Biological properties of two models of calcitonin gene related peptide with idealized amphiphilic alpha-helices of different lengths

Previous studies on calcitonin gene related peptide (CGRP) have demonstrated that it has the characteristics of an amphiphilic peptide, and from an examination of the sequence, we have proposed that it contains an amphiphilic alpha-helix. We have synthesized two analogues of CGRP which have different lengths of idealized amphiphilic alpha-helical secondary structure. The first model, CGRM-1, has been substituted with residues generating an idealized amphiphilic alpha-helix in the region between residues 8 and 25, equivalent to approximately five turns of an alpha-helix. This peptide is not an agonist in any of our bioassays, but it does bind with low affinity to rCGRP receptors in crude liver membranes. Our second model, CGRM-2, has an idealized amphiphilic alpha-helix between residues 8 and 18, which is equivalent to approximately three turns of an alpha-helix. In an in vitro rat vas deferens assay, this peptide is an agonist with a potency one-fourth that of the native hormone. However, the potency of CGRM-2 in an adenylate cyclase assay is much lower, only 1/140th the potency of CGRP. Both model peptides display amphiphilic characteristics commensurate with their design. We conclude that there is an amphiphilic alpha-helix in rCGRP between residues 8 and 18 and that this helix terminates in the vicinity of residue 18.     

Molecular structure of the rod domain of dictyostelium filamin

Dictyostelium discoideum filamin (ddFLN) is a two-chain F-actin crosslinking protein with an N-terminal actin-binding domain and a rod domain constructed from six tandem repeats of a 100 residue motif that has an immunoglobulin (Ig) fold. We report the 2.8 A resolution crystal structure of a homodimer of rod repeats 4, 5 and 6. The two chains are arranged in an antiparallel fashion and form an elongated element, which is shortened, however, compared to a fully extended, linear configuration because the long axis of each Ig domain is arranged at an angle to the long axis of the rod. Same arrangement of repeats should also be present in the rod domain of human FLNa, much longer than Dictyostelium FLN, which forms an extended structure able to crosslink F-actin chains over distances of more than 1000 A.     

Effect of environmental acidity and alkalinity on the physiology of Tilapia mossambica during acclimation

The pattern of carbohydrate metabolism in the muscle of the freshwater fish, Tilapia mossambica (Peters) varied according to the pH of the environmental medium. On acclimation to a more acidic water, the white muscle showed an elevated glycogen content with suppressed glycolysis. In contrast, the muscle tissue showed an accelerated glycolytic pathway with the accumulation of metabotic acids on acclimation to a basic water. In an acidic medium phosphorylase activity was inhibited with an elevated LDH activity, while the reverse pattern was observed in an alkaline medium. The oxidative pathway was elevated in both. Acclimation to acidic and basic environmental waters leads to an adaptive compensatory mechanisms providing increased resistance capacity to the fish under pH stress.     

Discovery of potent and selective thienopyrimidine inhibitors of Aurora kinases

In an effort to discover Aurora kinase inhibitors, an HTS hit revealed an amide containing pyrrolopyrimidine compound. Replacement of the pyrrolopyrimidine residue with a thienopyrimidine moiety led to a series of potent and selective Aurora inhibitors.     

Effect of Orientation of Fibers and Holes on the Radial Strain Amplification of Campaniform sensilla

##正## In this paper,a structural analysis is performed to gain insights on the synergistic mechanical amplification effect thatCampaniform sensilla have when combined in an array configuration.In order to simplify the analysis performed in this preliminaryinvestigation,an array of four holes in a single orthotropic lamina is considered.Firstly,a Finite Element Method(FEM) analysis is performed to discretely assess the influence that different geometrical parameters have on the mechanicalamplification properties of the array.Secondly,an artificial neural network is used to obtain an approximated multi-dimensionalcontinuous function,which models the relationship between the geometrical parameters and the amplification properties of thearray.Thirdly,an optimization is performed to identify the geometrical parameters yielding the maximum mechanical amplification.Finally,results are validated with an additional FEM simulation performed by varying geometrical parameters in theneighborhood of the identified optimal parameters.The method proposed in this paper can be fully automated and used to solvea wide range of optimization problems aimed at identifying optimal configurations of strain sensors inspired by Campaniformsensilla.     

Conjugation of cell-penetrating peptides leads to identification of anti-HIV peptides from matrix proteins

Compounds which inhibit the HIV-1 replication cycle have been found amongst fragment peptides derived from an HIV-1 matrix (MA) protein. Overlapping peptide libraries covering the whole sequence of MA were designed and constructed with the addition of an octa-arginyl group to increase their cell membrane permeability. Imaging experiments with fluorescent-labeled peptides demonstrated these peptides with an octa-arginyl group can penetrate cell membranes. The fusion of an octa-arginyl group was proven to be an efficient way to find active peptides in cells such as HIV-inhibitory peptides.     

