Phenotypic analyses of lymphokine-activated killer cells that release interferon γ and tumor necrosis factor α

Summary We recently reported that interleukin-2(IL-2)-activated peripheral blood lymphocytes and CD3⁺, lymphokine-activated killer (LAK) cell clones release tumor necrosis factor (TNF) and interferon (IFN) when stimulated with K562 erythroleukemia cells. We examined the phenotype of IL-2-activated peripheral blood leukocytes that secrete TNF and IFN when stimulated with K562 cells and demonstrated that TNF secretion is not due to the presence of contaminating mononuclear phagocytes. Further, we demonstrate that IL-2-activated natural killer (NK) cells release only IFN when stimulated with K562 cells while T lymphocytes exposed to monoclonal anti-CD3 and K562 cells secrete both TNF and IFN. However, T cells stimulated only with K562 cells did not release IFN or TNF while the admixture of these T cells with NK cells, when stimulated with K562 cells, released levels of TNF comparable to those produced by the unseparated cells. At present it is unclear whether only one or both effector cell types respond to K562 by releasing TNF or why the presence both cell types is needed.This work was supported by grants from the national Institutes of Health (CA 23074 and CA 17094) and the Arizona Disease Commission (8277-000000-1-0-YR-9301)     

Effect of α-amanitine on puffing and intranuclear RNA synthesis in Chironomus salivary glands

Incubation of Chironomus salivary glands with -amanitine in concentrations from 1 to 10 /ml results in the suppression of puffing and chromosomal ³H-uridine incorporation after 30 to 60 min in 80% of the cells. Nucleolar ³H-uridine incorporation remains completely unaffected. Even 4 h after the injection of high doses of -amanitine into living larvae, nucleolar incorporation is still pronounced. The distribution of resistant cells within the salivary glands suggests that the uptake of -amanitine is subject to physiological restrictions.—A puff typically induced during in vitro incubation of salivary glands was found to be less sensitive to -amanitine than the Balbiani rings.     

Characterization of phosphoinositide hydrolysis products induced by hexachlorocyclohexane isomers in rat brain cortex

Water-soluble inositol metabolites were separated by anion-exchange chromatogrphy in order to determine whether or not -hexachlorocyclohexane (-HCH, lindane) and related compounds affect phosphatidylinositol hydrolysis in rat brain cortex slices. Hydrolysis was increased by -and -HCH, while - and -HCH were inactive. Muscarinic receptor stimulation of rat cortical slices with carbachol increases inositol phosphates formation. The combined effect of carbachol and the hexachlorocyclohexane isomers together were approximately equal to the sum of the effect of each one separately. The results suggest that lindane stimulates phosphoinositide phospholipase C and/or inhibits the phosphases implicated in dephosphorylation of inositol phosphates.     

Agiotensin II induced cardiac hypertrophy in vivo is inhibited by cyclosporin A in adult rats

Recently, the calciumcalmodulindependent calcineurin pathway has been defined as a central pathway for the induction of cardiac hypertrophy. The purpose of this study was to determine if cardiac hypertrophy in animals chronically treated with angiotensin II (AngII), could be prevented by blocking this pathway with cyclosporin A (CsA). Female Wistar rats were treated with AngII by subcutaneous infusion and injected twice a day with CsA (25 mg/kg) for 7 days. In the AngII treated group there was a 30% increase in the heart/body weight ratio (p < 0.05 vs. control). The increase in heart weight was blocked with CsA. Substantial increases in ANF and MHC gene expression were detected in the AngII treated animals, which were either attenuated or blocked with CsA treatment. Thus, this study demonstrates that CsA does prevent the development of cardiac hypertrophy in Ang II treated rats, suggesting that the calciumcalmodulindependent calcineurin pathway is associated with angiotensin II induced hypertrophy in vivo.     

