Introduction of foreign genes into potato cultivars Bintje and Désirée using an Agrobacterium tumefaciens binary vector

Tuber discs of Solanum tuberosum cv Bintje and Désirée were cocultivated with an Agrobacterium tumefaciens binary vector, carrying both the neomycine phosphotransferase and the E. coli -glucuronidase gene fused to resp. the nopaline synthase and Cauliflower mosaic virus 35S promotor.Inoculated tuber discs produce transgenic shoots in selective media containing kanamycin. The transgenic plants are phenotypically normal and contain the euploid number of chromosomes. Both the neomycin phosphotransferase as well as the -glucuronidase gene are expressed conferring resp. kanamycin resistance and -glucuronidase activity to the plants.Abbreviations GUS -glucuronidase - NPT neomycin phosphotransferase - CaMV Cauliflower Mosaic Virus - BAP 6-benzylaminopurine - GA₃ gibberellic acid - NAA naphthalineacetic acid - LB Luria Broth - MU methylumbelliferone     

Hormonal regulation of gene expression in the “slender” mutant of barley (Hordeum vulgare L.)

The slender mutant of barley resembles a normal barley plant treated with high doses of gibberellic acid (GA₃). Expression of GA₃-regulated and abscisic acid (ABA)-regulated mRNAs was studied in the endosperm and roots of mutant and wild-type (WT) plants.Production of -amylase (EC 3.2.1.1) by WT embryoless half-grains was dependent on the presence of GA₃, and was prevented by ABA. In contrast, -amylase was produced by half-grains of the slender mutant in the absence of added GA₃, although it was still reduced by ABA. The spectrum of -amylase mRNAs in slender embryoless half-grains incubated in the absence of added GA₃ was the same as in WT endosperm half-grains incubated in the presence of GA₃. These results indicate that the endosperm of the slender mutant exhibits similar properties to WT endosperm treated with GA₃.In roots the expression of an ABA-inducible mRNA was similar in slender and WT seedlings either treated with exogenous ABA or exposed to dehydration. This result, and the effect of ABA on -amylase production by the endosperm, indicate that the slender plants retain sensitivity to ABA.Abbreviations ABA abscisic acid - AMV avian myeloblastosis virus - GA gibberellin - GA₁ gibberellin A₁ - GA₃ gibberellic acid - WT wild-type     

Micropropagation of Phytolacca dodecandra through shoot-tip and nodal cultures

A procedure for micropropagation of endod (Phytolacca dodecandra) is described. BA at 0.44 M produced 3.1 new shoots per expiant in six weeks using shoot tips. Nodal expiants, however, produced up to 4.7 shoots per explant on medium with 0.44 M BA and 0.27 M GA,. IBA at 0.49 M induced 90% rooting with minimal callus. Plantlets were successfully transferred to the greenhouse and some staminate clones produced flowers after six months.Abbreviations BA 6 benzylaminopurine - kinetin 6-furfurylaminopurine - 2iP N⁶-(2-isopentyl)adenine - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - GA₃ Gibberellic acid - IBA indole-3-butyric acid     

Further studies on the metabolism of gibberellins (GAs) A9, A20 and A29 in immature seeds of Pisum sativum cv. progress No. 9

Seed maturation of Pisum sativum cv. Progress No. 9 proceeds more slowly in winter than in summer even when the parent plants are grown in greenhouse conditions with light-and heat-supplementation. For parent plants grown under summer and winter conditions the metabolism of [³H]GA₉ in cultured seeds is qualitatively different in seeds of equivalent age and qualitatively the same in seeds of equivalent weight. 13-Hydroxylation of [³H]GA₉[³H]GA₂₀ is restricted to early stages of seed development. 2-Hydroxylation of [³H]GA₉2-OH-[³H]GA₉ has only been observed at a stage of development after endogenous GA₉ has accumulated. 2-OH-GA₉ has been shown to be endogenous to pea and is named GA₅₁. H₂-GA₃₁ and its conjugate have not been shown to be present in pea and may be induced metabolites of [³H]GA₉. The metabolism of GA₂₀GA₂₉ is used to illustrate a technique of feeding [²H][³H]GAs in order to distinguish a metabolite from the same endogenous compound. The in vitro conversion of [³H]GA₂₀[³H]GA₂₉, and the virtual non-metabolism of [³H]GA₂₉ have been confirmed for seeds in intact fruits. These results are discussed in relation to the apparent absence of conjugated GAs in mature pea seeds.Abbreviations GA_n gibberellin A_n - GC gas chromatography - GC-MS combined gas chromatography-mass spectrometry - GC-RC combined gas chromatography-radio counting - Me methyl ester - R_T etention time - SICM selected ion current monitoring - TLC thin layer chromatography - TMS trimethyl silyl ether The author is née Frydman     

