The Reduction of Nicotinamide Adenine Dinucleotide by Mitochondria Isolated from Helianthus tuberosus

Intact mitochondria isolated from Jerusalem artichoke tubersoxidize both malate and citrate. Both substrates were equallyeffective in reducing the endogenous pool of pyridine nucleotidesin the presence of respiratory inhibitors. Only malate, andnot citrate, was capable of reducing exogenous NAD⁺ under similarassay conditions. The rate at which malate reduced added NAD⁺was biphasic; the initial rapid phase was inhibited by the accumulationof oxaloacetate while the velocity of the second, slower, phasewas found to be insensitive to accumulated oxaloacetate. Theaddition of the detergent Decon 90 to intact mitochondria stimulatedboth the rapid and slow phases of NAD⁺ reduction and failedto convert the biphasic rate into a constant rate. Decon 90was found to cause inhibition of soluble malate dehydrogenase.Extensive efforts to purify the mitochondria using sucrose densitygradients failed to remove all of the soluble malate dehydrogenasefrom the preparation of mitochondria and approximately 3% ofthe total malate dehyrogenase present in the preparation appearedto behave as if it were outside the inner membrane.     

Early events in the cellular formation of proparathyroid hormone

Early events in the cellular synthesis and subsequent transfer into membrane-limited compartments of pre-proparathyroid hormone (pre-proPTH) and proparathyroid hormone (proPTH) were investigated by electrophoretic analyses of newly synthesized proteins in subcellular fractions of parthyroid gland slices pulse-labeled for 0.5-5 min with [(35)S] methionine. During these short times of incubation, both pre-proPTH and proPTH were confined to the microsomal fraction. Labeled pre-proPTH and proPTH were detected in a 30-s interval between 0.5 and 1.0 min of incubation. The radioactivity in proPTH became relatively constant between 3 and 5 min, whereas the radioactivity in ProPTH increased markedly over this period. When corrected for the known content of methionine in the prohormone and the prohormone, we found four times as much radiolabeled prohormone as prehormone between 0.5 and 1.0 min of synthesis. Sequestration of labeled prohomrone into endoplasmic reticulum compartments was shown by treatment of the microsomal fraction with chymotrypsin and trypsin, which resulted in the degradation of the prehormone but not of the prohormones. Approximately 50 percent of pre-prohormone and 25 percent of prohormone were released from the microsomes by their extraction with 1.0 M KCl, whereas 80-90 percent of both was released by treatment with Triton X-100. These results in intact cells support the signal hypothesis proposed by Blobel and his co-workers in studies utilizing cell-free systems, inasmuch as the results indicate transfer of prohormone into the cisternal space of the rough endoplasmic reticulum concomitant with the growth of the nascent polypeptide chain. Appearance of membrane-sequestered proPTH takes place without entry of pre-proPTH into the cisternal space, suggesting that proteolytic removal of the leader peptide occurs during transfer of the polypeptide through the lipid bilayer. Further evidence in support of this process is that pre-proPTH is only partly extracted from the microsomes by treatment with 1.0 M KCl, suggesting that a substantial fraction of the nascent pre-proPTH is integrally inserted into the membranes before it is cleaved to form proPTH.     

Sequelae of encephalitis; report of a study after the California epidemic

In a study of almost 500 patients to determine residual effects, the sequelae of both St. Louis and Western equine encephalitis were more prominent in the younger age group. Infants under three months with Western equine encephalitis had the greatest central nervous system damage. Forty-four per cent of this entire group had sequelae. In patients between one and four years of age, the incidence of sequelae was less. The Western equine infection was associated with the more disabling residual damage. Postencephalitic convulsions were fairly common in the younger patients with Western equine disease, but not in the St. Louis group. After the age of five the sequelae rate dropped. In all age groups the Western equine residual changes were more severe than the damage of St. Louis infection. Some infants, children and adults showed remarkable improvement from sequelae even as much as two years after the abnormalities occurred. With the longer period of follow-up, some late sequelae were noted in children and adults, primarily among those who had Western equine infection.     