The units of DNA replication in the mammalian chromosomes: Evidence for a large size of replication units

The replication of chromosomal DNA in human and Chinese hamster cell populations has been studied by means of the DNA fiber autoradiography. It was found that the rate of DNA replication for one fork in human cells varies from 0.2 to 0.9 m/min, the average being 0.6 m/min. In the Chinese hamster cells the rate of DNA replication is greater, varying from 0.3 to 1.2 m/min, the average being 0.8 m/min. There are no clusters containing a great number of replication units in human and Chinese hamster cells. Sequences consisting of two or three replicons which belong to single DNA molecule have been observed, but their frequency was relatively low. The distances between the initiation points in such sequences of replicons vary from 40 to 280 m, the average value being 130 m. This value represents the minimum size of the replication units which have completed the DNA synthesis within 3 h of the S-period. The DNA synthesis in most replication units fails to be accomplished within the three hours of labelling. The process can be completed only in the fragments of DNA molecules of 40 to 200 m (the average value being 100 m) in human cells, whereas in the Chinese hamster cells the fragments of 40 to 250 m (the average being about 140 m) are completely replicated. Provided that the replication is bidirectional the complete replicons are supposed to contain two such fragments. Consequently, the greater part of replication units in mammalian cells covers the pieces of a few hundred microns in DNA molecules. The relation between replication process at the DNA molecules level and that at the metaphase chromosome level is discussed.     

An approach for parameter estimation of biotechnological processes

An approach for parameter estimators design of biotechnological processes (BTP) is presented in case of lack of real time information about state variables. It is based on general reaction rate models and measurements of at least one reaction rate. A general parameter estimator of BTP is designed with the help of which specific rate estimators are synthesized. Stability and convergence of an estimator of specific growth rate for a class of aerobic batch processes are proved. Its effectiveness is illustrated by simulation results. The proposed on-line parameter estimation approach can be used for design of BTP on-line variable estimation algorithms (variable observers of BTP).List of Symbols X, S, P g/l biomass, substrate and product concentrations - C g/l oxygen concentration in the culture broth - C _sg/l saturation concentration of oxygen in the culture broth - C _in, C_outg/l oxygen concentrations in the input air flow and in the outlet gasphase - F _in, F_outl/h the input air flow in the fermenter and output air flow - OUR g/(lh) oxygen consumption rate - OUR _mg/(lh) measured values of OUR - V l volume - , , l/h specific growth, consumption and synthesis rates - K _La(o) l/h specific volumetric mass transfer coefficient - D l/h dilution rate - R _X, R_S, R_Pg/(lh) biomass growth, substrate consumption and product synthesis rates - K _b matrix of yield coefficients - H_b(), H() matrices of known functions of - H(R) matrix of known functions of R - and gain matrices - a vector of the state variables - () a reactions rates vector, describing qualitative relations among the components - R() a reactions rates vector, describing qualitative and quantitative relations among the components - F a feed rates vector - Q a gaseous outflow rates vector - _b () a vector of unknown functions of - ₁() a vector of functions - (t) a vector of unknown time-varying parameters - ₂(, ) an auxiliary vector-function of and - Y _X/S, Y_X/C, Y_X/P substrate, oxygen and product yield coefficients - b maintenence coefficient - k _i(i=1...6) kinetic coefficients - C _i(i=1,2) design parameters estimate     

