Enzyme linked immunosorbent assay (ELISA) for beta-endorphin and its antibodies

An Enzyme Linked Immunosorbent Assay (ELISA) is described for use in the determination of beta-endorphin antibody titers, as well as for the quantitation of naturally occurring levels of beta-endorphin in plasma and other bodily fluids. The ability of the assay to accommodate unpurified samples containing small concentrations of beta-endorphin was improved through the use of affinity purified antibodies in conjunction with a competitive inhibition ELISA. The problem of non-specific binding of beta-endorphin during competitive inhibition assays was circumvented through a two-step process in which the plate was first coated with BSA, followed by a second plate coating with poly-lysine (MW4000). The second coating with poly-lysine was found necessary in order to eliminate intermolecular void spaces following initial plate treatment with BSA. Following these procedures enabled quantitation of beta-EP at a level as low as 10 pmoles per microtitre plate well.     

Mirror-induced behavior in the magpie (Pica pica): evidence of self-recognition

Comparative studies suggest that at least some bird species have evolved mental skills similar to those found in humans and apes. This is indicated by feats such as tool use, episodic-like memory, and the ability to use one's own experience in predicting the behavior of conspecifics. It is, however, not yet clear whether these skills are accompanied by an understanding of the self. In apes, self-directed behavior in response to a mirror has been taken as evidence of self-recognition. We investigated mirror-induced behavior in the magpie, a songbird species from the crow family. As in apes, some individuals behaved in front of the mirror as if they were testing behavioral contingencies. When provided with a mark, magpies showed spontaneous mark-directed behavior. Our findings provide the first evidence of mirror self-recognition in a non-mammalian species. They suggest that essential components of human self-recognition have evolved independently in different vertebrate classes with a separate evolutionary history.     

Zinc(II)-phenoxyl radical complexes: Dependence on complexation properties of Zn-phenolate species

Zinc(II) complexes of N₃O-donor tripodal ligands, 2,4-di(tert-butyl)-6-{[bis(2-pyridyl)methyl]aminomethyl}phenol (HtbuL), 2,4-di(tert-butyl)-6-{[(6-methyl-2-pyridyl)methyl]-(2-pyridyl)methylaminomethyl}phenol (HtbuLMepy), and 2,4-di(tert-butyl)-6-{[bis(6-methyl-2-pyridyl)methyl]aminomethyl}phenol (HtbuL(Mepy)₂), [Zn(tbuL)Cl] · CH₃OH (1), [Zn(tbuLMepy)Cl] (2), and [Zn(tbuL(Mepy)₂)Cl] (3), respectively, were prepared and structurally characterized by the X-ray diffraction method. All the complexes were found to have a mononuclear structure with a coordinated phenolate moiety, the geometry of the Zn(II) center being 5-coordinate trigonal-bipyramidal. The Zn(II) binding ability of the ligands with and without 6-methyl-2-pyridylmethyl moieties was evaluated for similar ligands, which lacked the t-butyl groups at the 2- and 4-positions of the phenol moiety, by the stability constants determined by potentiometric titration at 25 °C (I = 0.1 M (KNO₃)). The stability of the complexes was found to be in the order L > LMepy > L(Mepy)₂, reflecting the steric hindrance of the 6-methyl group of the pyridine ring. Complexes 1, 2, and 3 were converted to the phenoxyl radicals upon oxidation with Ce(IV), giving a phenoxyl radical π-π∗ transition band at 394-407 nm. ESR and resonance Raman spectra established that the radical species had a Zn(II)-phenoxyl radical bond. The cyclic voltammograms showed similar quasi-reversible redox waves with E_1/2 = 0.68, 0.67, and 0.63 V (versus Ag/AgCl) for 1, 2, and 3, respectively, corresponding to the formation of the phenoxyl radical, which displayed a first-order decay. The half-lives, 58.6, 25.8, and 15.6 min at −40 °C for 1, 2, and 3, respectively, follow the order of the stability constants of the complexes, indicating that the metal(II)-phenoxyl radical stability is in close relationship with the complexation properties of the present series of N₃O-donor ligands.     

Courtship behavior of male white perch, Morone americana: evidence for control by androgens.

Courtship behaviors are androgen-dependent in many vertebrates and castration often decreases courtship. We examined the effectiveness of castration in reducing courtship behaviors and 11-ketotestosterone (KT) and testosterone (T) in restoring them in male white perch. Castrates were given implants containing KT, T or no hormone. Sham-operated males received implants without hormone. Three weeks later, males were exposed to an ovulated female for 1 h and two courtship behaviors were quantified. Attending behavior involves close and continuous following of a female with occasional contact. Circling involves rapid transits around the female in a circular pattern or back and forth in front of her. In plasma samples taken immediately after observations, KT and T were below detectable levels in castrated males but at high physiological levels in males implanted with KT or T. Castrated males given KT attended females more than castrated males given T implants or implants containing no hormone, but not more than sham-operated males. Circling was eliminated by castration but restored by implantation with T or 11-KT to values exhibited by sham-operated males. This is one of the few demonstrations that KT can regulate courtship behavior in a non-territorial and economically important fish species.     

