Comparison of the properties of five pox virus strains isolated from monkeys

Summary Five strains of monkey pox viruses were compared with respect to their cultural characteristics in primary and continuous cell cultures and the lesions developed in embryonated eggs and in rabbit skin as well as to their hemagglutinating activity.Four strains (Copenhagen 65-31 65-32 and 7-61) appeared to be similar in their properties. The cytopathogenic effect (CPE) was identical to that induced by vaccinia virus. There was no detectable virus multiplication in an pig kidney cell line (PEK). All four strains produced small, white, compact, hemorrhagic pock-like lesions on the chorioallantoic membrane.The strain 64–7275, isolated from healthy monkeys kidneys, had all properties of variola virus. It multiplied in the PEK cell line with a CPE. The lesions on the CAM were more compact without hemorrhage. In rabbit skin no detectable reaction occurred after infection with this strain.     

Rapid and quantitative in vitro measurement of cellular chemotaxis and invasion

An in vitro, fluorimetric method for cellular chemotaxis and invasion has been developed using a commercially available, disposable, 96-well chamber. This 4–18 hour microtiter chamber assay has a number of important advantages over existing methods. It does not require prior labeling of cells or radioactivity, and is rapid, automatable and quantitative. Cells are quantitated by a novel actin-based fluorescence tag as reported previously (Methods in Cell Science 17: 263–270, 1995). Following quantitation, cells are easily detectable by fluorescence microscopy. In addition, this assay conserves reagents due to its low volumes in the upper and lower chambers. The assay has been optimized using cultured human lung cancer cells to identify inhibitors or activators of directed cell migration. The effects of antibodies to V3, V5, and CD44 on the chemotaxis and invasion of A549 cultured lung tumor cells are reported.     

Tapasin Decreases Immune Responsiveness to a Model Tumor Antigen

The T-cell response against cancer is dependent on the cell surface presentation of tumor-associated or tumor-specific peptides by major histocompatibility complex (MHC) class I molecules. We found that tapasin, a chaperone protein that normally assists in the assembly of MHC class I molecules, is undetectable in an unstimulated pancreatic tumor cell line, Panc02, and only very weakly expressed after -interferon stimulation. Transfection of tapasin into the Panc02 cells did not quantitatively increase MHC class I surface expression or detectably affect MHC class I association with peptide and ₂-microglubulin (₂m). However, we found that transfected tapasin downregulated immune reactivity against a model tumor antigen, MUC1. Although tapasin has been previously shown by others to increase immune recognition of particular antigens, our results suggest that tapasin has a negative impact on the presentation of an immunodominant epitope from a specific model tumor antigen.     

IL-4 and TNF-α induce changes in integrin expression and adhesive properties and decrease the lung-colonizing potential of HT-29 colon carcinoma cells

The colon carcinoma cell line HT-29 was used to explore the potential of interleukin-4 (IL-4) and tumor necrosis factor (TNF-) to modify integrin expression and adhesive functions of tumor cells in vitro and to examine corresponding metastatic effects in vivo. Preincubation of HT-29 cells with 100 U/ml of IL-4 for 48 h downregulated the surface expression of the integrin subunits 2, 3, l and 4 after 48 h, whereas the 1 subunit was upregulated. In contrast, 100 U/ml of TNF-a selectively upmodulated the expression of av. Attachment to fibronectin of cells treated with IL-4 increased twofold (63.5% vs 32.4%). Adhesion to fibronectin (54.0% vs 32.4%) and vitronectin (37.9% vs 16.4%) was elevated in the case of TNF-a stimulation. Using an experimental metastasis model, HT-29 cells showed a significant reduction of their lung-colonizing potential in nude mice when preincubated with IL-4 for 48 h before intravenous injection. The decrease also observed for TNF--treated cells was less pronounced. The data indicate that the cytokines IL-4 and TNF- can act as direct regulators of adhesive mechanisms of tumor cells bearing adequate receptors, thus influencing lung-colony formation.     