Effect of short- and long-range interactions on protein folding

The relative importance of short- and long-range interactions is examined using a Monte Carlo simulation of protein folding on bovine pancreatic trypsin inhibitor. The model of the protein and the interaction energies were parametrized using X-ray structures of 30 native proteins. A nearest neighbor Ising model is used to determine the conformational state at each stage of the Monte Carlo procedure. Long-range interactions are simulated by contact free energies which become effective as two residues, separated by four or more residues along the chain, approach each other, and by disulfide-bond energies. Short-range interactions for residues separated by one, two, or three residues along the chain are also modeled by contact free energies and by -helical hydrogen bonds. A hard-sphere model is used to represent repulsive interactions. The ratios of short- to long-range interactions studied are 1:1, 2:1, 1:2, 0:1, and 1:0; e.g., for the 2:1 ratio, short-range interactions are weighted twice as much as long-range interactions, and for the 1:0 ratio, long-range interactions are omitted. For each ratio of short- to long-range interactions, a native conformation is found by a Monte Carlo procedure, a segment of 11 residues (residue numbers 1–11) is then rotated away from the rest of the molecule [breaking the 5–55 native disulfide bond, and moving this segment so that the distance between the sulfur atoms of the 5 and 55 cystine side chains (averaged for all native conformations) increases from 3.9 to 7.3 Å], and the Monte Carlo simulation is carried out (allowing the conformation of the whole molecule to change) until equilibrium is attained. For each ratio, the refolded conformation is compared to the native one using triangular distance maps and differential geometry distance criteria. With ratios of short- to long-range interaction energies of 1:1 and 0:1, the native disulfide bond could be re-formed; with ratios of 2:1 and 1:2 it did not; and with the 1:0 ratio, even a stable native conformation was not achieved. Therefore, long-range interactions (in addition to short-range ones) are required to bring remote parts of the protein together and to stabilize its native conformation.NIH Postdoctoral Fellow, 1977–1978.     

Coastal dune stability: The effect of meteorological conditions and vegetation

Quantitative investigations and research conducted along the north coast of New South Wales, Australia are evaluated with respect to coastal processes, coastal alignment, meteorological data, dune dimensions, dune vegetative cover, development on the dunal area and dune management.The data available covered an assessment of the extent of dune instabilities, an assessment of long term coastline movements, a study of the effect of a cyclone and storm surge, and an assessment and evaluation of a phase of extreme coastline erosion. Specific situations are described and evaluated within the above context. The evaluations are used to determine the extent of coastal dune areas required to be designated as buffer zone in land use planning. The extent of the zone required is dependent upon expected magnitude of wave erosion. Magnitude of wave erosion was found to be proportional to the interaction of coastal processes during periods of extreme erosive factors and the beach dune characteristics for the particular section of coastline. It was found that man's influence on this natural interaction can be a dominating factor in determining beach dune characteristics and therefore the magnitude of wave erosion. — management of the beach dunal area to maintain an acceptable dynamic equilibrium of the beach dune line by a vegetated sand dune of specific dimensions is possible despite weather conditions, if a designated buffer zone is maintained.Presented at the Seventh International Biometeorological Congress, 17–23 August 1975, College Park, Maryland, USA.     

X-ray microanalysis of calcium binding sites in Paramecium

Summary In Paramecium cells Ca⁺⁺-stimulated triggering of the exocytosis of secretory vesicles (trichocysts) was achieved by ionophores X-537 A or A 23187. Under triggering conditions electron dense deposits were present in some resting trichocysts and regularly in discharging trichocysts; upon subsequent fixation deposits occurred on the trichocyst membrane (on the inner side or within the membrane) and on the inner lamellar sheath from where deposits seemed to radiate into the secretory materials. Similar results were obtained with glutardialdehyde fixation alone which also triggers exocytosis but only at low concentrations. Element analysis by energy dispersive x-ray microanalysis ascertained the presence of Ca and P in deposits occurring in trichocysts. Those resting trichocysts which were devoid of deposits did not contain Ca or P enriched. Hence, an abrupt Ca⁺⁺-influx into individual trichocysts just before exocytosis seems to be involved in the triggering mechanism, possibly in combination with the sudden activation of an ATPase systemlocalized at those sites of the trichocysts which primarily contain the deposits. When paramecia were treated only with Ca⁺⁺ and then fixed with OsO₄ plus oxalate or merely with glutardialdehyde, electron scattering deposits were formed also on the inner side of the cell membrane and within the ciliary shaft (but rarely in trichocysts). Deposits obtained on cilia (including ciliary granule plaques) also contained Ca, P and S. Cells contain osmiophilic calcium-storing vacuoles which were selectively rich in Ca and S but devoid of P.     

