Serologic Changes Associated with the Encystment of Axenically Grown Hartmannella culbertsoni*

SYNOPSIS. Serologic reactions elicited by sonically ruptured trophic and cystic forms of Hartmannella culbertsoni were studied. The antigens of trophic amoebae reacted with their homologous rabbit antiserum showing multiple precipitin lines which could not be seen when the reacting antigens were treated with trypsin prior to application on the Ouchterlony plates. Antigens of trophic amoeba did not react with antiserum against cysts. Cyst antigens reacted with their homologous antiserum only after trypsin treatment. Antigens prepared from trophozoites excysting from cysts reacted positively with the antiserum against antigens of trophic amoebae. Antigens of trophic as well as cystic forms fixed guinea pig complement in presence of their homologous antisera. With the trophic form, this property was abolished after trypsin treatment. Non-specific complement fixation mediated by cyst antigens was abolished by treatment with cellulase. Antiserum against trophic amoebae immobilized trophozoites and, in the presence of guinea pig complement, led to their lysis.     

Ultrastructural Changes During Encystment of Schizopyrenus russelli*

Ultrastructural changes associated with the encystment of Schizopyrenus russelli have been studied by electron microscopy. Before encystment small “black bodies” appear in the cytoplasm and later migrate toward the periphery. The outer cyst wall is secreted at this stage as a thin discontinuous layer which thickens and subsequently becomes continuous. Concomitant with this, the endoplasmic reticulum surrounds the mitochondria. The inner cyst wall later appears as a multilayered structure which presumably is cast off from the plasma membrane. Between the inner and outer layers of the cyst wall, there is a middle, less electron-dense layer wherein extruded cytoplasmic material is found embedded at certain places.     

Acid Hydrolase Activity During Growth and Encystment in Acanthamoeba castellanii*

SYNOPSIS. The activity and sedimentation of acid phosphatase (APase), acid deoxyribonuclease (DNase), and acid ribonuclease (RNase) were investigated throughout growth and encystment in Acanthamoeba castellanii. The activities/mg protein of all 3 hydrolases are high in young cultures and decrease to constant levels in postlog cells. The RNase activity/ ameba decreases 50% during growth, whereas the activity/cell of both APase and DNase remains constant. The percent sedimentation at 20,000 g of all 3 enzymes gradually increases from about 40% in midlog to a plateau of 80% in postlog cells. During encystment, the sedimentation behavior of RNase differs from that of APase and DNase. Encystment is characterized by a differential decrease in the activity/cell of the 3 hydrolases, with RNase decreasing most rapidly and APase least rapidly. APase is unique in that a transient increase of its specific activity is noted during encystment, even though its activity/cell is decreasing.     

Synthesis of RNA,Protein, Cellulose,and Mucopolysaccharide and Changes in the Chemical Composition of Hartmannella culbertsoni During Encystment under Axenic Conditions*

Hartmannella culbertsoni trophozoites are transformed into viable cysts on exposure to a non-nutrient agar medium containing 15 mM MgCl₂ and 20 mM taurine. Amebae differentiating in this encystment medium incorporate more uracil-2-¹⁴C into RNA and more leucine-1-¹⁴C or valine-1-¹⁴C into proteins than controls. Encysting organisms incorporate significantly more glucose-U-¹⁴C into cellulose and glucosamine-1-¹⁴C into mucopolysaccharides. Incorporation of glucose-U-¹⁴C into cellulose and of glucosamine-1-¹⁴C into mucopolysaccharides are inhibited by actinomycin D or cycloheximide.     

