Development-Specific Incorporation of [14C]5-Aminolevulinate and [3H]Leucine into Cytochrome c and Biliprotein in the Butterfly, Pieris brassicae. Correlation with the Ecdysteroid Titer in the Pupa

1984 ◽  
Vol 39 (9-10) ◽  
pp. 948-957 ◽  
Author(s):  
Hartmut Kayser ◽  
Ute Krull-Savage
Keyword(s):  
Cytochrome C ◽  
Time Course ◽  
De Novo ◽  
Larval Instar ◽  
Preceding Paper ◽  
Pupal Stage ◽  
Adult Emergence ◽  
Protein S ◽  
Pieris Brassicae ◽  
Total Soluble Protein

Abstract Incorporation of [14C]5-aminolevulinate and [3H]leucine into cytochrome c, biliprotein and total soluble protein was followed from the last larval instar to the adult stage in Pieris brassicae. The titer of ecdysteroids during the pupal stage was determined with a radioimmunoassay to correlate synthesis of heme products and of protein(s) with adult differentiation. Incorporation of both precursors showed a developmental profile with high synthetic activities in feeding larvae and in pupae after the release of ecdysteroids. Variation of the hormone titer during pupal life differed significantly in males and females. Labeling of cytochrome c by both 14C and 3H was as expected from the variation of its concentration reported in a preceding paper; highest in corporation was around adult emergence. The results demonstrate that i) the accumulation of cytochrome c in the developing adult insect is primarily due to de novo synthesis of both heme and apocytochrome c, performed under coordinate control, and ii) the concentration of 5-aminolevulinate is not rate-limiting in the formation of cytochrome c. Biliverdin IXγ, the major tetrapyrrolic product in this insect, seems to be directly derived from (free) heme and relatively short-lived as deduced from a time-course study. Formation of the bilin, i.e. destruction of heme, increased concomitantly to the initiation of adult differentiation by ecdysteroids in the pupa but later decreased at adult emergence. Synthesis of cytochrome c takes place as a late event during terminal development. Thus, the pathways leading to the two major heme products seem to be differently regulated during development.

De novo Synthesis and Levels of Cytochrome c and a Biliprotein during Pupal-Adult Development of a Butterfly, Pieris brassicae

1984 ◽  
Vol 39 (9-10) ◽  
pp. 938-947 ◽  
Author(s):  
Hartmut Kayser
Keyword(s):  
Cytochrome C ◽  
Adult Development ◽  
De Novo ◽  
Larval Instar ◽  
Spectroscopic Method ◽  
Pieris Brassicae ◽  
Major Product ◽  
Exchange Chromatography ◽  
De Novo Synthesis ◽  
Large White

Abstract Using a sensitive pH-difference spectroscopic method in combination with a three-column procedure of ion-exchange chromatography (overall yield 94%) the levels of cytochrome c in the large white butterfly, Pieris brassicae, were determined from the last larval instar to the adult insect. In the larva cytochrome concentration reached a maximum at mid instar and sub­sequently decreased to very low values in the pupa. During adult development the level of cytochrome c increased 50-fold in males and 43-fold in females; this sexual difference was expressed only after adult emergence. Biliverdin IXγ which occurs as a specific biliprotein complex was accumulated during the last larval instar and also in young butterflies. De novo synthesis of heme c, biliverdin IXγ and the corresponding apoproteins was demonstrated in newly emerged butterflies by injections of radiolabeled 5-aminolevulinate, lysine, leucine, and succinate, respectively. Cycloheximide inhibited labeling of both apoproteins and of heme c to 90% but that of the bilin to only 25%. This suggests that in cytochrome c but not in the biliprotein formation of the holoprotein depended on a coordinated synthesis of both constituents. In­corporation of 5-aminolevulinate into the biliprotein exceeded that into cytochrome c sevenfold indicating that biliverdin IXγ is the major product of the heme pathway in P. brassicae. The results are discussed in relation to the formation of mitochondria and flight muscles during postembryonic development of insects.