Activity of the aldehyde and alcohol of gibberellins A12 and A14, two derivatives of gibberellin A15 and four decomposition products of gibberellin A3 in 13 plant bioassays

Summary The biological activities of the aldehyde and alcohol of gibberellins (GAs) A₁₂ and A₁₄, 3-OH-GA₁₅, 3-OH-GA₁₅ wrong lactone (i.e. GA₃₇ wrong lactone) and the four major decomposition products of GA₃ (isogibberellic, allogibberic, epiallogibberic and 9(11)-dehydroallogibberic acids) were tested over a wide range of concentrations on 13 plant bioassays in order to ascertain certain of the structural requirements for biological activity. Generally modification of the basic GA-molecule decreased its activity in all assays except for derivatives of GA₁₂ and GA₁₄ (suggesting conversion of these derivatives to more polar, active GAs). Modification of the 3-OH from the usual 3 to 3 configuration markedly reduced activity. Neither the presence of an inverted lactone ring (i.e. 3-OH-GA₁₅ wrong lactone) nor changes to the lactone ring of GA₃ (410) to form iso-GA₃ (42) appreciably reduce activity. Further decomposition of GA₃ to allogibberic and 9(11)-dehydroallogibberic acid reduced activity only slightly, but epimerization of allogibberic acid at C-9 essentially eliminated biological activity.     

Observations on the fine structure of the cercaria of Notocotylus attenuatus and formation of the cyst wall of the metacercaria

Summary The dorsal tegument of the mature cercaria of Notocotylus attenuatus is a syncytial, cytoplasmic layer, containing two types of secretory granule which are identifiable ultrastructurally. The type 1 secretory bodies are electron lucid, whereas most type 2 granules have a banded appearance. The ventral tegument contains granules which are secreted from the type 3 cells; the type 3 granules are membrane bound, electron dense, and consist of both an amorphous and a finely striated zone. The type 4 cells mainly contain cigar-shaped granules consisting of an amorphous core surrounded by concentric striations. The granules exhibit structural variability in shape and content. The type 4 cells undergo a cellular migration to the tegument during encystment. The structure of the posterior-lateral glands and mode of secretion of the granules are described. Possible functions of microtubules are discussed for each cell type. Details of some secretory processes involved in the formation of the hemispherical cyst wall are described. The layers of the cyst wall may be related to the granular contents of the various parenchymal cells of the cercaria. The tegument of the metacercaria originates primarily from the cytoplasm of the type 1, type 2, type 3 and type 4 cells.     

Micropropagation of Uniola paniculata L.

Uniola paniculata L. is a major sand-dune stabilizing grass which is being utilized to prevent shoreline erosion. In vitro cultured caryopses of U. paniculata produced callus on MS medium supplemented with 22.5 M 2,4-D, 4.4 M BA and 87.6 mM sucrose. Shoot induction occurred after these calli were inoculated onto the same medium without 2,4-D. Rooting of in vitro-derived shoots occurred when transferred to a one-half strength MS medium containing 43.8 mM sucrose and 14.7 M IBA. Plantlets were planted after the roots reached a length greater than 20 mm.Abbreviations BA N-(phenyl-methyl)-lH-purine-6-amine - IBA lH-indole-3-butanoic acid - MS Murashige and Skoog (1962) - 2,4-D (2,4-dichlorophenoxy) acetic acid - TC agar tissue culture agar, (K.C. Biologicals, Lenexa, KS) - Subdue methaxyl:N-(2,6-dimethylphenyl)-N-(methyoxyacetyl) alanine methyl ester (CIBA- GEIGY)     

Correction of the published sequence for the human proteolipid protein gene

Summary In the human proteolipid protein gene, the base sequence of the intronic region 5 to exon 6 was found to be 5-ctctttcattttcctgcag-3 and not 5-ctctttt-cattttcctgcag-3 as previously reported.     

