Control of corneal differentiation by extracellular materials. Collagen as a promoter and stabilizer of epithelial stroma production

The primary stroma of the cornea of the chick embryo consists of orthogonally arranged collagen fibrils embedded in glycosaminoglycan (GAG) produced by the epithelium under the early inductive influence of the lens. The experiments reported here were designed to test whether or not the collagen of the lens basement lamina is capable of stimulating corneal epithelium to produce primary stroma. Enzymatically isolated 5-day-old corneal epithelia were grown for 24 hr in vitro in the presence of ³⁵SO₄ or proline-³H on various substrata. Epithelia cultured on lens capsule synthesized 2.5 times as much GAG (as measured by incorporation of label into CPC precipitable material) and almost 3 times as much collagen (assayed by hot TCA extraction or collagenase sensitivity) as when cultured on Millipore filter or other noncollagenous substrata. A similar stimulatory response was observed when epithelium was combined with chemically pure chondrosarcoma collagen, NaOH-extracted lens capsule, vitreous humor, frozen-killed corneal stroma or cartilage, or tendon collagen gels; in the latter case, the magnitude of the effect can be shown to be related to concentration of the collagen in the gel. All of the collagenous substrata stimulate not only extracellular matrix production, but also polymerization of corneal-type matrix, as judged by ultrastructural criteria and by the association of more radioactivity with the tissue than the medium. Since purified chondrosarcoma collagen is as effective as lens capsule, the stimulatory effect on collagen and GAG synthesis by corneal epithelium is not specific for basal lamina (lens capsule) collagen.     

Ribonucleic Acid and protein synthesis in chick embryo cells infected with fowl plague virus

Fowl plague virus comprised four major protein components and several minor ones, two strains of the virus giving similar results. One of the components was identified as the nucleocapsid protein. Synthesis of the virion proteins could readily be detected in infected cells 3 hr after infection. The two subcellular fractions associated with viral ribonucleic acid (RNA) polymerase activity (nuclei and ribosomal pellet) were associated with the protein of the nucleocapsid and a second virion protein of unidentified function. Measurement of viral RNA and protein synthesis in cells infected with preparations of ultraviolet irradiated virus showed that the capacity to synthesise the RNA and protein species of highest molecular weight was lost most quickly, suggesting that the pieces of viral RNA function independently.     

The Endocommensal Ciliates of Venezuelan Sea Urchins

SYNOPSIS. Two hundred and fifty-nine specimens of the sea urchins Echinometra lucunter (201), Diadema antillarum (52), Tripneustes ventricosus (5) and Eucidaris tribuloides (1), collected on the littoral of the Federal District of Venezuela, were examined for intestinal ciliates.
Iron hematoxylin and silver-impregnation staining technics permitted determination of eight of the nine ciliates found: Anophrys aglycus, A. elongata, Cohnilembus caeci, Biggaria bermudensis, B. echinometris, Cyclidium rhabdotectum, Metopus circumlabens and M. rotundus . An unidentified species of Euplotes was also found.     

Cochlodinium heterolobatum n.sp.: Structure and Some Cytophysiological Aspects*

SYNOPSIS. Structure and morphogenesis, and cytochemical data on Cochlodinium heterolobatum, a new species of unarmored dinoflagellate, were derived from living and fixed material from culture. C. heterolobatum is characterized by the torsion of the girdle which descends in a left-hand spiral 1.8 turns; the sulcus having a torsion of 0.8 turn; a sulcus loop in the epicone; a tongue-shaped lobe in the right hypocone; nucleus in the epicone; and a stigma in the left epicone. Trichocysts and behavior of the nucleus during typical and atypical divisions are described in cells from cultures of different ages. A small form with the specific characters was found. Intracellular bacteria were seen and their growth followed in individuals from cultures of different ages. A possible relationship between those bacteria and the accumulation of metabolites inside old cells is discussed.     

Inhibition of glycosidases by aldonolactones of corresponding configuration: Preparation of (1-->5)-lactones by catalytic oxidation of pyranoses and study of their inhibitory properties

1. A method was devised for the preparation of (1→5)-lactones from pyranose sugars and uronic acids by platinum-catalysed oxidation with gaseous oxygen in aqueous solution at acid pH. It was applied to mannose, N-acetylglucosamine, N-acetylgalactosamine, glucuronic acid, galacturonic acid, galactose, l-arabinose and d-fucose. 2. Only the first three yielded products that could be obtained in the solid state without decomposition. In every case, however, the oxidation product in aqueous solution behaved as the aldono-(1→5)-lactone, and was more inhibitory towards the appropriate glycosidases than any aldonolactone preparation hitherto examined. 3. The stabilities of the oxidation products were studied, and their interconversion with the (1→4)-lactones was demonstrated. Ring-opening does not appear to be mandatory for this isomeric change, which in some instances is very rapid. 4. To explain all the inhibitory effects observed with aldonolactones on glycosidases of corresponding configuration, it is tentatively postulated that inhibition may be due entirely to the (1→5)-lactone, and that any inhibitory effect seen with the (1→4)-lactone is a measure of the extent and speed of its conversion into the (1→5)-lactone in aqueous solution.     

RECIPROCAL BEHAVIOR OF THE RACHIS AND PEDUNCLE IN COLONIES OF Veretillum cynomorium PALL.