Xylem ABA controls the stomatal conductance of field-grown maize subjected to soil compaction or soil drying

Stomatal conductance of individual leaves was measured in a maize field, together with leaf water potential, leaf turgor, xylem ABA concentration and leaf ABA concentration in the same leaves. Stomatal conductance showed a tight relationship with xylem ABA, but not with the current leaf water status or with the concentration of ABA in the bulk leaf. The relationship between stomatal conductance and xylem [ABA] was common for variations in xylem [ABA] linked to the decline with time of the soil water reserve, to simultaneous differences between plants grown on compacted, non-compacted and irrigated soil, and to plant-to-plant variability. Therefore, this relationship is unlikely to be fortuitous or due to synchronous variations. These results suggest that increased concentration of ABA in the xylem sap in response to stress can control the gas exchange of plants under field conditions.     

Influence of Ethylene and Kinetin on Tuberization and Enzyme Activity in Solanum tuberosum L.: Stolons Cultured in vitro

The response of potato stolons, cultured in vitro, to ethylenewas investigated utilizing 2-chloroethyl-phosphonic acid asthe source of ethylene. Concentrations of 0.067, 0.67, 6.7,and 67 µm did not promote tuber formation which occurredin the presence of 16 µm kinetin. In the presence of 2-chloroethyl-phosphonicacid stolon branching was promoted and they maintained a diageotropicgrowth habit in contrast to the negatively geotropic growthhabit of kinetin-treated cultures. The amount of soluble sugars accumulating at the stolon tipwas similar in the presence of kinetin and 2-chloroethylphosphonicacid. However, the level decreased in kinetin-treated stolonsas starch synthesis increased whereas a high specific activitywas maintained in the presence of 2-chloroethylphosphonic acidwhere no starch synthesis occurred. Higher levels of sucroseoccurred in kinetin-treated cultures. Both 2-chloroethylphosphonic acid and kinetin decreased invertaseactivity at the stolon tip. Peroxidase activity increased withtime in response to 2-chloroethylphosphonic acid whereas activityincreased appreciably in kinetin-treated stolons only afterday 5. Generally higher levels of 3'nucleotidase activity existedin 2-chloroethylphosphonic acid-treated stolons. RNase activitydecreased in kinetin treated stolons while activity increasedin the presence of 2-chloroethylphosphonic acid. It is suggestedthat ethylene may influence stolon growth but may not be directlyinvolved in tuber initiation.     

Biosynthesis of Choline and Ethanolamine Phospholipids in Crithidia fasciculata*

The incorporation of radioactivity from [1,2-³⁴C]choline, [1,2-³⁴C]ethanolamine, [3-¹⁴C]serine and [methyl-¹⁴C]methionine into lipids was studied in growing cultures of Crithidia fasciculata. Lecithin was formed both from choline and by the methylation of phosphatidylethanolamine. Mono- and dimethylphosphatidylethanolamines were present in no more than trace amounts. Growth of the protozoa in media containing choline (1 mM) did not decrease synthesis by the methylation pathway. Phosphatidylethanolamine was formed from ethanolamine. Radioactivity from serine also was present in both phosphatidylethanolamine and lecithin; however, the presumed intermediate, phosphatidylserine, could not be detected.     

The Influence of Exogenous Nicotinamide Adenine Dinucleotide on the Oxidation of Malate by Jerusalem Artichoke Mitochondria

Mitochond$$$a isolated from Jerusalem artichoke tubers oxidizedendogenous NADH by both a piericidin A-sensitive and -resistantdehydrogenase if the level of oxaloacetate was kept low. Inwashed mitochondria the addition of NAD⁺ stimulated respirationin the presence of a variety of NAD⁺ -linked substrates. Inmitochondria purified through a sucrose density gradient exogenousNAD⁺ caused a substantial stimulation of respiration only inthe presence of malate. The apparent K_m for malate was 20 mMin the absence of NAD⁺ and 2 mM in the presence of exogenousNAD⁺ The products of malate oxidation were altered by the additionof exogenous NAD⁺. Oxaloacetate and pyruvate were produced inequal amounts in the absence of added NAD⁺, but in the presenceof exogenous NAD⁺ more pyruvate was formed. The rapid oxidationof malate in the absence of added NAD⁺ required phosphate whilethe NAD⁺-stimulated component was not affected by the absenceof phosphate. Phenylsuccinate inhibited the reduction of exogenousNAD⁺ by malate; this compound was found to inhibit isolatedma!ate dehydrogenase and mahe enzyme. Several lines of evidencesuggest that electron flux through one of the NADH dehydrogenasesystems may directly affect the flow through the other dehydrogenases.     

