The stationary state of epithelia

A tissue is a geometrical, space-filling, random cellular network; it remains in this steady state while individual cells divide. Cell division is a local, elementary topological transformation which establishes statistical equilibrium of the structure. We describe the physical conditions to maintain stationary the epidermis (of mammals or of the cucumber), in spite of the fact that cells constantly divide and die. Specifically, we study the statistical equilibrium of the basal layer, a corrugated surface filled with cells, constituting a two-dimensional topological froth. Cells divide and detach from the basal layer, and these two topological transformations are responsible for the stationary state of the epidermis. The topological froth is capable of responding rapidly and locally to external constraints, and is a good illustration of the plasticity of random cellular networks.Statistical equilibrium is controlled by entropy, both as a measure of disorder and as information, and is characterized by observable relations between average cell shapes and sizes. The technique can be applied to any random cellular network in dynamical equilibrium. Mitosis as the dominating topological transformation and the fact that the distribution of cell shapes is very narrow are the only inputs specific to biology.

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Resume Un tissu est, à première vue, un pavage aléatoire d'une surface ou d'un volume par des polygones (polyèdres) topologiques, les cellules. Ce pavage reste dans un état stationnaire alors que les cellules se divisent constamment. Nous décrivons les conditions physiques nécessaires à l'état stationnaire de l'épiderme (des mammifères et du concombre), en dépit du fait que ses cellules se divisent et meurent. En particulier, nous étudions l'équilibre statistique de la couche basale, une surface couverte de cellules constituant une mousse topologique aléatoire. Les cellules se divisent et se détachent de la couche basale, et ces transformations topologiques sont responsable de l'état stationnaire de l'épiderme. Cette mousse topologique est capable de répondre rapidement et localement à des contraintes externes. C'est un exemple de plasticité de structures cellulaires aléatoires.L'équilibre statistique est contrôlé par l'entropie qui est ici à la fois une mesure du désordre et une quantité d'information. Il est caractérisé par des relations facilement observables entre les formes des cellules et leurs dimensions. Les seuls éléments spécifiques aux tissus biologiques sont la mitose comme transformation topologique dominante, et l'étroitesse de la distribution des formes de cellules.

     

Posterior dental size reduction in hominids: The Atapuerca evidence

In order to reassess previous hypotheses concerning dental size reduction of the posterior teeth during Pleistocene human evolution, current fossil dental evidence is examined. This evidence includes the large sample of hominid teeth found in recent excavations (1984–1993) in the Sima de los Huesos Middle Pleistocene cave site of the Sierra de Atapuerca (Burgos, Spain). The lower fourth premolars and molars of the Atapuerca hominids, probably older than 300 Kyr, have dimensions similar to those of modern humans. Further, these hominids share the derived state of other features of the posterior teeth with modern humans, such as a similar relative molar size and frequent absence of the hypoconulid, thus suggesting a possible case of parallelism. We believe that dietary changes allowed size reduction of the posterior teeth during the Middle Pleistocene, and the present evidence suggests that the selective pressures that operated on the size variability of these teeth were less restrictive than what is assumed by previous models of dental reduction. Thus, the causal relationship between tooth size decrease and changes in food-preparation techniques during the Pleistocene should be reconsidered. Moreover, the present evidence indicates that the differential reduction of the molars cannot be explained in terms of restriction of available growth space. The molar crown area measurements of a modern human sample were also investigated. The results of this study, as well as previous similar analyses, suggest that a decrease of the rate of cell proliferation, which affected the later-forming crown regions to a greater extent, may be the biological process responsible for the general and differential dental size reduction that occurred during human evolution. © 1995 Wiley-Liss, Inc.     

In vitro colonization ability of human colon mucosa by exogenous Lactobacillus strains

Abstract A 5.4 kb Hind III DNA fragment carrying the gene encoding raw starch-digesting α-amylase (RSDA), has been previously cloned from Bacillus circulans F-2 and expressed in Escherichia coli [Kim et al. (1990) Biochim. Biophys. Acta 1048, 2233–2238]. Interestingly, when the cell extract of E. coli harboring a plasmid carrying this fragment was incubated with l M NaCl, it exhibited about 10 times higher enzyme activity than when assayed without NaCl. Differential zymograms showed two different amylase activities: one for RSDA and the other for a salt-dependent a-amylase (SDA). Even though RSDA activity was detected without NaCl, SDA activity was detected only in high concentrations of NaCl. SDA activity was fully detected at above l M NaCl. Results from subcloning of the genes, fractionation analysis of cell extracts, and immunological assays clearly suggested that the two amylases are genetically distinct and that genes for both enzymes are closely linked on the 5.4 kb DNA fragment.     

