Microelectrodes for studying neurobiology

Microelectrodes have emerged as an important tool used by scientists to study biological changes in the brain and in single cells. This review briefly summarizes the ways in which microelectrodes as chemical sensors have furthered the field of neurobiology by reporting on changes that occur on the subsecond time scale. Microelectrodes have been used in a variety of fields including their use by electrophysiologists to characterize neuronal action potentials and develop neural prosthetics. Here we restrict our review to microelectrodes that have been used as chemical sensors. They have played a major role in many important neurobiological findings.     

Toward more effective gene delivery

A report on the symposium 'In vivo barriers to gene delivery', Cold Spring Harbor, USA, 26-29 November 2007.     

Conformational changes and molecular mobility in plasticized proteins

Most biopolymers exist in a plasticized state, whether it is naturally with water or unnaturally with glycerol or other suitable polyol, to make a flexible material. We have found that the extent to which a biopolymer can be plasticized is dependent on its molecular and higher order structures outside of simply molecular weight. Lactalbumin, ovalbumin, corn zein, wheat gluten, and feather keratin were plasticized with glycerol from very low to very high amounts. The conformation of the proteins was monitored with Fourier transform-infrared (FT-IR) spectroscopy and X-ray powder diffraction (XRD) and correlated with the tensile modulus. Protein conformational changes were pronounced for polar proteins with a low amount of cysteine. FT-IR showed that the conformational changes resulted in ordering of the protein at low to moderate plasticization levels. For proteins with little resistance to conformational changes, additional small-scale ordering occurred around the glass transition, as observed in XRD. Accurate comparison of plasticized proteins was dependent on knowing whether or not the protein was glassy or rubbery at room temperature as no differences arose in the glassy state. The transition from glassy to rubbery behavior with plasticization level can be found from modulus, FT-IR, and XRD data.     

Development of maximum metabolic rate and pulmonary diffusing capacity in the superprecocial Australian Brush Turkey Alectura lathami: an allometric and morphometric study.

The Australian Brush Turkey Alectura lathami is a member of the Megapodiidae, the mound-building birds that produce totally independent, "superprecocial" hatchlings. This study examined the post-hatching development of resting and maximal metabolic rates, and the morphometrically determined changes in pulmonary gas exchange anatomy, in chicks during 3.7 months of growth from hatchlings (122 g) to subadults (1.1 kg). Allometric equations of the form y=aM(b) related gas exchange variables (y) to body mass (M, g). Metabolic rates were measured with open-flow respirometry (mL O2 min(-1)) of chicks resting in the dark and running above the aerobic limit on a treadmill. Resting metabolic rate (RMR=0.02 M(0.99)) and maximal metabolic rate (MMR=0.05 M(1.07)) scaled with exponents significantly above those of interspecific allometries of adult birds. However MMR was below that expected for other species of adult birds in flapping flight, consistent with the Brush Turkey's ground-dwelling habits. Total lung volumes (mL) increased faster than isometrically (V(L)=0.0075 M(1.19)), as did the surface area (cm(2)) of the blood-gas barrier (S(t)=7.80 M(1.23)), but the data overlapped those of adult species. Harmonic mean thickness of the blood-gas barrier was independent of body size (mean tau(ht),=0.39 microm) and was about twice that expected for flying birds. Diffusing capacity (mL O2 min(-1) kPa(-1)) of the blood-gas tissue barrier increased faster than isometrically (Dto2=0.049 M(1.23)); in hatchling Brush Turkeys, it was about 30% expected for adult birds, but this difference disappeared when they became subadults. When compared to altricial Australian pelicans that hatch at similar body masses, superprecocial Brush Turkeys had higher MMR and higher Dto2 at the same body size. A parallel allometry between MMR and Dto2 in Brush Turkeys and pelicans is consistent with the concept of symmorphosis during development.     