Papillary mesothelioma of the tunica vaginalis propria testis

Summary Ultrastructural features of a papillary mesothelioma arising in a hydrocele-sack are reported. The tumour cells presented numerous microvilli, desmosomes, basement membranes and abundant bundles of microfilaments, which all are hallmarks of mesotheliomas. The predominant cell type was the clear epithelial cell, but transitional cells and degenerative forms (foamy cells) were also found. The morphology and differential diagnosis of mesothelial tumours arising in the tunica vaginalis propria testis are discussed.     

A method for simultaneous automatic analysis of many electrograms

Summary A new method is suggested for simultaneous automatic analysis of many electrograms.The method provides for transformation of continuous input signals into intermittents, their transference into the common tract, analysis of signals with regrad to strength, coding of signal strength and recording of analysis data on a single-track magnetic tape.The experimental findings are processed on a Ural-2 electronic computing machine which memorizes analysis data fed into it by means of a marking selector and a special input device.(Presented by Active Member AMN SSSR P. D. Gorizontov) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 61, No. 1, pp. 91–95, January, 1966     

Studies on in vitro neutralization of adenoviruses

Summary The incubation time titration method has been applied to a study of the neutralization of adenovirus by antibody. Following dilution of mixtures of virus and serum a marked reactivation phenomenon was apparent. The results indicate that this phenomenon was partly caused by remaining reactive antibody in the medium, partly by a faster and more extensive reaction between virus and serum in higher concentrations. Whether or not a real reactivation also contributed to the dilution effect cannot be definitely decided upon.The results indicated, furthermore, that antibody may influence the virus in another way than by a complete inactivation. This effect was correlated to the dose of inoculum. Some possibilities regarding the mechanism of the phenomenon are discussed.With suitable concentrations of virus the incubation time titration method has been found applicable for routine work.     

Interferon β1a Treatment Modulates TH1 Expression in γδ + T Cells from Relapsing-Remitting Multiple Sclerosis Patients

A paradigm exists that multiple sclerosis is causally related to dysregulation of T_H1 inflammatory cytokines and T_H2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, , CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon -1a (IFN1a) therapy and following 8–10 weeks of this therapy. The bioactivities of interferon (IFN), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN, IL10, and IL12 from each cell type did not change significantly with IFN1a treatment. The IL10 secreted by TCR+ cells strongly correlated with the IL12 secreted by the same TCR+ cells, supporting the paradigm. Furthermore, IFN1a therapy decreased the TCR+ cell secretion of T_H1 cytokines after 8–10 weeks of therapy.     

Immunological diagnosis in viral infections of the central nervous system: course of antibody titres against homo- and heterologous viruses

In clinical cases suspected for viral encephalitis or meningoencephalitis, the estimation of virus-specific antibodies especially in liquor requires high sensitivity as well as specificity. With enzyme immunoassays the sensitivity in detecting antibodies has increased compared to e.g., complement fixation tests. This report concerns the determination of virus-specific antibodies with a commercial enzyme-linked immunosorbent assay (ELISA) in paired liquor/serum samples of four patients with encephalitis or meningoencephalitis. Up to six virus-specific antibodies of the IgG and IgM classes have been determined [herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalo virus, mumps virus, measles virus, and rubella virus]. Additionally, serum samples from several patients suffering, or recovered from, diseases caused by HSV and VZV without CNS involvement have been included as controls. The results showed that besides the virus-specific antibody development (IgG and IgM) against the leading virus, i.e., principally concerned in the disease manifestation assumed to be primarily causing the disease, virus-specific antibodies of the IgG and IgM class against a heterologous virus (e.g., VZV) could also be measured with substantial titers. Cross-reacting antibodies to both HSV and VZV with the ELISA only appeared and were present in cases where the infection mainly affected the CNS: no such immunological cross-reactivity was observed in serum of individuals in clinically silent stages of both HSV and VZV infections. The same situation with no measurable cross-reacting antibodies was found in cases of acute HSV or VZV diseases where the CNS was not involved. These findings have been discussed with respect to the findings of common antigens, especially between HSV and VZV, and with respect to an unspecific stimulation of immunocompetent cells.Abbreviations and Definitions CSF Cerebrospinal fluid - ELISA enzyme-linked immunosorbent assay - CF-test complement fixation test - NT neutralization test - IFT immunofluorescence test - vIgG virus-specific IgG-antibody - vIgM virus-specific IgM-antibody - HSV herpes simplex virus - VZV varicella zoster virus - EEG electro-encephalogram - CCT cranial computer tomogram MEM, minim essential medium     

