Development of photochemical method for meniscal repair: a preliminary study

Visible light combined with naphthalimides has previously been shown to catalyze formation of physical bonds in avascular meniscal tissue. The first objective was to modify the existing in vitro testing method (i.e., adhesion testing using lap-jointed slices) to gain more sensitivity in detecting relative bonding strengths among candidate bonding agents. A repeated measures experimental design (RMED) was used to account for variability in properties among bovine menisci and was achieved by testing all treatments/controls on slices from each meniscus. Additionally, to make the method more clinically relevant in modeling a bucket-handle tear, the bovine meniscal slices were cut with collagen fibers parallel to the test slice's length. Peak stress was greater for the complete treatment group (light plus naphthalimide) than for the control or incomplete treatment groups (light only or napthalimide only). The second objective was to perform concentration and photoactivation time dose-response studies. In the concentration dose-response study, peak stress was greater for all treatments when compared with the control but not different among treatment groups; however, there was a trend of increased bonding strength with increased concentration. In the photoactivation time dose-response study, peak stress was greater for all treatments when compared with the control and greater for the 3-min treatment vs. the 6- and 9-min treatments. Peak stress was not different between the longer treatments. The RMED provided increased reproducibility and statistical sensitivity for detecting differences among treatments and will be used to test candidate bonding agents prior to in vivo testing.     

Song is a reliable signal of bill morphology in Darwin's small tree finch Camarhynchus parvulus, and vocal performance predicts male pairing success

The shape and movement of the vocal tract are known to influence bird song. Current theory predicts that large bill and body size are correlated with low frequency song and slow trill rate. It is also widely accepted that song characteristics are important for mate choice by females. We investigated the relationship between bill morphology, song characteristics, and pairing success in Darwin's small tree finch Camarhynchus parvulus , on the Galapagos Islands. Contrary to predictions from a previous cross-species study on Darwin's finches, we found that individuals with larger bill size produced songs with slow trill rate, high dominant frequency, and broad frequency bandwidth, indicating that song is a reliable signal of bill morphology. Vocal performance as indicated by the deviation from an upper performance limit was higher in paired than unpaired males. Pairing was not skewed in favour of a particular bill size, and both small and large billed males that sang high performance song had high pairing success. The reliable signalling function of song has implications for female choice and territorial defence, given that both females and conspecific competitors can assess the relative size of males' bills through song, while females may use vocal performance as a signal of male quality.     

Vive la variance: a functional oviposition theory for insect herbivores

We developed state-dependent life-history theory to explain the variance in clutch size decisions made by insect herbivores under a variety of ecological scenarios. An important aspect of our theory is explicit representation of the distribution of host quality and frequency of occurrence. Examination of the theory suggests that clutch size decisions can be highly non-linear with respect to host quality and variability. We then use our theory to explore the potential for bet-hedging strategies to evolve as a function of unpredictable catastrophic events that decimate entire clutches. Our analysis suggests that the benefits to employing such a strategy will frequently be outweighed by costs brought on by delayed oviposition.     

Individual Dopaminergic Neurons of Lamprey SNc/VTA Project to Both the Striatum and Optic Tectum but Restrict Co-release of Glutamate to Striatum Only

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Sensitive Detection of Gene Expression in Mycobacteria under Replicating and Non-Replicating Conditions Using Optimized Far-Red Reporters

Fluorescent reporter proteins have proven useful for imaging techniques in many organisms. We constructed optimized expression systems for several fluorescent proteins from the far-red region of the spectrum and analyzed their utility in several mycobacterial species. Plasmids expressing variants of the Discosoma Red fluorescent protein (DsRed) from the Mycobacterium bovis hsp60 promoter were unstable; in contrast expression from the Mycobacterium smegmatis rpsA promoter was stable. In Mycobacterium tuberculosis expression of several of the far-red reporters was readily visualised by eye and three reporters (mCherry, tdTomato, and Turbo-635) fluoresced at a high intensity. Strains expressing mCherry showed no fitness defects in vitro or in macrophages. Treatment of cells with antibiotics demonstrated that mCherry could also be used as a reporter for cell death, since fluorescence decreased in the presence of a bactericidal compound, but remained stable in the presence of a bacteriostatic compound. mCherry was functional under hypoxic conditions; using mCherry we demonstrated that the P_mtbB is expressed early in hypoxia and progressively down-regulated. mCherry and other far-red fluorescent proteins will have multiple uses in investigating the biology of mycobacteria, particularly under non-replicating, or low cell density conditions, as well as providing a novel means of detecting cell death rapidly.     

The Balance of Apoptotic and Necrotic Cell Death in Mycobacterium tuberculosis Infected Macrophages Is Not Dependent on Bacterial Virulence

Background

An important mechanism of Mycobacterium tuberculosis pathogenesis is the ability to control cell death pathways in infected macrophages: apoptotic cell death is bactericidal, whereas necrotic cell death may facilitate bacterial dissemination and transmission.

