Synthesis, Biological Properties, and Structure-Activity Relationships of Quinoxaline Angiotensin II Receptor Antagonists

Quinoxaline heterocycle containing angiotensin II receptor antagonist analogs were prepared. All five analogs reported here display potent antagonistic activities and most interestingly, quinozaline bis-N-oxide 10 exhibits very potent activities both in binding and functional assays.     

Combined 13C NMR- and mass-spectrometry for non-invasive monitoring of metabolism

Summary A method for the long-term incubation of cell suspensions at measured dissolved gas concentrations during acquisition of NMR spectra is described: a mass spectrometer probe in the cell suspension provides continuous monitoring of gases. Its usefulness is shown for Trichomonas vaginalis where fermentation products from glucose in 72 M O₂ are different from those under strictly anaerobic conditions.     

Intronic gene conversion in the evolution of human X-linked color vision genes

Human red and green visual pigment genes are X-linked duplicate genes. To study their evolutionary history, introns 2 and 4 (1,987 and 1,552 bp, respectively) of human red and green pigment genes were sequenced. Surprisingly, we found that intron 4 sequences of these two genes are identical and that the intron 2 sequences differ by only 0.3%. The low divergences are unexpected because the duplication event producing the two genes is believed to have occurred before the separation of the human and Old World monkey (OWM) lineages. Indeed, the divergences in the two introns are significantly lower than both the synonymous divergence (3.2% +/- 1.1%) and the nonsynonymous divergence (2.0% +/- 0.5%) in the coding sequences (exons 1-6). A comparison of partial sequences of exons 4 and 5 of human and OWM red and green pigment genes supports the hypothesis that the gene duplication occurred before the human-OWM split. In conclusion, the high similarities in the two intron sequences might be due to very recent gene conversion, probably during evolution of the human lineage.      

Somatostatin and lanthanum discriminate two Ca2+ mobilizing mechanisms regulated by bombesin receptors in peptic cells

Bombesin (BB)-stimulated pepsinogen secretion from frog esophageal peptic acini was inhibited 40% by somatostatin (SS) (IC50 = 1 nM), and by 30-50% by low concentrations (10(-7)-10(-4)M) of lanthanum chloride. SS inhibited basal secretion as well as both early (0-2 min) and late (2-30 min) BB-stimulated secretory phases. By contrast, LaCl3 selectively inhibited the late secretory phase and was without effect on basal secretion. SS (100 nM) and LaCl3 (30 microM) attenuated BB-stimulated 45Ca2+ uptake, and the combination resulted in additive inhibition. High K+ media decreased basal secretion, abolished SS but not LaCl3 inhibition of BB-stimulated secretion, and blocked SS inhibition of BB-mediated 45Ca2+ uptake. These findings suggest the existence on peptic cells of distinct La3+-sensitive, and somatostatin-sensitive, K+ dependent, Ca2+ mobilizing mechanisms which contribute to BB receptor-mediated secretion.     

Partial purification and characterization of phytases from pollen of lily (Lilium longiflorum Thunb.)

Summary Two phytases from lily pollen (Lilium longiflorum Thunb.) were partially purified and characterized. The first (pH optimum 5.0) was purified 40-fold from ungerminated pollen. The second (pH optimum 6.5) appeared during germination and was purified 68-fold from pollen germinated 2 h. Molecular weight of the first was 72 kD, and the second was 36 kD as determined by gel filtration. Both were active against phosphate esters other than phytate, although purification of the first reduced its activity against AMP and myo-inositol 2-P to 10% of activity against phytate. Phytase from germinated pollen (but not ungerminated) was inhibited by the sulfhydryl agent parahydroxy mercuribenzoate; P _i inhibited phytase from ungerminated but not germinated pollen. Such different catalytic and physical properties may reflect different biochemical functions.Abbreviations HPLC High performance liquid chromatography - DEAE diethyl aminoethyl - P _i orthophosphate - PP _i pyrophosphate - p-NPP para-nitrophenyl phosphate - pNP para-nitrophenol - MI myo-inositol - MI 2-P myo-inositol 2-P - MI penta P myo-inositol pentakisphosphate - PHMB para-hydroxy mercuribenzoate - PMSF phenyl methyl sulfonyl fluoride - AMP adenosine monophosphate - GMP guanosine monophosphate - EGTA ethylene glycol-bis (-aminoethyl ether) N, N, N, N-tetraacetic acid     

Surface shear viscometry as a probe of protein-protein interactions in mixed milk protein films adsorbed at the oil-water interface

