Ultrastable Silicon Cavity in a Continuously Operating Closed-Cycle Cryostat at 4 K.

We report on a laser locked to a silicon cavity operating continuously at 4 K with 1×10^{-16} instability and a median linewidth of 17 mHz at 1542 nm. This is a tenfold improvement in short-term instability, and a 10^{4} improvement in linewidth, over previous sub-10-K systems. Operating at low temperatures reduces the thermal noise floor and, thus, is advantageous toward reaching an instability of 10^{-18}, a long-sought goal of the optical clock community. The performance of this system demonstrates the technical readiness for the development of the next generation of ultrastable lasers that operate with an ultranarrow linewidth and long-term stability without user intervention.

Rodentolepis straminea (Cestoda: Hymenolepididae) in an urban population of Apodemus sylvaticus in the UK.

The presence of the cyclophyllidean cestode Rodentolepis straminea (Cestoda: Hymenolepididae), was confirmed by molecular DNA analysis from a wood mouse (Apodemus sylvaticus) population inhabiting urban woodland in Salford, Greater Manchester (UK) with a prevalence of 27.8%. It would appear that the only previously published record of this species in A. sylvaticus in the British Isles is that from south-west Ireland, where 24% of the wood mice examined were infected with R. straminea. This species has been recorded in studies on A. sylvaticus in continental Europe. The current report represents a new record for R. straminea on mainland Britain and a first study of helminth parasites in an urban wood mouse population.

Reduction of residual amplitude modulation to 1 × 10⁻6 for frequency modulation and laser stabilization.

Active control and cancellation of residual amplitude modulation (RAM) in phase modulation of an optical carrier is one of the key technologies for achieving the ultimate stability of a laser locked to an ultrastable optical cavity. Furthermore, such techniques are versatile tools in various frequency modulation-based spectroscopy applications. In this Letter we report a simple and robust approach to actively stabilize RAM in an optical phase modulation process. We employ a waveguide-based electro-optic modulator (EOM) to provide phase modulation and implement an active servo with both DC electric field and temperature feedback onto the EOM to cancel both the in-phase and quadrature components of the RAM. This technique allows RAM control on the parts-per-million level where RAM-induced frequency instability is comparable to or lower than the fundamental thermal noise limit of the best available optical cavities.

Inhibition of neointimal cell bcl-x expression induces apoptosis and regression of vascular disease.

We postulated that activation of a genetic program that tonically inhibits intimal cell death is a necessary condition for the pathogenesis of vascular disease. Studies of vascular lesions in humans and animal models documented increased expression of the anti-apoptotic gene product Bcl-xL within intimal cells. Downregulation of intimal cell bcl-xL expression with the use of antisense oligonucleotides induced apoptosis and acute regression of vascular lesions. These findings indicate that apoptosis regulatory genes such as bcl-xL are critical determinants of intimal lesion formation and that targeted apoptosis may be a novel therapy for intimal vascular disease.

A novel internal abdominal oblique muscle flap to close a major abdominal wall defect.

This report describes a novel technique for abdominal wall reconstruction using an internal abdominal oblique muscle flap in an Australian kelpie. En bloc resection of a chondrosarcoma and biopsy scar centred on the 13th rib was performed to include full thickness thoracic wall (12th rib, extending caudally) and lateral abdominal wall (including the vascular pedicle of the external abdominal oblique muscle). The diaphragm was advanced to close the thorax. A flap using the caudal internal abdominal oblique muscle with the base dorsally was elevated and rotated 90° to fill the dorsal defect. The ventral defect was closed using the composite ventral abdominal muscles. The skin was closed primarily. The dog developed a self-resolving seroma. Twelve months postoperatively, the dog was able to engage in agility competitions. A viable muscle flap using the internal abdominal oblique muscle provides a useful alternative to previously described techniques for autogenous closure of a large abdominal wall wound.

Transversus abdominis muscle as a gastric or intestinal on-lay flap in two dogs and one cat.

