cystatin A was often abundant in highly differentiated areas and almost absent in poorly differentiated ones. Cystatin A was found in the squamous epithelium in herpes and in condylomatous lesions. It was also found in the cytoplasm of neutrophils, but not in lymphocytes and plasma cells. In unspecific cervicitis, cystatin A was found extracytoplasmatically as small vesicles in the epithelial-stromal junction. The implications of cystatin A in neoplastic, virus, and inflammatory processes are discussed.


Cytochemical investigation of p-NPPase activity in rat cardiac muscle. The cytochemical demonstration of the atrial cardiac myocyte pumping activity has been made by detecting p-nitrophenylphosphatase (NPPase), which is used by investigating of Na(+)-K(+)-ATPase and H(+)-K(+)-ATPase activities. The fine ultrastructural localization of these enzymes was studied using cytochemical methods with cerium as a capturing agent. Na(+)-K(+)-ATPase was localized on the atrial muscle cell plasma membrane, T-tubule membrane, endothelial cell nuclear membrane, and cardiac myocyte nuclear membrane. H(+)-K(+)-ATPase was localized on the atrial muscle cells plasma membrane, T-tubule membrane, and sarcoplasmatic reticulum. The present findings indicate that the transporting metabolic activity of the heart as an endocrine organ is realized by the interaction between p-NPPases.


Expression of ras proto-oncogene related protein p21 in normal human skin and cutaneous tumours. The cellular proto-oncogene, ras, is known to play an important role in the regulation of cell growth and proliferation in normal and malignant conditions. The present study was undertaken to immunohistochemically examine the expression of ras protooncogene product p21 in normal human skin and some cutaneous tumours. In normal skin, the expression of p21 was found in sweat glands, sebaceous glands, capillary endothelium, and smooth muscles, while epidermis was devoid of reaction product. Keratoacanthoma and the granular cells of verruca vulgaris were immunoreactive to the antibody for p21. Bowen's disease and squamous cell carcinoma were positive for p21, but basal cell carcinoma and seborrheic keratosis were negative. In mammary and extramammary Paget's diseases, the immunoreactivity was inconsistent. The expression of p21 in malignant melanoma cells was intense, whereas normal melanocytes and nevus cells were devoid of the expression. These results suggest that the expression of p21 does not correlate with nuclear anaplasia and malignant behaviour of cutaneous tumours.


Cytochrome oxidase in rat ocular tissues with special reference to copper. The localization of cytochrome oxidase, one of the copper metalloenzymes, was determined histochemically and compared with that of copper. Copper and cytochrome oxidase coexisted in the corneal epithelium and endothelium, iris, ciliary body, lens epithelium, and retinal photoreceptor inner segment. In spite of the presence of copper, no cytochrome oxidase was demonstrated histochemically in the retinal pigment epithelium, choroid, sclera, or optic nerve. The coexistence of copper and cytochrome oxidase suggests that copper plays a role in this copper metalloenzyme, while the non-coincidence of localizations of copper and cytochrome oxidase may be attributed to histochemical problems or to some unknown function of copper.


De novo detrusor instability after Burch colposuspension. After successful surgery for stress incontinence, detrusor instability can develop in a previously stable bladder. A review of six studies with a total of 396 patients is presented. Sixty-eight of these patients (17%) developed de novo DI. The prevalence varied from 5 to 27% in the different studies. The highest cure rate was 98%; the lowest 64%. It is concluded that the prevalence of de novo DI may be dependent on the details of the cystometric and surgical technique as well as unknown patient characteristics. Since the exact pathophysiologic mechanism of this phenomenon is unknown, it is difficult to predict it preoperatively in the individual patient.


Expression of receptors for tumor necrosis factor in human placenta at term. The biological effects of tumor necrosis factor (TNF) are mediated through its interaction with high affinity receptors on target cells. Secretion of soluble cytokine receptors has been suggested as a mechanism of regulating cytokine activity in vivo. In a previous study we detected soluble TNF receptors (TNFRs) in amniotic fluid and urine samples from pregnant women, suggesting that secretion of soluble TNFRs may provide a mechanism for protection of the fetus against TNF action during pregnancy. In the present study, TNFR containing cells in cryostat sections from normal placentas at term were evaluated by monoclonal antibodies against the 55 kD--and the 75 kD TNFR in an indirect immunofluorescence technique. The 55 kD TNFR was expressed by the villous syncytiotrophoblasts, by vascular endothelial cells, by some decidual cells and by occasional cells in the placental stroma. Staining for the 75 kD TNFR was confined to the vascular endothelial cells, a relatively small number of stromal cells and decidual cells, whereas the villous syncytiotrophoblasts were negative. The abundant expression of TNFRs in placental tissue suggests: 1. That a considerable number of the placental cells are receptive to the regulatory activities of TNF; 2. That placental cells may be the cellular origin of soluble TNFRs secreted during pregnancy.