Arabidopsis 4-COUMAROYL-COA LIGASE 8 contributes to the biosynthesis of the benzenoid ring of coenzyme Q in peroxisomes

2019 ◽  
Vol 476 (22) ◽  
pp. 3521-3532
Author(s):  
Eric Soubeyrand ◽  
Megan Kelly ◽  
Shea A. Keene ◽  
Ann C. Bernert ◽  
Scott Latimer ◽  
...  
Keyword(s):  
Oxidative Metabolism ◽  
Time Course ◽  
Reverse Genetics ◽  
De Novo ◽  
Coenzyme Q ◽  
Control Level ◽  
Wild Type ◽  
Fluorescent Reporters ◽  
Coa Ligase ◽  
Peroxisomal Targeting

Plants have evolved the ability to derive the benzenoid moiety of the respiratory cofactor and antioxidant, ubiquinone (coenzyme Q), either from the β-oxidative metabolism of p-coumarate or from the peroxidative cleavage of kaempferol. Here, isotopic feeding assays, gene co-expression analysis and reverse genetics identified Arabidopsis 4-COUMARATE-COA LIGASE 8 (4-CL8; At5g38120) as a contributor to the β-oxidation of p-coumarate for ubiquinone biosynthesis. The enzyme is part of the same clade (V) of acyl-activating enzymes than At4g19010, a p-coumarate CoA ligase known to play a central role in the conversion of p-coumarate into 4-hydroxybenzoate. A 4-cl8 T-DNA knockout displayed a 20% decrease in ubiquinone content compared with wild-type plants, while 4-CL8 overexpression boosted ubiquinone content up to 150% of the control level. Similarly, the isotopic enrichment of ubiquinone's ring was decreased by 28% in the 4-cl8 knockout as compared with wild-type controls when Phe-[Ring-13C6] was fed to the plants. This metabolic blockage could be bypassed via the exogenous supply of 4-hydroxybenzoate, the product of p-coumarate β-oxidation. Arabidopsis 4-CL8 displays a canonical peroxisomal targeting sequence type 1, and confocal microscopy experiments using fused fluorescent reporters demonstrated that this enzyme is imported into peroxisomes. Time course feeding assays using Phe-[Ring-13C6] in a series of Arabidopsis single and double knockouts blocked in the β-oxidative metabolism of p-coumarate (4-cl8; at4g19010; at4g19010 × 4-cl8), flavonol biosynthesis (flavanone-3-hydroxylase), or both (at4g19010 × flavanone-3-hydroxylase) indicated that continuous high light treatments (500 µE m−2 s−1; 24 h) markedly stimulated the de novo biosynthesis of ubiquinone independently of kaempferol catabolism.

BioMime – A Sirolimus-eluting Coronary Stent System for the Treatment of Patients with De Novo Coronary Lesions – meriT-1 Report

2011 ◽  
Vol 6 (1) ◽  
pp. 39
Author(s):  
Keyword(s):  
Stent Thrombosis ◽  
Time Course ◽  
De Novo ◽  
Drug Eluting Stent ◽  
Cobalt Chromium ◽  
Cardiac Events ◽  
End Points ◽  
Safety And Efficacy ◽  
Binary Restenosis

