Gametophyte niche differences among sympatric tree ferns.

Forest community assembly is usually framed in terms of sporophyte dynamics; however, the recruitment and maintenance of fern populations, frequently influential in forest composition and structure, are initially determined by gametophytes. Sporophytes of three Cyathea tree fern species show habitat partitioning along gradients of phosphorus and light; we asked whether gametophyte niche differences parallel this pattern. To compare niche characteristics among taxa we compared growth rates to a size threshold (≥3 mm) of gametophytes under controlled conditions using a multi-factorial, multi-level (3 × 4) experiment, varying irradiance (5.4 ± 4.4; 59.1 ± 44.3; 107 ± 74.1 µmol m-2 s-1) and orthophosphate concentrations (5, 10, 20, 40 mg kg-1). Gametophytes of the pioneer species C. medullaris developed to the size threshold across a broad range of phosphate and irradiance treatments (more than 20% of gametophytes in ≥ 7 of the 12 treatments), peaking at 20 mg kg-1 P and 60 µmol m-2 s-1 irradiance. The growth rates of the forest understorey species C. dealbata and C. smithii also peaked at 60 µmol m-2 s-1 but varied across treatments, suggesting niche differentiation along irradiance and orthophosphate gradients. Our analysis suggests that gametophyte development is strategically aligned to the ecological habits of sporophytes and that forest community assembly is likely strongly influenced by the independent gametophyte life-stage.

Broadening the optical bandwidth of quantum cascade lasers using RF noise current perturbations.

We report on the broadening of the optical bandwidth of a distributed feedback quantum cascade laser (QCL) caused by the application of radio frequency (RF) noise to the injection current. The broadening is quantified both via Lamb-dip spectroscopy and the frequency noise power spectral density (PSD). The linewidth of the unperturbed QCL (emitting at ∼5.3 µm) determined by Lamb-dip spectroscopy is 680±170 kHz, and is in reasonable agreement with the linewidth of 460±40 kHz estimated by integrating the PSD measured under the same laser operating conditions. Measurements with both techniques reveal that by mixing the driving current with broadband RF noise the laser lineshape was reproducibly broadened up to ca 6 MHz with an increasing Gaussian contribution. The effects of linewidth broadening are then demonstrated in the two-color coherent transient spectra of nitric oxide.

Conduit dynamics and post explosion degassing on Stromboli: A combined UV camera and numerical modeling treatment.

Recent gas flux measurements have shown that Strombolian explosions are often followed by periods of elevated flux, or "gas codas," with durations of order a minute. Here we present UV camera data from 200 events recorded at Stromboli volcano to constrain the nature of these codas for the first time, providing estimates for combined explosion plus coda SO2 masses of ≈18-225 kg. Numerical simulations of gas slug ascent show that substantial proportions of the initial gas mass can be distributed into a train of "daughter bubbles" released from the base of the slug, which we suggest, generate the codas, on bursting at the surface. This process could also cause transitioning of slugs into cap bubbles, significantly reducing explosivity. This study is the first attempt to combine high temporal resolution gas flux data with numerical simulations of conduit gas flow to investigate volcanic degassing dynamics.

Enhancing the sensitivity of mid-IR quantum cascade laser-based cavity-enhanced absorption spectroscopy using RF current perturbation.

The sensitivity of mid-IR quantum cascade laser (QCL) off-axis cavity-enhanced absorption spectroscopy (CEAS), often limited by cavity mode structure and diffraction losses, was enhanced by applying a broadband RF noise to the laser current. A pump-probe measurement demonstrated that the addition of bandwidth-limited white noise effectively increased the laser linewidth, thereby reducing mode structure associated with CEAS. The broadband noise source offers a more sensitive, more robust alternative to applying single-frequency noise to the laser. Analysis of CEAS measurements of a CO(2) absorption feature at 1890 cm(-1) averaged over 100 ms yielded a minimum detectable absorption of 5.5×10(-3) Hz(-1/2) in the presence of broadband RF perturbation, nearly a tenfold improvement over the unperturbed regime. The short acquisition time makes this technique suitable for breath applications requiring breath-by-breath gas concentration information.

Pump and probe spectroscopy with continuous wave quantum cascade lasers.