Reproductive hormone genes in mothers of spontaneous dizygotic twins: an association study

There are important genetic influences on the tendency to dizygotic (DZ) twinning and it is a plausible hypothesis that these reside in one or more of the genes coding for the major reproductive hormones. We used Southern analysis of DNA from 50 young (<32) mothers of DZ twins, who also had a family history of DZ twinning, and 50 controls, to examine allele frequencies of five restriction fragment length polymorphisms (RFLPs) in four hormone genes coding for follicle stimulating hormone (FSH), chorionic gonadotropin (CGB), inhibin _B and gonadotropin releasing hormone (GnRH). Comparison of allele frequencies revealed no significant differences between DZ twin mothers and controls. However this does not rule out the role of these genes in the hereditary tendency of multiple ovulation in humans, since absence of linkage disequilibrium does not imply absence of linkage.     

Adaptation of inducible defense in Euplotes daidaleos (Ciliophora) to predation risks by various predators

The extent of induced morphological defense in Euplotes daidaleos correlates to this ciliate's predation risk from the defense-inducing predator species. Euplotes daidaleos responded by morphological transformation only to organisms that are able to feed on typically formed Euplotes cells (63 ± 5 m cell width in E. daidaleos). Three of those potential predator species caused defensive changes to various degrees (Student's t-test, P < 0.1 to P < 0.0001): Lembadion bullinum (Ciliata) induced 82 ± 6 m cell width in E. daidaleos; Chaetogaster diastrophus (Oligochaeta) induced 85 = 6 m width; and Stenostomum sphagnetorum (Turbellaria) induced 89 ± 8 m width (at a density of 10 predators per milliliter, respectively). At higher predator densities (50 or 100 organisms per milliliter), Euplotes developed a correspondingly larger width (to a maximum of 103 ± 10 m in the presence of S. sphagnetorum). Euplotes did not respond to organisms (e.g., Blepharisma japonicum, Colpidium campylum, Didinium nasutum, Paramecium caudatum, Spirostomum ambiguum, Stentor coeruleus) that cannot feed on this ciliate species. Daphnia longispina and Bursaria truncatella predators, which can feed on large prey of 125, or 200 m in diameter, respectively, also had no effect on the morphology of Euplotes. The extent of defense in Euplotes that was induced by 10 predators per milliliter during 24 h decreased the predation risk from those predators to 67% in the presence of S. sphagnetorum, to 50% with L. bullinum, and to 15% with C. diastrophus, compared to the typical form of Euplotes. In a natural population, the defensive form of E. daidaleos was found with average cell widths of 88 ± 8 m. The results indicate that predator-induced defense in natural Euplotes populations is beneficial to this prey and that it is adapted to the predation abilities of Euplotes predators, whereby energetical costs related to defensive changes may be saved.     

Changes in DNA content and chromosomal size during cell culture and plant regeneration of Scilla siberica: selective chromatin diminution in response to environmental conditions

The chromosomal complement and DNA content of cells of the monocotyledonous plant Scilla siberica were studied at various stages of growth, such as callus outgrowths from bulb tissue (bulb callus), callus grown from single protoplasts prepared from bulb callus (protoplast callus), regenerated plants kept for 2–4 years on agar medium and under constant climatic conditions, and regenerated plants grown first for 2 years on agar and then for 1 year in the garden. During callus culture, several different forms of chromatin loss were observed: (1) chromosome elimination early during cell culture, resulting in cells which were mostly diploid but still had large chromosomes similar to those of the original triploid plants (type 1 cells); from these cells no plants could be regenerated. (2) Dramatic reduction in heterochromatin containing the satellite DNA and, apparently subsequently, also of many other chromatin moieties, resulting in the formation of small chromosomes; frequent polyploidization in these cells, resulting in a variable number of chromosomes per cell (preferentially 30–40, in ca. 70% of the cells; type 2 cells). (3) Appearance of a large number of very small (< 1 m) Feulgenpositive chromatin particles (minute chromosomes; often arranged in metaphase-like arrays, suggesting that they were effectively distributed during mitosis). Such cells could survive and divide under cell culture conditions but did not regenerate plants (type 3 cells). In bulb calli, all three cell types were found whereas in protoplast calli only cells of type 2 and 3 were seen. Type 2 and 3 cells had lost, despite their frequent polyploidization, about 80% of their initial nuclear DNA content and an even higher proportion (>95%) of the satellite sequences. Southern DNA blot analysis revealed that sequences hybridizing with certain protein-coding genes such as that for chalcone synthase were also drastically reduced in copy number whereas the proportion of rDNA was even somewhat increased. In plants regenerated from type 2 cells of protoplast calli, which were aneuploid at near-pentaploidy to hexaploidy, further conspicuous changes in chromosomal and DNA content were not observed, as long as they were kept on agar medium and under constant climatic conditions. However, when such plants were grown in the garden for at least 1 year, the satellite DNA as well as the sequences hybridizing with the chalcone synthase gene were disproportionately increased to 30%–40% of their normal proportion, whereas the total DNA had increased by only approximately 15%. The chromosome numbers remained near-pentaploid to hexaploid as in the cell cultures from which these plants had been regenerated. These phenomena of selective loss and regain of chromatin in response to environmental conditions (cell culture, regenerated plants on agar, regenerated plants grown outdoors) are discussed in relation to other forms of chromatin loss, including developmentally controlled chromatin diminution in certain animals.     

