Differences in response patterns of labellar chemosensilla of the fleshfly Sarcophaga bullata

• 1.1. Electrophysiological responses to NaCl, sucrose, and a complex mixture containing NaCl, glucose, fructose, phenylalanine and valine were obtained from large, medium, and small hair sensilla on the proboscis of the fleshfly Sarcophaga bullata.
• 2.2. Responses from up to three cells in each sensillum were analysed and compared across the three types of sensilla.
• 3.3. We found qualitative differences in the patterns of responses from the different hair types.

     

Amino acid activating enzymes in Sarcophaga bullata

• 1.1. The fat-body soluble fraction from two-day Sarcophaga bullata larvae contain amino acid activating enzymes for nineteen amino acids.
• 2.2. The level of activity varies with the amino acid substrate.
• 3.3. The total ³²PP-ATP exchange activity of the pupae decreased with age for the first 6 days, then increased to a maximum one or two days prior to emergence of the adults.
• 4.4. The free amino acid concentration in the pupae decreased during the period when the amino acid activating activity increased.

     

Alterations in proteases,protease inhibitors and ecdysone levels: a profile of stress in insects

• 1.1. A comparison of proteolytic and protease inhibitory activity, and ecdysteroid levels in body fluids was made between normal larvae of the flesh fly, Sarcophaga bullata, and those that had been water-stressed for two days.
• 2.2. The course of proteolytic activity in water stressed flies decreases 6 hr after beginning the experiment and remains low in comparison with control flies.
• 3.3. The course of protease inhibitors exhibits a mirror image pattern to proteases.
• 4.4. Ecdysteroid pattern shows two peaks in control animals: minor at 24 hr and major at pupariation, in experimental animals: at 1 hr, at 6 hr and at white pupal stage.

     

Characterization of Colorado potato beetle lipophorin: A hydrocarbon-rich diacylgycerol-poor lipophorin

• 1.1. The lipid composition of lipophorin from the Colorado potato beetle, Leptinotarsa decemlineata Say, was analyzed.
• 2.2. This insect lipophorin contains 44% lipid and is characterized by large amounts of hydrocarbons and small amounts of diacylglycerol.
• 3.3. This is the first observation of a diacylglycerol-poor insect lipophorin in haemolymph.
• 4.4. Since the main energy source for flight in the Colorado potato beetle is proline, the low diacylglycerol content in lipophorin must be related to its peculiar flight metabolism.
• 5.5. This lipophorin, however, can still take up appreciable amounts of diacylglycerol from the locust fat body. Hydrocarbon uptake by this lipophorin was also demonstrated.
• 6.6. The main function of this lipophorin therefore seems to be transport of hydrocarbons from oenocytes to the cuticle.

     

GCMS-indentified steroids and steroid glucoronides in gonads and holding water of Trichogaster trichopterus (Anabantidae,Pallas 1770)

• 1.1. Analysis by gas chromatography—mass spectrometry was performed to identify steroids and steroid glucoronides in gonads of the tropical fish Trichogaster trichopterus and in the water in which the fish were maintained.
• 2.2. Full mass spectra of estradiol-17β (E₂), testosterone, 17α-hydroxyprogesterone, cholesterol, stigmasterol, 4β-methylcholesterol, estrone, 17α,20β-hydroxy-4-pregnen-3-one (17,20-P) and sitosterol were obtained.
• 3.3. The above steroids were detected in both female and male gonads, with the exception of estrone, which was detected only in the male, and 17,20-P, which was detected only in the female.
• 4.4. All steroids except 17,20-P were detected in the water in which the fish were maintained.

     

Phosphorylation in crayfish (Procambarus clarkii) eggs during hatching

• 1.1.|The high-energy phosphorylation metabolism in crayfish, Procambarus clarkii eggs during brooding and juvenile crayfish after hatching was studied by in vivo³¹P nuclear magnetic resonance (³¹P NMR) spectroscopy.
• 2.2.|Inorganic phosphoric acid (Pi) and adenosine-5′-triphosphate ATP(γ-,α-,β-) were detected in the dark brownish red eggs after oviposition.
• 3.3.|In orange unhatched eggs, only sugar phosphate (SP), Pi and resolved phosphometabolite from ATP were observed.
• 4.4.|Peaks of SP, Pi, arginine phosphate (Arg-P), and ATP (γ,α,β) appeared in larvae of crayfish after hatching (nauplius, zoea and juvenile crayfish).
• 5.5.|The high-energy phosphorylation metabolism changed to an anaerobic condition along with a decrease in the concentration of dissolved oxygen in fresh water.