Background:Since the first reported use of percutaneous transluminal coronary angioplasty, advances in the interventional cardiology arena have been fast paced. Developers and clinicians are adapting from the learning curve awarded by the time-course of drug-eluting stent (DES) evolution. BioMime™ sirolimus-eluting stent (SES) is a step towards biomimicry. The stent is built on a strut of ultra-low thickness (65μm), a cobalt–chromium platform using an intelligent hybrid of closed and open cells allowing for morphology-mediated expansion. It employs a well-known antiproliferative – sirolimus – that elutes from a known biodegradable copolymer formulation within 30 days. The resultant stent demonstrates almost 100% endothelialisation at 30 days in preclinical models.Methods:The meriT-1 was a prospective, single-arm, single-centre trial to evaluate the safety and efficacy of BioMime SES in 30 patients with a single de novo lesion in native coronary arteries. The primary safety and efficacy end-points were major adverse cardiac events (MACE) at 30 days and in-stent late lumen loss at eight months, as measured using quantitative coronary angiographic (QCA) method. Secondary safety and efficacy end-points included MACE at one and two years and angiographic binary restenosis at eight-month angiographic follow-up. Other end-points included the occurrence of stent thrombosis at acute, subacute, late and very late periods and the percentage of diameter stenosis by QCA.Results:No MACE were observed and the median in-stent late luminal loss in 20 (67%) subjects studied by QCA was 0.15mm, with 0% binary restenosis at eight-month follow-up. No stent thrombosis was observed up to one-year follow-up.Conclusions:In comparison to currently available DES, BioMime SES appears to have a considerable scientific basis for prevention of neointimal proliferation, restenosis and associated clinical events.

scholarly journals Lipid Metabolism Is Dysregulated before, during and after Pregnancy in a Mouse Model of Gestational Diabetes

2021 ◽  
Vol 22 (14) ◽  
pp. 7452
Author(s):  
Samuel Furse ◽  
Denise S. Fernandez-Twinn ◽  
Davide Chiarugi ◽  
Albert Koulman ◽  
Susan E. Ozanne
Keyword(s):  
Lipid Metabolism ◽  
Gestational Diabetes ◽  
Glucose Tolerance ◽  
Mouse Model ◽  
Time Course ◽  
De Novo ◽  
Lipid Analysis ◽  
Temporal Distribution ◽  
De Novo Lipogenesis ◽  
Analysis Tool

The aim of the current study was to test the hypothesis that maternal lipid metabolism was modulated during normal pregnancy and that these modulations are altered in gestational diabetes mellitus (GDM). We tested this hypothesis using an established mouse model of diet-induced obesity with pregnancy-associated loss of glucose tolerance and a novel lipid analysis tool, Lipid Traffic Analysis, that uses the temporal distribution of lipids to identify differences in the control of lipid metabolism through a time course. Our results suggest that the start of pregnancy is associated with several changes in lipid metabolism, including fewer variables associated with de novo lipogenesis and fewer PUFA-containing lipids in the circulation. Several of the changes in lipid metabolism in healthy pregnancies were less apparent or occurred later in dams who developed GDM. Some changes in maternal lipid metabolism in the obese-GDM group were so late as to only occur as the control dams’ systems began to switch back towards the non-pregnant state. These results demonstrate that lipid metabolism is modulated in healthy pregnancy and the timing of these changes is altered in GDM pregnancies. These findings raise important questions about how lipid metabolism contributes to changes in metabolism during healthy pregnancies. Furthermore, as alterations in the lipidome are present before the loss of glucose tolerance, they could contribute to the development of GDM mechanistically.

scholarly journals Pathogenicity and Side Effect of Indigenous Beauveria bassiana on Coccinella undecimpunctata and Hippodamia variegata (Coleoptera: Coccinellidae)

Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 42
Author(s):  
Samy Sayed ◽  
Sayed-Ashraf Elarrnaouty ◽  
Saad AlOtaibi ◽  
Mohamed Salah
Keyword(s):  
Beauveria Bassiana ◽  
Indirect Effect ◽  
Developmental Stages ◽  
Larval Instar ◽  
Pupal Stage ◽  
Tween 80 ◽  
Contact Method ◽  
Coccinella Undecimpunctata ◽  
Hippodamia Variegata ◽  
Application Methods