This paper details infra-red pump and probe studies on nitric oxide conducted with two continuous wave quantum cascade lasers both operating around 5 µm. The pump laser prepares a velocity selected population in a chosen rotational quantum state of the v = 1 level which is subsequently probed using a second laser tuned to a rotational transition within the v = 2 ← v = 1 hot band. The rapid frequency scan of the probe (with respect to the molecular collision rate) in combination with the velocity selective pumping allows observation of marked rapid passage signatures in the transient absorption profiles from the polarized vibrationally excited sample. These coherent transient signals are influenced by the underlying hyperfine structure of the pump and probe transitions, the sample pressure, and the coherent properties of the lasers. Pulsed pump and probe studies show that the transient absorption signals decay within 1 µs at 50 mTorr total pressure, reflecting both the polarization and population dephasing times of the vibrationally excited sample. The experimental observations are supported by simulation based upon solving the optical Bloch equations for a two level system.

Coherent transient spectroscopy with continuous wave quantum cascade lasers.

A high power continuous wave quantum cascade laser operating around 1900 cm(-1) has been used to conduct Lamb dip spectroscopy on a low pressure sample of NO. The widths of the Lamb dips indicate that the laser linewidth is 800 ± 60 kHz and the power sufficient to induce significant population transfer of up to 35%. While the Lamb dip signals are symmetric at low laser chirp rates, they become increasingly asymmetric as the chirp rate increases, further confirming the significant degree of population transfer. In addition rapid passage structure on the Lamb dip signal is observed after the weak probe beam is swept through the line center. This structure is sensitive to both the probe chirp rate and the underlying hyperfine structure of the rovibrational transition, and is accurately modeled using the optical Bloch equations.

Chromosomal mapping of genetic loci associated with non-insulin dependent diabetes in the GK rat.

Goto-Kakizaki (GK) rats are a well characterized model for non-insulin dependent diabetes mellitus (NIDDM). We have used a combination of physiological and genetic studies to identify quantitative trait loci (QTLs) responsible for the control of glucose homeostasis and insulin secretion in a F2 cohort bred from spontaneously diabetic GK rats. The genetic dissection of NIDDM allowed us to map up to six independently segregating loci predisposing to hyperglycaemia, glucose intolerance or altered insulin secretion, and a seventh locus implicated in body weight. QTLs implicated in glucose tolerance and adiposity map to the same region of rat chromosome 1, and may indicate the influence of a single locus. Our study demonstrates that distinct combinations of genetic loci are responsible for different physiological characteristics associated with the diabetic phenotype in the GK rat, and it constitutes an important step for directing the search for the genetic factors involved in human NIDDM.

A radiation hybrid map of 506 STS markers spanning human chromosome 11.

We present a high resolution radiation hybrid map of human chromosome 11 using 506 sequence tagged sites (STSs) scored on a panel of 86 radiation hybrids. The 506 STSs fall into 299 unique positions (average resolution of about 480 kilobases (kb)) that span the whole chromosome. A subset of 260 STSs (143 positions) form a framework map that has a resolution of approximately 1 megabase between adjacent positions and is ordered with odds of at least 1,000:1. The centromere was clearly defined with pericentric markers unambiguously assigned to the short or long arm. The map contains most genes (125) and expressed sequence tags (26) currently assigned to chromosome 11 and more than half of the STSs are polymorphic microsatellite loci. These markers and the map can be used for high resolution physical and genetic mapping.

A major quantitative trait locus influences hyperactivity in the WKHA rat.

The syndrome of hyperactivity describes behavioural disorders existing mainly in children and characterized by increased levels of motor activity, inattention and impulsivity. Overall the aetiology is poorly understood due to the heterogeneity of the pathology although psychological, biological and social factors acting singly or in concert are generally thought to be involved. In animal studies the observed hyperactivity phenotype results from relative participation of exploration, emotionality and general activity. Studies using brain lesions, neuropharmacology and gene knock-out strategies have shown that specific elements of the brain dopaminergic system can subserve hyperactivity. Evidence of a genetic contribution comes from family and twin studies but also from the ability to select divergent animal lines on the basis of their differential activity. The Wistar-Kyoto (WKY) and Wistar-Kyoto hyperactive (WKHA) rats are such strains--distinct for their low and high activity scores in a novel environment, respectively. Here, we report the detection of a major hyperactivity-related QTL on chromosome 8, explaining 29% of the variance of an intercross between these strains. This study represents the first behavioural QTL analysis in rat and provides a new starting point for biologically categorizing different forms of hyper-activity.