PathVisio-MIM: PathVisio plugin for creating and editing Molecular Interaction Maps (MIMs)

MOTIVATION: A plugin for the Java-based PathVisio pathway editor has been developed to help users draw diagrams of bioregulatory networks according to the Molecular Interaction Map (MIM) notation. Together with the core PathVisio application, this plugin presents a simple to use and cross-platform application for the construction of complex MIM diagrams with the ability to annotate diagram elements with comments, literature references and links to external databases. This tool extends the capabilities of the PathVisio pathway editor by providing both MIM-specific glyphs and support for a MIM-specific markup language file format for exchange with other MIM-compatible tools and diagram validation. AVAILABILITY: The PathVisio-MIM plugin is freely available and works with versions of PathVisio 2.0.11 and later on Windows, Mac OS X and Linux. Information about MIM notation and the MIMML format is available at http://discover.nci.nih.gov/mim. The plugin, along with diagram examples, instructions and Java source code, may be downloaded at http://discover.nci.nih.gov/mim/mim_pathvisio.html.     

Courtship behavior of male white perch, Morone americana: evidence for control by androgens

Courtship behaviors are androgen-dependent in many vertebrates and castration often decreases courtship. We examined the effectiveness of castration in reducing courtship behaviors and 11-ketotestosterone (KT) and testosterone (T) in restoring them in male white perch. Castrates were given implants containing KT, T or no hormone. Sham-operated males received implants without hormone. Three weeks later, males were exposed to an ovulated female for 1 h and two courtship behaviors were quantified. Attending behavior involves close and continuous following of a female with occasional contact. Circling involves rapid transits around the female in a circular pattern or back and forth in front of her. In plasma samples taken immediately after observations, KT and T were below detectable levels in castrated males but at high physiological levels in males implanted with KT or T. Castrated males given KT attended females more than castrated males given T implants or implants containing no hormone, but not more than sham-operated males. Circling was eliminated by castration but restored by implantation with T or 11-KT to values exhibited by sham-operated males. This is one of the few demonstrations that KT can regulate courtship behavior in a non-territorial and economically important fish species.     

Azobenzene-modified poly(l-glutamic acid) (AZOPLGA): its conformational and photodynamic properties

Azobenzene-modified poly(l-glutamic acid) (AZOPLGA) polymers with 22 and 35 mol % of azo chromophores in the side chains have been synthesized by condensing 4-methoxy-4'-aminoazobenzene and poly(l-glutamic acid). These polymers have been characterized by NMR, FT-IR, and UV-visible spectroscopic techniques. The conformational features of the polymer backbone chains in the films that were cast from the polymer solutions prepared in different solvents have been investigated by circular dichroism spectroscopy. Experimental data suggested that the thermal cis-trans relaxation and photoinduced birefringence, which are related to the azo chromophores in the side chains of polymer, are not affected by the conformations of polymer backbones. However, the modulations of the surface relief gratings, the result of photoinduced mass transport process, recorded on these polymers are sensitive to polymer main chain conformation, as well as the degree of functionalization.     

Dependence of mechanical behavior of the murine tail disc on regional material properties: a parametric finite element study

In vivo rodent tail models are becoming more widely used for exploring the role of mechanical loading on the initiation and progression of intervertebral disc degeneration. Historically, finite element models (FEMs) have been useful for predicting disc mechanics in humans. However, differences in geometry and tissue properties may limit the predictive utility of these models for rodent discs. Clearly, models that are specific for rodent tail discs and accurately simulate the disc's transient mechanical behavior would serve as important tools for clarifying disc mechanics in these animal models. An FEM was developed based on the structure, geometry, and scale of the mouse tail disc. Importantly, two sources of time-dependent mechanical behavior were incorporated: viscoelasticity of the matrix, and fluid permeation. In addition, a novel strain-dependent swelling pressure was implemented through the introduction of a dilatational stress in nuclear elements. The model was then validated against data from quasi-static tension-compression and compressive creep experiments performed previously using mouse tail discs. Finally, sensitivity analyses were performed in which material parameters of each disc subregion were individually varied. During disc compression, matrix consolidation was observed to occur preferentially at the periphery of the nucleus pulposus. Sensitivity analyses revealed that disc mechanics was greatly influenced by changes in nucleus pulposus material properties, but rather insensitive to variations in any of the endplate properties. Moreover, three key features of the model-nuclear swelling pressure, lamellar collagen viscoelasticity, and interstitial fluid permeation-were found to be critical for accurate simulation of disc mechanics. In particular, collagen viscoelasticity dominated the transient behavior of the disc during the initial 2200 s of creep loading, while fluid permeation governed disc deformation thereafter. The FEM developed in this study exhibited excellent agreement with transient creep behavior of intact mouse tail motion segments. Notably, the model was able to produce spatial variations in nucleus pulposus matrix consolidation that are consistent with previous observations in nuclear cell morphology made in mouse discs using confocal microscopy. Results of this study emphasize the need for including nucleus swelling pressure, collagen viscoelasticity, and fluid permeation when simulating transient changes in matrix and fluid stress/strain. Sensitivity analyses suggest that further characterization of nucleus pulposus material properties should be pursued, due to its significance in steady-state and transient disc mechanical response.     