Dietary habits, prepregnancy weight, and weight gain during pregnancy. Risk of pre term rupture of amniotic sac membranes. Premature rupture of amniotic sac membranes (PROM) is associated with perinatal morbidity and mortality. A matched case-control study was undertaken to determine whether dietary habits, prepregnancy weight, and weight gain during pregnancy were associated with increased risks for PROM. Three study groups were defined, consisting of women delivering pre term with PROM (PP), full term with PROM (FP), and pre term without PROM (PWP). Cases were singly matched by race, age, and parity with women having full term deliveries without PROM. Data were collected by face-to-face structured interviews with eligible subjects and by medical records abstraction. Conditional multiple logistic regression indicated that during pregnancy, PP cases were significantly more likely to have gained less than 21 pounds (9.5 kg) (OR = 2.7, CI = 1.14-6.36) and were only half as likely as controls to have improved their diet (OR = 0.43, CI = 0.18-0.99) while controlling for cigarette smoking, urinary tract infection, chorioamnionitis, chlamydia, and a history of previous PROM. FP cases were significantly less likely than controls to have gained 31-40 pounds (14-18 kg) while pregnant (OR = 0.56, CI = 0.33-0.94) and were significantly more likely than controls to have a somewhat inadequate pregnancy diet (OR = 2.05, CI = 1.11-3.77) while controlling for a history of previous PROM. PWP cases were significantly less likely to have gained 31-40 pounds (14-18 kg) (OR = 0.42, CI = 0.21-0.84) or greater than 40 pounds (18 kg) (OR = 0.37, CI = 0.17-0.80) while pregnant or to have had adequate dairy products intake (OR = 0.60, CI = 0.36-0.99) while controlling for cigarette usage. Maternal dietary habits, weight gain during pregnancy, and supplement intake are associated with the occurrence of both pre term and full term PROM and pre term delivery without PROM.


Conservative treatment of mild and moderate hypertension in pregnancy. In a prospective study regarding conservative treatment of mild to moderate hypertension in pregnancy 5244 women were evaluated at delivery. Two hundred and fifty-eight (4.9%) were registered as hypertensive during pregnancy with a blood pressure (BP) greater than or equal to 140/90 mmHg. One hundred and ninety-six of these continued their pregnancy without medication and 96/196 were defined as preeclamptic (PE), 45/196 as chronic hypertensive (CH), and 55/196 as having gestational hypertension (GH). In 62/258 women antihypertensive treatment was initiated in the mean 6.5 (+/- 8.7) days after onset of hypertension, due to a BP greater than or equal to 150/100 mmHg. There was a later onset of hypertension in the untreated group, and BP at delivery differed in the untreated groups (p less than 0.001) with the lowest BP in women with mild GH. There was no difference in cesarean section rate in the mild hypertensive group as compared to the normal population. Birth weight and length of pregnancy were significantly lower in the untreated mild hypertensive group as compared to normal pregnant women (p less than 0.05). But in the subgroup with mild GH pregnancy length and birth weight did not differ from normal pregnancy. The conclusion from our study is that women with mild hypertension in pregnancy might refrain from antihypertensive therapy if they are closely observed during pregnancy and delivery, especially if there has been no hypertension before pregnancy and no proteinuria develops.


Organisation of obstetric ultrasound in Denmark 1990. To provide an overview of the current organisational status of obstetric ultrasound in Denmark, a questionnaire was sent in spring 1990 to all 59 departments responsible for the hospital antenatal care program and to the 25 imaging diagnostic departments providing ultrasound services for those of the 59 departments that had either insufficient or no ultrasound facilities. The response rate was 100%. Obstetric ultrasound was performed in 25 obstetric, 11 surgical, 22 radiology, 2 clinical physiology departments and 1 ultrasound laboratory (total 61). The departments had a total of 110 ultrasound scanners. The median year of purchase of these scanners was 1985 (range 1976-1990). 488 persons performed ultrasound examinations; the examiners were mainly doctors (72%). 40% of the pregnant population was routinely offered an ultrasound scan in the 15-20th week of gestation. Of these 2/5 were offered screening for fetal malformations. One department offered examinations in both the 19th and 34th week. The scanning capacity, as judged by the departments themselves, was also assessed through the study.