Functional analysis of a complex oncogene arrangement in biotype III Agrobacterium tumefaciens strains

The ubiquitous grapevine-associated octopine/cucumopine Ti plasmids of biotype III Agrobacterium tumefaciens strains carry two T regions, TA and TB, with a complex oncogene arrangement. Within the octopine/cucumopine group, two main strain types were identified: large TA strains with a TA region resembling the TL region of the biotype I octopine strain Ach5 and small TA strains with a similar T region organization as the large TA strains but with a large internal TA deletion. Structural and functional studies of the representative large TA strain Tm4 revealed six oncogenes. Each oncogene was inserted in a disarmed vector and tested for biological activity using the corresponding oncogenes of Ach5 as standards. Five Tm4 oncogenes, TA-iaaM, T-ipt, T-6b, TB-iaaH and TB-iaaM, were shown to be active, the IS-interrupted TA-iaaH gene was inactive. To study the role of each gene in the pTiTm4 context, several single and multiple pTiTm4 mutations were constructed. It was shown that whereas TA-iaaM and TB-iaaH are essential for tumour formation on grapevine, T-ipt, T-6b and TB-iaaM are not. The avirulence of the TA-iaaM ^- mutant was shown to be due to an inhibitory effect of the T-ipt gene, since a TA-iaaM ^- /T-ipt ^- double mutant was fully virulent. We conclude that the TA-iaaM gene of large TA strains is specifically required to counteract the tumour growth inhibiting activity of the T-ipt gene. Both TA-iaaM and T-ipt are absent from the small TA strains. A model on the roles and interactions of the different oncogenes in large TA and small TA strains is presented.     

17 β-Estradiol Enhances the Outgrowth and Survival of Neocortical Neurons in Culture

Results of this investigation demonstrate that exposure to 17 -estradiol differentially and significantly regulates cortical nerve cell outgrowth depending on the cortical region. Parietal and occipital neurons treated with 1 nM 17 -estradiol showed a greater magnitude of neuronal outgrowth whereas outgrowth of temporal cortex neurons was decreased in the presence of 1 nM 17 -estradiol. Frontal cortex neurons showed a consistent enhancement of neuronal outgrowth that did not reach statistical significance. The dose response profile for 17 -estradiol regulation of the macromorphological features exhibited a bimodal dose response relationship whereas the dose response profile for 17 -estradiol regulation of the micromorphological features exhibited a dose response more characteristic of an inverted V-shaped function. An antagonist to the NMDA receptor antagonist, AP5, abolished the growth promoting effect of 17 -estradiol whereas the nuclear estrogen receptor antagonist ICI 182,780 did not. Lastly, neocortical neurons exposed to 17 -estradiol exhibited greater viability and survival than control neurons over a two week period. These data indicate that 17 -estradiol can enhance the growth and viability of select populations of neocortical neurons and that the growth promoting effects of 17 -estradiol can be blocked by an antagonist to the NMDA glutamate receptor and not by an antagonist to the estrogen nuclear receptor.     