This study aimed to estimate the virulence of an indigenous Beauveria bassiana on all developmental stages of two indigenous coccinellids; Coccinella undecimpunctata and Hippodamia variegata through three application methods; direct spray, contact method, and feeding on aphids treated with the fungus (ingestion). Also, indirect effect on all developmental stages resulted from 1st larval instar treated with these application methods. All treatments were done with a concentration of 1 × 105 which was recommended in previous studies for different aphid species with a control of 0.02% Tween 80 (v/v). The mortality of 1st larval instar of both H. variegata and C. undecimpunctata and pupal stage of C. undecimpunctata were significantly increased with spray method only. Also, contact method achieved significantly higher mortality on 1st larval instar of C. undecimpunctata only. Regard to indirect effect, except of mortality of 1st larval instar of both predators and 2nd larval instar of H. variegata, other developmental instars/stages of both predators were not affected by B. bassiana through the three tested application methods in the mortality, duration, survival, cumulative survival male and female longevity, and fecundity. Therefore, both tested predatory coccinellids could be compatible with this indigenous isolate of B. bassiana where, in general, there are no negative effects of the fungus on both predators.

Differential expression of TNF-α, IL-6, and IGF-1 by graded mechanical stress in normal rat myocardium

2002 ◽  
Vol 282 (3) ◽  
pp. H926-H934 ◽  
Author(s):  
Emiliano A. Palmieri ◽  
Giulio Benincasa ◽  
Francesca Di Rella ◽  
Cosma Casaburi ◽  
Maria G. Monti ◽  
...  
Keyword(s):  
Time Course ◽  
De Novo ◽  
Mechanical Stretch ◽  
Progressive Increase ◽  
Compensatory Hypertrophy ◽  
Current Data ◽  
Rat Myocardium ◽  
Tnf Α ◽  
Normal Rat ◽  
Necrosis Factor

An isovolumic normal rat heart Langendorff model was used to examine the effects of moderate (15 mmHg) and severe (35 mmHg) mechanical stretch on the time course (from 0 to 60 min) of myocardial expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and insulin-like growth factor (IGF)-1 and their cognate receptors. After 10 min of moderate stretch, TNF-α was de novo expressed, whereas constitutive IL-6 and IGF-1 levels were slightly upregulated; no further changes occurred up to 60 min. In comparison, severe stretch resulted in a higher and progressive increase in TNF-α, IL-6, and IGF-1 expression up to 20 min. After 20 min, whereas TNF-α expression further increased, IL-6 and IGF-1 levels progressively reduced to values lower than those observed under moderate stretch and in unstretched (5 mmHg) control myocardium (IL-6). Mechanical stretch did not significantly alter the expression of the cognate receptors. Indeed, the TNF-α receptor (p55) tended to be progressively upregulated under severe stretch over time. The current data provide the first demonstration that TNF-α, IL-6, and IGF-1 ligand-receptor systems are differentially expressed within the normal rat myocardium in response to graded mechanical stretch. Such findings may have potential implications with regard to compensatory hypertrophy and failure.

scholarly journals Detection of G1 proteins in Chinese hamster cells synchronized by isoleucine deprivation or mitotic selection.

1975 ◽  
Vol 66 (1) ◽  
pp. 95-101 ◽  
Author(s):  
K D Ley
Keyword(s):  
Time Course ◽  
Sodium Dodecyl ◽  
Chinese Hamster ◽  
Protein S ◽  
S Phase ◽  
Supernatant Fraction ◽  
G1 Phase ◽  
Differential Centrifugation ◽  
Chinese Hamster Cells ◽  
Soluble Supernatant