     

Sterol composition of the “living fossil” crinoid Gymnocrinus richeri

• 1.1. The composition of sterol mixture from the “living fossil” crinoid Gymnocrinus richeri collected off Nouméa (New Caledonia) was investigated.
• 2.2. The free 3β-OH sterol mixture was found to contain 14 components, Δ⁵ and ring saturated stanols, identified by GC-MS.
• 3.3. Cholest-4-en-3-one, cholesta-1, 4-dien-3-one (this latter firstly isolated from a marine source), 5α-8α-epidioxy sterols, and 5α-ergosta-7,22-diene-3β,5,6β-triol were also present, their characterization being accomplished by EI-MS and ¹H-NMR. The methanol extract also contained sterol sulphates, which were identified by GC-MS after solvolysis to remove the sulphate group.

     

Steroid metabolites of the far eastern Holothurian Stichopus japonicus Selenka

• 1.1. Sulphated and etherified sterols were isolated from the far eastern holothurian Stichopus japonicus S. The sterol composition of both fractions was determined using gas-liquid chromatography and mass-spectroscopic methods. The structures of individual sterols were proved on the basis of mass-spectrometry and ¹H-NMR-spectroscopy data.
• 2.2. The structures of 29 sterols were established.
• 3.3. Sterols (22E, 24R)-23,24-dimethyl-5α-cholest-22-en-3β-ol, 23,24-dimethyl-cholesta-5,22-dien-3β-ol, 24-methyl-cholesta-5,24(28)-dien-3β-ol, (24Z)-24-ethyl-cholesta-5,24(28)-dien-3β-ol, 24-nor-cholesta-5,22-dien-3β-ol, 24-ethyl-cholesta-5,25-dien-3β-ol were described for holothurians for the first time.
• 4.4. Δ⁵-sterols were shown to be the main components of the sulphated alcohol fractions (67.61%), while the saturated and Δ⁷-sterols were there in less quantities (14.72 and 9.52%, respectively).
• 5.5. The etherified sterols were represented, mainly, by saturated and Δ⁷-sterols (37.82% and 33.95%, respectively). Δ⁵-sterols were 19%.
• 6.6. The sensitivity of liposomal membranes, containing steroid metabolites of the holothurian St. japonicus (Δ⁷-, sulphated and glycosilated sterols) to the action of endotoxin-stichoposide A, was studied.

     

Sterol composition of two actiniaria

• 1.1. The sterol composition of Condylactis aurantiaca and Cereus pedunculatus (phylum Coelenterata, class Anthozoa, order Actiniaria) was investigated by silver nitrate-silica gel column chromatography, combined gas chromatography-mass spectrometry and NMR.
• 2.2. Sea anemones contained Δ³-sterols accompanied by small amounts of Δ^5.7-sterols and ring saturated sterols.
• 3.3. Sterols with 27 carbon atoms are predominant and cholesterol is the principal sterol, followed by 24-methylenecholesterol.
• 4.4. A 4-methyl ring saturated sterol, identified as 4,24-dimethyl-5α-cholest-24(28)-en-3β-ol, occurs in small amount in Actiniaria, accompanied by the corresponding 4-demethylstanol.

     

Chemical intermediates in α-methylnoradrenaline oxidation by tyrosinase—I. Spectral properties and stoichiometry

• 1.1. A pathway for a-methylnoradrenaline oxidation to α-methylnoradrenochrome, by tyrosinase, is proposed. Characterization of intermediates in this oxidative reaction and stoichiometry determination have both been performed.
• 2.2. It has been possible to detect spectrophotometrically o-quinone-H⁺ as the first intermediate in this pathway after oxidizing α-methylnoradrenaline with mushroom tyrosinase or sodium periodate in a pH range from 5 to 6.
• 3.3. The steps for α-methylnoradrenaline transformation into its aminochrome could be: α-methylnoradrenaline → o -α-methylnoradrenaline — H⁺ → o — α -methylnoradrenalinequinone → leuko — α — methylnora — drenochrome→α-methylnoradrenochrome.
• 4.4. No participation of oxygen was detected in the conversion of leuko-α-mehtylnoradrenochrome into α -methylnoradrenochrome.
• 5.5. Matrix analysis of the spectra obtained with a rapid scan spetrophotometer verified that o-quinone-H⁺ was transformed into aminochrome in a constant ratio.
• 6.6. The stoichiometry for this conversion followed the equation: 2 α-methylnoradrenalinequinone-H⁺ → α-methylnoradrenaline + α-methylnoradrenochrome.