Time-dependent surface viscosities are reported for films adsorbed from binary mixtures of the proteins alpha-lactalbumin, beta-lactoglobulin and beta-casein. The measurements were made at a planar interface between n-tetradecane and various protein solutions (10(-3) wt% of each protein, pH 7, 25 degrees C) using a Couette-type torsion-wire surface viscometer operating at very low shear-rate. Differences in behaviour between simultaneous and sequential exposure of the pairs of proteins to the interface were investigated. Some experiments were performed with chemically modified beta-lactoglobulin samples whose disulphide bonds had been cleaved and blocked. Displacement of one protein by another (e.g. alpha-lactalbumin by beta-casein) is indicated by a sudden drop in surface viscosity immediately after addition of the second protein. In systems containing beta-lactoglobulin, the long-time surface viscosity is very sensitive to the adsorption time of beta-lactoglobulin prior to addition of the second protein. Blocking the disulphide bonds in beta-lactoglobulin leads to a much faster approach to a steady-state surface viscosity. This is interpreted in terms of a much more rapid unfolding of the disordered molecules of modified beta-lactoglobulin at the oil-water interface. We conclude that surface viscosity experiments give useful and sensitive information about competitive adsorption and cooperative interactions in mixed protein films.     

Chromosomal localization of seven members of the murine TGF-beta superfamily suggests close linkage to several morphogenetic mutant loci

Chromosomal locations have been assigned to seven members of the TGF-beta superfamily using an interspecific mouse backcross. Probes for the Tgfb-1, -2, and -3, Bmp-2a and -3, and Vgr-1 genes recognized only single loci, whereas the Bmp-2b probe recognized two independently segregating loci (designated Bmp-2b1 and Bmp-2b2). The results show that the seven members of the TGF-beta superfamily map to eight different chromosomes, indicating that the TGF-beta family has become widely dispersed during evolution. Five of the eight loci (Tgfb-1, Bmp-2a, Bmp-2b1, Bmp-2b2, Vgr-1) mapped near mutant loci associated with connective tissue and skeletal disorders, raising the possibility that at least some of these mutations result from defects in TGF-beta-related genes.     

A comparison of methods to partition selection acting via components of fitness: Do larger male bullfrogs have greater hatching success?

We contrast three methods for measuring selection at sequential fitness components (here called the additive, changing variance, and independent methods). The independent method (Koenig and Albano, 1987; Conner, 1988) describes the relationship between a phenotypic character and one fitness component independent of other components. This method is appropriate when the question is whether or not a character has fitness consequences independent of selection at other stages. The additive (Arnold and Wade, 1984a) and changing variance (Kalisz, 1986; Koenig and Albano, 1987) methods measure selection via one component of fitness, taking into consideration constraints imposed by selection via earlier components in the sequence. These methods therefore more accurately track selection over a sequence of fitness components. Of these latter two methods, the changing variance method yields erratic results in simulation studies and is not recommended in its unmodified form. The additive method (equivalent to the changing variance method weighted as described in Wade and Kalisz [1989]) explicitly partitions selection into additive components and is useful for measuring selection taking into account the constraints imposed by selection acting via prior fitness components. The methods often yield very different estimates of the relative degree to which the mean of a character is changed by selection acting via a particular component of fitness (the “strength” of selection). However, neither the additive nor independent method is inherently superior to the other; rather, these measures are complementary.     

Major muscle systems in the larval caenogastropod,Ilyanassa obsoleta,display different patterns of development

This study describes the anatomical and developmental aspects of muscular development from the early embryo to competent larval stage in the gastropod Ilyanassa obsoleta. Staining of F‐actin revealed differential spatial and temporal patterns of several muscles. In particular, two major muscles, the larval retractor and pedal retractor muscles originate independently and display distinct developmental patterns similar to observations in other gastropod species. Additionally, together with the larval retractor muscle, the accessory larval muscle developed in the embryo at the trochophore stage. Therefore, both these muscles develop prior to ontogenetic torsion. The pedal retractor muscle marked the most abundant growth in the mid veliger stage. Also during the middle stage, the metapodial retractor muscle and opercular retractor muscle grew concurrently with development of the foot. We show evidence that juvenile muscles, such as the buccal mass muscle and siphon muscle develop initially during the late veliger stage. Collectively, these findings substantiate that larval myogenesis involves a complex sequence of events that appear evolutionary conserved within the gastropods, and set the stage for future studies using this model species to address issues concerning the evolution and eventual fates of larval musculature in molluscs. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

Protein evolution of ANTP and PRD homeobox genes

Background  

Although homeobox genes have been the subject of many studies, little is known about the main amino acid changes that occurred early in the evolution of genes belonging to different classes.