Changes in Enzymes of Sucrose Metabolism and the Activation of Certain Chloroplast Enzymes During Wheat Leaf Senescence

Relatively little change was observed in the partitioning ofphotosynthetically fixed carbon between sucrose and starch duringsenescence of vegetative leaves of wheat (Triticum aestivumL. cv. Roy). The maximum activities of sucrose phosphate synthaseand cytoplasmic fructose-1, 6-bisphosphatase in leaf extractsapproximated photosynthetic sucrose formation in leaf segmentsand the three parameters declined together with age. Changesin activities of sucrose synthase and uridine-5'-diphosphatase,and in leaf phosphate content did not parallel changes in sucroseformation with age. Chloroplast fructose-1, 6-bisphosphatase activity, activatedin vivo with light and in vitro with dithiothreitol, declinedcontinually throughout senescence of the second vegetative leaf.The ability of light to activate chloroplast fructose-1, 6-bisphosphatasedid not, however, appear to change with age. Ribulose-l, 5-bisphosphatecarboxylase activity also declined continually with age, butin vivo-activated ribulose-l, 5-bisphosphate carboxylase wasa minimum of three fold greater than needed to account for photosyntheticactivity. The percentage of ribulose-l, 5-bisphosphate carboxylaseactivated in vivo was essentially constant during senescence.In general, activities of soluble chloroplast enzymes declinedcontinually from the point of full leaf expansion throughoutsenescence, whereas the activities of cytoplasmic enzymes thatwere studied remained relatively constant until the latter stagesof senescence. Key words: Wheat leaf senescence, Sucrose metabolism enzymes, Chloroplast enzymes     

顽拗性黄皮种子脱水过程中活性氧清除酶活性的变化

以正常性种子花生为对照,研究了顽拗性黄皮种子脱水过程中活性氧清除酶、膜脂过氧化作用以及电解质渗漏率的变化。随着含水量的下降,黄皮胚的电解质渗漏率和膜脂过氧化产物丙二醛(MDA)含量均显著增加;当黄皮胚含水量下降至40％后,SOD活性开始急剧下降,而POD和CAT活性在胚含水量下降过程中呈现出缓慢下降的趋势。花生胚在含水量从45％降至14％的过程中,电解质渗漏率没有明显增加,MDA含量只有少量增加;当含水量降至14％后,电解质渗漏率出现少量增加。花生胚脱水初期,活性氧清除酶活性明显增加,并在整个脱水过程中维持较高的水平。以上结果表明顽拗性种子黄皮的脱水敏感性与活性氧清除酶相对活性变化有关。脱水引起黄皮胚活性氧清除酶活性降低,活性氧清除能力下降,膜脂过氧化作用加强,膜透性增大。黄皮胚的膜系统可能是脱水伤害的靶位之一。     

Changes in Activity of Glyoxylate Cycle Enzymes During Myxospore Development in Myxococcus xanthus

Activities of the glyoxylate cycle enzymes isocitrate lyase (EC 4.1.3.1) and malate synthase (EC 4.1.3.2) were assayed in extracts prepared at different stages of myxospore formation in liquid cultures of Myxococcus xanthus. Activities of both enzymes attained peak values during conversion of rods to spheres. Isocitrate lyase activity decreased after reaching its peak value. Malate synthase activity also declined but at a much slower rate. The loss of isocitrate lyase activity could be prevented by the addition of chloramphenicol to cultures early in myxospore formation (during the initial rise in enzyme activity), but not by such addition at later stages of myxospore formation. The increase in glyoxylate cycle enzymes was not observed in a mutant unable to form myxospores in liquid culture under conditions suitable for morphological conversion of the wild type, or in wild-type cells incubated in the absence of an inducer for myxospore formation. It is concluded that the changes in the glyoxylate cycle enzymes represent regulatory phenomena associated with the development of the myxospore.     

Changes in the Activity of Certain Enzymes of Acer pseudoplatanus L. Cells at four Stages of Growth in Suspension Culture

The activities of twelve enzymes were studied in Acer psendoplatanum L. cells grown in suspension cultures for 6, 12, 19 and 22 days. The enzyme activities of cells (6 and 12 days) in the phase of rapid cell division were rather different from that of cells in the stationary phase (19, 22 days). The activity of aldolasc was highest at the first two stages of growth, when the amount of RNA and protein per cell was highest, and after this decreased steeply. The curves of specific activities of several phosphatases (glucosc-6-phosphatasc, α-glycerophosphatase, acid phosphatase and ATPase) and ribonuclease were almost the reverse of the curve of aldolase, showing a minimum in 12-day-old cells and a maximum in cells 19 and 22 days old. The activity of peroxidase was also high in ageing cells. Glutamate: oxaloacetate transaminase and aminopeptidases (leucine and alanine) had synchronous maxima (6 and 19 days) of specific activity but the activity of aminopeptidases decreased gradually during ageing of the suspension, if the enzyme activities are calculated per 10⁷ cells. Glutamate: pyruvate transaminasc activity was very low and no dcoxyribonuclease activity could be detected.     