Historically, omentalisation and serosal patching have been used to augment gastrointestinal wall defects or incisions where gastrointestinal viability is of concern. This report describes the novel use of a transversus abdominis muscle on-lay flap to augment and provide support to compromised intestine in three cases. The muscle flap was used to support: the ileum following reduction of an intussusception in a dog (case 1), the gastric wall following gastric strangulation, dilation and volvulus in a diaphragmatic rupture in a cat (case 2) and the jejunum following enterotomy and full-thickness ulcer resection with primary repair in a dog (case 3). All animals were discharged within 4 days postoperatively with no short- or long-term complications reported by either the referring veterinary surgeons on routine postoperative examination or on telephone follow-up with the owners (case 1: 10 months, case 2: 30 months, case 3: 6 weeks). The creation of a flap of the transversus abdominis muscle may provide a useful alternative or adjunct to previously described techniques for supporting the gastrointestinal tract when gastrointestinal tissue viability is questionable or resection is not feasible. Further prospective clinical evaluation studies would be indicated to determine whether the muscle flap remains viable or compare whether this technique should be recommended over conventional methods.

Evidence for neuroendocrine function of a unique splicing form of TCF7L2 in human brain, islets and gut.

AIMS/HYPOTHESIS: Variants in the TCF7L2 gene remain the strongest genetic associations with increased risk of type 2 diabetes. Recently, we identified a unique splicing form of TCF7L2 expressed in pancreatic islets, pancreas and colon and detected by assay 'ex13-13b'. The expression of ex13-13b strongly correlated with proinsulin in glucose-stimulated pancreatic islets, suggesting a potential role for this form in the development of type 2 diabetes. The goal of this study was to further characterise this unique TCF7L2 splicing form in human tissues. METHODS: We used a panel of 34 human tissues and 80 human cell lines to measure the expression of assay ex13-13b with use of quantitative RT-PCR. RESULTS: The highest expression of assay ex13-13b was detected in several areas of the brain (hypothalamus/thalamus, occipital lobe) and in neuronal cell line SHS5Y5. Low expression was confirmed in pancreatic islets, small intestine, pancreas and colon, while no expression was detected in other human tissues and cell lines. The expression of assay ex13-13b correlated with the gene for cocaine- and amphetamine-regulated transcript (CART, also known as CARTPT) in a panel of human tissues (n = 12, r = 0.85, p = 0.00046), pancreatic islets (n = 23, r = 0.62, p = 0.0016) and colon (n = 98, r = 0.54, p < 0.0001). CONCLUSIONS/INTERPRETATION: The significant correlation between expression of a unique splicing form of TCF7L2, named here TCF7L2-NE, and CART, the gene for an anorexigenic neurohormone expressed in the central and peripheral nervous system, suggests that these transcripts may share neuroendocrine functions important for brain, gut and pancreatic islets.

Expression analysis of loci associated with type 2 diabetes in human tissues.

AIMS/HYPOTHESIS: Genetic mapping has identified over 20 loci contributing to genetic risk of type 2 diabetes. The next step is to identify the genes and mechanisms regulating the contributions of genetic risk to disease. The goal of this study was to evaluate the effect of age, height, weight and risk alleles on expression of candidate genes in diabetes-associated regions in three relevant human tissues. METHODS: We measured transcript abundance for WFS1, KCNJ11, TCF2 (also known as HNF1B), PPARG, HHEX, IDE, CDKAL1, CDKN2A, CDKN2B, IGF2BP2, SLC30A8 and TCF7L2 by quantitative RT-PCR in human pancreas (n = 50), colon (n = 195) and liver (n = 50). Tissue samples were genotyped for single nucleotide polymorphisms (SNPs) associated with type 2 diabetes. The effects of age, height, weight, tissue and SNP on RNA expression were tested by linear modelling. RESULTS: Expression of all genes exhibited tissue bias. Immunohistochemistry confirmed the findings for HHEX, IDE and SLC30A8, which showed strongest tissue-specific mRNA expression bias. Neither age, height nor weight were associated with gene expression. We found no evidence that type 2 diabetes-associated SNPs affect neighbouring gene expression (cis-expression quantitative trait loci) in colon, pancreas and liver. CONCLUSIONS/INTERPRETATION: This study provides new evidence that tissue-type, but not age, height, weight or SNPs in or near candidate genes associated with increased risk of type 2 diabetes are strong contributors to differential gene expression in the genes and tissues examined.