Translational arrest in hypoxic potato tubers is correlated with the aberrant association of elongation factor EF-1α with polysomes

Translation elongation factor EF-1 became stably associated with potato tuber polysomes at the onset of hypoxia, coincident with a sharp rise in lactate and decrease in tissue pH. This aberrant association of EF-1 with polysomes also occurred when aerobic tuber extracts were acidified in vitro. Upon resumption of protein synthesis, an increase in the steady-state levels of EF-1, and expression of an EF-1/GUS transgene was observed. These results indicate that translational arrest results from to the failure of EF-1 to dissociate from ribosomes during the elongation cycle, and that restoration of protein synthesis is coordinated with expression of EF-1.     

γ-Glutamylamine cyclotransferase

Summary -Glutamylamine cyclotransferase, an enzyme found in a number of animal tissues and cells, catalyzes the conversion of -(L--glutamyl)-L-lysine to free lysine and 5-oxo-L-proline as well as the release of free amines and the formation of 5-oxo-L-proline from a variety of other L--glutamylamines. Among its substrates are both the mono- and di--glutamyl derivatives of putrescine, spermidine and spermine, and a derivative of -(L--glutamyl)-L-lysine in which both the -amino group and the carboxyl group of the lysine moiety are blocked. The enzyme does not act on most -glutamyl--amino acids, nor is it active toward the -lysyl derivatives of L-aspartic acid or D-glutamic acid. Derivatives of -(L--glutamyl)-L-lysine in which the -amino or the -carboxyl function of the glutamyl moiety is blocked also do not serve as substrates. The specificity of -glutamylamine cyclotransferase is in accordance with the proposal that it functions biologically in the latter stages of the catabolism of products of the action of transglutaminases. Some suggestions as to the manner in which -glutamylamine cyclotransferase serves this function are made based on present knowledge of protein degradation.     

Utilization and degradation of lindane by soil microorganisms

Of 147 microorganisms isolated from a loamy sand, 71 showed good growth with lindane (-1,2,3,4,5,6-hexachlorocyclohexane) and produced chloride in an aqueous medium. Thirteen soil microorganisms were selected to study the utilization of lindane. Lindane was metabolized by the microbes to -2,3,4,5,6-pentachloro-1-cyclohexene (-PCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), and pentachlorobenzene (PCB). Cells of Pseudomonas sp. No. 62 grown on lindane simultaneously adapted to -PCCH, -TCCH, -TCCH, -TCCH, PCB, 1,2,3,4-tetrachlorobenzene (1,2,3,4-TCB) and 1,2,4,5-tetrachlorobenzene (1,2,4,5-TCB). The bacteria degraded each of these chemicals at least partially as indicated by an increased rate of oxygen consumption.Abbreviations Lindane -1,2,3,4,5,6-hexachlorocyclohexane - -PCCH -2,3,4,5,6-pentachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - PCB pentachlorobenzene - 1,2,3,4-TCB 1,2,3,4-tetrachlorobenzene - 1,2,3,5-TCB 1,2,3,5-tetrachlorobenzene - 1,2,4,5-TCB 1,2,4,5-tetrachlorobenzene - 1,2,3-tCB 1,2,3-trichlorobenzene - 1,2,4-tCB 1,2,4-trichlorobenzene - 1,3,5-tCB 1,3,5-trichlorobenzene - 1,2-DCB 1,2-dichlorobenzene - 1,3-DCB 1,3-dichlorobenzene - 1,4-DCB 1,4-dichlorobenzene - MCB monochlorobenzene Contribution No. 631, Research Institute, Agriculture Canada, University Sub Post Office, London, Ontario N6A 5B7     