Monitoring endometrial adenocarcinoma with a four tumor marker combination. CA 125, squamous cell carcinoma antigen, CA 19.9 and CA 15.3. The combined value of four tumor markers, in the follow-up of endometrial adenocarcinoma, is analyzed. Cancer antigen 125 (CA 125), squamous cell carcinoma antigen (SCC), carbohydrate antigen 19.9 (CA 19.9) and carbohydrate antigen 15.3 (CA 15.3) were used in 213 evaluations from 105 patients. Sensitivity as regards recurrence or progression of disease was 45% (CA 125), 9% (SCC), 51% (CA 19.9) and 21% (CA 15.3). Specificities as regards the 'no evidence of disease' ranged from 95% to 99%. Single tumor marker efficiency ranged from 90% for CA 125 to 84% for SCC (p = 0.08). With the two tumor marker combination sensitivity increased up to 77% achieved with CA 125-CA 19.9, but efficiency increased only slightly (92.0% for CA 125-SCC). In the best three tumor marker combination, a sensitivity of 85% was achieved (CA 125-CA 19.9-CA 15.3), and an efficiency of 92.2%. The simultaneous use of the four tumor markers did not improve assay results. The possibility of recurrence or progression of disease in some combinations was very low (4.6% when CA 125 and CA 19.9 negative, 3% when CA 125, CA 19.9 and CA 15.3 negative), a fact to be considered in order to avoid aggressive management in such cases. The tumor markers were of limited value for the prediction of recurrences. The suggestion of recurrence when the increase in tumor markers was the only finding was confirmed in only 7%, while confirmation was made in 100% when there was another pathological finding.


Clinical evaluation of contact Nd.YAG laser conization for cervical intraepithelial neoplasia of the uterus. Contact Nd.YAG laser conization (laser cone) was performed in 500 patients with preoperatively diagnosed cervical intraepithelial neoplasia (CIN) since September 1983. The patients treated were followed for 12 to 96 months (39.2 months on average). Average operation time was 11.4 minutes. A cure with laser cone (single treatment) was achieved in 98.6%, including cases with microinvasive and invasive carcinoma, post-operatively. There were only 7 (1.4%) true residual cases of 47 cases (9.4% of total cases) with incomplete excision. High spontaneous cure rates (85.1%) in cases with incomplete excision are considered to be a characteristic of laser cone treatment. The contact laser cone method also got good specimens for histologic examinations. They gave a large amount of information for our follow-up. The results indicate that contact Nd.YAG laser conization for CIN is an excellent conservative therapy from the point of cure rate, safety, indication, operation time and cone specimen, even compared with CO2 laser conization.


Comparison of low molecular weight heparin vs. unfractionated heparin in gynecological surgery. II: Reduced dose of low molecular weight heparin. In a double blind, randomized trial the hemorrhagic complications of a reduced dose of low molecular weight heparin (LMWH) (Fragmin, KabiPharmacia) were compared to those of the conventional dose of unfractionated heparin (UH). 2500 anti-XaU of LMWH was given once daily and UH in a dose of 5000 anti-XaU twice daily. During a one year period 141 patients undergoing gynecological surgery were included in this study. The patients were examined clinically for hematomas and for deep venous thrombosis (DVT) on the third and fifth day. Venography was performed when DVT was suspected. No patients developed clinical DVT. One woman in the LMWH group had pulmonary embolism 3 days after the prophylaxis was stopped. Two women in the LMWH group died, one from a stroke on day 2, one from cancer on day 39. There was no significant difference in serious bleeding complications between the two regimens, 20% in the LMWH group and 14% in the UH group. Even with the reduced dose of LMWH the mean plasma concentration of heparin in the LMWH group was higher (mean 0.14 anti-XaU/ml) than in the UH group (0.029 anti-XaU/ml) 3 hours after injection on the 2nd postoperative day. A reduced dose of LMWH (2500 anti XaU once daily) does not cause more bleeding complications than the conventional heparin regimen to prevent thrombosis, as was the case in our previous study with 5000 anti XaU of LMWH once daily.


Fetal abdominal cavernous hemangioma diagnosed by duplex Doppler velocimetry. A fetal abdominal wall mass was detected by ultrasound and analyzed by duplex Doppler showing a low resistance to blood flow through its vessels. The antenatal diagnosis of cavernous hemangioma was confirmed after birth.


Choriocarcinoma of the cervix.

Choriocarcinoma of the cervix. The incidence of primary cervical choriocarcinoma is extremely low, with only approximately 60 cases reported. We present a case of primary cervical choriocarcinoma, which was originally mis-diagnosed as a cervical pregnancy. After failure of conservative treatment with methotrexate, the patient underwent a hysterectomy, followed by chemotherapy with actinomycin-D. After twenty months of follow-up, the patient is in good condition and her serum beta-hCG levels are normal.


Leiomyomatosis peritonealis disseminata with adipocytic differentiation. We report a rare autopsy case of leiomyomatosis peritonealis disseminata in a woman, who had been operated upon for ovarian granulosa cell tumor two years prior to her death. The deposits showed both myofibroblastic and adipocytic differentiation.


