Changes in trigonelline (N-methylnicotinic acid) content and nicotinic acid metabolism during germination of mungbean (Phaseolus aureus) seeds

Changes in trigonelline content and in biosynthetic activity were determined in the cotyledons and embryonic axes of etiolated mungbean (Phaseolus aureus) seedlings during germination. Accumulation of trigonelline (c. 240 nmol per pair of cotyledons) was observed in the cotyledons of dry seeds; trigonelline content decreased 2 d after imbibition. Trigonelline content in the embryonic axes increased with seedling growth and reached a peak (c. 380 nmol per embryonic axis) at day 5. Trigonelline content did not change significantly during the differentiation of hypocotyls, and the concentration was greatest in the apical 5 mm. Nicotinic acid and nicotinamide were better precursors for pyridine nucleotide synthesis than quinolinic acid, but no great differences were found in the synthesis of trigonelline from these three precursors. Trigonelline synthesis was always higher in embryonic axes than in cotyledons. Activity of quinolinate phosphoribosyltransferase (EC 2.4.2.19), nicotinate phosphoribosyltransferase (EC 2.4.2.11), and nicotinamidase (EC 3.5.1.19) was found in cotyledons and embryonic axes, but no nicotinamide phosphoribosyltransferase (EC 2.4.2.12) activity was detected. It follows that quinolinic acid and nicotinic acid were directly converted to nicotinic acid mononucleotide by the respective phosphoribosyltransferases, but nicotinamide appeared to be converted to nicotinic acid mononucleotide after conversion to nicotinic acid. Trigonelline synthase (nicotinate N-methyltransferase, EC 2.1.1.7) activity increased in the embryonic axes, but decreased in cotyledons during germination. [14C]Nicotinic acid and trigonelline absorbed by the cotyledons were transported to the embryonic axes during germination. Trigonelline had no effect on the growth of seedlings, but nicotinic acid and nicotinamide significantly inhibited the growth of roots. Based on these findings, the role of trigonelline synthesis in mungbean seedlings is discussed.     

Pyridine nucleotide cycle and trigonelline (N-methylnicotinic acid) synthesis in developing leaves and fruits of Coffea arabica

We examined the biosynthesis of trigonelline in leaves and fruits of Arabica coffee ( Coffea arabica ) plants. [³H]Quinolinic acid, which is an intermediate of de novo pyridine nucleotide synthesis, and [¹⁴C]nicotinamide and [¹⁴C]nicotinic acid, which are degradation products of NAD, were converted to trigonelline and pyridine nucleotides. These tracer experiments suggest that the pyridine nucleotide cycle, nicotinamide → nicotinic acid → nicotinic acid mononucleotide (NaMN) → nicotinic acid adenine dinucleotide (NaAD) → NAD → nicotinamide mononucleotide (NMN) → nicotinamide, operates in coffee plants, and trigonelline is synthesized from nicotinic acid formed in the cycle. Trigonelline accumulated up to 18 µmol per leaf in developed young leaves, and then decreased with age. Although the biosynthetic activity of trigonelline from exogenously supplied [¹⁴C]nicotinamide was observed in aged leaves, the endogenous supply of nicotinamide may be limited, reducing the contents in these leaves. Trigonelline is synthesized and accumulated in fruits during development. The trigonelline synthesis in pericarps is much higher than that in seeds, but its content in seeds is higher than pericaps, so that some of the trigonelline synthesized in the pericarps may be transported to seeds. Trigonelline in seeds may be utilized during germination, as its content decreases. Trigonelline synthesis from [¹⁴C]nicotinamide was also found in Theobroma cacao plants, but instead of trigonelline, nicotinic acid-glucoside was synthesized from [¹⁴C]nicotinamide in Camellia sinensis plants.     

