Conjugate Position Research Articles

Encoding of Eye Position in the Goldfish Horizontal Oculomotor Neural Integrator

Monocular organization of the goldfish horizontal neural integrator was studied during spontaneous scanning saccadic and fixation behaviors. Analysis of neuronal firing rates revealed a population of ipsilateral (37%), conjugate (59%), and contralateral (4%) eye position neurons. When monocular optokinetic stimuli were employed to maximize disjunctive horizontal eye movements, the sampled population changed to 57, 39, and 4%. Monocular eye tracking could be elicited at different gain and phase with the integrator time constant independently modified for each eye by either centripetal (leak) or centrifugal (instability) drifting visual stimuli. Acute midline separation between the hindbrain oculomotor integrators did not affect either monocularity or time constant tuning, corroborating that left and right eye positions are independently encoded within each integrator. Together these findings suggest that the "ipsilateral" and "conjugate/contralateral" integrator neurons primarily target abducens motoneurons and internuclear neurons, respectively. The commissural pathway is proposed to select the conjugate/contralateral eye position neurons and act as a feedforward inhibition affecting null eye position, oculomotor range, and saccade pattern.

Synthesis and characterization of naphthalene diimide (NDI)-based near infrared chromophores with two-photon absorbing properties

A new series of 2,7-bis-(2-ethylhexyl)-benzo[ lmn][3,8]-phenanthroline-1,3,6,8-tetraone (NDI) based pseudo-quadrupolar molecules ( 1– 6) is presented and their two-photon absorption (2PA) cross-sections measured with the Z-scan method. The spectral properties of these compounds can be fine-tuned via modification of the donor segments. The corresponding 2PA cross-section ( σ 2) values at the most readily available 800 nm excitation range from 229±15 to 1092±59 GM owing to differences in conjugation length and/or position of substitution.

Phase II Metabolism of Hesperetin by Individual UDP-Glucuronosyltransferases and Sulfotransferases and Rat and Human Tissue Samples

Phase II metabolism by UDP-glucuronosyltransferases (UGTs) and sulfotransferases (SULTs) is the predominant metabolic pathway during the first-pass metabolism of hesperetin (4'-methoxy-3',5,7-trihydroxyflavanone). In the present study, we have determined the kinetics for glucuronidation and sulfonation of hesperetin by 12 individual UGT and 12 individual SULT enzymes as well as by human or rat small intestinal, colonic, and hepatic microsomal and cytosolic fractions. Results demonstrate that hesperetin is conjugated at positions 7 and 3' and that major enzyme-specific differences in kinetics and regioselectivity for the UGT and SULT catalyzed conjugations exist. UGT1A9, UGT1A1, UGT1A7, UGT1A8, and UGT1A3 are the major enzymes catalyzing hesperetin glucuronidation, the latter only producing 7-O-glucuronide, whereas UGT1A7 produced mainly 3'-O-glucuronide. Furthermore, UGT1A6 and UGT2B4 only produce hesperetin 7-O-glucuronide, whereas UGT1A1, UGT1A8, UGT1A9, UGT1A10, UGT2B7, and UGT2B15 conjugate both positions. SULT1A2 and SULT1A1 catalyze preferably and most efficiently the formation of hesperetin 3'-O-sulfate, and SULT1C4 catalyzes preferably and most efficiently the formation of hesperetin 7-O-sulfate. Based on expression levels SULT1A3 and SULT1B1 also will probably play a role in the sulfo-conjugation of hesperetin in vivo. The results help to explain discrepancies in metabolite patterns determined in tissues or systems with different expression of UGTs and SULTs, e.g., hepatic and intestinal fractions or Caco-2 cells. The incubations with rat and human tissue samples support an important role for intestinal cells during first-pass metabolism in the formation of hesperetin 3'-O-glucuronide and 7-O-glucuronide, which appear to be the major hesperetin metabolites found in vivo.