PHOSPHOINOSITIDE KINASES IN CHICK BRAIN AND SCIATIC NERVE, A DEVELOPMENTAL STUDY

Phosphatidylinositol kinase and diphosphoinositide kinase activities were measured in homogenates of brain and sciatic nerve of developing chick embryos and chicks. Characteristics of the chick nervous system enzymes were similar to those reported for rat brain. Diphosphoinositide kinase was inhibited by high concentrations of ATP and by low concentrations of triphosphoinositide. Both activities were greatly enhanced by the non-ionic detergent, Cutscum, and the ratio of detergent to protein in the reaction mixture was important. Optimum phosphatidylinositol kinase activity required a ratio of 7 : 1 for both tissues. The optimum ratio for diphosphoinositide kinase was 3:1 for nerve homogenates and 0.6:1 for brain. Cutscum increased the concentration of diphosphoinositide that is required for maximum diphosphoinositide kinase activity. Developmental changes were the same for both kinase activities, which were low in unmyelinated brain and sciatic nerve. The activities correlated with the concentration of polyphosphoinositides in chick brain where they increased 4-5 fold during the period of active myelination and remained high in the mature brain. The kinase activities correlated with the rate of triphosphoinositide deposition in sciatic nerve. Following a 2-3 fold increase during the initial phase of myelination the activities declined to values as low as those of embryonic nerve.     

Phylogenetic relationships within the Alcidae (Charadriiformes: Aves) inferred from total molecular evidence

The Alcidae is a unique assemblage of Northern Hemisphere seabirds that forage by "flying" underwater. Despite obvious affinities among the species, their evolutionary relationships are unclear. We analyzed nucleotide sequences of 1,045 base pairs of the mitochondrial cytochrome b gene and allelic profiles for 37 allozyme loci in all 22 extant species. Trees were constructed on independent and combined data sets using maximum parsimony and distance methods that correct for superimposed changes. Alternative methods of analysis produced only minor differences in relationships that were supported strongly by bootstrapping or standard error tests. Combining sequence and allozyme data into a single analysis provided the greatest number of relationships receiving strong support. Addition of published morphological and ecological data did not improve support for any additional relationship. All analyses grouped species into six distinct lineages: (1) the dovekie (Alle alle) and auks, (2) guillemots, (3) brachyramphine murrelets, (4) synthliboramphine murrelets, (5) true auklets, and (6) the rhinoceros auklet (Cerorhinca monocerata) and puffins. The two murres (genus Uria) were sister taxa, and the black guillemot (Cepphus grylle) was basal to the other guillemots. The Asian subspecies of the marbled murrelet (Brachyramphus marmoratus perdix) was the most divergent brachyramphine murrelet, and two distinct lineages occurred within the synthliboramphine murrelets. Cassin's auklet (Ptychoramphus aleuticus) and the rhinoceros auklet were basal to the other auklets and puffins, respectively, and the Atlantic (Fratercula arctica) and horned (Fratercula corniculata) puffins were sister taxa. Several relationships among tribes, among the dovekie and auks, and among the auklets could not be resolved but resembled "star" phylogenies indicative of adaptive radiations at different depths within the trees.      

Distribution, prevalence and intensity of earthworm populations in arable land and grassland in Scotland

The earthworms of arable and pasture fields from 100 randomly chosen arable farms were identified and counted and their relationship with soil factors examined. Thirteen species of earthworm were recorded including Lumbricus friendi from Scotland for the first time. The most prevalent and numerous species were Aporrectodea longa, Aporrectodea caliginosa and Lumbricus terrestris. All earthworm species had cosmopolitan distributions apart from Aporrectodea nocturna and Lumbricus festivus which were confined to southern/central Scotland. Four soil characteristics (% moisture, % sand, % organic matter and pH) shared no relationship with species recorded but tillage may have had a detrimental effect on species composition and size of population. It is suggested that the magnitudes of the earthworm populations found, particularly in permanent pasture, were important in maintaining soil structure and fertility, and the spread of the New Zealand flatworm (Artioposthia triangulata), an obligate predator of earthworms, could have a considerable impact on agricultural productivity and wildlife in Scotland.