Noradrenergic mechanisms appear not to be involved in cocaine-induced seizures and lethality

In the present study we have investigated the effects of compounds which increase synaptic levels of noradrenaline on cocaine-induced seizures and lethality in mice. The noradrenaline uptake blocker desipramine (0.3, 3, 30 mg/kg i.p.; 1h pretreatment) and the alpha 2-antagonists idazoxan (0.05, 0.5, 5 mg/kg i.p.; 15 min) and RX811059A (0.01, 0.1, 1 mg/kg i.p.; 15 min) neither reduced nor increased the number of animals having convulsions in the 10 min following administration of cocaine (45, 60 mg/kg i.p.). None of these drugs increased lethality when assessed 10 minutes after 60 mg/kg cocaine and the alpha 2-antagonists did not protect against the lethal effects of a 90 mg/kg dose. On the other hand, desipramine significantly reduced the number of animals dying after this high dose of cocaine. These results suggest that noradrenergic mechanisms do not promote cocaine-induced convulsions and lethality - an important observation in light of the growing use of desipramine for initiation of abstinence in cocaine-dependent outpatients.     

Antibody responses to trinitrophenyl (TNP)-l-glutamic acid60-l-alanine30-l-tyrosine10 (GAT) in microcultures: Anti-hapten and anti-carrier responses appear to be under separable control

We have previously reported that the IgM anti-hapten plaque-forming cell (PFC) response to trinitrophenyl (TNP)-conjugated l-glutamic acid⁶⁰-l-alanine³⁰-l-tyrosine¹⁰ (GAT) is not under conventional Ir gene control in an in vitro microculture system. To explore this phenomenon, we examined both the anti-hapten and anti-carrier antibody responses to TNP-GAT in the microculture supernatants using a solid-phase radioimmunoassay (RIA). We show that splenocytes from GAT-responder BALB/c mice produce anti-hapten and anti-carrier antibody responses to TNP-GAT in microcultures, while splenocytes from GAT-nonresponder DBA/ 1 mice produce anti-hapten but not anti-carrier responses to TNP-GAT in these culture conditions. We further demonstrate that supernatants from rat splenocyte cultures stimulated by concanavalin A (Con A) similarly drive unprimed, T-lymphocyte-depleted splenocytes to produce anti-hapten but not anti-carrier antibody responses to TNP-GAT in microcultures. This observation suggests that T-cell-B-cell contact may not be required for the generation of anti-hapten responses by splenocytes to TNP-GAT in microcultures and may explain the absence of conventional Ir gene control of such responses in these cultures.     

Unexpected systematic affinities and geographic expansion of a marine alien hydroid (Cnidaria: Hydrozoa)

Biological systematics provides taxonomic information and expertise for biogeography and biological invasion research and management. However, the systematic classification of most taxa, including some alien and invasive species, relies only on morphology. This applies for example to Sertularella tongensis Stechow, 1919 Stechow, E. (1919). Zur Kenntnis der Hydroidenfauna des Mittelmeeres, Amerikas und anderer Gebiete, nebst Angaben über einige Kirchenpauer'sche Typen von Plumulariden. Zoologische Jahrbuecher (Systematik), 42, 1–172. [Google Scholar] (Cnidaria: Sertulariidae), an alien hydrozoan recently reported in the Mediterranean. Its genus affiliation is still unclear and controversial. Historically, it has been assigned to several existing genera. However, molecular data (COI, 16S, 18S, 28S) provided here do not support these previous assignments. Instead, integrative analyses combining morphological and molecular data support reassigning this species to a new genus, Bicaularia gen. nov. A biogeographic review revealed that this species is widely distributed in the mid- and low-latitude waters of the Indo-Pacific region, and is spreading to some tropical and temperate regions including the Mediterranean and Caribbean Seas, as well as Korea and Japan. This expansion may be attributable to human activity such as shipping through the Suez and the Panama Canals. However, the new record attached to a floating Sargassum algal fragment in the surface water of the open Pacific, might suggest a neglected cryptic spread pathway with the assistance of floating biological organisms. Our findings shed light on the complexity of the phylogeny and spread pathway of the investigated hydrozoan species. We regard the fine systematics as the first step towards future global sampling for molecular studies aiming to elucidate the origins and pathways for alien and invasive species.

http://zoobank.org/urn:lsid:zoobank.org: pub: 45AB0849-1E3B-477FB643-1B3B4603C1FF     

Is homarine a morphogen in the marine hydroid Hydractinia?