Benign mixed Müllerian tumor of the ovary. A case of benign Müllerian tumor of the ovary in a 53-year old woman is reported. The stromal element consisted of rhabdomyoblasts which clearly indicate that this tumor belongs to the Müllerian family. She remains free of disease 12 months following an abdominal hysterectomy and bilateral salpingo-oophorectomy.


Preparation and in vitro antiviral activity of liposomes of lipophilic esters of acyclovir. The long chain acyclovir such as the acyclovir laurate and acyclovir palmitate were prepared directly from acyclovir by application of the usual esterification methods with appropriate acyl chlorides. The lipophilic prodrugs were found to be retained easier by liposomes whereas acyclovir escaped readily from liposomes. When assayed in African green monkey cell cultures against herpes simplex virus type I strain, the acyclovir palmitate liposomes proved to be more active compared with the parent drug and its liposome, suggesting an enhanced compatibility between the ester and liposomal lipids and an increased uptake of encapsulated prodrug by infected cells.


Influence of angiotensin II, alpha- and beta-adrenoceptors on peripheral noradrenergic neurotransmission in canine gracilis muscle in vivo. Interactions between angiotensin II and adrenoceptor-mediated effects on peripheral sympathetic neurotransmission were investigated in constant flow blood-perfused canine gracilis muscle in situ, without and with pretreatment by non-competitive alpha-adrenoceptor blockade. Angiotensin converting enzyme (ACE)-inhibition by benazeprilat increased nerve stimulation (2 Hz, 4 min)-evoked noradrenaline (NA) overflow (+ 21 +/- 5%) with alpha-adrenoceptors intact, but reduced NA overflow (- 18 +/- 6%) when alpha-adrenoceptors were blocked. Vasoconstrictor responses were slightly reduced by benazeprilat. Subsequent infusion of angiotensin II (Ang II, 20 and 500 ng kg-1 min-1 i.v., raising arterial concentrations from 0.6 +/- 0.2 pM to 1390 +/- 240 and 25,110 +/- 3980 pM, respectively) failed to increase NA overflow or to enhance stimulation-evoked vasoconstriction. Adrenaline (0.4 nmol kg-1 min-1 i.v.) did not change evoked NA overflow before or after benazeprilat, either with or without alpha-adrenoceptor blockade, despite high concentrations (approximately 10 nM) in arterial plasma. Following benazeprilat, propranolol reduced NA overflow (- 24 +/- 3%) only if the alpha-adrenoceptors were blocked. In conclusion, benazeprilat reduced evoked NA overflow in the presence of alpha-adrenoceptor blockade to a similar degree as previously shown in the presence of neuronal uptake inhibition in this model. However, contrasting to our previous findings, benazeprilat enhanced NA overflow and reduced the post-junctional response to nerve stimulation in the absence of alpha-adrenoceptor blockade. This could be related to bradykinin accumulation during ACE-inhibition, in addition to the reduction of Ang II generation. Our data are not compatible with facilitation of NA release by circulating Ang II even at pharmacological dose levels. Although activation of prejunctional beta-adrenoceptors may facilitate evoked NA overflow in this model, circulating adrenaline is ineffective under physiological conditions even after alpha-adrenoceptor blockade. Also, beta-adrenoceptor-mediated prejunctional effects do not seem to involve Ang II in canine skeletal muscle in vivo.


Activation of the Na-K pump by intracellular Na in rat slow- and fast-twitch muscle. Experiments were performed on isolated rat soleus (slow-twitch) and extensor digitorum longus (EDL) (fast-twitch) muscle of 4-week-old rats. In soleus muscle, electrical simulation at 2 Hz for 5 min increased the ouabain-suppressible 86Rb+ uptake by 138%, without significant changes in intracellular Na+ content or Na+/K+ ratio. In EDL muscle, the ouabain-suppressible 86Rb+ uptake was stimulated by only 58%, whereas intracellular Na+ content and Na+/K+ ratio were increased by around 70%. Na(+)-loading of the muscles by exposure to K(+)-free or K(+)-Ca(2-)-Mg(2+)-free buffer stimulated the ouabain-suppressible 86Rb+ uptake in the two muscles to roughly the same extent, but in EDL muscle this was associated with a more than twofold larger increase in Na+/K+ ratio. When the Na+ influx was increased by exposure to veratridine similar results were obtained. Graded variation in intracellular Na+ content was achieved by exposure to monensin. In soleus muscle, a 25% increase in intracellular Na+/K+ ratio resulted in a doubling of the ouabain-suppressible 86Rb+ uptake, whereas a doubling of the Na(+)-K+ transport rate in EDL muscle required a 140% increase in Na+/K+ ratio. The results indicate that in soleus muscle the Na+/K+ pump is much more sensitive to changes in intracellular Na+ content than in EDL muscle. This might explain the larger activation of the Na(+)-K+ pump in slow-twitch muscle during electrical stimulation and might be of significance for the activation of the Na(+)-K+ pump in vivo during work.