The Chlamydomonas FLA10 gene encodes a novel kinesin-homologous protein.

Many genes on the uni linkage group of Chlamydomonas affect the basal body/flagellar apparatus. Among these are five FLA genes, whose mutations cause temperature-sensitive defects in flagellar assembly. We present the molecular analysis of a gene which maps to fla10 and functionally rescues the fla10 phenotype. Nucleotide sequencing revealed that the gene encodes a kinesin-homologous protein, KHP1. The 87-kD predicted KHP1 protein, like kinesin heavy chain, has an amino-terminal motor domain, a central alpha-helical stalk, and a basic, globular carboxy-terminal tail. Comparison to other kinesin superfamily members indicated striking similarity (64% identity in motor domains) to a mouse gene, KIF3, expressed primarily in cerebellum. In synchronized cultures, the KHP1 mRNA accumulated after cell division, as did flagellar dynein mRNAs. KHP1 mRNA levels also increased following deflagellation. Polyclonal antibodies detected KHP1 protein in Western blots of purified flagella and axonemes. The protein was partially released from axonemes with ATP treatment, but not with AMP-PNP. Western blot analysis of axonemes from various motility mutants suggested that KHP1 is not a component of radial spokes, dynein arms, or the central pair complex. The quantity of KHP1 protein in axonemes of the mutant fla10-1 was markedly reduced, although no reduction was observed in two other uni linkage group mutants, fla9 and fla11. Furthermore, fla10-1 was rescued by transformation with KHP1 genomic DNA. These results indicate that KHP1 is the gene product of FLA10 and suggest a novel role for this kinesin-related protein in flagellar assembly and maintenance.

Stabilized lasers and precision measurements.

This article traces the development of stabilized lasers from the Massachusetts Institute of Technology passive-stabilization experiments of the early 1960's up through the current epoch of highly stabilized helium-neon and carbon dioxide and continuous wave dye lasers. The utility, present performance, and limitations of stabilized lasers as standards of length or frequency for precision measurements are discussed. Examples considered of laser applications to physical measurements of outstanding scientific interest include determination of the speed of light, redefinition of the meter, resolution of the photon recoil-induced spectral doubling, use of optical "Ramsey" interference fringes from ultrahigh-resolution spectroscopy, and two improved tests of special relativity.

Detection of daily clouds on Titan.

We have discovered frequent variations in the near-infrared spectrum of Titan, Saturn's largest moon, which are indicative of the daily presence of sparse clouds covering less than 1% of the area of the satellite. The thermodynamics of Titan's atmosphere and the clouds' altitudes suggest that convection governs their evolutions. Their short lives point to the presence of rain. We propose that Titan's atmosphere resembles Earth's, with clouds, rain, and an active weather cycle, driven by latent heat release from the primary condensible species.

Phase-coherent optical pulse synthesis from separate femtosecond lasers.

We generated a coherently synthesized optical pulse from two independent mode-locked femtosecond lasers, providing a route to extend the coherent bandwidth available for ultrafast science. The two separate lasers (one centered at 760 nanometers wavelength, the other at 810 nanometers) are tightly synchronized and phase-locked. Coherence between the two lasers is demonstrated via spectral interferometry and second-order field cross-correlation. Measurements reveal a coherently synthesized pulse that has a temporally narrower second-order autocorrelation width and that exhibits a larger amplitude than the individual laser outputs. This work represents a new and flexible approach to the synthesis of coherent light.

Novel application of internal obturator and semitendinosus muscle flaps for rectal wall repair or reinforcement.