Metal swap between Zn7-metallothionein-3 and amyloid-beta-Cu protects against amyloid-beta toxicity

Aberrant interactions of copper and zinc ions with the amyloid-beta peptide (Abeta) potentiate Alzheimer's disease (AD) by participating in the aggregation process of Abeta and in the generation of reactive oxygen species (ROS). The ROS production and the neurotoxicity of Abeta are associated with copper binding. Metallothionein-3 (Zn(7)MT-3), an intra- and extracellularly occurring metalloprotein, is highly expressed in the brain and downregulated in AD. This protein protects, by an unknown mechanism, cultured neurons from the toxicity of Abeta. Here, we show that a metal swap between Zn(7)MT-3 and soluble and aggregated Abeta(1-40)-Cu(II) abolishes the ROS production and the related cellular toxicity. In this process, copper is reduced by the protein thiolates forming Cu(I)(4)Zn(4)MT-3, in which an air-stable Cu(I)(4)-thiolate cluster and two disulfide bonds are present. The discovered protective effect of Zn(7)MT-3 from the copper-mediated Abeta(1-40) toxicity may lead to new therapeutic strategies for treating AD.     

A catalog of neutral and deleterious polymorphism in yeast

The abundance and identity of functional variation segregating in natural populations is paramount to dissecting the molecular basis of quantitative traits as well as human genetic diseases. Genome sequencing of multiple organisms of the same species provides an efficient means of cataloging rearrangements, insertion, or deletion polymorphisms (InDels) and single-nucleotide polymorphisms (SNPs). While inbreeding depression and heterosis imply that a substantial amount of polymorphism is deleterious, distinguishing deleterious from neutral polymorphism remains a significant challenge. To identify deleterious and neutral DNA sequence variation within Saccharomyces cerevisiae, we sequenced the genome of a vineyard and oak tree strain and compared them to a reference genome. Among these three strains, 6% of the genome is variable, mostly attributable to variation in genome content that results from large InDels. Out of the 88,000 polymorphisms identified, 93% are SNPs and a small but significant fraction can be attributed to recent interspecific introgression and ectopic gene conversion. In comparison to the reference genome, there is substantial evidence for functional variation in gene content and structure that results from large InDels, frame-shifts, and polymorphic start and stop codons. Comparison of polymorphism to divergence reveals scant evidence for positive selection but an abundance of evidence for deleterious SNPs. We estimate that 12% of coding and 7% of noncoding SNPs are deleterious. Based on divergence among 11 yeast species, we identified 1,666 nonsynonymous SNPs that disrupt conserved amino acids and 1,863 noncoding SNPs that disrupt conserved noncoding motifs. The deleterious coding SNPs include those known to affect quantitative traits, and a subset of the deleterious noncoding SNPs occurs in the promoters of genes that show allele-specific expression, implying that some cis-regulatory SNPs are deleterious. Our results show that the genome sequences of both closely and distantly related species provide a means of identifying deleterious polymorphisms that disrupt functionally conserved coding and noncoding sequences.     

Anti-HIV-1 entry optimization of novel imidazopiperidine-tropane CCR5 antagonists

A novel series of imidazopiperidine-tropane CCR5 antagonists is described. The series was optimized for anti-HIV-1 potency using a set of phenotypic viral entry assays. This strategy resulted in the identification of several very potent (IC(50)<10nM) inhibitors of HIV-1 entry. One compound (40) was further profiled and was found to have attractive selectivity, pharmacokinetic, and antiviral properties.     

Novel 2-imidazoles as potent, selective and CNS penetrant alpha1A adrenoceptor partial agonists

A novel series of central nervous system (CNS) penetrant indane 2-imidazoles have been identified as potent, partial agonists of the alpha(1A) adrenergic receptor, having good selectivity over the alpha(1B), alpha(1D) and alpha(2) sub-types. A key structural motif to impart selectivity is a methylene spacer between the indane and a pendant substituent, which includes heterocycles, sulphones and ethers. Introduction of an ortho-halogen to this group led to a lowering of intrinsic efficacy (E(max)).     

Release characteristics of reattached barnacles to non-toxic silicone coatings

Release mechanisms of barnacles (Amphibalanus amphitrite or Balanus amphitrite) reattached to platinum-cured silicone coatings were studied as a function of coating thickness (210-770 microm), elastic modulus (0.08-1.3 MPa), and shear rate (2-22 microm s(-1)). It was found that the shear stress of the reattached, live barnacles necessary to remove from the silicone coatings was controlled by the combined term (E/t)(0.5) of the elastic modulus (E) and thickness (t). As the ratio of the elastic modulus to coating thickness decreased, the barnacles were more readily removed from the silicone coatings, showing a similar release behavior to pseudobarnacles (epoxy glue). The barnacle mean shear stress ranged from 0.017 to 0.055 MPa whereas the pseudobarnacle mean shear stress ranged from 0.022 to 0.095 MPa.