Summary Homogenate of coelenterate tissue interferes with metamorphosis in Hydractinia and pattern formation in both Hydractinia, and Hydra. From the extracts two fractions comprising low-molecular-weight compounds with strong metamorphosis-inhibiting activity were separated. One of these contains, as the active compound, homarine (N-methyl picolinic acid). Homarine concentrations down to 10^–6 mol/l stop or retard metamorphosis. High concentrations block the continuation of metamorphosis as long as they are maintained in the culture medium and treatment with homarine during metamorphosis influences the proportioning of the future polyp's body pattern. Most of the homarine found in Hydra tissue derives from Artemia given as food. It is not identical with inhibitor I, an activity partially purified from Hydra tissue, which prevents head and foot formation in Hydra.     

Chinese hamster ovary K2 cell lipid droplets appear to be metabolic organelles involved in membrane traffic

The principal lipids in animal cell lipid droplets are cholesterol, cholesterol ester, and triglyceride, but the protein composition of this compartment is largely unknown. Here we report on the proteomic analysis of lipid droplets. Using a combination of mass spectrometry and immunoblotting, we identify nearly 40 specifically associated proteins in droplets isolated from Chinese hamster ovary K2 cells grown in normal medium. The proteins fall in to five groups: structural molecules of the droplet-like adipose differentiation-related protein; multiple enzymes involved in the synthesis, storage, utilization, and degradation of cholesterol esters and triglycerides; multiple, different Rab GTPases known to be involved in regulating membrane traffic; signaling molecules such as p50RhoGAP; and a group of proteins that do not fit any classification but include proteins often found in caveolae/rafts such as caveolin-1 and 2 and flotillin-1. The proteome of droplets isolated from cells grown in the presence of oleate is largely the same except for an increase in the amount of adipose differentiation-related protein, caveolin-1, and a protein thought to be involved in phospholipid recycling called CGI-58. Based on the protein profile, the lipid droplet appears to be a complex, metabolically active organelle that is directly involved in membrane traffic and possibly phospholipid recycling. We propose the name adiposome for this organelle.     

Extracellular cytochrome nanowires appear to be ubiquitous in prokaryotes

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Antitrypanosomal activity of isololiolide isolated from the marine hydroid Macrorhynchia philippina (Cnidaria,Hydrozoa)

Marine invertebrates are a rich source of small antiparasitic compounds. Among them, Macrorhynchia philippina is a chemically underexplored marine cnidarian. In the search for candidates against the neglected protozoan Chagas disease, we performed a bio-guided fractionation to obtain active compounds. The structural characterization of the active compound was determined using NMR analysis and MS and resulted in the isololiolide, a compound described for the first time in this species. It showed in vitro activity against both trypomastigote and intracellular amastigotes of Trypanosoma cruzi, with IC₅₀ values of 32 µM and 40 µM, respectively, with no mammalian cytotoxicity (>200 µM). The lethal action was investigated in T. cruzi using different fluorophores to study: (i) mitochondrial membrane potential; (ii) plasma membrane potential and (iii) plasma membrane permeability. Our results demonstrated that isololiolide caused disruption of the plasma membrane integrity and a strong depolarization of the mitochondrial membrane potential, rapidly leading the parasite to death. Despite being considered a possible covalent inhibitor, safety in silico studies of isololiolide also considered neither mutagenic nor genotoxic potential. Additionally, isololiolide showed no resemblance to interference compounds (PAINS), and it succeeded in most filters for drug-likeness. Isololiolide is a promising candidate for future optimization against Chagas disease.     