Comparative analysis of DNA alkylation by conjugates between pyrrole–imidazole hairpin polyamides and chlorambucil or seco-CBI

We investigated sequence-specific DNA alkylation using conjugates between the N-methylpyrrole (Py)- N-methylimidazole (Im) polyamide and the DNA alkylating agent, chlorambucil, or 1-(chloromethyl)-5-hydroxy-1,2-dihydro-3 H-benz[ e]indole ( seco-CBI). Polyamide–chlorambucil conjugates 1– 4 differed in the position at which the DNA alkylating chlorambucil moiety was bound to the Py–Im polyamide. High-resolution denaturing polyacrylamide gel electrophoresis (PAGE) revealed that chlorambucil conjugates 1– 4 alkylated DNA at the sequences recognized by the Py–Im polyamide core moiety. Reactivity and sequence specificity were greatly affected by the conjugation position, which reflects the geometry of the alkylating agent in the DNA minor groove. Polyamide– seco-CBI conjugate 5 was synthesized to compare the efficacy of chlorambucil with that of seco-CBI as an alkylating moiety for Py–Im polyamides. Denaturing PAGE analysis revealed that DNA alkylation activity of polyamide– seco-CBI conjugate 5 was similar to that of polyamide–chlorambucil conjugates 1 and 2. In contrast, the cytotoxicity of conjugate 5 was superior to that of conjugates 1– 4. These results suggest that the seco-CBI conjugate was distinctly active in cells compared to the chlorambucil conjugates. These results may contribute to the development of more specific and active DNA alkylating agents.

Jensen–Shannon divergence in conjugate spaces: The entropy excess of atomic systems and sets with respect to their constituents

The disorder of a composite system or of a set of different systems is always higher than the mere sum of the internal disorder of its constituents. One of the most widely used functionals employed for measuring the randomness of a single distribution is the Shannon entropy, from which the increase of disorder within the composite system with respect to those of the constituents is quantified by means of the Jensen–Shannon Divergence (JSD). In this work two different applications of the JSD in the study of the information content of atomic electron densities are carried out: (i) finding the contribution for a given atom of its composing subshells to the total atomic information; (ii) and similarly for selected sets of atoms, such as periods and groups throughout the Periodic Table as well as isoelectronic series. In both cases, the analysis is performed in the two conjugate position and momentum spaces, and the results are interpreted according to physically relevant quantities such as the ionization potential.

Molecular design of sequence specific DNA alkylating agents

Sequence-specific DNA alkylating agents have great interest for novel approach to cancer chemotherapy. We designed the conjugates between pyrrole (Py)-imidazole (Im) polyamides and DNA alkylating chlorambucil moiety possessing at different positions. The sequence-specific DNA alkylation by conjugates was investigated by using high-resolution denaturing polyacrylamide gel electrophoresis (PAGE). The results showed that polyamide chlorambucil conjugates alkylate DNA at flanking adenines in recognition sequences of Py-Im polyamides, however, the reactivities and alkylation sites were influenced by the positions of conjugation. In addition, we synthesized conjugate between Py-Im polyamide and another alkylating agent, 1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benz[e]indole (seco-CBI). DNA alkylation reactivies by both alkylating polyamides were almost comparable. In contrast, cytotoxicities against cell lines differed greatly. These comparative studies would promote development of appropriate sequence-specific DNA alkylating polyamides against specific cancer cells.

On the dynamical and geometrical symmetries of Keplerian motion

The dynamical symmetries of classical, relativistic and quantum-mechanical Kepler systems are considered to arise from geometric symmetries in PQET phase space. To establish their interconnection, the symmetries are related with the aid of a Lie-algebraic extension of Dirac's correspondence principle, a canonical transformation containing a Cunningham–Bateman inversion, and a classical limit involving a preliminary canonical transformation in ET space. The Lie-algebraic extension establishes the conditions under which the uncertainty principle allows the local dynamical symmetry of a quantum-mechanical system to be the same as the geometrical phase-space symmetry of its classical counterpart. The canonical transformation converts Poincaré-invariant free-particle systems into ISO(3,1) invariant relativistic systems whose classical limit produces Keplerian systems. Locally Cartesian relativistic PQET coordinates are converted into a set of eight conjugate position and momentum coordinates whose classical limit contains Fock projective momentum coordinates and the components of Runge–Lenz vectors. The coordinate systems developed via the transformations are those in which the evolution and degeneracy groups of the classical system are generated by Poisson-bracket operators that produce ordinary rotation, translation and hyperbolic motions in phase space. The way in which these define classical Keplerian symmetries and symmetry coordinates is detailed. It is shown that for each value of the energy of a Keplerian system, the Poisson-bracket operators determine two invariant functions of positions and momenta, which together with its regularized Hamiltonian, define the manifold in six-dimensional phase space upon which motions evolve.