Distribution of cortisol among its free and protein-bound fractions in rainbow trout (Oncorhynchus mykiss): evidence of control by sexual maturation.

1. Total cortisol concentrations did not differ in sexually matured female, male, and immature rainbow trout. 2. The per cent cortisol bound to a corticosteroid binding protein was greater in mature female fish (48.2%) than in mature male (16.0%) and immature fish (19.5%). 3. The mature female fish exhibited a lower percentage of free cortisol (21.8%) compared to mature males (44.8%) and immature fish (43.2%). 4. Parallel aspects of the teleostean cortisol binding-protein and the mammalian counterpart are compared and commented upon.     

Further evidence of joint time-place control of rats' behavior: results from an 'open hopper' test

Rats were trained on an interval time-place task. Food was intermittently available on each of four levers for 4 min in a 16-min session. After baseline training the rats received 'open hopper' sessions in which food was available on all levers for all of the 16-min sessions. Despite the absence of any contingencies for doing so, the rats continued to press the levers in the 'correct' sequence, for roughly the 'correct' amount of time. This confirms that the rat behavior was controlled, in part, by a representation of an elapsed interval of time. The rats responding was more variable in 'open hopper' sessions and error increased (in an exponential fashion) as the session proceeded. This finding suggests that the rats may have used shifts in the location of food availability to minimize the accumulation of error throughout baseline sessions.     

Oviposition behavior of Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) on its natural host

Phymastichus coffea LaSalle is an African endoparasitoid of adults of the coffee berry borer, Hypothenemus hampei (Ferrari) that was introduced into Mexico for biological control purposes. The objective of this study was to investigate the oviposition behavior of P. coffea on its host under laboratory conditions. The results indicate that P. coffea females exhibited 12 behavioral acts during and after oviposition, such as, walking, cleaning, flying, surface landing, resting, host landing, host antennation, touching the host with forelegs, host recognition, walking on the host, oviposition and cleaning after oviposition. The sequence of each behavior is described in an ethogram.     

Dihydrolipoamide transacetylase from Escherichia Coli: evidence for internal gene duplication.

The dihydrolipoamide transacetylase component of the pyruvate dehydrogenase complex from $\text{Escherichia}$ coli consists of identical subunits of 89.000 daltons. During the purification procedure it is partially cleaved into active fragments of 82.000 daltons, 37.000 daltons and 35.000 daltons. Fingerprint analysis shows at least 80% similarity of the fragments compared with the native component. This provides evidence for two large homologous domains within same polypeptide chain.     

黑角直缘跳甲的生物学及其防治

黑角直缘跳甲是五倍子夏寄主盐肤木和红肤杨的重要食叶害虫,一年一代,以卵及卵块在寄主枝干上越冬,次年4月当寄主幼芽萌发达3cm,叶片尚未完全展开时孵化。幼虫期4—5月,蛹期5—6月,成虫期6—10月,卵期9至次年4月。幼虫始见期随不同年份和不同海拨而异,物候期明显。数量消长与天敌和降雨密切相关。本文首次报道了该虫的生物学特性,并提出人工灭虫和保护天敌的防治措施。     

Placental estrogen synthetase (aromatase): evidence for phosphatase-dependent inactivation

The acute regulation of estrogen synthetase (aromatase), the cytochrome P450 enzyme system responsible for estrogen production, is not well explored. We report here that aromatase, but not NADPH-cytochrome c (P450) reductase, activity from human term placental microsomes decreased when incubated in phosphate-free buffer at 37 degrees C. Aromatase activity was stabilized by phosphate buffer or by the phosphatase inhibitors tartaric acid or EDTA, but not NaF, in phosphate-free buffer. Alkaline phosphatase also inhibited aromatase in phosphate-free buffer relative to phosphate buffer, but the inactivation appears to be due primarily to proteolytic solubilization of NADPH-cytochrome c reductase from the microsomes by proteases within the alkaline phosphatase preparation. Based on these data, we suggest that the cytochrome P450 component of aromatase may be regulated acutely by phosphorylation-dependent processes.