Examination of labeling patterns of proteins in Chinese hamster cells(line CHO) revealed the presence of a class of protein(s) that is synthesized during G1 phase of the cell cycle. Cells arrested in G1 by isoleucine (Ile) deprivation were prelabeded with [14-C]Ile, induced to traverse G1 by addition of unlabeled Ile, and labeled with [3-H]Ile at hourly intervals. Cells were fractionated into neclear and cytoplasmic portions, and proteins were separated by sodium dodecyl sulfate-polyacrylamide get electrophoresis. Gel profiles of proteins in the 45,000-160,000 mol wt range from the cytoplasm of cells in G1 were similar to those from cells arrested in G1 except for the presence of a mojor peak of [1-H]Ile incorporated into a protein(s) of approximately 80,000 mol wt. Peaks of net [3-H]Ile incorporation were not detected in neclear preparations. Cellular fractionation by differential centrifugation showed the peak I protein was located in the soluble supernatant fraction of the cytoplasm. Time-course studies showed that synthesis of this protein began 1-2 h after initiation of G1 traverse; the protein reached maximum levels in 4-6 h and was reduced to undetectable levels by 9 h. A cytoplasmic protein with similar electrophoretic mobility was found in G1 phase of cells synchronized by mitotic selection. This class of proteins is synthesized by cells before entry into S phase and may be involved in initiation of DNA synthesis.

The Post-embryonic Development of the Tracheal System in Drosophila melanogaster

1957 ◽  
Vol s3-98 (41) ◽  
pp. 123-150
Author(s):  
JOAN M. WHITTEN
Keyword(s):  
Drosophila Melanogaster ◽  
Adult Stage ◽  
Larval Instar ◽  
Pupal Stage ◽  
Tracheal System ◽  
True Nature ◽  
The Third ◽  
Air Sacs ◽  
Abdominal Region ◽  
Pupal Cuticle

The fate of the tracheal system is traced from the first larval instar to the adult stage. The basic larval pattern conforms to that shown for other Diptera Cyclorrhapha (Whitten, 1955), and is identical in all three instars. According to previous accounts the adult system directly replaces the larval: the larval system is partly shed, partly histolysed, and the adult system arises from imaginal cell clusters independently of the preceding larval system. In contrast, it is shown here that in the cephalic, thoracic, and anterior abdominal region there is a definite continuity in the tracheal system, from larval, through pupal to the adult stage, whereas in the posterior abdominal region the larval system is histolysed, and the adult system is independent of it in origin. Moreover, in the pupal stage this region is tracheated by tracheae arising from the anterior abdominal region and belonging to a distinct pupal system. Moulting of the tracheal linings is complete at the first and second larval ecdyses, but incomplete at the third larval-pupal and pupal-adult ecdyses. In consequence, in both pupal and adult systems there are tracheae which are secreted around preexisting tracheae, others formed as new ‘branch’ tracheae, and those which have been carried over from the previous instar. In the adult the newly formed tracheae of the posterior abdominal region fall into a fourth category. Most of the adult thoracic air sacs correspond to new ‘branch’ tracheae of other instars. The pre-pupal moult and instar are discussed with reference to the tracheal system and tentative suggestions are made concerning the true nature of the pre-pupal cuticle. There is no pre-pupal tracheal system. Events traced for Drosophila would seem to be general for Cyclorrhapha, both Acalypterae and Calypterae. The separate fates of the anterior and posterior abdom inal systems, in contrast with the straightforward development in Dipterc Nematocera, would appear to mark a distinct step in the evolution of the system in Diptera.

Glucocorticoid-regulated compartmentalization of cell surface-associated glycoproteins in rat hepatoma cells: evidence for an independent response that requires receptor function and de novo RNA synthesis

1987 ◽  
Vol 7 (4) ◽  
pp. 1508-1517
Author(s):  
O K Haffar ◽  
A K Vallerga ◽  
S A Marenda ◽  
H J Witchel ◽  
G L Firestone
Keyword(s):  
Cell Surface ◽  
Time Course ◽  
Rna Synthesis ◽  
De Novo ◽  
Receptor Occupancy ◽  
Receptor Function ◽  
Intracellular Fraction ◽  
Culture Cells ◽  
Viral Glycoproteins ◽  
Rat Hepatoma