Not lipoteichoic acid but lipoproteins appear to be the dominant immunobiologically active compounds in Staphylococcus aureus

Lipoteichoic acid (LTA) derived from Staphylococcus aureus is reported to be a ligand of TLR2. However, we previously demonstrated that LTA fraction prepared from bacterial cells contains lipoproteins, which activate cells via TLR2. In this study, we investigated the immunobiological activity of LTA fraction obtained from S. aureus wild-type strain, lipoprotein diacylglycerol transferase deletion (delta lgt) mutant, which lacks palmitate-labeled lipoproteins, and its complemented strain and evaluated the activity of LTA molecule. LTA fraction was prepared by butanol extraction of the bacteria followed by hydrophobic interaction chromatography. Although all LTA fractions activated cells through TLR2, the LTA from delta lgt mutant was 100-fold less potent than those of wild-type and complemented strains. However, no significant structural difference in LTA was observed in NMR spectra. Further, alanylation of LTA molecule showed no effect in immunobiological activity. These results showed that not LTA molecule but lipoproteins are dominant immunobiologically active TLR2 ligand in S. aureus.     

Population increases in obesity appear to be partly due to genetics

Studies have documented substantial increases in obesity throughout most of the industrialized world in recent decades. The majority of explanations for these increases have centred around environmental factors such as the increasing availability of high-fat, high-carbohydrate foods and sedentary lifestyles. This study sought to determine if genetic factors might be contributing to the increases in the proportions of North Americans who are obese and overweight. The body mass index (BMI) for a large sample of two generations of United States and Canadian subjects was correlated with family fertility indicators. Small but highly significant positive correlations were found between the BMIs of family members and their reproduction rates, especially in the case of women. For instance, mothers in the sample (most of whom were born in the 1940s and 50s) who were in the normal or below normal range had an average of 4.3 siblings and 3.2 children, compared with 4.8 siblings and 3.5 children for mothers who were overweight or obese. When combined with evidence from twin and adoption studies indicating that genes make substantial contributions to obesity, this study suggests that recent increases in obesity are partially the result of overweight and obese women having more children than is true for average and underweight women. It is speculated that improvements in medical treatments for conditions associated with obesity--particularly diabetes and heart disease--are making it possible for overweight women to live longer and to be more fertile than was true historically.     

Numerical simulations of the effects of retinoids on pattern formation in a hydroid

Summary Retinoids have been shown to influence pattern formation in hydroid polyps (Hydractinia echinata) at various levels. These effects are counteracted by an inhibitor isolated from Hydra. The present study provides a theoretical attempt to elucidate the role of retinoids by computer simulations based on a simple model of pattern formation in Hydractinia. The elements of this model are morphogens of the Gierer-Meinhardt type, namely a long-range inhibitor and a short-range activator. From the calcualtions, a reducing effect of retinoids on the propagation of the inhibitor seems most probable.     

Tissue compatibility in regenerating explants from the colonial marine hydroid Hydractinia echinata (Flem.)

Artificially maintained colonies of the colonial marine hydroid Hydractinia echinata are endogenously limited in lateral growth and usually develop in a circular pattern with a closed periderm. Further lateral growth consists of free stolons. Preliminary studies of tissue compatibility in regenerating explants indicate that fusion generally fails to occur if the opposing explants are of differing sex, from different individuals of the same sex, or if peripheral explant contact occurs after free stolons have begun to form. Of the three aforementioned causes of fusion failure, the most important factor appears to be the timing of peripheral explant contact. If regenerating explants contact each other before the endogenous circular growth pattern is achieved, fusion will occur regardless of sex and/or individuality. Incompatibility between explants from the same individual may be because of periderm development inhibition in some instances. However, the failure of free stolon fusion between explants from the same individual suggests the development of “temporal specificity” once the endogenous limit of lateral growth is achieved.     