Modulation of Ca2+ release and Na+ channel function in skeletal muscle by fatty acids. Ca(2+)- and halothane-induced Ca(2+) release and Na+ currents are modulated by free fatty acids (FAs). FA modulation of ion currents may have important implications for general muscle physiology and skeletal muscle disorders, including malignant hyperthermia (MH).


Ag(+)-induced inward current on frog skeletal muscle. Ag+ caused an inward current on voltage-clamped skeletal muscle. The current was carried by Ca2+, Mg2+, and Na+, and was blocked by Cd2+ or Ni2+ but not by nifedipine or D600. Channel gating is supposed to be modified by Ag+.


Ca2+ influx through voltage- and purinoceptor-operated channels estimated from [Ca2+]c signals (myocytes from guinea-pig urinary bladder). The rise in free cytosolic calcium was studied by a combination of the techniques of microspectrofluorometry and whole-cell patch clamp. By comparing the membrane currents with their effect on [Ca2+]c, the relative importance of Ca2+ influx could be quantified for both L-type Ca2+ channels and non-selective channels activated by extracellular ATP.


Citrate decreases Ca current and changes selectivity of cardiac Ca channels. Previous studies using citrate as a low affinity Ca buffer have shown that this agent inhibits myocardial contractions and neurotransmitter release. Using several different low affinity Ca buffers we have demonstrated that these effects are unlikely to be attributable to Ca buffering per se. Here we demonstrate that citrate can directly inhibit Ca current (ICa). The decrease in ICa appears to be due to shifts in channel gating (due to surface charge effects) and also to a decrease in channel selectivity for Ca over monovalent ions.


Effects of diaphragmatic fatigue on phosphoinositide hydrolysis. We evaluated the effects of diaphragmatic fatigue on muscle phosphatidylinositol phosphate (PIP) metabolism. Results revealed that the total PIP pool was 76% greater in fatigued rat diaphragms compared to controls, which suggests that fatigue was associated with inhibition of sarcolemmal membrane bound PIP hydrolysis.


The influence of superoxide scavenging compound CTC 23 on type II collagen-induced arthritis in mice. The anti-arthritic activity of the superoxide scavenging compound CTC 23 was tested in murine type II collagen-induced arthritis (CIA). CTC 23 demonstrated a dose-dependent prevention of the onset of CIA, with a significant reduction of disease incidence in mice receiving prophylactic administration of CTC 23 from 2.5-5 mg/kg/day. CTC 23 also delayed the development of the anti-collagen immune response, causing a significant decrease in antibody levels 14 days after immunization. CTC 23 reduced the arthritis score and number of involved limbs in mice with established CIA. The results suggest that CTC 23 acts as a disease modifying anti-arthritic compound to prevent the progression of the experimental arthritis.


Effects of nedocromil sodium on the binding of N-formyl-methionyl-leucyl-phenylalanine in human neutrophils. In the present study the inhibition by nedocromil sodium of the specific receptor binding of FMLP was evaluated in human neutrophils (PMNs) using a FMLP-(3H) binding assay. The time course of the binding was markedly influenced by nedocromil sodium used at a concentration of 300 microM. No significant inhibition was obtained when the cells were treated with nedocromil sodium 3 microM or with sodium cromoglycate 300 microM. FMLP binding is essentially eliminated by the highest dose of nedocromil sodium. The biologic meaning of this effect in asthmatic patients should be further evaluated.


Protease and protease inhibitor secretion by postsurgical macrophages following in vitro exposure to tolmetin. Tolmetin sodium was shown previously to reduce the level of adhesions formed after peritoneal surgery. In addition, when tolmetin was administered to rats at the time of surgery, the level of macrophage activity (phagocytosis, superoxide anion production and tumoricidal activity) was elevated. In this study, the level of neutral protease activity in the culture supernatant of peritoneal macrophages from postsurgical rabbits. after in vitro exposure to tolmetin was examined. The level of collagenase, elastase and plasminogen activator inhibitor activity in the culture supernatant of macrophages was modulated after peritoneal surgery. Collagenase activity in the culture supernatant of macrophages from nonsurgical and postsurgical rabbits up to 48 h was reduced after co-culture with tolmetin. In contrast, elastase secretion was elevated to maximal levels (which plateaued 24 h after surgery in cultures of control macrophages) in cultures of macrophages from nonsurgical and early postsurgical (6 and 12 h) rabbits by in vitro exposure to tolmetin. Alternatively, the level of plasminogen activator inhibitor activity in the culture supernatants of postsurgical macrophages was reduced after in vitro exposure to tolmetin. These data indicate that tolmetin, a nonsteroidal anti-inflammatory agent, modulates the levels of collagenase, elastase and plasminogen activator inhibitor activity in culture supernatants of postsurgical macrophages.