Simulation of human sleep: ultradian dynamics of electroencephalographic slow-wave activity

The typical declining trend of electroencephalographic (EEG) slow-wave activity (SWA) within a sleep period is represented in the two-process model of sleep regulation by an exponentially decaying process (Process S). The model has been further elaborated to simulate not only the global changes of SWA, but also the dynamics within non-rapid-eye-movement (non-REM) sleep episodes. In this new model, the initial intraepisodic buildup of SWA is determined by the combined action of an exponentially increasing process and a saturation process, whereas its fall at the end of an episode is due to an exponentially decreasing process. The global declining trend of SWA over consecutive episodes results from the monotonic decay of the intraepisodic saturation level. In contrast to Process S in the two-process model, this decay is not represented by an exponential function, but is proportional to the momentary level of SWA. REM sleep episodes are triggered by an external function. The model allows one to simulate the ultradian pattern of SWA for baseline nights as well as changes induced by a prolonged waking period, a daytime nap, a partial slow-wave sleep deprivation, or an antidepressant drug.     

Study of bone remodeling of two models of femoral cementless stems by means of DEXA and finite elements

Background  

A hip replacement with a cemented or cementless femoral stem produces an effect on the bone called adaptive remodelling, attributable to mechanical and biological factors. All of the cementless prostheses designs try to achieve an optimal load transfer in order to avoid stress-shielding, which produces an osteopenia.     

Accumulation of adenine and thymine in agroE-homologous operon of an intracellular symbiont

Summary As a result of the nucleotide sequence analysis of an aphid endosymbiont's operon homologous to theEscherichia coli groE, we noted that directional base substitutions tending toward an increase of A + T content represent an obvious evolutionary trend in this prokaryotic operon, housed for a long period by an eukaryotic cell. This result, when taken together with previous reports, raised the possibility that genomic DNA of prokaryotes residing in an eukaryotic cell is subject to A/T-biased directional mutation pressure and/or both negative and positive selection operating under conditions specific to the intracellular environments.Offprint requests to: H. Ishikawa     

Extension of the range of DNA sequences available for triple helix formation: stabilization of mismatched triplexes by acridine-containing oligonucleotides.

Triple helix formation usually requires an oligopyrimidine*oligopurine sequence in the target DNA. A triple helix is destabilized when the oligopyrimidine*oligopurine target contains one (or two) purine*pyrimidine base pair inversion(s). Such an imperfect target sequence can be recognized by a third strand oligonucleotide containing an internally incorporated acridine intercalator facing the inverted purine*pyrimidine base pair(s). The loss of triplex stability due to the mismatch is partially overcome. The stability of triplexes formed at perfect and imperfect target sequences was investigated by UV thermal denaturation experiments. The stabilization provided by an internally incorporated acridine third strand oligonucleotide depends on the sequences flanking the inverted base pair. For triplexes containing a single mismatch the highest stabilization is observed for an acridine or a propanediol tethered to an acridine on its 3'-side facing an inverted A*T base pair and for a cytosine with an acridine incorporated to its 3'-side or a guanine with an acridine at its 5'-side facing an inverted G*C base pair. Fluorescence studies provided evidence that the acridine was intercalated into the triplex. The target sequences containing a double base pair inversion which form very unstable triplexes can still be recognized by oligonucleotides provided they contain an appropriately incorporated acridine facing the double mismatch sites. Selectivity for an A*T base pair inversion was observed with an oligonucleotide containing an acridine incorporated at the mismatched site when this site is flanked by two T*A*T base triplets. These results show that the range of DNA base sequences available for triplex formation can be extended by using oligonucleotide intercalator conjugates.     

Aerosol transmission of a viable virus affecting swine: Explanation of an epizootic of pseudorabies

A Gaussian diffusion model was applied to an epizootic of pseudorabies in ten swine herds located in Decatur County, Indiana, USA to test the hypothesis that the virus can be spread via aerosol. The epizootic occurred during January to March, 1988, spreading through ten farms across an area of about 150 km². The model included a receptor component that provided an estimate of viruses received by the pig within an enclosed barn. Results show that the diffusion model can explain the spread of the virus during the epizootic for all nine farms to which the virus spread.     

Mathematical model of aneurysm of the circle of Willis: II. a qualitative analysis of the equation of Austin

Periodic solutions of the equation derived by Austin [1,1e] to describe an aneurysm of the circle of Willis are studied by qualitative methods which are extensions of the method of Poincaré. The results obtained reinforce strongly Austin's conclusion concerning the danger of increased pulse pressure; they suggest the possibility of the danger of an increased pulse rate (this also agrees with Austin's conclusions) or a decreased pulse rate. The conclusions provide a theoretical justification for the treatment of intracranial aneurysms by hypotensive methods [14] and also an explanation of why there does not seem to be an immediate precipitating cause of rupture (i.e., rupture seems to be just as likely to occur during relaxation as during physical activity).     

Synthesis and evaluation of phosphoramidate amino acid-based inhibitors of sialyltransferases

Several phosphoramidate analogues of CMP-N-acetylneuraminic acid were prepared for evaluation as inhibitors of alpha-2,3- and alpha-2,6-sialyltransferase. Central to the synthesis was the oxidative coupling of an amino acid ester with an H-phosphonate to construct the phosphoramidate linkage. All compounds synthesized were weak inhibitors of both of the sialyltransferases as determined by an HPLC-based inhibition assay.     