A hemolysin of Vibrio mimicus (VMH) stimulates cells to produce ATP and cyclic AMP which appear to be secretory mediators

The hemolysin of Vibrio mimicus(VMH) is a pore-forming toxin with both enterotoxic and hemolytic activity. The hemolysis by VMH is induced by creation of pores in the membrane of erythrocyte; however, the mechanism for the enterotoxic action of VMH has remained unclear. In order to clarify the mechanism, we incubated T84 cells (a human colon carcinoma cell line) with VMH and found that the levels of ATP and cyclic AMP of culture medium increased after exposure of the cells to VMH. Subsequently, we found that the fluid accumulating activity of VMH in a mouse internal loop assay was reduced by administration of glibenclamide, an inhibitor of cyclic AMP-dependent chloride channels, into the intestinal loop. These results suggest that the stimulation of cells to produce nucleotides by VMH is linked to the enterotoxic activity of the toxin.     

Human glucosamine-6-phosphate isomerase, a homologue of hamster oscillin, does not appear to be involved in Ca2+ release in mammalian oocytes

Injections of cytosolic preparations from mammalian sperm into oocytes have been shown to trigger calcium [Ca2+]i oscillations and initiate activation of development. Recently, a protein isolated from hamster sperm has been suggested to be involved in the generation of these oscillations and it was named "oscillin." The human homologue of hamster oscillin is glucosamine 6-phosphate isomerase (GPI, EC no. 5.3.1.10), an enzyme so far described to be involved in hexose phosphate metabolism. To assess the role of GPI on Ca2+ signaling, a human recombinant protein was generated in a prokaryotic system and injected into fura-2-dextran-loaded metaphase II (MII) mouse oocytes. Injection of recombinant GPI failed to induce Ca2+ responses in 12/12 injected MII oocytes despite the fact that the recombinant GPI was active as assessed by an enzymatic assay. Injection of buffer (0/6 oocytes) or fructose-6-phosphate, a product of GPI enzymatic reaction (0/5 oocytes), also failed to initiate Ca2+ responses. Conversely, injections of sperm cytosolic factor induced [Ca2+]i oscillations in all 17/17 oocytes. In addition, injection of recombinant GPI or GPI mRNA failed to induce parthenogenetic activation (0/30 oocytes). Immunofluorescence studies using an anti-GPI polyclonal antibody (GK) resulted in localization of GPI to the sperm's equatorial region. Incubation of the GK antibody with sperm extracts failed to block the [Ca2+]i responses induced by these extracts. Moreover, near complete depletion of GPI from sperm fractions by immunoprecipitation did not impair the ability of these fractions to induce [Ca2+]i oscillations. In summary, our results support the role of a sperm cytosolic component(s) in the generation of [Ca2+]i oscillations during mammalian fertilization, although a protein other than GPI/oscillin is likely to be the active calcium releasing factor.     

Immature euphausiids do not appear to be prey for humpback whales (Megaptera novaeangliae) during spring and summer in Southeast Alaska

Humpbacks whales (Megaptera novaeangliae) have shown a remarkable recovery in the North Pacific, raising concerns regarding their impact on marine communities. In Southeast Alaska, humpbacks feed heavily on euphausiids; however, it remains unclear whether they target immature individuals despite evidence that they do so elsewhere. I evaluate the hypothesis that humpbacks target immature euphausiids in late spring‐summer in Southeast Alaska. Plankton samples were collected at random sites (n = 44) and near whales (n = 53) between 8 June and 9 September 2008 in Frederick Sound and Stephens Passage. The proportion of samples containing immature euphausiids, and immature euphausiid abundance within those samples, were compared between the two sample types. Similar analyses were conducted for adult euphausiids (prey) and calanoid copepods (nonprey) for comparison. I found no statistical difference between the whale and random samples with respect to the occurrence or numerical density of immature euphausiids, which is consistent with the hypothesis that whales did not target them in 2008. Smaller size, insufficient numerical densities and lower energy density of immature euphausiids are suggested as possible reasons. These findings can assist in resolving regional humpback abundance and distribution patterns, and can contribute to an understanding of the trophic interactions characterizing the local ecosystem.