Internal obturator and/or semitendinosus muscle flaps were used to reinforce primary appositional rectal wall repair in three dogs and one cat in this case series. All three dogs incurred rectal wall compromise during surgical excision of anal sac tumours. The cat sustained bite wounds to the perianal region resulting in abscessation and a rectal tear. Our results indicate that application of an internal obturator and/or semitendinosus muscle flap can reduce the risk of rectal wall dehiscence after primary repair, and consequently the risk of pararectal abscess or rectocutaneous fistula formation.

Urinary incontinence in male dogs under primary veterinary care in England: prevalence and risk factors.

OBJECTIVE: To estimate prevalence and identify demographic risk factors for urinary incontinence in male dogs. METHODS AND METHODS: The study population included all dogs within the VetCompass database from September 1, 2009 to July 7, 2013. Electronic patient records were searched for urinary incontinence cases; demographic and clinical information were extracted and analysed. RESULTS: Of 109,428 male dogs attending 119 clinics in England, there were an estimated 1027 dogs diagnosed with urinary incontinence, giving a prevalence of 0.94% (95% confidence interval: 0.88 to 1.00). Breeds with highest odds of incontinence in male dogs (compared with mixed breed dogs) included the bull mastiff (odds ratio: 17.21, 95% confidence interval: 6.65 to 44.56, case=5, non-case=314, P<0.001), Irish red setter (odds ratio: 12.79, 95% confidence interval: 4.83 to 33.84, case=5, non-case=142, P<0.001), fox terrier (odds ratio: 9.60, 95% confidence interval: 3.68 to 25.05, case=5, non-case=176, P < 0.001), bulldog (odds ratio: 5.72, 95% confidence interval: 2.24 to 14.59, case=5, non-case=929, P<0.001) and boxer (odds ratio: 3.65, 95% confidence interval: 1.84 to 7.25, case=10, non-case=1470, P<0.001). Increased odds of urinary incontinence were associated with greater age (age 9 to 12 years, odds ratio: 10.46, 95% confidence interval: 6.59 to 16.62, n=12,348, P<0.001) and being insured (odds ratio: 1.96, 95% confidence interval: 1.53 to 2.51, n=26,202, P<0.001). There was no association with castration or bodyweight using multi-variable analysis. CLINICAL SIGNIFICANCE: The overall prevalence of urinary incontinence in male dogs is approximately 1%, which may be higher than expected given the sparsity of reports describing this problem. In contrast to bitches, neutering and bodyweight were not associated with greater odds of urinary incontinence, which is important when giving neutering advice.

Identification of two human beta-tubulin isotypes.

The sequence of a human beta-tubulin cDNA clone (D beta-1) is described; our data revealed 95.6% homology compared with the sequence of a human beta-tubulin processed pseudogene derived by reverse transcription of a processed mRNA (Wilde et al., Nature [London] 297:83-84, 1982). However, the amino acid sequence encoded by this cDNA showed less homology with pig and chicken beta-tubulin sequences than the latter did to each other, with major divergence within the 15 carboxy-terminal amino acids. On the other hand, an independently isolated, functionally expressed genomic human beta-tubulin sequence (5 beta) possessed a very high degree of homology with chicken and pig beta-tubulins in this region. Thus, human cells appear to contain two distinct beta-tubulin isotypes. Both the intact beta-tubulin cDNA clone and a subclone containing only the 3' untranslated region detected two mRNA species in HeLa cells; these mRNAs were 1.8 and 2.6 kilobases long and were present in about equal amounts. Two independently subcloned probes constructed from the 3' untranslated region of the 5 beta genomic sequence also detected a 2.6-kilobase beta-tubulin mRNA. However, the 3'-untranslated-region probes from the cDNA clone and the genomic sequence did not cross-hybridize. Thus, at least two human beta-tubulin genes, each specifying a distinct isotype, are expressed in HeLa cells, and the 2.6-kilobase mRNA band is a composite of at least two comigrating beta-tubulin mRNAs.

Urinary incontinence in bitches under primary veterinary care in England: prevalence and risk factors.