Novel optical accelerometer based on Fresnel diffractive micro lens

This paper presents an innovative optical accelerometer based on an integration of a Fresnel diffractive micro lens with a reflecting membrane. The reflecting membrane mounted on an inertial mass is placed parallel behind the diffractive micro lens. The light intensity at the conjugate position of an applied monochromatic point source is highly sensitive to the displacement of the membrane. The working principle is specifically discussed. In addition, this sensor makes use of a quad beam structure to allow the flat displacement of the inertial mass. Mechanical simulations using finite element method show the designed structure has a wide range of operation. A prototype accelerometer has been successfully developed. Dynamic tests indicate that the accelerometer has promising performance. This accelerometer can be fabricated using microelectromechanical systems (MEMS) technology, with relatively few process steps, so it is suitable for batch production. By optimizing structural designs, it can be a competitive candidate for a number of application fields ranging from vibration monitoring to inertial navigation.

Influence of the metabolic profile on the in vivo antioxidant activity of quercetin under a low dosage oral regimen in rats

Flavonoids are known to possess a broad set of pharmacological effects, some of which have been attributed to their antioxidant properties and, more recently, to cell signalling modulation. Nevertheless, flavonoids are extensively metabolized and their metabolites are the potential bioactive forms in vivo. Therefore, a first and crucial step to understand the mechanisms underlying potential health benefits of flavonoids is knowledge of their metabolites and their biological activities. To approximate a human dietary pattern of intake of flavonoids, regular rat chow was supplemented with 0.02% quercetin and fed to Sprague-Dawley rats over 3 weeks. Plasma samples were analysed by HPLC and electrospray tandem mass spectrometry, and plasma antioxidant capacity was measured by the 2,2'-azino-bis(3-ethylbenzothiazoline sulphonate) assay. Major metabolites were 3'-methylquercetin (isorhamnetin) glucuronide sulphate conjugates, the most plausible conjugation positions being at the 3-, 5- and 7-hydroxyl positions. Isorhamnetin conjugates are methylated at the 3'-OH position, which decreases the high antioxidant activity of quercetin and its metabolites and their contribution to plasma antioxidant potential. This metabolic pattern differs from that observed after a single high-dose administration, where the major metabolites were quercetin conjugates at 5- and 7-hydroxyl positions and a significantly increased plasma antioxidant activity was observed. These data show altogether that the different metabolic patterns obtained under a prolonged low-dosage regimen or after a single high dose, crucially affected the antioxidant potential of plasma in treated animals. Our data also allow for the establishment of structure-antioxidant activity relationships for quercetin metabolites.

Modulation of Human Multidrug Resistance Protein (MRP) 1 (ABCC1) and MRP2 (ABCC2) Transport Activities by Endogenous and Exogenous Glutathione-Conjugated Catechol Metabolites

Members of the multidrug resistance protein (MRP/ABCC) subfamily of ATP-binding cassette proteins transport a wide array of anionic compounds, including sulfate, glucuronide, and glutathione (GSH) conjugates. The present study tested the ATP-dependent vesicular transport of leukotriene C(4) and 17beta-estradiol 17-(beta-d-glucuronide) (E(2)17betaG) mediated by the MRP1 and MRP2 transporters in the presence of six potential modulators from three different classes of GSH-conjugated catechol metabolites: the ecstasy metabolite 5-(glutathion-S-yl)-N-methyl-alpha-methyldopamine (5-GS-N-Me-alpha-MeDA), the caffeic acid metabolite 2-(glutathion-S-yl)-caffeic acid (2-GS-CA), and four GSH conjugates of 2-hydroxy (OH) and 4-OH estrogens (GS estrogens). MRP1-mediated E(2)17betaG transport was inhibited in a competitive manner with a relative order of potency of GS estrogens (IC(50) <1 microM) > 2-GS-CA (IC(50) 3 microM) > 5-GS-N-Me-alpha-MeDA (IC(50) 31 microM). MRP2-mediated transport was inhibited with a similar order of potency, except the 2-hydroxy-4-(glutathion-S-yl)-estradiol and 4-hydroxy-2-(glutathion-S-yl)-estradiol conjugates were approximately 50- and 300-fold less potent, respectively. Transport activity was unaffected by N-acetylcysteine conjugates of N-Me-alpha-MeDA and CA. The position of GSH conjugation appears important as all four GS estrogen conjugates tested were potent inhibitors of MRP1 transport, but only the 2-hydroxy-1-(glutathion-S-yl)-estradiol and 2-hydroxy-1-(glutathion-S-yl)-estrone conjugates were potent inhibitors of MRP2-mediated transport. In conclusion, we have identified three new classes of MRP1 and MRP2 modulators and demonstrated that one of these, the estrogen conjugates, shows unanticipated differences in their interactions with the two transporters.