Lipoxygenase- and cyclooxygenase-reaction products and incorporation into glycerolipids of radiolabeled arachidonic acid in the bovine retina

The metabolism of radiolabeled arachidonic acid (AA) by the intact bovine retina has been studied. Synthesis of prostaglandins (PGs) and hydroxyeicosatetraenoic acids (HETEs), and incorporation of AA into glycerolipids has been measured by reverse-phase and straight-phase high performance liquid chromatography with flow scintillation detection, and by thin-layer chromatography. AA was actively acylated into glycerolipids, particularly triglycerides, phosphatidylcholine and phosphatidylinositol. AA was also converted to the major PGs, PGF_2α, PGE₂, PGD₂, 6-keto-PGF_1α and TXB₂, and to the lipoxygenase reaction products, 12-HETE, 5-HETE, and other monohydroxy isomers. Approximately 6% of the radiolabeled AA was converted to eicosanoids. The synthesis of HETEs was inhibited in a concentration-dependent manner (IC₅₀ = 8.3 NM) by nordihydroguaiaretic acid (NDGA). PG synthesis was inhibited by aspirin (10 μM), indomethacin (1 μM) and NDGA (IC₅₀ = 380 nM). Metabolism of AA via lipoxygenase, cyclooxygenase and activation-acylation was inhibited by boiling retinal tissue prior to incubation. These studies demonstrate an active system for the uptake and utilization of AA in the bovine retina, and provide the first evidence of lipoxygenase-mediated metabolism of AA, resulting in the synthesis of mono-hydroxyeicosatetraenoic acids, in the retina.     

Etude olfactométrique de l'action de diverses substances allélochimiques végétales dans la recherche de l'hote par Diadromus pulchellus (Hymenoptera,Ichneumonidae)

Résumé La recherche de l'habitat d'un hôte potentiel par D. pulchellus, endoparasite considéré comme spécialiste des nymphes d'A. assectella, est abordée chez les adultes femelles et mâles, en olfactométrie, par l'étude de leur comportement locomoteur. L'activité locomotrice et l'attraction sont ainsi analysées en présence de substances allélochimiques issues ou non de végétaux pouvant abriter un hôte. Que les composés soufrés testés soient stables ou non, ils ne déclenchent pas d'attraction chez les hyménoptères quel que soit leur sexe et le type d'olfactomètre utilisé, contrairement aux imagos du phytophage-hôte, A. assectella, attirés par les composés soufrés instables spécifiques du poireau. L'activité locomotrice des hyménoptères des deux sexes est stimulée par divers composés volatils soufrés de synthèse ou émis naturellement par les Crucifères et par les Allium consommés par les larves d'A. assectella. Les substances efficaces sont généralément stables et sont caractérisées par le groupement actif R-S-, à condition que R ne soit pas un allyle. Par ailleurs, des composés volatils non soufrés contenus dans des végétaux autres que les Allium et les Crucifères ne stimulent pas le comportement locomoteur des hyménoptères. Ces différentes observations permettent d'envisager que D. pulchellus est un spécialiste inféodé à des microlépidoptères s'alimentant sur des végétaux soufrés et non sur les seuls Allium.

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Summary Searching the host habitat by the endoparasite wasp Diadromus pulchellus, considered to be a specialist of Acrolepiopsis assectella nymphs, is studied in females and males by analysis of locomotory behaviour in olfactometers. Thus, locomotor activity and attraction are studied in the presence or absence of air-borne volatiles emitted by plants able to shelter a potential host-moth. Stable and unstable sulphur compounds used do not release attraction in female and male ichneumonids whatever the type of olfactometer used. By contrast, phytophagous host adults are attracted by unstable specific sulphur compounds of the leek. Locomotor activities of D. pulchellus females and males are stimulated by various volatile sulphur compounds of synthetic origin or naturally occurring in Allium eaten by A. assectella larvae and in Crucifera. The effective compounds generally are stable and characterized by the active group R-S — in which the alkyl moiety must not be an allyl one. On the other hand, volatiles without sulphur, emitted by plants which do not belong to Cruciferae or Allium, do not stimulate the hymenopteran locomotor activity. These observations suggest that D. pulchellus is oligophagous, attracted to microlepidoptera developing on plants containing sulphur compounds and not only on Allium.