     

Preparation and characterization of biotinylated probes for the β-interferon receptor

• 1.1. Recently we described the isolation of the β-interferon receptor [Zhang et al. (1986) J. biol. Chem. 261, 8017–8021]. A highly purified product was obtained but in low quantities.
• 2.2. The use ofbiotinylated β-interferon as a ligand represents an alternate approach to receptor isolation.
• 3.3. We have prepared and characterized the derivatives N-(biotinyl)- and N-(biotinyl-ϵ-aminocaproyl)-recombinant human [Ser¹⁷-interferon β (B- and BC-recHulFNβ).
• 4.4. Biotin incorporation does not result in any loss of antiviral activity, demonstrating the recognition of the derivative by the cell receptor.
• 5.5. The biotinylated recHuIFNβ binds specifically and reversibly to succinoylavidin or guanidine thiocyanate-stripped succinoylavidin linked to a Sepharose matrix.
• 6.6. Comparison of the competition curves obtained with [¹⁴C]biotin and [³H]biotinyl recHuIFN, in the presence of increasing concentrations of biotin suggests that the IFN moiety of the derivative has little effect on the affinity of biotin for avidin.
• 7.7. Biotinylated recHuIFNβ derivatives represent useful probes for the β-IFN receptor.

     

Strain-specific differences in the regulation of the hepatic cytoplasmic 17β-hydroxysteroid dehydrogenase activity of the rat

• 1.1. The effect of prepuberal gonadectomy of Sprague-Dawley/NIH/HAN rats on cytoplasmic 17β-hydroxysteroid dehydrogenase was examined on day 30, 45, 60, 75, 90 and 105 of life.
• 2.2. The activity in male rats was not significantly affected by gonadectomy, whereas the activity in females showed an age-dependent oestrogen dependency.
• 3.3. This age-dependent oestrogen dependency could also be demonstrated in 5α-dihydrotestosterone treated intact females.
• 4.4. Cytoplasmic 17β-hydroxysteroid dehydrogenase activity of female Chbb:THOM rats also showed an age-dependent oestrogen dependency, whereas the enzyme activity of male rats of this strain showed a distinct androgen dependency absent in Sprague-Dawley/NIH/HAN rats.
• 5.5. On the basis of previous investigations it is concluded that the androgen dependency of the enzyme activity of male Chbb:THOM rats has been bred into this strain in the period 1974–1977.

     

The occurrence of brassicasterol and epibrassicasterol in the chromophycota

• 1.1. Sterols were identified from eight isolates of five species in the Chromophycota that were cultured axenically and harvested in the stationary phase.
• 2.2. Analyses were performed on four strains from the Prymnesiophyceae, two strains from the Cryptophyceae and one from the Bacillariophyceae. Most strains examined contained only one major sterol, 24-methyl-22-dehydrocholesterol.
• 3.3. Analysis by capillary GC, HPLC, and in one instance NMR, showed that the two strains provisionally identified as Isochrysis contained brassicasterol (24β-methyl-22-dehydrocholesterol); whereas, all other species examined contained primarily epibrassicasterol (24α-methyl-22-dehydrocholesterol).
• 4.4. Stigmasterol (24α-ethyl-22-dehydrocholesterol) accompanied epibrassicasterol in Pleurochrysis carterae.
• 5.5. Analyses of C-24 alkyl isomers in these algae may provide useful information concerning their taxonomic placement.
• 6.6. The occurrence of both isomers of 24-methyl-22-dehydrocholesterol in oysters is explained by the occurrence of both isomers among algae which are probably dietary sources for oysters.

     

Deterioration of chum salmon (Oncorhynchus keta) muscle during spawning migration—III. Changes in protein composition and protease activity of juvenile chum salmon muscle upon treatment with sex steroids

• 1.1. Changes in protein composition and protease activity of juvenile chum salmon muscle upon treatment with sex steroids were investigated.
• 2.2. A slight breeding color was observed on chum salmon following the oral administration of 17α-methyltestosterone. Sarcoplasmic protein significantly decreased, while ninhydrin-positive substances from protein-free fractions significantly increased upon treatment with 17α-methyltestosterone. Autolytic activity of the fish treated with 17α-methyltestosterone drastically increased.
• 3.3. Estradiol-17β did not significantly influence the protein composition and autolytic activity.
• 4.4. These results indicate that androgen is closely related to the deterioration of chum salmon muscle.

     

Evoked effects of oestradiol on hepatic cholesterol 7α-hydroxylase and drug oxidase in castrated rats

• 1.1. Oestradiol administration in castrated rats resulted in an increased activity of the cholesterolα-hydroxylase and a decreased activity of the drug oxidase enzyme systems.
• 2.2. Aqueous solutions of oestradiol (up to 25·10⁻⁶M) incubated in vitro with microsomes, binds into the microsomal membrane framework reducing the activity of both enzyme systems.
• 3.3. The specific activity of cholesterol 7α-hydroxylase. drops after 3 hr preincubation with oestradiol to at least 70% of its original value.
• 4.4. Actinomycin D and cycloheximide administration reduced the oestradiol-induced and control cholesterol 7α-hydroxylase activity to the same level, 6 hr after the injections.