N-acetylcysteine enhances receptor-mediated phagocytosis by human neutrophils. The effect of the sulphur compound N-acetylcysteine (NAC) on certain receptor-mediated cellular functions [chemiluminescence (CL), phagocytosis and degranulation] in human neutrophils was studied, to evaluate how a scavenger of certain toxic oxygen product can protect the phagocyte and the bystander tissue cells from oxidative damage. When using IgG-opsonized yeast particles as stimulating agent, preincubating the neutrophils with NAC (0.25 mg/ml = 1.5 mM) increased both the CL response and phagocytosis. Higher concentrations of NAC (0.50-1.00 mg/ml = 3-6 mM) decreased the CL response, whereas the phagocytic capacity was still enhanced. This effect was more pronounced with adherent neutrophils than with neutrophils in suspension. No increased CL or phagocytic activity was, however, induced by NAC when C3bi-opsonized particles were used as a prey. From the fact that NAC (i) inhibited extracellularly localized myeloperoxidase dependent activities, and (ii) had no effect on neutrophils from patients with chronic granulomatous disease (CGD), we conclude that the scavenger effect of NAC not only reduces the accumulation of oxidative metabolites per se, but also enhances receptor-mediated phagocytosis by protecting Fc(IgG)-receptors from oxidative damage mediated by myeloperoxidase (MPO) and hydrogen peroxide (H2O2). Since NAC can increase phagocytosis and reduce the extracellularly produced oxidative metabolites, we furthermore conclude that NAC possesses some ideal properties as an anti-inflammatory agent.


[Two cases of mucinous adenocarcinoma of the prostate]. We report two cases of mucinous adenocarcinoma of the prostate. A 56-year-old man underwent subcapsular prostatectomy under the diagnosis of benign prostatic hyperplasia in 1968, and was found to have mucinous adenocarcinoma of the prostate, which proved to be prostatic acid phosphatase (PAP) and prostate specific antigen (PSA) positive, and carcinoembryonic antigen (CEA) negative by immunohistochemical staining. Subsequently he received 70 Gy of irradiation to the prostate, but died in 1976, when serum PAP was elevated. Autopsy revealed metastases to the liver, lungs, bone, peritoneum, spleen, pancreas, lymph nodes, and no primary gastrointestinal adenocarcinoma. The other case was a 57-year-old man, who underwent transurethral resection (TUR) for papillary tumor located just lateral to the verumontanum in 1982. The tumor was misdiagnosed as adenomatous polyp, and was PSA and PAP negative, and CEA positive. After 3 TURs of the recurrent tumor on the prostatic urethra, he underwent prostatourethrectomy, pelvic lymphadenectomy, and cystostomy for radical cure in 1985. The specimen proved to be mucinous adenocarcinoma of the prostate. He suffered recurrence of the tumor in the retrovesical space in 1987, and died in 1990. Autopsy revealed no evidence of metastasis except the local recurrence and no primary gastrointestinal adenocarcinoma.


CT of portal venous occlusion.

CT of portal venous occlusion. Thrombosis of a portion of the portal venous system can be directly imaged by contrast-enhanced CT as a low-attenuation lesion within the involved portal venous segment with or without expansion of the vessel or enhancement at the margin of the thrombus. Collateral venous pathways are often evident, which provide supporting evidence of the occlusion. Alterations in portal venous blood flow lead to metabolic disturbances in the liver and to abnormalities in parenchymal enhancement during dynamic CT scanning, and these changes are manifested as abnormalities in hepatic parenchymal density. The detection of portal venous thrombosis or occlusion, collateral veins, or abnormal liver enhancement should initiate a search for the diseases that cause these abnormalities.