A radiation hybrid map of the rat genome containing 5,255 markers.

A whole-genome radiation hybrid (RH) panel was used to construct a high-resolution map of the rat genome based on microsatellite and gene markers. These include 3,019 new microsatellite markers described here for the first time and 1,714 microsatellite markers with known genetic locations, allowing comparison and integration of maps from different sources. A robust RH framework map containing 1,030 positions ordered with odds of at least 1,000:1 has been defined as a tool for mapping these markers, and for future RH mapping in the rat. More than 500 genes which have been mapped in mouse and/or human were localized with respect to the rat RH framework, allowing the construction of detailed rat-mouse and rat-human comparative maps and illustrating the power of the RH approach for comparative mapping.

Beetle adhesive hairs differ in stiffness and stickiness: in vivo adhesion measurements on individual setae.

Leaf beetles are able to climb on smooth and rough surfaces using arrays of micron-sized adhesive hairs (setae) of varying morphology. We report the first in vivo adhesive force measurements of individual setae in the beetle Gastrophysa viridula, using a smooth polystyrene substrate attached to a glass capillary micro-cantilever. The beetles possess three distinct adhesive pads on each leg which differ in function and setal morphology. Visualisation of pull-offs allowed forces to be measured for each tarsal hair type. Male discoidal hairs adhered with the highest forces (919 ± 104 nN, mean ± SE), followed by spatulate (582 ± 59 nN) and pointed (127 ± 19 nN) hairs. Discoidal hairs were stiffer in the normal direction (0.693 ± 0.111 N m(-1)) than spatulate (0.364 ± 0.039 N m(-1)) or pointed (0.192 ± 0.044 N m(-1)) hairs. The greater adhesion on smooth surfaces and the higher stability of discoidal hairs help male beetles to achieve strong adhesion on the elytra of females during copulation. A comparison of pull-off forces measured for single setae and whole pads (arrays) revealed comparable levels of adhesive stress. This suggests that beetles are able to achieve equal load sharing across their adhesive pads so that detachment through peeling is prevented.

Biallelic mutations in the ATM gene in T-prolymphocytic leukemia.

Ataxia-telangiectasia (AT) is an autosomal recessive disorder characterized by cerebellar ataxia, oculocutaneous telangiectasia, immune deficiency, genome instability and predisposition to malignancies, particularly T-cell neoplasms. The responsible gene, designated ataxia-telangiectasia mutated (ATM), was recently identified by positional cloning in the chromosomal region 11q22.3-23.1 (ref. 4, 5) ATM is 150 kb in length, consists of 66 exons and encodes a nuclear phosphoprotein of approximately 350 kDa (ref. 4-9). Although ATM is considered to be a tumorigenic factor in several human cancers, it has not yet been found mutated in tumors of non-AT patients. Given the marked predisposition of AT patients to develop neoplasms of the T-cell lineage, we analyzed a series of T-cell leukemias (T-prolymphocytic leukemia, or T-PLL) in non-AT patients in search of genomic changes associated with the development of this disease. Among the recurrent aberrations identified, deletion of the chromosome arm 11q was very frequent. Subsequent molecular cytogenetic analyses allowed us to define a small commonly deleted segment at 11q22.3-23.1 in 15 of 24 T-PLLs studied. Since this critical region contained ATM, we further analyzed the remaining copy of the gene in six cases showing deletions affecting one ATM allele. In all six cases, mutations of the second ATM allele were identified, leading to the absence, premature truncation or alteration of the ATM gene product. Thus, our study demonstrates disruption of both ATM alleles by deletion or point mutation in T-PLL, suggesting that ATM functions as a tumor-suppressor gene in tumors of non-AT individuals.

Genome-wide association for major depressive disorder: a possible role for the presynaptic protein piccolo.