     

Nonsense RNA: a tool for specifically inhibiting the expression of a gene in vivo

We describe a general technique to inhibit gene expression in eukaryotic cells. The gene we chose to inhibit was the E. coli LacZ gene (encoding beta-galactosidase), which has previously been cloned into a eukaryotic expression vector [1]. This plasmid is called pCH110. We constructed a variant of pCH110 in which we flipped a 2566 base pair 5' fragment of the LacZ gene into the antiparallel orientation. The plasmid containing this mutated LacZ gene is called pNSLacZ (NS signifies non-sense coding sequence). When equal amounts of pCH110 and pNSLacZ are co-transfected into 3T6 mouse fibroblasts, the beta-galactosidase activity is decreased by approximately a factor of ten. Increasing the ratio of pNSLacZ to pCH110 above 1:1 does not appreciably increase the level of inhibition. Next, we prove the specificity of the inhibition by adding a third gene to the transfection mixture. For this purpose, we used pSVneo beta, a plasmid which expresses a phosphotransferase. We found that even when the beta-galactosidase activity was diminished by a factor of 10, the phosphotransferase activity was unaffected. Therefore, we have demonstrated that: the presence of an antiparallel copy of the LacZ gene results in a significant and specific diminution of the LacZ gene's expression; only a fraction of the LacZ gene needs to be in the antiparallel orientation in order to observe this effect. These results suggest that this technique can serve as a tool to decrease the level of gene expression in order to study the function of specific genes, or as a therapeutic manoeuvre in the treatment of disorders of abnormal gene expression.     

Interaction of peanut agglutinin,a lectin specific for nonreducing terminal d-galactosyl residues,with embryonal carcinoma cells

Peanut agglutinin (PNA), a lectin specific for terminal d-galactosyl residues, was found to react with embryonal carcinoma cells, but not with their differentiated derivatives. Receptors for PNA were detectable at the surface of all cells of the quasinullipotent F9 line and on only 50% of the multipotent PCC3/A/1 line. The fraction of the PCC3/A/1 population which expresses the F9 antigen was found to be included in the subpopulation carrying the PNA receptors. PNA⁺ and PNA^? subpopulations of PCC3/A/1 were separated by a PNA-mediated reversible agglutination of PNA⁺ cells with rabbit erythrocytes. These subpopulations were essentially F9⁺ and F9^?, respectively.     

Autolytic activities of the cell wall in rice coleoptiles. Effects of nojirimycin

Oryza sativa L. var. bahia coleoptile cell walls show sufficient autolytic activity for the release into the surrounding medium of amounts up to 60 μg of sugars per mg of dry weight of cell wall. The products released elute in Bio-gel P.2 as mono- and polysaccharides with glucose as the sole component. The polysaccharide component releases tri- and tetrasaccharides on treatment with a glucanase specific for β (1–3) (1–4) linkages in the same proportion as that of the mixed glucan of the cell wall. This supports the hypothesis that the polysaccharide component originates from the cell wall glucan and that autolysis is therefore related to the processes of the loss of rigidity of the cell wall. Nojirimycin (a specific glucanase inhibitor and inhibitor of auxin-induced elongation) decreases autolytic activity of the cell walls, reducing it to 30% of its normal value. Bio-gel P. 2 elution of the products released in autolysis in the presence of nojirimycin shows that only the monosaccharide fraction was affected.     

Role of pancreatic enzymes in the intraluminal transport and ileal absorption of vitamin B 12 (Cb 1)

The present paper outlines the classical concepts of transport and absorption of vitamin B12 and discusses findings which provide new insight into the important role of pancreatic enzymes in the absorption of the vitamin B12. In vivo experiments with healthy subjects and patients with exocrine pancreatic insufficiency demonstrate that the pancreatic enzymes do not activate "the precursor" intrinsic factor molecule but solely dissociate vitamin from the inactive R type proteins with a consequent coupling to the biologically active intrinsic factor.