青花菜生育过程中叶片的几种酶活性变化（简报）

青花菜生长发育过程中,叶片的硝酸还原酶（NR）活性高峰出现在花序分化后2周,超氧化物歧化酶（SOD）活性高峰比NR早1周,过氧化物酶（POD）活性变化规律大致与SOD相同,苯丙氨酸解氨酶（PAL）活性在花序分化前逐渐降低,花序分化后逐渐回升,现苗时达到高峰,花球形成过程中逐渐下降,蛋白酶活性在花序分化前逐渐上升,花序分化后一度下降,花球形成过程中持续上升。     

Structure of Cyst Wall Precursors and Kinetics of Their Appearance During the Encystment of Laurentiella acuminata (Hypotrichida,Oxytrichidae)1

The encystment of Laurenliella acuminata was divided into five stages: stage A (precystic semitransparent cell with dark-globules), stage B (precystic transparent cell), stage C (precystic pigmented cell), stage D (spherical shape without cyst wall) and stage E (young resting cyst), on the basis of observations of changes in morphology and pigmentation during encystment. The duration of these stages was also established. Observations by electron microscopy confirmed that the cyst wall, composed of four layers, is derived from different kinds of precursors which are synthesized “de novo.” The ectocyst precursors are composed of stacks of between 5 and 12 small thin plates or discs; these stacks are about 0.9 μm in length and 0.06 μm in height. The mesocyst precursors are fibrillar bodies of variable shapes, about 2.4 μm in maximum length and 0.12–0.16 μm in diameter. These precursors appear in the cytoplasm of the precystic cell during the first precystic stage (stage A). The endocyst precursors are rounded bodies surrounded by a fine membrane, and their contents appeared similar to the endocyst. The granular layer precursors are spherical bodies about 0.1–0.2 μm in diameter, surrounded by a double membrane presenting ribosomes adhering to its outer membrane. Both endocyst and granular layer precursors are observed in the precystic cytoplasm from stage B. On the basis of ultrastructural studies, a formation and growth model of the cyst wall of the hypotrichous ciliate Laurentiella acuminata is proposed.     

Changes in Esophageal Gland Activity During the Life Cycle of Nacobbus aberrans (Nemata: Pratylenchidae)

Electron and light microscopy were used to study the dorsal gland (DG) and the two subventral glands (SvG) of seven developmental phases of Nacobbus aberrans: pre-parasitic second-stage juveniles (J2), parasitic J2, third- (J3) and fourth- (J4) stages, migratory females, young sedentary females, and mature sedentary females. In each developmental phase the level of esophageal gland activity, was estimated by the abundance of organelles associated with secretory pathways, including endoplasmic reticulum, ribosomes, Golgi, multivesicular bodies, and secretory granules. All esophageal glands were metabolically active in all J2 examined, although only in parasitic J2 were there numerous secretory granules in the esophageal gland extensions and ampullae. No evidence of secretory activity was observed in the esophageal glands of the coiled and relatively inactive J3 and J4, nor in migratory females; these stages apparently do not feed. Observations suggest that reserves stored by J2 sustain three ecdyses and the migratory female''s search for a feeding site and induction of a syncytium. Feeding activity is resumed in young and mature sedentary females, in which the DG is highly active and enlarged. The SvG are metabolically active, but with little synthesis of secretory granules, suggesting that in sedentary females the SvG may have physiological roles other than digestion.     