Major depressive disorder (MDD) is a common complex trait with enormous public health significance. As part of the Genetic Association Information Network initiative of the US Foundation for the National Institutes of Health, we conducted a genome-wide association study of 435 291 single nucleotide polymorphisms (SNPs) genotyped in 1738 MDD cases and 1802 controls selected to be at low liability for MDD. Of the top 200, 11 signals localized to a 167 kb region overlapping the gene piccolo (PCLO, whose protein product localizes to the cytomatrix of the presynaptic active zone and is important in monoaminergic neurotransmission in the brain) with P-values of 7.7 x 10(-7) for rs2715148 and 1.2 x 10(-6) for rs2522833. We undertook replication of SNPs in this region in five independent samples (6079 MDD independent cases and 5893 controls) but no SNP exceeded the replication significance threshold when all replication samples were analyzed together. However, there was heterogeneity in the replication samples, and secondary analysis of the original sample with the sample of greatest similarity yielded P=6.4 x 10(-8) for the nonsynonymous SNP rs2522833 that gives rise to a serine to alanine substitution near a C2 calcium-binding domain of the PCLO protein. With the integrated replication effort, we present a specific hypothesis for further studies.

Evidence for self-cleaning in fluid-based smooth and hairy adhesive systems of insects.

Insects possess adhesive organs that allow attachment to diverse surfaces. Efficient adhesion must be retained throughout their lifetime even when pads are exposed to contamination. Many insects groom their adhesive structures, but it is possible that self-cleaning properties also play an important role. We measured attachment forces of insect pads on glass after contamination with microspheres and found that both smooth pads (stick insects: Carausius morosus) and hairy pads (dock beetles: Gastrophysa viridula) exhibit self-cleaning. Contaminated pads recovered high levels of adhesion after only eight simulated steps; this was accompanied by the deposition of spheres. Self-cleaning was strongly enhanced by shear movements, and only beetle pads showed the ability to self-clean during purely perpendicular pull-offs. Hairy pads also self-cleaned more efficiently than smooth pads for both large (45 microm) and small (1 microm) particle sizes. However, the beetles' self-cleaning was not superior to smooth pads when contaminated with 10-microm beads. This limitation of self-cleaning is explained by the coincidence of bead diameter and inter-seta distance, which caused beads to remain trapped in between setae.

Homodyne locking of a squeezer.

We report on the successful implementation of an approach to locking the frequencies of an optical parametric oscillator (OPO)-based squeezed-vacuum source and its driving laser. The technique allows the simultaneous measurement of the phase shifts induced by a cavity, which may be used for the purposes of frequency locking, as well as the simultaneous measurement of the sub-quantum-noise-limited (sub-QNL) phase quadrature output of the OPO. The homodyne locking technique is cheap, easy to implement, and has the distinct advantage that subsequent homodyne measurements are automatically phase locked. The homodyne locking technique is also unique in that it is a sub-QNL frequency discriminator.

A whole genome association study of neuroticism using DNA pooling.

We describe a multistage approach to identify single nucleotide polymorphisms (SNPs) associated with neuroticism, a personality trait that shares genetic determinants with major depression and anxiety disorders. Whole genome association with 452 574 SNPs was performed on DNA pools from approximately 2000 individuals selected on extremes of neuroticism scores from a cohort of 88 142 people from southwest England. The most significant SNPs were then genotyped on independent samples to replicate findings. We were able to replicate association of one SNP within the PDE4D gene in a second sample collected by our laboratory and in a family-based test in an independent sample; however, the SNP was not significantly associated with neuroticism in two other independent samples. We also observed an enrichment of low P-values in known regions of copy number variations. Simulation indicates that our study had approximately 80% power to identify neuroticism loci in the genome with odds ratio (OR)>2, and approximately 50% power to identify small effects (OR=1.5). Since we failed to find any loci accounting for more than 1% of the variance, the heritability of neuroticism probably arises from many loci each explaining much less than 1%. Our findings argue the need for much larger samples than anticipated in genetic association studies and that the biological basis of emotional disorders is extremely complex.

Effect of the BDNF V166M polymorphism on working memory in healthy adolescents.

Brain-derived neurotrophic factor (BDNF) may play a role in modulating memory function and there is growing evidence that the BDNF V166M polymorphism may influence episodic memory in humans. However, previous association studies examining this polymorphism and working memory are inconsistent. The current study examined this association in a large sample of adolescent twin-pairs and siblings (785 individuals from 439 families). A range of measures (event-related potential, general performance and reaction time) was obtained from a delayed-response working-memory task and total association was examined using the quantitative transmission disequilibrium tests (QTDT) program. Analyses had approximately 93-97% power (alpha= 0.05) to detect an association accounting for as little as 2% of the variance in the phenotypes examined. Results indicated that the BDNF V166M polymorphism is not associated with variation in working memory in healthy adolescents.