Mechanism of RNA Cleavage Catalyzed by Sequence Specific Polyamide Nucleic Acid-Neamine Conjugate

In earlier studies, we found that a conjugate of neamine-polyamide nucleic acid targeting transactivation response element of HIV-1 RNA genome (HIV-1 TAR) displayed anti-HIV-1 activity and sequence-specific cleavage of the target RNA in vitro. Here we show that both the position of conjugation of polyamide nucleic acid (PNA) on neamine and the length of the spacer are critical parameters for conferring cleavage activity to the conjugate. The conjugation of PNA via a spacer incorporating 11 atoms to the 5-position of ring I of the neamine core conferred sequence-specific RNA cleavage activity on the conjugate, while conjugation to the 4'-position of ring II abolished this activity. Similarly, 5-neamine PNA complementary to TAR sequence of HIV-1 genome (PNA(TAR)) conjugates having either a 23-atom spacer or a bulky dansyl group between PNA and the neamine core also resulted in complete loss of cleavage activity. Based on these observations, we propose a mechanism for the observed RNA cleavage catalyzed by the conjugate involving unprotonated and protonated amino groups at the 3-position of ring I and the 6'-position of ring II of the neamine core, respectively.

Direct Ring Conjugation of Catecholamines and Their Immunological Interactions

Catecholamine derivatives were synthesized with potential applications as coating antigens in biosensors or in the raising of specific antibodies. Thioether-bridged derivatives of the catecholamines dopamine, norepinephrine, and epinephrine that attach carboxylic acid functionalities directly to the aromatic ring via an easily incremented linker chain were synthesized by an electrochemical method. These derivatives were purified by convenient ion-exchange chromatography, exact positions of conjugation determined by NMR, and a dopamine derivative immobilized in situ in a BIAcore surface plasmon resonance (SPR) biosensor and its antibody binding studied in comparison with immobilization via the catecholamine primary amine. Binding of an antibody raised to an amine-conjugated protein conjugate showed clear distinction between conjugations at different positions on the catecholamine, illustrating the importance of rational conjugate design in immunosensing of the catecholamines.

Metabolic Profile of FYX-051 (4-(5-Pyridin-4-yl-1H-[1,2,4]triazol-3-yl)pyridine-2-carbonitrile) in the Rat, Dog, Monkey, and Human: Identification of N-Glucuronides and N-Glucosides

FYX-051, 4-(5-pyridin-4-yl-1H-[1,2,4]triazol-3-yl)pyridine-2-carbonitrile, is a novel xanthine oxidoreductase inhibitor that can be used for the treatment of gout and hyperuricemia. We examined the metabolism of FYX-051 in rats, dogs, monkeys, and human volunteers after the p.o. administration of this inhibitor. The main metabolites in urine were pyridine N-oxide in rats, triazole N-glucoside in dogs, and triazole N-glucuronide in monkeys and humans, respectively. Furthermore, N-glucuronidation and N-glucosidation were characterized by two types of conjugation: triazole N(1)- and N(2)-glucuronidation and N(1)- and N(2)-glucosidation, respectively. N(1)- and N(2)-glucuronidation was observed in each species, whereas N(1)- and N(2)-glucosidation was mainly observed in dogs. With regard to the position of conjugation, N(1)-conjugation was predominant; this resulted in a considerably higher amount of N(1)-conjugate in each species than N(2)-conjugate. The present results indicate that the conjugation reaction observed in FYX-051 metabolism is unique, i.e., N-glucuronidation and N-glucosidation occur at the same position of the triazole ring, resulting in the generation of four different conjugates in mammals. In addition, a urinary profile of FYX-051 metabolites in monkeys and humans was relatively similar; triazole N-glucuronides were mainly excreted in urine.

