Thiamine and Cholinergic Transmission in the Electric Organ of Torpedo

The electric organ of Torpedo marmorata was found to contain as much as 120 +/- 24 nmol of thiamine per g of fresh tissue. The vitamin was distributed as nonesterified thiamine (32%), thiamine monophosphate (22%), thiamine diphosphate (8%), and an important proportion of thiamine triphosphate (38%). A high level of thiamine triphosphate was found in synaptosomes isolated from the electric organ. In contrast, the synaptic vesicles did not show any enrichment in thiamine, whereas they contained a marked peak of acetylcholine (ACh) and ATP. Thus thiamine seems to be very abundant in cholinergic nerve terminals; its localization is apparently extravesicular, either in the axoplasm or in association with plasma membrane. When calcium was reduced and magnesium increased in the external medium, the efficiency of transmission was diminished, owing to inhibition of ACh release; in a parallel manner the degree of thiamine phosphorylation was found to increase--this condition is known to modify the repartition of ACh between vesicular and extravesicular compartments. Electrical stimulation, which causes periodic variations of the level of ACh and ATP, also caused significant changes in thiamine esters. In addition, related changes of the vitamin and the transmitter were observed under other conditions, suggesting a functional link between the metabolism of thiamine and that of ACh in cholinergic nerve terminals.     

L'encapsulation hémocytaire expérimentale chez le lépisme Thermobia domestica

The introduction of inert foreign objects into the thorax of the thysanuran Thermobia domestica provoked the formation of a cellular capsule, the development and fine structure of which were examined.Encapsulation at first simply results from the accumulation of blood cells around the implant. It is possible to distinguish 48 hr later four regions in the cellular capsule: (1) An exterior layer including normal haemocytes. (2) An intermediate layer formed by homogeneous intercellular electron-dense material and by stretched haemocytes. These haemocytes have numerous microtubules, without any granular particles, and are linked together by desmosomes. (3) An interior layer of cells in the process of necrosis and rich in lysosomes. (4) A very thin limiting layer tentatively interpreted as melanin.The large number of haemocytes devoid of the specific features of the fibroblasts and the very important reduction of the acellular material without collagen fibrils distinguish clearly the cellular capsules of the Insecta from the granuloma of the Vertebrata and other groups.     

Modification of Oudin’s method of single immunodiffusion in agar gel for detection of IgA in bronchoalveolar lavage of white mice

Oudin’s principle of single immunodiffusion in agar gel was modified for quantitative determination of IgA in bronchoalveolar lavage (BAL) of normal 20–25 g mice. The reaction took place at 25 °C in 0.3 % agarose with 16.7 % pig serum against mouse IgA, and was evaluated on the basis of a relationship between the progress of the precipitin zone and the square root of time. The linear dependence of the derived constantk on the logarithmic concentration of antibody in the sample permitted to express the results as titre, corresponding to a dilution wherek = 0. Examination of seven samples of pooled blood serum of normal mice shoved taht (1) the IgA level was practically constant, (2) serum IgA possessed under given conditions similar properties as IgA from the bronchoalveolar secretion; it is therefore possible to employ pooled sera as a reliable control of the immunodiffusion system in ease of lack of reference standards with defined IgA content. Examination of 82 individual BAL samples of normal mice revealed that the mean IgA concentration in 2.5 mL samples was almost 1000 times lower than in blood serum.     

Recherches sur la conservation du virus de la densonucléose

Résumé Le virus de la densonucléose (VDN) conserve une partie de sa virulence en solution aqueuse entre 0°C et 30°C pendant deux mois. Son pouvoir infectieux se maintient à—10°C pendant deux mois mais est supprimé après dix jours de dessication. On peut done considérer ceParvovirus d’Insectes comme un virus relativement résistant aux températures que l’on rencontre ordinairement dans la nature et au laboratoire.

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Summary The persistence of DNV is described. This virus retains a part of its infectivity in aqueous solution between 0°C and 30°C. The log LD₅₀ initially 7,15 drops to 2 after two months. The virus keeps its infectivity at 10°C but it loses all its pathogenicity after 10 days in dry conditions.

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Ce travail a été réalisé en partie dans le cadre d’une convention I.N.R.A.-D.R.M.E.     

Distinct role for CD8 T cells toward cutaneous tumors and visceral metastases

The growth of immunogenic tumors in immunocompetent individuals is one of the oldest conundrums in tumor immunology. Although the ability of mouse CD8+ T cells to control transplanted tumors is well documented, little is known about their impact on autochthonous tumors. To gain insight into the role of CD8+ T cells during the course of cancer development, we produced a novel model of spontaneous melanoma. The metallothionein (MT)-ret/AAD mouse is transgenic for the RET oncogene and the chimeric MHC molecule AAD (alpha1-alpha2 domains of HLA-A2 linked to alpha3 domain of H2-Dd). This model recapitulates the natural history of human melanoma, and expression of the AAD molecule makes it suitable for analyzing CD8+ T cell responses directed against peptide Ags that have been previously identified in HLA-A2+ melanoma patients. We found that, as tumors grow, mice develop a broad melanoma-specific CD8+ T cell response. Occurrence of cutaneous nodules is not affected by CD8+ T cell depletion, showing that although CD8+ T cells are functional, they have no effect on established cutaneous tumors. However, depleted mice die from visceral disease much earlier than controls, showing that CD8+ T cells control metastasis spreading and disease progression. Antigenic modulation is observed in visceral metastases, suggesting that visceral nodules may be subject to immunoediting. Our data demonstrate that growth of melanoma in the MT-ret/AAD model involves several tolerance mechanisms sequentially. They also reveal a different role for CD8+ T cells toward early stage of cutaneous tumors and late visceral metastatic stage of the disease.     