Inactivation of a transfected gene in human fibroblasts can occur by deletion, amplification, phenotypic switching, or methylation.

Plasmids containing the bacterial gpt gene under control of the simian virus 40 promoter were transfected into a simian virus 40-transformed human fibroblast line. Two transfectants, E2 and C10, which contain stably integrated single copies of the gpt gene, were isolated. These two lines produce Gpt- variants spontaneously with a frequency of about 10(-4). We carried out a detailed molecular analysis of the spectrum of alterations which gave rise to the Gpt- phenotype in these variants. DNA from 14 of 19 Gpt- derivatives of one of the cell lines (E2) contains deletions or rearrangements of gpt-containing sequences. In four of the remaining five lines, the Gpt- phenotype was correlated with reduced levels of expression rather than with changes in the gross structure of the gpt gene, and it was possible to reactivate the gpt gene. In one Gpt- line, gpt mRNA was present at normal levels, but no active enzyme was produced. Spontaneous Gpt- derivatives of the other cell line (C10) produced a completely different spectrum of alterations. Very few deletions were found, but several derivatives contained additional extrachromosomal gpt sequences, and, remarkably, in two other Gpt- lines, gpt-containing sequences were amplified more than 100-fold. The phenotypes of the majority of the Gpt- derivatives of C10 could be attributed to alterations in gene expression caused by methylation.

Stable correction of a genetic deficiency in human cells by an episome carrying a 115 kb genomic transgene.

Persistent expression of a transgene at therapeutic levels is required for successful gene therapy, but many small vectors with heterologous promoters are prone to vector loss and transcriptional silencing. The delivery of genomic DNA would enable genes to be transferred as complete loci, including regulatory sequences, introns, and native promoter elements. These elements may be critical to ensure prolonged, regulated, and tissue-specific transgene expression. Many studies point to considerable advantages to be gained by using complete genomic loci in gene expression. Large-insert vectors incorporating elements of the bacterial artificial chromosome (BAC) cloning system, and the episomal maintenance mechanisms of Epstein-Barr virus (EBV), can shuttle between bacteria and mammalian cells, allowing large genomic loci to be manipulated conveniently. We now demonstrate the potential utility of such vectors by stably correcting a human genetic deficiency in vitro. When the complete hypoxanthine phosphoribosyltransferase (HPRT) locus of 115 kilobases (kb) was introduced into deficient human cells, the transgene was both maintained as an episome and expressed stably for six months in rapidly dividing cell cultures. The results demonstrate for the first time that gene expression from an episomal genomic transgene can correct a cell culture disease phenotype for a prolonged period.

Jumping without slipping: leafhoppers (Hemiptera: Cicadellidae) possess special tarsal structures for jumping from smooth surfaces.

Many hemipteran bugs can jump explosively from plant substrates, which can be very smooth. We therefore analysed the jumping performance of froghoppers (Philaenus spumarius, Aphrophoridae) and leafhoppers (Aphrodes bicinctus/makarovi, Cicadellidae) taking off from smooth (glass) and rough (sandpaper, 30 µm asperity size) surfaces. On glass, the propulsive hind legs of Philaenus froghoppers slipped, resulting in uncontrolled jumps with a fast forward spin, a steeper angle and only a quarter of the velocity compared with jumps from rough surfaces. By contrast, Aphrodes leafhoppers took off without their propulsive hind legs slipping, and reached low take-off angles and high velocities on both substrates. This difference in jumping ability from smooth surfaces can be explained not only by the lower acceleration of the long-legged leafhoppers, but also by the presence of 2-9 soft pad-like structures (platellae) on their hind tarsi, which are absent in froghoppers. High-speed videos of jumping showed that platellae contact the surface briefly (approx. 3 ms) during the acceleration phase. Friction force measurements on individual hind tarsi on glass revealed that at low sliding speeds, both pushing and pulling forces were small, and insufficient to explain the recorded jumps. Only when the tarsi were pushed with higher velocities did the contact area of the platellae increase markedly, and high friction forces were produced, consistent with the observed jumps. Our findings show that leafhoppers have special adhesive footpads for jumping from smooth surfaces, which achieve firm grip and rapid control of attachment/detachment by combining anisotropic friction with velocity dependence.