Aldose reductase inhibitory effect by tectorigenin derivatives from Viola hondoensis

Aldose reductase (AR), the key enzyme of the polyol pathway, is known to play important roles in the diabetic complication. The inhibitors of AR, therefore, would be potential agents for the prevention of diabetic complication. The AR inhibition activity of several isoflavonoids was evaluated in vitro against rat lens. Tectoridin-4′- O-β- d-glucoside exhibited strong AR inhibition activity on rat lens with an IC 50 of 0.54 μM. Similar activities were recorded for the natural tectorigenin and tectoridin. In contrast, tectoridin-4′- O-β- d-glucoside showed a stronger inhibitory activity than tectorigenin and tectoridin. Our results indicate that glucose conjugation position in this type of isoflavonoids may be required for the activity.

Differentiation of estriol glucuronide isomers by chemical derivatization and electrospray tandem mass spectrometry

This paper describes a way of differentiating between the three isomers of estriol glucuronide by the use of chemical derivatization and liquid chromatography/electrospray tandem mass spectrometry (MS/MS). In their native form, these isomers gave rise to almost identical product ion spectra, involving the neutral loss of 176 Da (i.e. monodehydrated glucuronic acid), which made it impossible to determine the position of conjugation by MS/MS alone. In order to change the fragmentation pathways, positive charges were introduced into the analytes by chemical derivatization. The following reagents were tested: 2-chloro-1-methylpyridinium iodide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 2-picolylamine. Interestingly, derivatization using a combination of all three reagents gave a selective fragmentation pattern that could differentiate between the isomers estriol-16-glucuronide and estriol-17-glucuronide. Estriol-3-glucuronide, which lacks a free phenolic group, could be differentiated through a different type of reaction product when exposed to 2-chloro-1-methylpyridinium iodide. Furthermore, in order to assist structural assignment of the fragments, their accurate masses were determined using a hybrid quadrupole time-of-flight mass spectrometer and fragmentation pathways were elucidated by the use of MS3 on an ion trap mass spectrometer.

Olfactory responses to steroids in an African mouth‐brooding cichlid, Haplochromis burtoni(Günther)

Underwater electro‐olfactogram (EOG) recordings involving 150 steroids and eight prostaglandins were used to determine which of these potential odorants are detected by the olfactory organ of an African cichlid, Haplochromis burtoni. In initial EOG tests at 10−9 M, H. burtoni did not respond to unconjugated steroids or prostaglandins, but did respond to 17 conjugated steroids, 11 of which (17β‐oestradiol‐17β‐glucuronide; 17β‐oestradiol‐3‐sulphate; 17β‐oestradiol‐3,17β‐disulphate; epiandrosteron‐3β‐sulphate; etiocholanolone‐3α‐glucuronide; testosterone‐17β‐sulphate; dehydroepiandrosterone‐3β‐sulphate; 5α‐pregnan‐3β‐ol‐20‐one‐3β‐sulphate; 5β‐pregnan‐3α,17‐diol‐20‐one‐3α‐glucuronide; 5β‐pregnan‐3α,17,21‐triol‐11,20‐dione‐3α‐glucuronide; pregnenolone‐3β‐sulphate) were selected for EOG concentration‐response, cross‐adaptation and binary mixture tests. The EOG detection thresholds ranged from 10−11 to 10−9 M in all but one instance (female threshold to pregnenolone‐3β‐sulphate; 10−8 M), and males and females exhibited only minor differences in EOG threshold or response magnitude. Results of EOG cross‐adaptation tests, which were supported by results of binary mixture tests, indicated that the response to the 11 steroid conjugates is mediated by five putative olfactory receptor mechanisms characterized by specificity for conjugate position and type: 3‐sulphate, 17‐sulphate, 3,17‐disulphate, 3‐glucuronide, 17‐glucuronide. Although there is no evidence that H. burtoni releases, or exhibits biological response to, the steroids shown to be detected in this study, the present results are suggestive of a complex pheromone system utilizing steroid conjugates.