Methods

We examine M.tuberculosis control of spontaneous and chemically induced macrophage cell death using automated confocal fluorescence microscopy, image analysis, flow cytometry, plate-reader based vitality assays, and M.tuberculosis strains including H37Rv, and isogenic virulent and avirulent strains of the Beijing lineage isolate GC1237.

Results

We show that bacterial virulence influences the dynamics of caspase activation and the total level of cytotoxicity. We show that the powerful ability of M.tuberculosis to inhibit exogenously stimulated apoptosis is abrogated by loss of virulence. However, loss of virulence did not influence the balance of macrophage apoptosis and necrosis – both virulent and avirulent isogenic strains of GC1237 induced predominantly necrotic cell death compared to H37Rv which induced a higher relative level of apoptosis.

Conclusions

This reveals that macrophage necrosis and apoptosis are independently regulated during M. tuberculosis infection of macrophages. Virulence affects the level of host cell death and ability to inhibit apoptosis but other strain-specific characteristics influence the ultimate mode of host cell death and alter the balance of apoptosis and necrosis.     

Natural variation in the sexually selected feather ornaments of crested auklets (Aethia cristatella) does not predict future survival

We evaluated whether sexually selected crest and auricular plumefeather ornaments of crested auklet (Aethia cristatella) adultscovaried with individual local survival over 11 years (1991–2001).Crested auklets (n = 364 total) were captured near breedingsites, marked with color rings, and local survival estimateswere based on color ring resightings at a breeding colony. Survivalestimates and relationships among local survival and crest length,auricular plume length, mass and tarsus were evaluated usingthe program MARK. The best models included four groups, definedby sex and ease of resighting, that differed in resighting rate(p) but not local survival rate (). This model structure effectivelyexplained sources of variation in local survival and resightabilityamong individuals. The best fitting model showed local survivalrate varying annually, while accounting for differences in resightabilityof marked individuals between the sexes and groups ([t], p[sex*easeof resighting]). Annual local survival varied from 0.940 ±0.029 (SE) in 1993–1994 to 0.767 ± 0.034 in 1997–1998and averaged 0.859 ± 0.019. We found no evidence thatcrested auklet local survival covaried with continuous variationin individuals' ornaments. Simulations indicated that our dataset was sufficient to detect a relationship between local survivaland a covariate that equaled or exceeded a range of 8%. Theimplications for competing sexual selection mechanisms of empiricallymeasured survival–ornament relationships are controversial,but our study emphasizes that survival estimates for such investigationsmust control for confounding factors such as resighting rateas well as have sufficient statistical power and time scaleto be biologically meaningful. Our results are most consistentwith the idea that the conspicuous variation in crested auklet'sshowy ornaments is arbitrary with respect to individual viabilityas quantified by their long-term survival.     

Characterization of moorland vegetation and the prediction of bird abundance using remote sensing

Aims To characterize and identify upland vegetation composition and height from a satellite image, and assess whether the resulting vegetation maps are accurate enough for predictions of bird abundance. Location South‐east Scotland, UK. Methods Fine‐taxa vegetation data collected using point samples were used for a supervised classification of a Landsat 7 image, while linear regression was used to model vegetation height over the same image. Generalized linear models describing bird abundance were developed using field‐collected bird and vegetation data. The satellite‐derived vegetation data were substituted into these models and efficacy was examined. Results The accuracy of the classification was tested over both the training and a set of test plots, and showed that more common vegetation types could be predicted accurately. Attempts to estimate the heights of both dwarf shrub and graminoid vegetation from satellite data produced significant, but weak, correlations between observed and predicted height. When these outputs were used in bird abundance–habitat models, bird abundance predicted using satellite‐derived vegetation data was very similar to that obtained when the field‐collected data were used for one bird species, but poor estimates of vegetation height produced from the satellite data resulted in a poor abundance prediction for another. Conclusions This pilot study suggests that it is possible to identify moorland vegetation to a fine‐taxa level using point samples, and that it may be possible to derive information on vegetation height, although more appropriate field‐collected data are needed to examine this further. While remote sensing may have limitations compared with relatively fine‐scale fieldwork, when used at relatively large scales and in conjunction with robust bird abundance–habitat association models, it may facilitate the mapping of moorland bird abundance across large areas.     

Parallel tandem: a program for parallel processing of tandem mass spectra using PVM or MPI and X!Tandem

A method for the rapid correlation of tandem mass spectra to a list of protein sequences in a database has been developed. The combination of the fast and accurate computational search algorithm, X!Tandem, and a Linux cluster parallel computing environment with PVM or MPI, significantly reduces the time required to perform the correlation of tandem mass spectra to protein sequences in a database. A file of tandem mass spectra is divided into a specified number of files, each containing an equal number of the spectra from the larger file. These files are then searched in parallel against a protein sequence database. The results of each parallel output file are collated into one file for viewing through a web interface. Thousands of spectra can be searched in an accurate, practical, and time effective manner. The source code for running Parallel Tandem utilizing either PVM or MPI on Linux operating system is available from http://www.thegpm.org. This source code is made available under Artistic License from the authors.