Identification and quantitation of cortisol metabolites in rat liver homogenate and -slices

Summary Cortisol-1, 2-H³ was incubated with rat liver homogenate and/or rat liver slices in the presence of a NADPH-generating system. The following metabolites could be identified in adult male rats: -cortol, allo--cortol, 3-allo--cortol, 20-hydroxy-cortisol, 11, 17, 20, 21-tetrahydroxy-5-pregnan-3-one, 3-allotetrahydrocortisol, tetrahydrocortisol, trace amounts of allotetrahydrocortisol and two highly polar metabolites only partly identified. In female rats only tetrahydrocortisol, allotetrahydrocortisol and allodihydrocortisol could be detected in significant amounts.The radioactive metabolites mentioned above were localized and quantitated on paper chromatograms by a 4-radiochromatogram scanner. A nearly perfect correlation was found between these results so obtained and those given by liquid-scintillation counting of each metabolite after its elution from the paper.Part of this work was supported by grant n° 695 of the National Fonds voor Wetenschappelijk Geneeskundig Onderzoek.Stagiair of the Nationaal Fonds voor Wetenschappelijk Onderzoek.     

Der gegenwärtige Stand der Mastzell-Forschung

Zusammenfassung Seit der Erstbeschreibung der MZ durchEhrlich (1877) ist die Kenntnis über die biologische Rolle dieser Zellen wesentlich erweitert worden; diese, mit metachromatischen Granula erfüllten Zellen haben nicht nur die Eigenschaft, Substanzen zu speichern (Mastzellen), sie haben auch die Fähigkeit, biologisch aktive Substanzen zu bilden, diese an das Erfolgsgewebe abzugeben (sekretorische Zellen, Heparinocyten, Histaminocyten) und sind sowohl in morphologischer als auch in funktioneller Hinsicht in den Bauplan der vegetativen nervösen Peripherie mit eingeschlossen (neurohormonale Zellen).     

The pig mesonephros

Summary The ultrastructure of the proximal tubule of mature mesonephric nephrons was studied in perfusion-fixed pig embryos of the 41st gestational day. Despite its 8–12 mm long course, the proximal tubule possesses no cytologically different subsegments except its very last cells at the abrupt transition into the distal tubule. The first brush-bordered proximal tubule cells stand considerably within Bowman's capsule, abuting its attenuated cells. In SEM specimens, the average luminal cell diameters are 8×13 m. The cells are 6–11 m in height with overlying microvilli 2–4 long. Lateral faces of perfectly disjoined cells exhibit plate-like interdigitating processes projecting more than 5 m deep into the neighboring cells. The basal cell face is completely covered with microvilli. The TEM pictures reveal an endocytic apparatus largely matching its metanephric counterpart. Mitochondria account for 23% of the cytoplasm and together with the many basolateral cell membranes indicate a high capacity for energy-dependent transport processes. Small basal lipid droplets represent a species peculiarity. Freeze-fracture specimens show an electrocoupling of the cells by gap junctions. The tight junction, with only 1–2 strands, characterizes a leaky epithelium. In most features, this tissue is as mature as its metanephric counterpart.Supported by the Deutsche Forschungsgemeinschaft.     

Untersuchungen zur Ultrastruktur lympho-epithelialer Thymustumoren unter besonderer Berücksichtigung der sog. „Emperipolesis“