Growth stimulation ofEuphorbia lathyris L. by GA4+7 and BA

Container-grownEuphorbia lathyris plants were treated with foliar sprays of various combinations of BA and GA₄₊₇ or 0–3600 mg L^–1 Promalin (11 BA + GA₄₊₇) in separate experiments. GA₄₊₇ and Promalin stimulated plants to grow taller. BA and Promalin promoted axillary shoot growth. Multiple applications of Promalin stimulated branching more than single treatments. Dry weight accumulation was stimulated only if the growth regulators were applied to 28–33-cm and not to 56-cm tall plants. Chemical names used: (1, 2, 4a, 4b, 10)-2,4a,7-trihydroxy-1-methyl-8-methylenegibb-3-ene-1,10-dicarboxylic acid 1,4a-lactone (GA₄₊₇),N-(phenylmethyl)-H-purin-6-amine (BA), and Promalin [11 (wt/wt) GA₄₊₇ and BA].The use of the name Promalin or other trade names does not imply endorsement to the exclusion of other products or vendors that may also be suitable.     

Zur Histochemie von Hydrolasen im Hoden des Hundes und der Katze

Zusammenfassung Wir untersuchten das Verteilungsmuster von unspezifischer Esterase, alkalischer Phosphatase, Adenosintriphosphatase, 5-Nucleotidase und -D-Glucuronidase im Hoden von Hund und Katze. Besonders hervorzuheben sind eine starke Aktivität der unspezifischen Esterase in den Sertolizellen der Katze, der Reichtum der Membrana propria aller Hodentubuli an alkalischer Phosphatase und Adenosintriphosphatase sowie die kräftige Reaktion auf -D-Glucuronidase in den Leydigzellen beider Tierarten.Die Befunde werden diskutiert.

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Summary The localization of unspecific esterase, alkaline phosphatase, adenosine triphosphatase, 5-nucleotidase, and -D-glucuronidase in the testes of cat and dog was demonstrated by histochemical means. We observed a strong esterase activity in the Sertoli cells of the cat and high amounts of alkaline phosphatase and adenosine triphosphatase in the membrana propria of all seminiferous tubules. In both species the principal site of -D-glucuronidase was in the Leydig cells. Our findings obtained being discussed.

     

Relationship of embryogenic competence in maize (Zea mays L.) leaves to mitotic activity,cell cycle and nuclear DNA content

Axillary bud explants of 11 selected mature waratah clones were established in vitro on a modified Murashige & Skoog medium. Adequate proliferation of axillary shoots was achieved by optimisation of the growth regulator status of the culture medium. For the majority of clones, a three to six times rate of proliferation was achieved with 1.25 M BA and 1.0 M GA₃ without the occurrence of abnormalities. The white flowering clone did not respond favourably to the addition of GA₃ to the medium.Abbreviations BA benzyladenine - GA₃ gibberellic acid - IBA indole-3-butyric acid - LSD least significant difference - MS Murashige & Skoog medium     

In vitro regeneration through organogenesis of Meconopsis simplicifolia — An endangered ornamental species

Meconopsis simplicifolia (D.Don) Walp. could be propagated by induction of adventitious shoots from callus produced on hypocotyl, cotyledon and rosette leaf explants of 4-month-old seedlings. Callus was initiated on agar-solidified Murashige and Skoog medium supplemented with 1 M kinetin +10 M -naphthaleneacetic acid (NAA). Shoots formed when the callus was subcultured on medium supplemented with kinetin or benzyladenine (BA) in combination with NAA, indoleacetic acid, indolebutyric acid or gibberellic acid. Excised shoots were rooted on medium containing auxin with 10 M NAA producing the best rooting (55%).Abbreviations BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxy-acetic acid - FAA formalin-acetic acid-alcohol - GA₃ gibberellic acid - IAA indole-3-acetic acid - IBA -indolebutyric acid - NAA -naphthaleneacetic acid     

