Study of plasmids isolated from saccharolytic Clostridia: Construction of hybrid plasmids and transfer toEscherichia coli andBacillus subtilis

Summary Small plasmids ofClostridium acetobutylicum and related strains were isolated and studied. Their restriction maps were established and different hybrid plasmids were constructed by ligation with plasmid pHV33.     

Variants of the anti-Müllerian hormone gene in a compound heterozygote with the persistent Müllerian duct syndrome and his family

Summary The human genome contains a large number of interspersed simple repeat sequences that are variable in length and can therefore serve as highly informative, polymorphic markers. Typing procedures include conventional multilocus and single locus probing, and polymerase chain reaction aided analysis. We have identified simple sequences in a cosmid clone stemming from the human Y chromosome and consisting of (gata)_n repeats. We have compared these with two equivalent simple repeat loci from chromosome 12. After amplifying the tandemly repeated motifs, we detected between four and eight different alleles at each of the three loci. Codominant inheritance of the alleles was established in family studies and the informativity of the simple repeat loci was determined by typing unrelated individuals. The polymorphisms are suitable for application in linkage studies, practical forensic case work, deficiency cases in paternity determination, and for studying ethnological questions. The mutational mechanisms that bring about changes in simple repeats located both on the autosomes and on the sex chromosomes, are discussed.Professor Dr. Otto Prokop (Humboldt-Universität Berlin) on the occasion of his 70th birthday     

Effect of monoclonal antibodies (MoAb) to class I and class II HLA antigens on lectin- and MoAb OKT3-induced lymphocyte proliferation

We have examined the effect of several monoclonal antibodies (MoAb) to monomorphic determinants of class II HLA antigens, and MoAb to monomorphic determinants of class I HLA antigens and to beta-2-microglobulin (beta 2-mu) on lectin- and MoAb OKT3-induced proliferation of human peripheral blood mononuclear cells (PBMNC) and cultured T cells (CTC). Some, but not all, anti-class II HLA MoAb inhibited the proliferative response of PBMNC to MoAb OKT3 and pokeweed mitogen (PWM). The degree of inhibitory effect varied considerably. This effect was not limited to anti-class II HLA MoAb since anti-class I HLA MoAb and anti-beta 2-mu MoAb also inhibited MoAb OKT3- or PWM-induced proliferative responses. In contrast, the response of PBMNC to phytohemagglutinin (PHA) and concanavalin A (Con A) was not blocked by any anti-class II HLA MoAb. However, some anti-class II HLA MoAb also inhibited the proliferative response of CTC plus allogeneic peripheral blood adherent accessory cells (AC) to PHA or Con A as well as to MoAb OKT3 or PWM. This may be attributable to the substantially greater class II HLA antigen expression by CTC than by fresh lymphocytes. Pretreatment of either CTC or AC with anti-class II HLA MoAb inhibited OKT3-induced proliferation. In contrast, pretreatment of CTC, but not AC, with anti-class I HLA MoAb inhibited the proliferative response of CTC to OKT3. Pretreatment of CTC with anti-class I HLA MoAb inhibited PHA-, Con A and PWM-induced proliferation, to a greater degree than the anti-class II HLA MoAb. It appears as if lymphocyte activation by different mitogens exhibits variable requirements for the presence of cells expressing major histocompatibility determinants. Binding of Ab to membrane markers may interfere with lymphocyte-AC cooperation, perhaps by inhibiting binding of mitogens to their receptors or by interfering with lymphocyte and AC function. We also have examined the role of class II HLA antigens on CTC by depleting class II HLA-positive cells. As expected, elimination of class II HLA-positive AC with anti-class II HLA MoAb plus complement caused a decrease in proliferation of CTC in response to all the mitogens tested. In contrast, elimination of class II HLA-positive CTC was shown to clearly increase proliferation of CTC, perhaps because this may deplete class II HLA-positive suppressor cells.     

Réactions observées chez l'Anguille maintenue dans un milieu privé d'électrolytes,en particulier au niveau du système hypothalamo-hypophysaire

Résumé 45 Anguilles sont maintenues (32 h. à 46 jours) en eau déminéralisée (désionisée ou distillée). On note un assombrissement de la pigmentation, une hypertrophie importante de la vésicule biliaire et des modifications du système hypothalamo-hypophysaire.Dans la pars distalis rostrale, les cellules à prolactine subissent une régression, ne semblant plus élaborer de granulations. Les cellules corticotropes sont faiblement stimulées, partiellement dégranulées, en accord avec la légère activation de l'interrénal. Les cellules thyréotropes apparaissent souvent peu actives après 46 jours; cependant, l'examen préliminaire des thyroïdes ne montre pas d'hypoactivité. Dans la pars distalis proximale, les cellules gonadotropes ne sont pas modifiées; les cellules somatotropes sont stimulées: hypertrophie nucléaire et nucléolaire, développement de l'ergastoplasme, sans dégranulation complète. Leur rôle dans l'osmorégulation est discuté. Dans le lobe intermédiaire, les cellules du type 1, hématoxyline au plomb positives, prédominent chez les témoins; elles subissent une stimulation avec dégranulation partielle correspondant peut-être à l'assombrissement de la pigmentation. Le type 2, PAS positif, s'hypertrophie et s'hyperplasie rapidement en eau déminéralisée. Cette hyperactivité indiquerait un rôle dans l'osmorégulation. Le neuroséorétat s'accumule, puis tend à diminuer, la neurohypophyse étant de volume réduit. Le noyau préoptique paraît peu actif. L'hypothèse d'une faible utilisation des produits élaborés par l'axe hypothalamo-neurohypophysaire est envisagée. En plus de l'interrénal et des corpuscules de Stannius, l'hypothalamus et l'hypophyse interviennent donc dans la réponse de l'Anguille à une déminéralisation du milieu ambiant.