The role of glucocorticoid hormones in the compartmentalization of cell surface-associated mouse mammary tumor virus (MMTV) glycoproteins was examined in M1.54, a cloned line of MMTV-infected rat hepatoma tissue culture cells. The expression of cellular [2-3H]mannose-labeled and cell surface 125I-labeled MMTV glycoproteins was examined throughout a time course of exposure to dexamethasone, a synthetic glucocorticoid. Posttranslational localization of cell surface MMTV glycoproteins required 6 h of exposure to hormone and occurred approximately 4 h after their initial production in an intracellular fraction. This regulated localization to the cell surface correlated with glucocorticoid receptor occupancy and was inhibited by exposure to RU 38486, a powerful antagonist of glucocorticoid-mediated responses. Cell surface immunoprecipitation demonstrated that actinomycin D, an inhibitor of de novo RNA synthesis, prevented regulated expression of cell surface viral glycoproteins, suggesting that newly synthesized cellular components mediate this process. The localization of cell surface MMTV glycoproteins appeared normal in a transcriptional variant (CR1) that produces basal levels of MMTV RNA and glycoprotein precursors in the presence of dexamethasone. Thus, regulated compartmentalization of viral glycoproteins is not an obligate consequence of a critical precursor concentration. Taken together, our results suggest that posttranslational trafficking of cell surface-destined MMTV glycoproteins resulted from an independent glucocorticoid hormone response that required receptor function and de novo RNA synthesis.

scholarly journals Breaking the rule: Five larval instars in the podonomine midge Trichotanypus alaskensis Brundin from Barrow, Alaska

2018 ◽  
Author(s):  
Alec R. Lackmann ◽  
Malcolm G. Butler
Keyword(s):  
Life Cycle ◽  
Larval Instar ◽  
Adult Emergence ◽  
Head Capsule ◽  
The Arctic ◽  
Larval Instars ◽  
Daily Monitoring ◽  
Field Samples ◽  
Standard Life ◽  
Capsule Size

Except for one unconfirmed case, chironomid larvae have been reported to pass through four larval instars between egg and pupal stages. We have observed a fifth larval instar to be a standard life-cycle feature of the podonomine Trichotanypus alaskensis Brundin 1966 in tundra ponds on the Arctic Coastal Plain near Barrow, Alaska. T. alaskensis has a one-year life cycle in these arctic ponds. Adults emerge in June ~2-3 weeks after pond thaw, then mate and oviposit; most newly-hatched larvae reach instar IV by October when pond sediments freeze. Overwintering larvae complete instar IV within a few days of thaw, then molt again to a fifth larval instar. Imaginal discs, normally seen only during instar IV in Chironomidae, develop across both instars IV & V prior to pupation and adult emergence. While monitoring larval development post-thaw in 2014, we noticed freshly-molted T. alaskensis larval exuviae a week or more prior to any pupation by that species. In 2015-16 we reared overwintering instar IV larvae from single pond sources, individually with daily monitoring, through molts to instar V, pupa, and adult. Some overwintering instar II and III larvae were reared as well, but were few in number. During 2016 we also reared T. alaskensis progeny (from eggs) through instar II, thus documenting head capsule size ranges for all five instars in a single pond’s population. Without individual rearings, the fifth larval instar was not readily apparent for two reasons: 1) The molt itself occurs immediately after thaw and is so synchronous it is difficult to discern in daily field samples. 2) The head capsule size increment between instars IV-V is much lower than the ratio predicted by the Brooks-Dyar Rule. Up through instar IV, the Brooks-Dyar ratio for T. alaskensis ranged 1.30-1.61, but during the IV-V molt head capsule dimensions (sexes pooled) increased by a ratio of 1.09 – comparable to the magnitude of sexual dimorphism in head capsule size within each of the final two larval instars. Individual rearings coupled with 2014-2016 field surveys in nine other ponds suggest that five larval instars is an obligatory trait of this species at this location. As this is the first confirmed case of five larval instars in a chironomid, the phylogenetic uniqueness of this trait needs further investigation.

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