Establishing micropropagation conditions for five Cotoneaster genotypes

Micropropagation from shoot tips of five Cotoneaster genotypes C. franchetii, C dammeri var. Radicans, C. dammeri Skogholm, C. dammeri Eichholz N° 1 and C. dammeri Streib's Findling) is described. For culture initiation and multiplication, a medium based on Murashige and Skoog basal salts was suitable for all genotypes. For multiplication, 4.4 M 6-benzyladenine provided the optimum results, while auxins proved to be essential for rooting in vitro. However, direct acclimatization of unrooted microcuttings was more efficient than in vitro rooting and acclimatization of rooted plants.Abbreviations BA 6-benzyladenine - GA₃ gibberellic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige & Skoog (1962) basal medium - NAA -naphthaleneacetic acid     

Degradation pathways from n-tridecane to α, ω-tridecanedioic acid in a mutant of Candida tropicalis

Summary The metabolic formation of ,-tridecanedioic acid via n-tridecanoic acid and via ,-tridecanediol from n-tridecane in the mutant S 76 of Candida tropicalis was studied. It was found that resting cells of S 76 produced ,-tridecanediol from n-tridecane.With n-tridecanol as substrate, the ,-diol could also be detected. The mutant S 76 was able to produce ,-tridecanedioic acid using either n-tridecanol or n-tridecanoic acid as the sole carbon source. Quantitative changes in the concentration of -hydroxy tridecanoic acid and other intermediates were recorded during the formation of ,-dioic acid.The results confirm the existence of two metabolic pathways mentioned above in the course of ,-dioic acid formation from odd n-alkane in the mutant S 76 of C. tropicalis.     

(Aminooxy)acetic acid inhibits petunia growth and gibberellin- and cytokinin-stimulated growth in bioassays

(Aminooxy)acetic acid (AOA) was applied to greenhouse-grown petunias and was used in bioassays for three plant growth hormones so that its growth regulator properties could be studied. In greenhouse studies foliar sprays of 4.8–12 mm AOA inhibited vegetative growth of petunia seedlings (Petunia xhybrida Vilm. White Flash). When gibberellin A ₃ (GA₃) was applied to shoot tips previously treated with AOA, plant growth was stimulated, but there was no AOA x GA₃ interaction. Some changes in petunia leaf morphology induced by AOA were reversed by GA₃. AOA inhibited elongation of corn coleoptile segments (Zea mays L. B73 x Mol7) whether or not 10 m indole-3-acetic acid (IAA) was present, but there was no AOA x IAA interaction. AOA reduced lettuce hypocotyl (Lactuca sativa L. Grand Rapids) elongation induced by GA₃ and radish cotyledon (Raphanus sativus L. Champion) expansion induced by benzyladenine (BA). We propose that AOA interferes with postsynthetic metabolism of plant hormones during cell elongation induced by GA₃ and cell expansion induced by BA.Abbreviations AOA (aminooxy)acetic acid - GA₃ gibberellin A₃ - IAA indole-3-acetic acid - BA benzyladenine     

The biological activities of some new gibberellins (GAs) in six plant bioassays

The biological activities of GA₄₀, GA₄₃, GA₄₆, GA₄₇, GA₅₁ and GA₄ 20,4-lactone were tested over a wide range of concentrations in six plant bioassays. GA₄ 20,4-lactone showed the highest activity. Of the two 2-hydroxylated compounds GA₄₇ showed moderately high activity, and GA₄₀ was slightly active. The 2-hydroxylated compunds GA₄₃, GA₄₆ and GA₅₁ were virtually inactive.Abbreviations GA_n gibberellin A_n - GC-MS gas chromatography-mass spectrometry - Me methyl - TLC thin layer chromatography - TMS trimethylsilyl The author is née Frydman     

Phagostimulation of the mediterranean fruit fly, Ceratitis capitata by ribonucleotides and related compounds

Females of the medfly, Ceratitis capitata, prefer sucrose solutions containing ribonucleotides to sucrose solutions without them. The order of preference for the nucleotides was: 5GMP>GTP>5CMP>5IMP >dGMP>5UMP>5AMP>5XMP=ATP=2 & 3GMP=RP>3AMP.2AMP, guanosine, inosine, adenine and 5TMP produced no significant stimulation. Females sterilized by irradiation showed reduced attraction to 5GMP as compared to non-irradiated females.Optimal molecular configuration for phagostimulation includes: phosphorylation at the 5 position of the ribose, free hydroxyl groups at 2 and 3 on the ribose, and an NH₂ group at the 2 position of the aromatic ring of purine.It is proposed that the 5GMP in yeast hydrolyzate can be used as a measure of the suitability of the hydrolyzate as a bait.