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Summary 45 male silver eels were kept from 32 hours to 46 days in demineralized (distilled or deionized) water. They show a darkening of the skin, a hypertrophy of the gall-bladder and some modifications of the hypothalamo-neuro-adenohypophysial system.In the rostral pars distalis, the prolactin cells regressed and appeared chromophobic. The corticotrophic cells are slightly stimulated and partly degranulated, in correlation with some activation of the interrenal, previously described. The thyrotrophic cells do not appear active at the end of the experiment, however the thyroid glands are not often inactive.In the proximal pars distalis, the gonadotrophic cells remain undifferentiated. The somatotrophic cells are strongly stimulated: a cellular, nuclear and nucleolar hyperactivity, without degranulation and a development of the ergastoplasm are observed. Their role in osmoregulation and metabolism of potassium is discussed.The pars intermedia is composed of two cell types: one of which is predominant, leadhematoxylin positive. It could elaborate the MSH (intermedin). In demineralized water, it is partly degranulated and progressively stimulated, perhaps in correlation with the darkening of the pigmentation. The other cell type is PAS positive, and not abundant in the control fish. In the treated fish, rapid hyperplasia and hypertrophy occur, with cytoplasmic degranulation and mitotic activity, so that it becomes predominant in this lobe after a month or more in demineralized water. Cytologic criteria indicate a great hyperactivity which seems to play a role in the processes of the osmoregulation; this hypothesis is discussed.The neurosecretory material tends to increase, as a storage, then to decrease in the neurohypophysis which is of a reduced volume. The preoptic nucleus does not appear active. In addition to the modifications of the interrenal and the corpuscles of Stannius previously described, the changes in the hypothalamus and the pituitary constitute evidence of their important role in the reaction of the eel to a demineralized environment.

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Ce travail a été réalisé avec la collaboration technique de Mademoiselle Jacqueline Olivereau, du C.N.R.S., que nous remercions bien vivement: elle nous a beaucoup aidée au cours de ces expériences, en particulier lors des changements quotidiens de l'eau distillée ou désionisée, et a réalisé toutes les préparations histologiques et les microphotographies illustrant cet article.     

Interaction between cells and elastin fibers: An ultrastructural and immunocytochemical study

Mesenchymal cells (fibroblasts, smooth muscle cells) and endothelial cells were shown to interact with elastin fibers. The strong adhesion of elastin fibers to these cells is mediated by a cell membrane complex with a major glycoprotein component of 120 kDa designated as elastonectin. This interaction was studied by transmission electron microscopy (TEM) and immunocytochemical techniques using antibodies raised against the elastin adhesive proteins. When fibroblasts and smooth muscle cells were cultured in presence of elastin fibers, TEM showed an adhesion mechanism that takes place over several sites along the plasma membrane of these cells. Endothelial cells showed a very close association with elastin, emitting “pseudopodia” that embody the fibers. TEM, indirect immunofluorescence, immunoperoxidase, and confocal microscopy showed the presence and localization of cell membrane components synthesized in large quantities when cells were incubated in presence of elastin. Cells without elastin fibers barely revealed the adhesive membrane complex. These results confirm and extend previous findings concerning the presence of an inducible cell membrane complex that mediates the adhesion of elastin fibers to these cell types. © 1994 Wiley-Liss, Inc.     

Ozone-induced microscopical changes and quantitative carbohydrate contents of hybrid poplar (Populus × euramericana)

Summary Cuttings of hybrid poplar (Populus × euramericana var. Dorskamp) were exposed to ozone (80 g/m³ from 2100 hours to 0700 hours, 180 g/m³ from 0700 hours to 2100 hours) for 3 months. Ozone reduced the starch content in leaves and stem bark, whereas starch granules accumulated in bundle sheath cells along small leaf veins. At the same time, sucrose and inositol content increased in the leaves. Mesophyll cells in the vicinity of the stomata were injured first, and droplet-like material appeared on their walls. In the sieve plates of fumigated trees, the pores showed a higher degree of narrowing than those of the control treatment. Cell collapse in the leaves was accompanied by water loss and an increase in air space. In the stems, the ozone treatment led to a reduced radial width, particularly in the xylem tissue. These results are discussed in relation to reduced or inhibited phloem loading and ozone-induced drought stress. The plants injured by ozone showed quite distinct patterns of metabolite responses as well as enzyme activities (PEP- and RubP-carboxylase) in the leaves from the top to the bottom. There were also remarkable differences in the reaction of sucrose and inositol between leaves and stem bark. Future research should therefore increasingly follow a whole-plant approach for a better understanding of complex plant reactions.