Detection of malignant tumors in end-stage cirrhotic livers: efficacy of sonography as a screening technique. Patients with hepatic cirrhosis are at an increased risk of developing primary malignant tumors of the liver. If these tumors are discovered early, current therapies may be curative. We conducted a prospective study to assess the accuracy of sonographic screening for the detection of malignant tumors in cirrhotic livers as determined by correlation with resected whole livers. A total of 200 prospectively interpreted preoperative sonograms from 200 patients with cirrhosis who underwent hepatic transplantation were correlated with specimens of freshly resected whole livers. The results were analyzed to determine the sensitivity and specificity of sonography in identifying patients with malignant tumors and detecting individual tumors in each patient. Pathologic examination showed 80 malignant lesions in 34 patients (28 with hepatocellular carcinoma, three with cholangiocarcinoma, two with metastases, and one with non-Hodgkin's lymphoma) and three hemangiomas in two patients. Sonography correctly showed malignant tumors in 17 of the 34 patients, for a sensitivity of 50%. Sonograms were false-positive for malignant tumors in three patients, two of whom had a total of three hemangiomas. Sonography correctly showed 36 of the 80 malignant lesions, for a lesion sensitivity of 45% and specificity of 98%. Of the 44 missed lesions, 24 were 1 cm or less, 12 were between 1 and 3 cm, and eight were more than 3 cm in diameter. Our results show that sonography is highly insensitive in the detection of malignant lesions in end-stage cirrhotic livers and thus is not a reliable screening technique. However, because of sonography's very high specificity, any sonographically identified lesion in a cirrhotic liver should be considered malignant until proved otherwise.


Differential diagnosis of hepatic tumors (hepatoma, hemangioma, and metastasis) with CT: value of two-phase incremental imaging. Two-phase dynamic incremental CT is a technique in which CT scans are obtained 45 sec and 6 min after commencing the rapid bolus injection of contrast medium. We analyzed the contrast enhancement patterns of three types of hepatic tumors (72 hepatomas, 39 hemangiomas, and 28 metastases) in 139 patients to determine if any differences in the patterns are useful in the differential diagnosis of these lesions. Dynamic incremental CT scanning was performed after 100 ml of iodinated contrast material was administered i.v. with a power injector at a rate of 2 ml/sec. A 1-sec scanning time was used with a 1.6-sec inter-scan delay, which allowed table motion between scans. CT scans (eight to 16 sections) were obtained 45-110 sec (early phase) and 6-7 min (delayed phase) after commencing the injection of contrast medium. The enhancement patterns of hepatomas were as follows: 32% were totally hyperdense in the early phase and totally hypodense in the delayed phase, while 24% were totally hypodense in both phases. Most of the hepatomas (88%) appeared as totally hypodense lesions in the delayed phase. In the case of hemangiomas, 56% were peripherally hyperdense in the early phase; in the delayed phase, 36% were isodense and 31% were totally hyperdense. Most hemangiomas (85%) were not totally hypodense in the early phase, and no hemangioma was totally hypodense in both phases. In the early phase, 61% of metastases were hypodense. In the delayed phase, 57% were hypodense. Metastases most commonly were totally hypodense in both phases (43%). We conclude that contrast enhancement patterns of hepatomas, hemangiomas, and metastases seen on two-phase dynamic incremental CT scans are useful in the differential diagnosis of these tumors.


CT arterial portography: causes of technical failure and variable liver enhancement. We studied the causes of technical failure and enhancement variability encountered during CT arterial portography. CT arterial portograms and digital arteriograms were obtained via the superior mesenteric artery before partial liver resection in 43 patients with malignant tumors. These studies were reviewed for causes of technical failure and variable enhancement. Eleven (26%) of 43 procedures were technical failures. Causes of failure included aortic injection after catheter dislodgement (four), dense hyperenhancement associated with laminar flow in the portal vein produced by rapid venous return from a selective injection into a proximal branch vessel of the superior mesenteric artery (two), premature scanning beginning at the iliac crest (two), reflux into a replaced right hepatic artery (one), hepatic arterial enhancement via the pancreaticoduodenal arcade (one), and portal hypertension (one). Of the 32 remaining studies, 28 showed areas of parenchymal hypoenhancement or hyperenhancement. Causes of variable enhancement included impaired portal vein perfusion from mass effect of the tumor, laminar flow in the portal vein, and focal fatty infiltration. Technical failures and enhancement variability are common in CT arterial portography. Factors leading to technical failure include catheter choice and position, portal hypertension, and operator error.


Focal hyperdense areas in endometriomas: a characteristic finding on CT. To assess the value of a hyperdense focus seen on CT scans of endometrial cysts in the differential diagnosis of a lesion. The preoperative CT scans of 328 patients with 410 ovarian masses (54 patients with 62 pathologically proved endometriomas and 274 patients with 348 pathologically proved other ovarian masses) were retrospectively reviewed in a random fashion without knowledge of the pathologic findings to determine whether a hyperdense focus was visible inside a cyst. In nine of 62 endometrial cysts (sensitivity, 15%), CT scans showed a hyperdense round or crescent-shaped focus, measuring 2 to 15 mm. This focus was located close to the inner border of the cyst in eight cases and in the central part of the cyst in one case. A hyperdense focus was not seen on CT scans of 348 other ovarian masses (specificity, 100%). An in vitro CT study of two specimens showed that this hyperdense area corresponded to a blood clot next to the inner wall of the cyst. This hyperdense area appeared as a nonspecific hyperechogenic focus on sonograms in five of nine cases and as a hypointense signal on T1- and/or T2-weighted MR images in four of five cases. The finding on CT scans of a hyperdense focus inside an ovarian cyst is suggestive of endometrioma and should help distinguish endometrioma from other pelvic masses.