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Résumé La femelle de la mouche méditerranéenne des fruits, Ceratitis capitata, préfère les solutions de sucrose contenant des ribonucléotides aux simples solutions de sucrose. Lórdre de préférence pour les nucléotides est le suivant: 5GMP>GTP>5CMP>5IMP >dGMP>5UMP>5AMP>5XMP=ATP =2 & 3GMP=RP>3AMP.Le 2AMP, la guanosine, l'inosine, l'adénine et le 5TMP provoquent une stimulation significative. Les femelles montrent aprés stérilisation par irradiation une attirance réduite pour le 5GMP par comparaison avec les femelles non-irradiées.La configuration moléculaire optimale pour la phagostimulation comprend: la phosphorylation en position 5 du ribose; des groupes hydroxyles libres en 2 et 3 sur le ribose; et un groupe NH₂ en position 2 sur le noyau aromatique.Nous proposons que le 5GMP dans l'hydrolysat de levure puisse être utilisé pour mesurer la capacité de l'hydrolysat comme appât.

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Abbreviations 5AMP Adenosine 5-monophosphate - 3AMP Adenosine 3-monophosphate - 2AMP Adenosine 2-monophosphate - dAMP 2-deoxyadenosine 5-monophosphate - ADP Adenosine 5-diphosphate - ATP Adenosine 5-triphosphate - 5GMP Guanosine 5-monophosphate - 2GMP Guanosine 2-monophosphate - 3GMP Guanosine 3-monophosphate - dGMP 2-deoxyguanosine 5-monophosphate - GDP Guanosine 5-diphosphate - GTP Guanosine 5-triphosphate - 5IMP Inosine 5-monophosphate - IDP Inosine 5-diphosphate - ITP Inosine 5-triphosphate - 5XMP Xanthosine 5-monophosphate - 5CMP Cytidine 5-monophosphate - dCMP 2 deoxycytidine 5-monophosphate - CTP Cytidine 5-triphosphate - 5UMP Uridine 5-monophosphate - 5TMP Thymidine 5-monophosphate - RP Ribose 5 monophosphate     

Abbau von 14C-, 3H- und 36Cl-markiertem γ-Hexachlorcyclohexan durch anaerobe Bodenmikroorganismen

Zusammenfassung Durch eine anaerobe Mischflora aus Ackerboden wurde -Hexachlorcyclohexan (-HCH) in 4–5 Tagen zu 90% abgebaut. Dabei erfolgte eine schnelle Abspaltung des Chlors in Form von Chloridionen und danach eine Freisetzung des C- und H-Anteiles in Form flüchtiger Verbindungen, in denen kein Chlor und auch kein CO₂ nachzuweisen war.Die Verwendung von ¹⁴C/³H- und ³⁶Cl/³H-doppelmarkiertem -HCH zeigte, daß die Cl- und H-Abspaltung nicht im Verhältnis von 1:1 erfolgte, sondern mehr Cl als H abgespalten wurde. Die flüchtigen Verbindungen enthielten andererseits höhere ¹⁴C- als ³H-Anteile. Gaschromatographische Untersuchungen zeigten ebenfalls eine rasche Verminderung des -HCH und die Bildung verschiedener Metabolite. Es wurde jedoch kein -Pentachlorcyclohexen nachgewiesen. Bei steigenden O₂-Gehalten in der Gasphase verminderte sich der -HCH-Abbau. Jedoch fanden auch noch bei 5% O₂ Chlorabspaltung und die Freisetzung flüchtiger Metabolite statt.-HCH wurde ebenfalls, jedoch langsamer, durch die anaerobe Mischflora abgebaut. Auch hier wurde Chlorid abgespalten, und es traten ebenfalls flüchtige Verbindungen auf, die kein Chlor enthielten.