OBJECTIVES: To estimate prevalence and demographic risk factors for urinary incontinence in bitches under primary veterinary care in England. METHODS: The study population included all bitches within the VetCompass database from September 1, 2009 to July 7, 2013. Electronic patient records were searched for urinary incontinence cases and additional demographic and clinical information was extracted. RESULTS: Of 100,397 bitches attending 119 clinics in England, an estimated 3108 were diagnosed with urinary incontinence. The prevalence of urinary incontinence was 3·14% (95% confidence intervals: 2·97 to 3·33). Medical therapy was prescribed to 45·6% cases. Predisposed breeds included the Irish setter (odds ratio: 8·09; 95% confidence intervals: 3·15 to 20·80; P< 0·001) and Dobermann (odds ratio: 7·98; 95% confidence intervals: 4·38 to 14·54; P< 0·001). Increased odds of a diagnosis of urinary incontinence were associated with: (1) weight at or above the mean adult bodyweight for the breed (odds ratio: 1·31; 95% confidence intervals: 1·12 to 1·54; P< 0·001), (2) age 9 to 12 years (odds ratio: 3·86; 95% confidence intervals: 2·86 to 5·20, P< 0·001), (3) neuter status (odds ratio: 2·23; 95% confidence intervals: 1·52 to 3·25, P< 0·001) and (4) being insured (odds ratio: 1·59; 95% confidence intervals: 1·34 to 1·88, P< 0·001). CLINICAL IMPACT: Clinical Impact: Urinary incontinence affects just over 3% of bitches overall but affects more than 15% of bitches in high-risk breeds including the Irish setter, Dobermann, bearded collie, rough collie and Dalmatian. These results provide an evidence base for clinicians to enhance clinical recommendations on neutering and weight control, especially in high-risk breeds.

Hyperglycemia inhibits vascular smooth muscle cell apoptosis through a protein kinase C-dependent pathway.

We hypothesized that the pathogenesis of diabetic vasculopathy involves the abnormal regulation of vascular smooth muscle cell (VSMC) apoptosis. In nondiabetic mice, a reduction in carotid artery blood flow resulted in a significant loss of medial VSMCs via apoptosis (normal flow 84+/-1 viable VSMCs, reduced flow 70+/-5 viable VSMCs; n=12, P:<0.01). In contrast, flow-induced VSMC apoptosis was markedly attenuated in streptozotocin-induced diabetic mice (normal flow 85+/-2 viable VSMC, reduced flow 82+/-4 viable VSMC; n=13, NS). In accord with our in vivo findings, the exposure of cultured rat and human VSMCs to high glucose (17.5 mmol/L) significantly attenuated the induction of apoptosis in response to serum withdrawal (rat VSMCs in normal [5.5 mmol/L] glucose 28+/-1%, high D-glucose 19+/-2%; P:<0.0001). High glucose also inhibited apoptosis induced by Fas ligand (100 ng/mL) (normal 23+/-2%, high D-glucose 13+/-2%; P:<0.006). Supplementation with the nonmetabolized enantiomer L-glucose had no effect. We confirmed reports that high glucose activates protein kinase C (PKC) and demonstrated that PKC blockade with long-term phorbol ester treatment or calphostin C prevented the antiapoptotic effect (P:<0. 001). Moreover, the upregulation of either PKCalpha or PKCbetaII expression was sufficient to inhibit serum withdrawal-induced apoptosis (control 25+/-2%, PKCalpha 11+/-2%, PKCbetaII 8+/-2%; P:<0. 0001), whereas the upregulation of PKCdelta had no significant effect. Taken together, these findings demonstrate that hyperglycemia inhibits VSMC apoptosis via a PKC-dependent pathway.

Overexpression of Ref-1 inhibits hypoxia and tumor necrosis factor-induced endothelial cell apoptosis through nuclear factor-kappab-independent and -dependent pathways.

