Mouse models of deep vein thrombosis.

The pathogenesis of venous thromboembolism (VTE) is still not completely understood. Experimental animals in which human deep vein thrombosis can be modeled are useful tools to investigate the pathogenesis of VTE. Besides the availability of transgenic and genetically modified mice, the use of high frequency ultrasound and intravital microscopy plays an important role in identifying thrombotic processes in mouse models. In this article, an overview about the application of various new technologies and existing mouse models is provided, and the impact of venous side branches on deep vein thrombosis in the mouse model is discussed.

Cofactor-independent human antiphospholipid antibodies induce venous thrombosis in mice.

UNLABELLED: Essentials Cofactor-independent antiphospholipid antibodies (CI-aPL) are generally considered non-pathogenic. We analyzed the effects of human monoclonal CI-aPL in a mouse model of venous thrombosis. As shown in vitro, CI-aPL induce a procoagulant state in vivo by activation of endosomal NADPH-oxidase. Contrary to common belief, CI-aPL induce venous thrombosis in vivo. SUMMARY: Background There is general consensus that the antiphospholipid syndrome (APS) is caused by antiphospholipid antibodies (aPL) with antibodies against ß2-glycoprotein-I being the most relevant. aPL that bind phospholipids in the absence of protein cofactors are generally considered pathogenetically irrelevant. We showed that cofactor-independent human monoclonal aPL isolated from APS patients induce proinflammatory and procoagulant cellular responses by activating endosomal NADPH-oxidase 2 (NOX2). Similar aPL were detected in all IgG fractions from APS patients analyzed. Objectives We aimed to clarify if cofactor-independent aPL can be thrombogenic in vivo and, if so, whether these effects are mediated via activation of NOX2. Methods Two cofactor-independent human monoclonal aPL, HL5B and RR7F, were tested in a mouse model of venous thrombosis. Genetically modified mice and in vitro assays were used to delineate the mechanisms underlying thrombus induction. Results HL5B and RR7F dramatically accelerate thrombus formation in this mouse model. Thrombus formation depends on tissue factor activation. It cannot be induced in NOX2-deficient mice. Bone marrow chimeras of C57BL/6J mice reconstituted with NOX2-deficient bone marrow showed that leukocyte activation plays a major role in thrombus formation. Neither TLR4 signaling nor platelet activation by our aPL is required for venous thrombus formation. Conclusions Cofactor-independent aPL can induce thrombosis in vivo. This effect is mainly mediated by leukocyte activation, which depends on the previously described signal transduction via endosomal NOX2. Because most APS patients have been shown to harbor aPL with similar activity, our data are of general relevance for the APS.

Improvement of Sporobolomyces ruberrimus carotenoids production by the use of raw glycerol.

The red yeast Sporobolomyces ruberrimus H110 was able to use glycerol as a carbon source. The highest concentration (0.51gL(-1)) and productivity (0.0064gL(-1)h(-1)) of carotenoids were achieved when raw glycerol from biodiesel production, containing around 1gL(-1) of fatty acids, was used as the carbon source, which represented increases of 27% and 1.5×, respectively, in relation to pure glycerol. Mass spectrometry analysis led to the identification of four carotenoids in the fermented samples, torularhodin, torulene, ß-carotene and γ-carotene. The use of raw glycerol also enhanced the proportion of torularhodin (69% against 59% in pure glycerol). The addition of individual fatty acids (palmitic, stearic, oleic and linoleic acids) to pure glycerol resulted in increases between 15% and 25% in maximum concentration and between 1.6× and 2.0× in productivity of carotenoids. The presence of palmitic and oleic acids increased the torularhodin proportion to 66%.

A novel indirect ELISA based on glycoprotein Gn for the detection of IgG antibodies against Rift Valley fever virus in small ruminants.