Epstein-Barr virus-associated anaplastic large cell lymphoma in renal transplant patients. A novel association, Epstein-Barr virus-positive Ki-1+/CD30+ anaplastic large cell non-Hodgkin's lymphoma of B-cell phenotype in immunosuppressed renal transplant recipients is reported. Case 1 involved an aggressive clinical evolution, whereas case 2 followed a more "benign" clinical course. Both lymphomas were Epstein-Barr virus-positive as assessed by in situ hybridization, Southern blot, polymerase chain reaction, and immunohistochemical analysis. Both lymphomas contained a single clonal Epstein-Barr virus terminal-repeat fragment. In case 1, clonality was confirmed by the detection of bi-allelic immunoglobulin (Ig) heavy chain gene rearrangement. Case 2 showed germline Ig genes at presentation and oligoclonal Ig heavy chain gene rearrangements at relapse. These results are consistent with the notion that anaplastic large cell lymphoma might arise in a B cell transformed by Epstein-Barr virus at a very early stage, before Ig gene rearrangement. The latter may occur later in the course of clonal evolution, thus permitting investigators to trace intermediate and late stages within a process of multistep lymphomagenesis and/or tumor progression.


Influence of Epstein-Barr virus genomes on patient survival in Hodgkin's disease. In previous studies, Epstein-Barr virus was considered a possible etiologic factor in Hodgkin's disease. Two hundred twenty-nine cases of Hodgkin's disease were investigated for the presence of Epstein-Barr virus DNA using the polymerase chain reaction technique on formalin-fixed, paraffin-embedded lymph node tissue to clarify the clinical importance of the incidence of this genome. In 42 cases (18.3%), genomic DNA was not amplifiable. The remaining 187 cases included the following subtypes: lymphocyte-predominant type (n = 13), nodular sclerosis type (n = 98), mixed cellularity type (n = 68), and lymphocyte-depleted type (n = 8). Sixty-six cases (35.2%) were positive for Epstein-Barr virus DNA. In the statistical analysis of available follow-up data from 130 patients, no influence of a positive Epstein-Barr virus DNA finding on length of survival time was revealed. This was true within the cohort of all patients and within the histologically defined subtypes of Hodgkin's disease. In this investigation, detection of Epstein-Barr virus DNA by polymerase chain reaction showed no prognostic relevance for patients with Hodgkin's disease.


Primary Epstein-Barr virus infection with clonal T-cell lymphoproliferation. A case of fatal Epstein-Barr virus infection in a previously healthy girl who was first found to have severe infectious mononucleosis with spontaneous recovery is reported. Because an abnormal immune response to the virus persisted, the disease relapsed, manifesting in cutaneous and pulmonary lesions associated with hemophagocytic syndrome responsible for death. Pathologic findings were characterized by polymorphous atypical lymphoid infiltrate, prominent necrosis, and histiocytic hyperplasia. Lymphoid cells displayed CD8 phenotype and clonal T-cell receptor gene rearrangement. Viral genome was detected in lesions by Southern blot and located in nuclei of lymphoid cells by in situ hybridization. Pathologic findings suggested fatal infectious mononucleosis; however, phenotype and genotype favored a malignant diagnosis. Clonality was demonstrated to have arisen during primary infection. Virologic examination indicated that Epstein-Barr virus was a causative agent. Such a process belongs to the recently recognized spectrum of Epstein-Barr virus-related T-cell lymphoproliferative disorders that might overlap fatal infectious mononucleosis in patients who are especially vulnerable to the virus.


Simian virus 40-transformed human ovarian surface epithelial cells escape normal growth controls but retain morphogenetic responses to extracellular matrix. We aimed to improve the availability of experimental models for the study of human ovarian surface epithelium. Low-passage cultures of human ovarian surface epithelium were transfected with SV40 large- T antigen and the resulting lines were characterized. Three immortalized lines were obtained. They formed epithelial monolayers resembling ovarian surface epithelium in serum-free medium, expressed large-T antigen and overexpressed p53, produced laminin, and were CA 125 negative. Two lines expressed keratin. On plastic surfaces, the growth rate and growth potential of immortalized ovarian surface epithelium were increased over the growth of ovarian surface epithelium, but on extracellular matrices their growth and morphologic features resembled ovarian surface epithelium. The lines were not tumorigenic in Nu/Nu mice. The immortalized ovarian surface epithelium lines resemble cells early in neoplastic progression and should be useful to study ovarian carcinogenesis.