Summary The ultrastructure of eleven thymomas with lymphocytic predominance, one epitheloid cell thymoma and two normal human thymuses is described with special reference to Emperipolesis. All patients have had myasthenia gravis.The normal human thymus consists of three parts: outer cortex, inner cortex, and medulla. The outer cortex contains mainly lymphoblasts and Metcalf's macrophages within the so-called Clark-packet's. The inner cortex consists mainly lymphocytes and interdigitating reticulum cells, and the medulla of epithelial cells, lymphocytes and Hassall's corpuscles.In all cases of lympho-epithelial thymoma and in normal human thymuses there are enormous interdigitations between epithelial (tumor) cells, lymphocytes and macrophages. The epitheloid cell thymomas also show findings which suggest an epithelial cell interaction. We have not found intact lymphocytes inside the cytoplasm of normal and/or tumor epithelial cells, macrophages or interdigitating reticulum cells.The intracellular existence of intact lymphocytes has been termed Emperipolesis by Humble, Jayne, and Pulvertaft, meaning internal wandering. These investigations indicate that Emperipolesis is not an adequate term for cellular interaction in normal human thymuses and thymomas. A false impression of intraepithelial location of thymic lymphocytes is created by two-dimensional sections of complex thymic structure. These ultrastructural studies revealed damage to lymphocytes only in macrophages with lymphocytolysis within these cells and accumulation of numerous heterophagic vacuoles containing fragments of lymphocytic debris within them.

FrÄulein C. Schürmann danke ich für die gute technische Assistenz, Herrn Priv.-Doz. Dr. med. R. W. Ch. Janzen, Neurologische Klinik der UniversitÄt Hamburg, für die klinischen Daten der Myasthenie-Patienten     

Facilitation by 5-hydroxytryptamine of ATP-activated current in rat pheochromocytoma cells

The effects of 5-hydroxytryptamine (5-HT) on an inward current activated by extracellular ATP were investigated in rat pheochromocytoma PC12 cells. Under whole-cell voltage-clamp conditions 5-HT (10 M) reversibly enhanced the amplitude of the current activated by 30 M ATP. The enhancement may not be due to an increase in the number of functional channels because the current activated by 300 M ATP was not remarkably augmented compared with the current activated by 30 M ATP. The current enhancement by 100 M 5-HT was less obvious than that by 10 M 5-HT. When the current kinetics were compared, activation of the ATP-evoked current was accelerated to the same extent by either 10 or 100 M 5-HT, whereas deactivation was largely more accelerated by 100 M 5-HT. Propranolol (10 M), a 5-HT₁ receptor antagonist, or LY53857 (10 M), a 5-HT₂ receptor antagonist, exerted an agonistic effect: the ATP-activated current was facilitated by these compounds. Metoclopramide (10 M), a 5-HT₃ receptor antagonist, neither facilitated the ATP-activated current, nor blocked the current facilitation by 5-HT. Guanosine 5-O-(2-thiodiphosphate) (GDP[S]) (2 mM), the non-hydrolysable analog of guanosine 5-triphosphate (GTP), or K-252a (2 M), a protein kinase inhibitor, did not affect the facilitation by 5-HT of the ATP-activated current when they were included in the intracellular solution. The ATP-activated current pre-facilitated by 10 M dopamine was not enhanced by 10 M 5-HT. Similarly, the pre-facilitation by 5-HT attenuated the current enhancement by dopamine. The results suggest that 5-HT facilitates the ATP-activated channels through receptors that are not readily classified into conventional subclasses of 5-HT receptors. The reciprocal masking between the current facilitation by 5-HT and that by dopamine, combined with their sensitivities to the compounds involved in the intracellular solution, indicates that the facilitation by 5-HT may share not all, but some, common cellular mechanism with that by dopamine.     

Cationic proteins of human granulocytes Effects on human platelet aggregation and serotonin release

Immune-aggregate and thrombin-mediated [³H]serotonin release from human platelets are shown to be enhanced when platelets are preincubated with the antibacterial chymotrypsin-like cationic protein isolated from human granulocytes. The enhancement is dose dependent and inhibited by heating of the cationic protein. Release with chymotrypsin-like cationic protein alone was not observed, although the protein was shown to micro-aggregate platelets irreversibly by an ADP-dependent reaction. Platelet macro-aggregation induced by immune-aggregate was also enhanced by chymotrypsin-like cationic protein whereas platelet macro-aggregation induced by thrombin was inhibited competitively. Platelet micro-aggregation induced by chymotrypsin-like cationic protein was inhibited when preincubated for more than 5 min with a 2-fold molar excess of-1-antitrypsin. Chymotrypsin-like cationic protein interaction with several platelet reactions suggests a close relationship between neutrophils and platelets in the inflammatory process.     