Effect of the Rhizobium leguminosarum252 Agglutinins on the Activity of Certain Enzymes in Plant Cells

The incubation of pea seedling roots with the surface agglutinins R₁and R₂of Rhizobium leguminosarum252 brought about an increase in the activity of proteases, -glucosidase, and, especially, succinate dehydrogenase in the roots. The data presented show that rhizobial agglutinins play an important part in the formation and functioning of legume–rhizobial associations.     

Changes in Activity of Mitochondrial Enzymes during the Development of Xenopus laevis

Changes in activity of mitochondrial enzymes were studied during the embryonic development of Xenopus laevis.
The following enzymes were determined: malate dehydrogenase (MDH), isocitrate dehydrogenase (NAD⁺-dependent) (IDH), aspartate aminotransferase (GOT), cytochrome oxidase (COX), succinate dehydrogenase (SDH), rotenone-insensitive NADH cytochrome c reductase (NADH-red) and monoamine oxidase (MAO). IDH is constant throughout the period studied. COX and SDH, two enzymes of the inner membrane, are constant in pregastrula stages, and subsequently decrease significantly. MDH and NADH-red are highly active in the pregastrula stages and decline thereafter, while MAO is undetectable during early development and increases significantly only in the larvae. GOT increases during the cleavage stages, being most active in the gastrula stages, and decreases subsequently.
The results are discussed in the sense of mitochondrial differentiation during the early development of the amphibian embryo.     

Changes in the Cell Volume of Didinium nasutum During Population Increase*

SYNOPSIS. In 3 species of carnivorous protozoa, the rate of individual food intake per generation declines with an increase in the density of the population. In all, the rate of division remains constant. Three hypotheses may be proposed to explain these phenomena: the individual size decreases, thus bringing about a decline in food need; the individuals in the earlier stages of population growth consume excess food which is passed on to later generations to supplement their food intake; an increase in density within the carnivore population decreases the activity rate, thus reducing the food requirement. Experiments using Didinium nasutum as the carnivore and Paramecium aurelia, syngen 4, as the food source were conducted to test these 3 hypotheses. The results contradicted the 1st hypothesis and part of the 2nd. A new working hypothesis based on the remaining hypotheses is proposed.     

Metabolism Changes During Aging in the Hippocampus and Striatum of Glud1 (Glutamate Dehydrogenase 1) Transgenic Mice

The decline in neuronal function during aging may result from increases in extracellular glutamate (Glu), Glu-induced neurotoxicity, and altered mitochondrial metabolism. To study metabolic responses to persistently high levels of Glu at synapses during aging, we used transgenic (Tg) mice that over-express the enzyme Glu dehydrogenase (GDH) in brain neurons and release excess Glu in synapses. Mitochondrial GDH is important in amino acid and carbohydrate metabolism and in anaplerotic reactions. We monitored changes in nineteen neurochemicals in the hippocampus and striatum of adult, middle aged, and aged Tg and wild type (wt) mice, in vivo, using proton (¹H) magnetic resonance spectroscopy. Significant differences between adult Tg and wt were higher Glu, N-acetyl aspartate (NAA), and NAA + NAA–Glu (NAAG) levels, and lower lactate in the Tg hippocampus and striatum than those of wt. During aging, consistent changes in Tg and wt hippocampus and striatum included increases in myo-inositol and NAAG. The levels of glutamine (Gln), a key neurochemical in the Gln-Glu cycle between neurons and astroglia, increased during aging in both the striatum and hippocampus of Tg mice, but only in the striatum of the wt mice. Age-related increases of Glu were observed only in the striatum of the Tg mice.     

豇豆叶片衰老过程中多胺氧化酶活性的变化（简报）

豇豆幼苗的连体和离体叶片衰老过程中多胺氧化酶（PAO）活性与叶绿素和蛋白质的含量都呈下降趋势,它们的降幅依次是：PAO活性大于叶绿素含量大于蛋白质含量;用氨基胍抑制离体叶圆片PAO活性,不能延缓叶绿素和蛋白质的含量下降。