Some integrated patterns of behavior have been studied in coloniesof Veretillum cynomorium, whose various parts often act as ifthey were those of an individual consisting of the whole colony.The entire colony is involved in slow rhythmical movements,in which phases of contraction alternate with expansion. Thisbehavior corresponds to a permanent reciprocity (or duality)between the zooid-bearing rachis and the peduncle; overall behavioris determined by the number of "functionally active units" whicheither contains at any given time. This number is influencedby the excitatory or restraining effects of external factorssuch as light intensity or the entry of water. In this way,when contraction proceeds, activity of the rachis predominates,whereas during swelling of the colony, activity of the peduncleprevails. Observations on intact colonies and on specimens without peduncles,experiments using light and electrical stimuli, and the studyof peristalsis, all suggest that the mutually reciprocal natureof activity in the rachis and peduncle may be attributed tothe differential influence of external factors on the functionalstate of centers of spontaneous activity or of conduction pathwaysin the colony. Zooids present on the rachis are responsibleboth for the reciprocal activity of the colony and for its morphologicalpolarity.     

The Respiratory Metabolism of Strelitzia juncea Ait. Seeds. The Effect of Dormancy Release by Oxygen on certain Glycolytic Enzyme Activities and Metabolite Concentrations

The activity of the glycolytic enzymes PFK, PFP, PK and aldolaseas well as the content of glucose, fructose, glucose-6-phosphateand fructose-6-phosphate were compared in the embryos of airand oxygen-incubated seeds of Strelitzia juncea. Determinationswere made during the first 4 d of incubation, prior to radicleemergence, which commences on day five for oxygen-treated seeds. No difference in PFK activity was found for the two treatments,and for both treatments PFK tended to increase with the incubationperiod. The fr2, 6P₂-stimulated PFP activity was slightly higherfor oxygen-incubated seeds, and showed a significant increasein activity over the 4 d incubation period for both treatments.No significant change in the general trend of PK and aldolaseactivity resulted from incubating the seeds in oxygen. The almost equimolar glucose and fructose contents of the embryoswere lower after 1 d of oxygen incubation of the seeds, andthe content decreased sharply during the incubation period.It is concluded that a moderate increase in the glycolytic capacityof embryos resulted from oxygen treatment of S. juncea seeds. Key words: Dormancy, glycolysis, Strelitzia juncea     

Spikelet Sterility and Flowering Response of Rice to Water Stress at Anthesis

Water deficits at the anthesis stage of rice (Oryza sativa L.)induce a high percentage of spikelet sterility and reduce grainyield. This study attempted to elucidate the direct effectsof water stress on panicle exsertion, spikelet opening, andspikelet desiccation leading to spikelet sterility. A well-wateredtreatment and two water stress levels were imposed in pot-grownplants of IRAT 13 (upland cultivar) and IR20 (lowland cultivar)at the time of flowering under greenhouse conditions A cultivar difference was observed in the flowering responseto water stress with a high sensitivity in IR20. The time courseof panicle exsertion showed an inhibitory effect due to thelow panicle water status. Low panicle water potentials significantlyreduced the number of opened spikelets. Spikelet opening wascompletely inhibited at panicle water potentials below –1·8MPa and –2·3 MPa in IR20 and IRAT 13, respectively.However, the peak spikelet opening time in a day was not influencedby the stress treatment. Spikelets in stressed panicles wereobserved to remain open for a longer period than in the well-wateredpanicles. The role of turgor in spikelet opening is also discussedin the study. At low panicle water potentials, severe desiccationof spikelets and anthers was noted. The deleterious effectsof water deficits on spikelet opening and spikelet water losscontributed to reduced spikelet fertility Oryza sativa L., rice, spikelet sterility, flowering, water stress, panicle water potentials, turgor potentials, desiccation     

Complete primary structure of statherin, a potent inhibitor of calcium phosphate precipitation, from the saliva of the monkey, Macaca arctoides

Human saliva, which is supersaturated with respect to basic calcium phosphate salts, is stabilized primarily by the presence of two classes of phosphoproteins, statherin and the acidic proline-rich proteins (PRP). These molecules act by inhibiting both primary (spontaneous) precipitation of calcium phosphates in saliva and secondary (surface induced) precipitation of these salts onto dental enamel. The complete amino-acid sequences of several human PRP and the N-terminal sequence of PRP from saliva of M. arctoides have been determined. Similarly, the complete sequence of statherin from human and M. fascicularis saliva is known. We now report the complete structure of statherin from the saliva of the stump-tailed monkey, M. arctoides. The structure was determined by gas-phase sequencing of intact statherin, elucidating positions 1-26, and sequencing an unpurified mixture of tryptic peptides which elucidated the remaining positions through the C-terminus (residue 42) of the molecule. This latter degradation produced an eight amino-acid overlap with that of intact statherin and was confirmed by C-terminal analysis and amino-acid composition of native statherin. The complete amino-acid sequence of M. arctoides statherin is: NH2-Asp-PSer-PSer-Glu-Glu5-Lys-Phe-Leu-Arg-Arg10 -Leu-Arg-Arg-Phe-Asp15-Glu- Gly-Arg-Tyr-Gly20-Pro-Tyr-Gln-Pro-Phe25-Val-Pro-Pro- Pro29Leu30-Tyr- Pro-Gln-Pro-Tyr35-Gln-Pro-Tyr-Gln-Pro40-Gln-Tyr-COOH This sequence differs from human statherin at positions 11, 12, 15, 16, 18, 25-27, 38-40 and from M. fascicularis statherin at positions 26 and 28.