The inner magnetosphere of Saturn: Cassini RPWS cold plasma results from the first encounter

We present new results from the inner magnetosphere of Saturn obtained by the Radio and Plasma Wave Science (RPWS) investigation onboard Cassini around the period of the Saturn orbit injection (July 1, 2004). Plasma wave electric field emissions, voltage sweeps by the Langmuir probe (LP) and radio sounder data were used to infer the cold plasma (&lt;100 eV) characteristics within 20 RS of Saturn. A dense (&lt;150 cm−3) and cold (&lt;7 eV) plasma torus was found just outside the visible F‐ring. This torus of partly dusty plasma does not perfectly co‐rotate with Saturn, which suggests the cold plasma is electro‐dynamically coupled to the charged ring‐dust particles. The spacecraft potential was a few volts negative above the E‐ and G‐rings, indicating the dust‐particles were likewise negatively charged. The cold ion characteristics changed near the magnetically conjugate position of Dione, indicating release of volatile material from this icy moon.

Smart Polymer‐Streptavidin Conjugates

AbstractFor Abstract see ChemInform Abstract in Full Text.

Identification of phenolic compounds in strawberries by liquid chromatography electrospray ionization mass spectroscopy

Strawberry ( Fragaria x ananassa Duch.) fruits contain phenolic compounds that have antioxidant, anticancer, antiatherosclerotic and anti-neurodegenerative properties. Identification of food phenolics is necessary since their nature, size, solubility, degree and position of glycosylation and conjugation influence their absorption, distribution, metabolism and excretion in humans. Freeze-dried whole strawberry fruit powder and strawberry fruit extracts were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC–ESI–MS) methods. Phenolics were identified as ellagic acid (EA), EA-glycosides, ellagitannins, gallotannins, anthocyanins, flavonols, flavanols and coumaroyl glycosides. The anthocyanidins were pelargonidin and cyanidin, found predominantly as their glucosides and rutinosides. The major flavonol aglycons were quercetin and kaempferol found as their glucuronides and glucosides. LC–ESI–MS/MS methods differentiated EA from quercetin conjugates since both aglycons have identical molecular weights (302 g/mol). The identification of strawberry phenolics is necessary to generate standardized materials for in vitro and in vivo studies and for the authentication of strawberry-based food products.

Seismic structure of the Carnegie ridge and the nature of the Galápagos hotspot

The Galápagos volcanic province (GVP) includes several aseismic ridges resulting from the interaction between the Galápagos hotspot (GHS) and the Cocos–Nazca spreading centre (CNSC). The most prominent are the Cocos, Carnegie and Malpelo ridges. In this work, we investigate the seismic structure of the Carnegie ridge along two profiles acquired during the South American Lithospheric Transects Across Volcanic Ridges (SALIERI) 2001 experiment. Maximum crustal thickness is ∼19 km in the central Carnegie profile, located at ∼85°W over a 19–20 Myr old oceanic crust, and only ∼13 km in the eastern Carnegie profile, located at ∼82°W over a 11–12 Myr old oceanic crust. The crustal velocity models are subsequently compared with those obtained in a previous work along three other profiles over the Cocos and Malpelo ridges, two of which are located at the conjugate positions of the Carnegie ones. Oceanic layer 2 thickness is quite uniform along the five profiles regardless of the total crustal thickness variations, hence crustal thickening is mainly accommodated by layer 3. Lower crustal velocities are systematically lower where the crust is thicker, thus contrary to what would be expected from melting of a hotter than normal mantle. The velocity-derived crustal density models account for the gravity and depth anomalies considering uniform and normal mantle densities (3300 kg m−3), which confirms that velocity models are consistent with gravity and topography data, and indicates that the ridges are isostatically compensated at the base of the crust. Finally, a two-dimensional (2-D) steady-state mantle melting model is developed and used to illustrate that the crust of the ridges does not seem to be the product of anomalous mantle temperatures, even if hydrous melting coupled with vigorous subsolidus upwelling is considered in the model. In contrast, we show that upwelling of a normal temperature but fertile mantle source that may result from recycling of oceanic crust prior to melting, accounts more easily for the estimated seismic structure as well as for isotopic, trace element and major element patterns of the GVP basalts.