We hypothesized that a redox-sensitive transcription factor, redox factor-1 (Ref-1) (HAP1, APE, and APEX), was critical in the regulation of endothelial cell survival in response to hypoxia and cytokines, including tumor necrosis factor (TNF)-alpha. Hypoxia resulted in a significant decrease in Ref-1 protein expression in both human umbilical vein endothelial cells and calf pulmonary artery endothelial cells. The hypoxia-induced decrease in Ref-1 expression was followed by a significant induction of apoptosis as measured by caspase 3 activity and nuclear morphology. Transient upregulation of Ref-1 significantly inhibited hypoxia-induced apoptosis. However, deletion of the redox-sensitive domain of Ref-1 abolished the antiapoptotic effect. We postulated that the antiapoptotic effects of Ref-1 were mediated through nuclear factor-kappaB (NF-kappaB). However, blockade of NF-kappaB with a dominant-negative IkappaB (S32A/S36A) expression vector had no effect on Ref-1-mediated survival under hypoxic conditions. The second aim of this study was to test the cytoprotective ability of Ref-1 upregulation in response to TNF-induced apoptosis. Ref-1 inhibition of TNF-induced death was associated with a significant potentiation of NF-kappaB activity. Deletion of the redox-sensitive domain of Ref-1 significantly inhibited TNF-induced NF-kappaB activation. Moreover, loss of the redox-sensitive domain also abolished the antiapoptotic effect of Ref-1 in response to TNF. To test whether Ref-1 induced activation of NF-kappaB was necessary to promote survival, we blocked NF-kappaB activity with a dominant-negative IkappaB (S32A/S36A). Indeed, blockade of NF-kappaB activity abolished the ability of Ref-1 to rescue TNF-induced apoptosis. In conclusion, upregulation of Ref-1 promotes endothelial cell survival in response to hypoxia and TNF through NF-kappaB-independent and NF-kappaB-dependent signaling cascades, respectively. Moreover, it seems that Ref-1 may act as a critical cofactor, mediating the TNF-induced NF-kappaB response in the vascular endothelium.

Emerging mechanisms for heavy metal transport in plants.

Heavy metal ions such as Cu(2+), Zn(2+), Mn(2+), Fe(2+), Ni(2+) and Co(2+) are essential micronutrients for plant metabolism but when present in excess, these, and non-essential metals such as Cd(2+), Hg(2+) and Pb(2+), can become extremely toxic. Thus mechanisms must exist to satisfy the requirements of cellular metabolism but also to protect cells from toxic effects. The mechanisms deployed in the acquisition of essential heavy metal micronutrients have not been clearly defined although a number of genes have now been identified which encode potential transporters. This review concentrates on three classes of membrane transporters that have been implicated in the transport of heavy metals in a variety of organisms and could serve such a role in plants: the heavy metal (CPx-type) ATPases, the natural resistance-associated macrophage protein (Nramp) family and members of the cation diffusion facilitator (CDF) family. We aim to give an overview of the main features of these transporters in plants in terms of structure, function and regulation drawing on information from studies in a wide variety of organisms.

Cloning of a cDNA coding for an amino acid carrier from Ricinus communis (RcAAP1) by functional complementation in yeast: kinetic analysis, inhibitor sensitivity and substrate specificity.

A cDNA for the amino acid permease gene RcAAP1 has been isolated from Ricinus communis by yeast complementation and subjected to a detailed kinetic analysis. RcAAP1 cDNA is 1.5 kb with an open reading frame that codes for a protein with 486 amino acids and a calculated molecular mass of 53.1 kDa. RcAAP1-mediated histidine uptake was pH dependent with highest transport rates at acidic pH; it was sensitive to protonophores and uncouplers and the Km for histidine uptake was 96 microM. The substrate specificity was investigated by measuring the levels of inhibition of histidine uptake by a range of amino acids. The basic amino acids (histidine, lysine and arginine) showed strongest inhibition of uptake whereas acidic amino acids competed less effectively. Alanine was the most efficient competitor of the neutral amino acids. Glutamine, serine, asparagine, methionine and cysteine showed moderate inhibition whereas threonine, isoleucine, leucine, phenylalanine, tyrosine and tryptophan showed only low levels of inhibition. Glycine, proline and citrulline caused slight stimulation. More detailed competition kinetics indicated that both lysine and arginine showed simple competitive inhibition of histidine uptake. When direct uptake measurements were carried out, both lysine and arginine were found to be effective substrates for RcAAP1.