Formation of a new receptor-operated channel by heteromeric assembly of TRPP2 and TRPC1 subunits

Although several protein-protein interactions have been reported between transient receptor potential (TRP) channels, they are all known to occur exclusively between members of the same group. The only intergroup interaction described so far is that of TRPP2 and TRPC1; however, the significance of this interaction is unknown. Here, we show that TRPP2 and TRPC1 assemble to form a channel with a unique constellation of new and TRPP2/TRPC1-specific properties. TRPP2/TRPC1 is activated in response to G-protein-coupled receptor activation and shows a pattern of single-channel conductance, amiloride sensitivity and ion permeability distinct from that of TRPP2 or TRPC1 alone. Native TRPP2/TRPC1 activity is shown in kidney cells by complementary gain-of-function and loss-of-function experiments, and its existence under physiological conditions is supported by colocalization at the primary cilium and by co-immunoprecipitation from kidney membranes. Identification of the heteromultimeric TRPP2/TRPC1 channel has implications in mechanosensation and cilium-based Ca(2+) signalling.     

Inherited variation at the epigenetic level: paramutation from the plant to the mouse

In contrast with a wide definition of the 'epigenetic variation', including all changes in gene expression that do not result from the alteration of the gene structure, a more restricted class had been defined, initially in plants, under the name 'paramutation'. It corresponds to epigenetic modifications distinct from the regulatory interactions of the cell differentiation pathways, mitotically stable and sexually transmitted with non-Mendelian patterns. This class of epigenetic changes appeared for some time restricted to the plant world, but examples progressively accumulated of epigenetic inheritance in organisms ranging from mice to humans. Occurrence of paramutation in the mouse and possible mechanisms were then established in the paradigmatic case of a mutant phenotype maintained and hereditarily transmitted by wild-type homozygotes. Together with the recent findings in plants indicative of a necessary step of RNA amplification in the reference maize paramutation, the mouse studies point to a new role of RNA, as an inducer and hereditary determinant of epigenetic variation. Given the known presence of a wide range of RNAs in human spermatozoa, as well as a number of unexplained cases of familial disease predisposition and transgenerational maintenance, speculations can be extended to possible roles of RNA-mediated inheritance in human biology and pathology.     

DING proteins; novel members of a prokaryotic phosphate-binding protein superfamily which extends into the eukaryotic kingdom

PstS proteins are the cell-bound phosphate-binding elements of the ubiquitous bacterial ABC phosphate uptake mechanisms. Primary and tertiary structures, characteristic of pstS proteins, are conserved in proteins, which are expressed in secretory operons and induced by phosphate deprivation, in Pseudomonas species. There are two subsets of these proteins; AP proteins, which are alkaline phosphatases, and DING proteins, named for their N-terminal sequence, which are phosphate-binding proteins. Both form elements of a proposed phosphate-scavenging system in pseudomonads. DING proteins have also been isolated from many eukaryotic sources, and are associated with both normal and pathological functions in mammals. Their phosphate-binding function suggests a role in biomineralization, but the ability to bind other ligands may be related to signal transduction in eukaryotes. Though it has been claimed that all such proteins may originate from pseudomonads, many eukaryotic DING proteins have unique features which are incompatible with a bacterial origin.     

Comparative evaluation of Strepto B ID chromogenic medium and Granada media for the detection of Group B streptococcus from vaginal samples of pregnant women

Two types of selective media, the chromogenic medium Strepto B ID® and two non-chromogenic media Strepto B agar® and the Granada® medium, were tested and compared to blood agar plates (BAP) for screening of Group B streptococcus vaginal colonization in pregnant women. All tested media were comparable in terms of sensitivity however, their use in routine laboratories may markedly facilitate the rapid detection of GBS in vaginal samples.     

Serological survey of T-lymphocyte differentiation antigens in wild mice

Allelic distributions of Thy-1, Ly-l, and Ly-2 antigens in wild mice are characteristic of each Mus musculus subspecies. Eastern mice (M.m.molossinus, M.mmusculus, M.m.castaneus, M.m.bactrianus) express the Thy-1.1 antigen, whereas Western mice (M.m. domesticus, M.m.brevirostris) express the Thy-1.2. All mice from wild populations examined in this survey express the Ly-1.2. The Ly-2.1 is distributed in Eastern mice and some Western mice, and the Ly-2.2 is found in the remaining Western mice. Allelic distributions of these antigens were also examined in two other species, Mus spretus and Mus spicilegus. Allelic constitutions of Thy-1 and Ly-1 in these species are similar to those of Eastern mice. Some M.spicilegus, however, express the Ly-1.1 antigen. This antigenic type is not found in M.musculus. Some Eastern mice related to M.m.castaneus react weakly to Ly-1.2-specific and Ly-2.1-specific monoclonal antibodies in both the complement-mediated cytotoxicity test and the absorption test. These results suggest that M.m.castaneus has unique alleles in the Ly-1 and Ly-2 loci.