Assessment of meaning and the detection of unreliable signals by vervet monkeys

Free-ranging vervet monkeys, Cercopithecus aethiops, who had learned to ignore playbacks of one type of call by an unreliable signaller subsequently also ignored playback of an acoustically different call by the same individual if the calls had similar referents. Such transfer did not occur if either the identity of the signaller changed or if the two calls had different referents. After repeatedly being played an intergroup call in the absence of other groups, vervets also ignored an acoustically different intergroup call given by the same individual. The monkeys did not transfer habituation, however, if the new call was given by a different individual. In tests where the referents of two calls were different, the monkeys also failed to transfer habituation across call types. Vervet monkeys who had learned to ignore an unreliable leopard alarm call did not later ignore an eagle alarm call, even when the signaller remained the same. Results suggest that vervets, like humans, process information at a semantic, and not just an acoustic, level.     

Stabilization of a protein by removal of unfavorable abnormal pKa: substitution of undissociable residue for glutamic acid-35 in chicken lysozyme.

Glu35 in chicken lysozyme has an abnormally high pKa (6.1) partly due to the hydrophobic environment provided by Trp108. The relationship between protein stability and abnormal pKa was investigated in detail by using mutant lysozymes in which Glu35 was replaced by undissociable residues and an oppositely ionizable residue. It was found that lysozyme was stabilized at alkaline pH range by the replacement of Glu35 with an undissociable residue, Gln (E35Q lysozyme) or Al (E35A lysozyme). On the other hand, when Glu35 was replaced by His (E35H lysozyme), which could have an opposite charge to Glu by ionization, the introduced His35 was found to have an abnormally low pKa (3.6), leading to the destabilization of lysozyme at acidic pH. These observations are completely consistent with the situation that the environment around Glu35 is highly hydrophobic and therefore the placement of either a positive or negative charge in such an environment leads to destabilization of lysozyme. These observations also indicate that the replacement of an acidic residue having abnormally high pKa or a basic residue having abnormally low pKa by an undissociable residue is a very efficient and general method for stabilization of a protein.     

Relative stability of the central mechanisms that determine the depth and frequency of respiration

Not only an intensified respiration, but also an increase in inspiratory discharges can be caused by hypercapnia in vagotomized cats. With the deepening of anesthesia an increase of the inspiratory discharges persists, whereas an increased frequency of respiration disappears. This testifies to the presence of two different central mechanisms for the regulation of the rate and depth of respiration; the first is subject to the suprabulbar influences more than the second.     

The relationship of post-stimulus time and interval histograms to the timming characteristics of spike trains.

PST (post-stimulus time) and interval histograms computed from recorded spike trains are related to an average timing characteristics of the spike train. The exact nature of this relationship varies with recording parameters, interfering signals, the histogram bin width, and the duration of the measurement interval. This work describes the conditions under which a PST histogram can serve as an unbiased estimate of the ensemble average of a spike train's intensity and an interval histogram can serve as an unbiased estimate of the probability density function of the interspike intervals. Simulation studies are used to confirm the validity of the theoretical results. As an example of an application, these results are used to analyze recordings of singleunit activity in the eight cranial nerve.     

Mode of action of tetrahydrolipstatin: a derivative of the naturally occurring lipase inhibitor lipstatin

Tetrahydrolipstatin is a specific lipase inhibitor derived from lipstatin, a lipid produced by Streptomyces toxytricini. In addition to pancreatic lipase, it is shown in the present study that tetrahydrolipstatin also inhibits human gastric lipase, carboxyl ester lipase (cholesterol esterase) of pancreatic origin and the closely related bile-salt-stimulated lipase of human milk. It does not inhibit the exocellular lipase from Rhizopus arrhizus or a lipase recently isolated from Staphylococcus aureus. In the presence of a water-insoluble substrate, such as tributyrin, the inhibition has the characteristics of an irreversible inactivation of the uncompetitive type, thus indicating that an enzyme.substrate.inhibitor complex is formed, which cannot undergo further reaction to yield the normal product. This reaction probably takes place at the aqueous/oil interface of the substrate. In aqueous solution, in the absence of substrate, the inhibition of carboxyl ester lipase by tetrahydrolipstatin has the characteristics of being reversible, and finally becomes of a temporary nature analogues to the trypsin-trypsin inhibitor system. It is suggested that an enzyme-inhibitor complex of an acyl-enzyme type is formed that is slowly hydrolysed, with water as the final acceptor, leaving an intact enzyme and an inactive form of the inhibitor. The enzyme thus consumes the inhibitor, which undergoes a chemical conversion, as indicated by a change in mobility in an appropriate thin-layer chromatographic system, indicating an increase in hydrophilicity. Evidence is presented that the reaction product is an acid and that the functional group of tetrahydrolipstatin is the beta-lactone reacting with the active site of the enzyme.