Antigenic relationships between avian paramyxoviruses

Summary The suggested model of antigenic kinship between related paramyxoviruses is based on another concept of antigenic determinant, as compared to the previously suggested combinatorial mathematical model by the authors. According to it, antigenic changes of any determinant do not proceed by leaps but can be changedgradually. Such changed determinant can induce a corre-spondingly changed type of antibodies which still preserve a certain kinship to the original type of the determinant (before its changing) revealed by cross reaction serological tests. Accordingly, there can be families of the determinants differing by degree of relatedness to (or, reversely, by antigenic distance from) the original (ancestor) determinant.In addition to another interpretation of the antigenic kinship, the new mathematical model was used as an approach for revealing phylogenetic relationships between antigenically related viruses.     

Suppression of temperature sensitive mutations in oncogene-related CDC genes in Saccharomyces cerevisiae by catabolite repression resistance and cytoplasmic petite mutations

Summary The start cell division control genes CDC36 and CDC28 have been reported to contain a certain sequence homology to tissue oncogenes (ets and some protein kinase encoding oncogenes respectively). Here we report that temperature sensitive mutations in these genes are suppressed in cytoplasmic petite mutants and catabolite repression resistant mutants.     

Prostate specific antigen and prostate specific acid phosphatase in adenocarcinoma of Skene's paraurethral glands and ducts

An autopsy case of adenocarcinoma of Skene's paraurethral gland co-incident with renal cell carcinoma is described. The adenocarcinoma showed distinct prostate specific antigen and prostate specific acid phosphatase pointing to the equivalence between the male prostate and Skene's paraurethral glands and ducts. Skene's gland are the homologue of the prostate in females and tumours arising from them are immunohistochemically similar to male prostate carcinoma.In the title and text the authors used the official term of Nomina Anatomica paraurethral (Skene's) glands and ducts. Nevertheless recently published data on cross-antigenicity between the male prostate and Skene's glands and the newly discovered exocrine and neuroendocrine parameters of the prostate homologue in the female, comparable with the male prostate (Zaviai 1987), support the use of the same term — the prostate — for prostatic tissue in both sexes (Zaviai 1987, Zaviai et al. 1985). The designations female prostate homologue or female prostate equivalent are a compromise between terms the female prostate and Skene's paraurethral glands.     

The maturation and release of infectious polio and Coxsackie viruses in individual tissue cultured cells

Summary Examination of the intracellular and extracellular virus yields from individual FL and primary human amnion cells of uniform size, infected at a high multiplicity and incubated for specified time intervals, showed that the time of the onset of the appearance of mature infectious virus and the amount of accumulated infectious virus were variable from cell to cell. The duration of the major accumulation of mature virus by individual cells was found to be from 4 to 6 hours. The cells in the FL population, infected with Type 1 poliovirus, fell into two classes in regard to the accumulation and release of virus. The early releasers released some virus during the period of accumulation, yet they continued to accumulate additional virus which they subsequently released. The late releasers, represented by the majority of the cells, released virus at some time after accumulation had ceased. All cells in the primary human amnion cultures, infected with Coxsackie A-9 virus studied fell into the late releasing class.Aided by American Cancer Society Grant, E-82, and United States Public Health Service Grant, C-2245.     

Further observations on the morphogenesis of human rotavirus in MA 104 cells

Summary Human rotavirus KUN strain was cultivated in a fetal rhesus monkey kidney cell line, MA 104 cells. Four types of virus particles in cells infected with KUN strain were clearly identified: nucleoid cores, single-shelled particles, double-shelled particles, and membrane band, enveloped particles. Enveloped particles were found only in the thin sections of infected cells. When first visible, the virus precursors appeared at the ribosome free membrane of rough endoplasmic reticulum (RER), increasing in size while simultaneously being coated with nucleocapsid, inner shell. Single-shelled particles were also synthesized within bundles of filaments of viroplasm in the cytoplasma. During subsequent virus maturation two types of budding processes were observed. Double-shelled particles arising at the RER membrane entered the cisternae of the RER through an exocytosis-like process. In contrast, the enveloped particles developed in the cisternae by being completely enclosed with RER membrane, and later during cytolysis released the single-shelled particles. These enveloped virus particles appeared to be the result of inefficient virus maturation at the last stage of outer capsid formation.With 11 Figures