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Degradation of ¹⁴C-, ³H- and ³⁶Cl-labelled -hexachlorocyclohexane by anaerobic soil microorganisms

Up to 90% of the -Hexachlorocyclohexane (-HCH) applied to an anaerobic mixed bacterial flora enriched from an arable soil were degraded within 4–5 days. Degradation resulted in a rapid release of chloride and in formation of chlorine-free volatile metabolites. CO₂ formation from the molecule was not detected.Investigations with ¹⁴C/³H- and ³⁶Cl/³H double-labelled -HCH indicated that the release of Cl and H did not occur in the ratio of 1:1. More Cl than H was split off. The volatile compounds contained more ¹⁴C than ³H. Gas chromatographic studies also showed the rapid decrease of -HCH and the formation of several metabolites. -Pentachlorocyclohexene was not detected. Increasing O₂-contents in the gas phase of cultures resulted in decreases of the compound's degradation. Release of chloride and of volatile metabolites were observed with O₂ contents in the gas phase up to 5%.-HCH was also, but more slowly as with -HCH, degraded by the anaerobic mixed flora. Chloride was released and volatile, chlorine-free metabolites were found.

     

α-Amylase production and leaf protein synthesis in a gibberellin-responsive dwarf mutant of ‘Himalaya’ barley (Hordeum vulgare L.)

A dwarf mutant, M117, was isolated following sodium-azide mutagenesis of barley (Hordeum vulgare L. Himalaya). Treatment of the mutant with gibberellic acid (GA₃) restored growth to levels of the tall parent, -Amylase production was examined in germinated grains of the dwarf mutant and in Himalaya plants treated with gibberellin (GA) biosynthesis inhibitors. The mutant showed reduced -amylase activity relative to the parent when grains were germinated on water, but activities were equivalent to the parent following germination on GA₃ solution. Germination of normal or mutant grains in the presence of GA biosynthesis inhibitors led to reduced -amylase activity levels, but normal levels were restored if GA₃ was included in the inhibitor solution. These data are consistent with a model in which -amylase production in the germinated grain is regulated by the supply of active GAs. Treatment of M117 with GA₃ increased the length, fresh weight, dry weight, volume, cell number, and protein content of the first leaf. Proteins being synthesized in the first leaf were labelled with [³⁵S]methionine and fractionated by two-dimensional electrophoresis. No reproducible qualitative or quantitative differences in protein profiles were detected in response to GA₃ treatment. In contrast, first leaves from seedlings exposed to dehydration stress had profiles clearly distinguishable from those of control seedlings. Stem sections from dwarf plants maintained on 10 M GA₃ in the presence of sucrose elongated significantly more than controls without GA₃, but two-dimensional analysis of the [³⁵S]methionine-labelled radioactive polypeptides again revealed no GA₃-induced differences. It was concluded that enhanced elongation rates of leaves or stem segments were not associated with major changes in gene expression.Abbreviations 2D two-dimensional - GA gibberellin - GA₃ gibberellic acid - PB paclobutrazol We would like to thank Dr Barbara Read (Agricultural Research Institute, Wagga Wagga, Australia) for assistance with growth of barley plants, and Tony Carter, Alison McInnes, and Mark Cmiel for skilled technical assistance.     

Über das Wachstum der Pankreaszellen von Säugetieren als Monolayer Cultures

Zusammenfassung Glucose und Coffein erhöhen den Gehalt 4 und 7 Tage alter Zellen an immunologisch meßbarem Insulin (Ausgangsmaterial: Ratten- und Schweinepankreas). Morphologisch (Aldehydfuchsin, Pseudoisocyanin) sind bei coffeinbehandelten Zellen in der Intensität der Anfärbung und in der Granuladichte zeitabhängige Unterschiede zu beobachten, die an 4 Tage alten Zellen deutlicher als an 7 Tage alten zu erkennen sind. Coffein erhöht die Verfettung der Zellen. Weiter nimmt der Gehalt an sauren Mucopolysacchariden (Eisenbindungsreaktion) und an PAS-positivem Material zu. Diese Erhöhung läuft mit dem Anstieg des Insulingehaltes im Nährmedium parallel.

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Monolayer cultures of pancreatic tissueIII. Insulin release induced by coffein and glucose

Summary Glucose and caffeine increase the immunologically measurable insulin content of four and seven-day old cells (starting material: rats' or pigs' pancreas). In morphological examination (aldehyde fuchsin, pseudoisocyanin), time-dependent differences in the intensity of staining and granular density are observable in caffeine-treated cells; these phenomena are more distinct in four-day old cells than in seven-day old ones. Caffeine increases fatty degeneration of the cells. Furthermore, the content of acid mucopolysaccharides (iron-binding reaction) and of PAS positive material increases. This increase runs parallel with the rise in insulin content of the nutritive medium.