     

Reabsorption and accumulation of α-amino-iso-butyric acid in the nephridia of Sabella pavonina Sa vigny (Annelida polychaeta)

• 1.l. High amino acid concentrations were found in the anterior coelomic fluid of a Polychaeta (Sabella pavonina Savigny).
• 2.2. The concentrations being much higher in the fluid which penetrates the nephrostomia into the nephridia lumen than in the final urine indicates that the nephridia reabsorbs large amounts of amino acids.
• 3.3. Nephridial perfusion experiments showed that an amino acid analogue (α-amino-iso-butyric acid, AIB) is transported by the nephidia.
• 4.4. The transport took place across the nephridial wall owing to the presence of a carrier-mediated transport system and a diffusion system.
• 5.5. For the carrier-mediated transport, the V_max was 0.234 ± 0.025 nmol·min⁻ and the K_m 3.715 ± 0.315mmol·l⁻.
• 6.6. AIB accumulated in the nephridial cells up to a maximum rate of 01.17 nmol·min⁻.
• 7.7. Intracellular accumulation stopped increasing when the V_max for reabsorption was reached.
• 8.8. These results indicate that the carrier-mediated transport of AIB is located at the apical membrane of the nephridial cell, and that AIB transport by simple diffusion takes place through the paracellular pathway.

     

Possible involvement of adenylylation in the modification of a 26 kDa protein in rat parotid acinar cells

• 1.1. Adenylylation, a posttranslational modification of proteins, was investigated in saponin-permeabilized acinar cells of the rat parotid gland.
• 2.2. When cells were incubated with [2,8-³H]ATP, several proteins, including a 26 kDa protein in the particulate fraction, were labeled.
• 3.3. Upon incubation of cells with [α-³²P]ATP in the presence of cAMP and 3-isobutyl-lmethylxanthine, ³²P-labeling of the 26 kDa protein was observed.
• 4.4. After treatment with snake venom phosphodiesterase, [³²P]AMP was released from the 26kDa protein. Such release was not observed when cells were labeled with [γ-³²P]ATP.
• 5.5. The ³²P-labeling pattern of proteins with [α-³²P]ATP was clearly different from that with [adenylate-³²P]NAD⁺.
• 6.6. The results suggest that the 26 kDa protein is one of the adenylylation substrates in rat parotid acinar cells.

     

Co-localization of locustamyotropin- and pheromone biosynthesis activating neuropeptide-like immunoreactivity in the central nervous system of five insect species

• 1.1. Myotropin I of Locusta and Pheromone Biosynthesis Activating Neuropeptide (PBAN) of Heliothis share the same carboxyterminal pentapeptide FSPRL-amide.
• 2.2. Immunostaining revealed colocalization in cells and axons in the central nervous system, especially in the suboesophageal ganglion, of Locusta, Periplaneta, Leucophaea, Neobellieria (Sarcophaga) and Mamestra.
• 3.3. Following preabsorption with synthetic FSPRL-amide, the PBAN antiserum continued to immunostain cells in Mamestra and Neobellieria only. The preabsorbed Lom-MT I antibody did not yield any positive reaction.
• 4.4. Our results indicate that the functional epitope FXPRL-amide is widespread among insect orders. Its distribution in the nervous system seems to be rather similar in all investigated species.

     

Insect acetyl-CoA carboxylase: Activity during the larval,pupal and adult stages of insect development

• 1.1. The activity of the lipogenic enzyme, acetyl-CoA carboxylase, was investigated in four insect species; Bombyx mori (Lepidoptera), Tenebrio molitor (Coleoptera), Glossina morsitans and Sarcophaga nodosa (Diptera).
• 2.2. Acetyl-CoA carboxylase activity in larval, pupal and adult forms was compared with the saponifiable lipid mass at each stage of the life-cycle, and found to follow similar patterns except for Tenebrio molitor.
• 3.3. The results are examined in relation to known metabolic requirements for each insect.

     

Catalytic and inhibitory properties of a major molecular form of acetylcholinesterase isolated from Lygus hesperus knight (Hemiptera: Miridae)

• 1.1. The major form of acetylcholinesterase (AChE) from Lygus hesperus demonstrated a greater affinity to selected substrates than unresolved AChE.
• 2.2. The turnover numbers of the native AChE were 7000 min⁻¹ for acetylthiocoline, 4800 for acetyl-(β-methyl) thiocholine, 3000 for propionylthiocholine, and 390 for S-butyrylthiocholine.
• 3.3. Each molecule of the major form had two active sites and each subunit had one active site.
• 4.4. Paraoxon or dichlorvos had a higher affinity to the major AChE form than to the unresolved AChE, resulting in a higher potency for the inhibition.
• 5.5. Some references of comparison are also made with AChE from other animal species.