Heterogeneity of epithelial cells in the human thymus

Summary To evaluate interrelationships among epithelial cells, and between morphology and function in the microenvironment, we studied the ultrastructural morphology of epithelial cells in sections of human thymus from donors aged 2 months to 31 years. Six types of epithelial cells were observed: subcapsular-perivascular (type 1); pale (type 2); intermediate (type 3); dark (type 4); undifferentiated (type 5); and large-medullary (type 6). Cells of types 2, 3 and 4 were found throughout the organ. The type-2 to -4 epithelial cells may represent various stages in a differentiation process. In this, type-2 cells are very active and type-4 cells are possibly degenerating elements. Type-4 cells can also contribute to Hassall's corpuscles. Type-5 cells were located mainly in the cortico-medullary region and showed the morphological characteristics of undifferentiated elements. Type-6 cells were located exclusively in the medulla and displayed characteristics of cellular activity. Small Hassall's corpuscles consisted of type-6 epithelial cells; in larger corpuscles many nuclei of type-6 cells were found. Cells of types 2 and 6 contained tubular structures (diameter approximately 20 nm).Concerning the function of thymus epithelial cells, the features associated with protein synthesis observed in cellular types 2 and 6 make them likely candidates for humoral factor-producing and/or secreting elements. In addition, type-2 and -3 cells in the cortex appear to contribute to a special pattern of epithelium-lymphocyte interaction (thymic nurse cells), as demonstrated by the intracytoplasmic location of lymphocytes in the epithelial cells. The various steps in intrathymic T-cell maturation occur at locations in a microenvironment composed of morphologically distinct epithelial cells.     

<Emphasis Type="Italic">In vitro</Emphasis> screening of sugarcane to evaluate smut susceptibility

Tissue-cultured plantlets of three sugarcane (Saccharum spp.) cultivars having a known field smut reaction were screened for susceptibility to Ustilago scitaminea H&P Sydow. Plantlets were inoculated with 0.5 l of a suspension of equally mixed quantities of plus and minus mating type sporidia of U. scitaminea at concentrations ranging from 1×10¹ to 1×10⁶ cells. Fungal sori (whips) were produced in cultivar N12 (intermediate) 6weeks following inoculation with 1×10⁵ mixed sporidia and thereafter in cultivar NCo310 (susceptible) but not in cultivar N19 (resistant). Sori bearing teliospores were produced up to 3months following inoculation and incubation at 26°C. No sori were produced at mixed sporidial concentrations lower than 1×10⁵cells. The in vitro soral production in cultivars N19, N12 and NCo310 was 0, 27.5 and 47.5% respectively. Plantlets inoculated with 1×105sporidia of only one mating-type did not produce sori in any of the three cultivars tested. Blind scoring of an unknown sugarcane cultivar by this method corresponded exactly with its field smut rating.     

Comparative studies of the development of endosperm-degrading enzymes in malting sorghum and barley

The endosperm cell walls of barley are degraded extensively during malting whilst those of sorghum are not. Malting barley produced endo--1,3:1,4-glucanase, endo--1,3-glucanase and pentosanase in large quantities. In contrast, malting sorghum developed mainly endo--1,3-glucanase and pentosanase. Although the limited break-down of the endosperm cell walls of sorghum may reflect sub-optimal activities of -glucanases, such as endo--1,3:1,4-glucanases, it is possible that the highly intractable nature of the cell walls and their high protein content (approx. 60%) may contribute to the low susceptibility of sorghum endosperm cell walls to enzymic degradation during malting.

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Résumé Les parois cellulaires endospermiques de l'orge sont fortement dégradées pendant le maltage, tandis que celles du sorghum ne le sont pas. L'orge en maltage produit l'endo--1,3:1,4-glucanase, l'endo--1,3-glucanase et la pentosanase en grandes quantités. Par contre, le sorghum en maltage dévéloppe principalement l'endo--1,3-glucanase et la pentosanase. Bien que la destruction limitée des parois cellulaires endospermiques puisse réflecter des activités sub-optimales des -glucanases, comme l'endo--1,3:1,4-glucanase, il n'en est pas molns possible que la nature hautement intractile des parols cellulaires et leur contenu élevé en protéines (approximativement 60%) pulsse contribuer à la faible susceptibilite des parois cellulaires endospermiques du sorghum à la dégradation enzymatique durant le maltage.