Rift Valley fever virus (RVFV) is an emerging zoonotic pathogen that causes high morbidity and mortality in humans and livestock. In this paper, we describe the cloning, expression and purification of RVFV glycoprotein Gn and its application as a diagnostic antigen in an indirect ELISA for the specific detection of RVF IgG antibodies in sheep and goats. The performance of this Gn based ELISA is validated using a panel of almost 2000 field samples from sheep and goats from Mozambique, Senegal, Uganda and Yemen. All serum samples were also tested by virus neutralization test (VNT), the gold standard method for RVFV serological testing. Compared to the VNT results the Gn based ELISA proved to have an excellent sensitivity (94.56%) and specificity (95.57%). Apart from establishing this new diagnostic assay, these results also demonstrate a close correlation between the presence of RVFV Gn and neutralizing antibodies.

Molecular and serological studies on the Rift Valley fever outbreak in Mauritania in 2010.

Rift Valley fever virus (RVFV) is a vector-borne RNA virus affecting humans, livestock and wildlife. In October/November 2010, after a period of unusually heavy rainfall, a Rift Valley fever outbreak occurred in northern Mauritania causing clinical cases in cattle, sheep, goats and camels, 21 of which were of lethal outcome. The aim of this study was to obtain further information on the continuation of RVF virus activity and spread in animal species in Mauritania after this outbreak. We therefore tested sera from small ruminants, cattle and camels for the presence of viral RNA and antibodies against RVFV. These sera were collected in different parts of the country from December 2010 to February 2011 and tested with three different ELISAs and an indirect immunofluorescence assay. The results show a high seroprevalence of RVFV IgM and IgG antibodies of about 57% in all animals investigated. Moreover, in four camel sera, viral RNA was detected emphasizing the important role camels played during the latest RVF outbreak in Mauritania. The study demonstrates the continuous spread of RVFV in Mauritania after initial emergence and highlights the potential role of small ruminants and camels in virus dissemination.

Spatially variable retardation plate for beam brightness enhancement in a high-power laser.

We present a simple and robust method for brightness enhancement, efficiently transforming a radially polarized LG (0,1)(*) mode into a nearly Gaussian beam of much higher quality. We use for this a spatially variable retardation plate and a spatial filter. The analysis shows that the transformation yields an increase in brightness by a factor of 3.4. In the experiment, we transformed a high-power Nd:YAG radially polarized (0,1)(*) LG beam with power of 70 W and M(2)=2.6 into a nearly Gaussian beam with M(2)=1.36. This resulted in brightness enhancement by a factor of 2.6.

Efficient extracavity generation of radially and azimuthally polarized beams.

We demonstrate an efficient transformation of a linearly polarized Gaussian beam to a radially or an azimuthally polarized doughnut (0,1)* Laguerre-Gaussian beam of high purity. We use a spatially variable retardation plate, composed of eight sectors of a lambda/2 retardation plate, to transform a linear polarization distribution to radial/azimuthal distribution. We transformed an Nd:YAG Gaussian beam with M(2)=1.3 to a radially and azimuthally polarized (0,1)* Laguerre-Gaussian beams with M(2)=2.5 and degree of radial/azimuthal polarization of 96-98%.

Efficient conversion of a radially polarized beam to a nearly Gaussian beam.

We demonstrate an efficient method for transformation of a radially polarized Laguerre-Gaussian beam to a nearly Gaussian beam with much higher beam quality. The method is based on separation of the radially polarized mode into two degenerate modes and coherent addition of the modes after phase flattening. We transformed a high-power Nd:YAG radially polarized (0,1)(*) Laguerre-Gaussian beam with M(2)=2.52 and power of 30 W into a nearly Gaussian beam with M(2)=1.3. As a result, the brightness increased by a factor of approximately 2.5.

Prognostic value of genomic alterations in minimal residual cancer cells purified from the blood of breast cancer patients.

The prognostic value of disseminated tumour cells derived from 353 breast cancer patients was evaluated. Disseminated tumour cells were purified from blood using a newly established method and nucleic acids were subsequently isolated. We investigated genomic imbalances (GI) such as mutation, amplification and loss of heterozygosity of 13 tumour suppressor genes and 2 proto-oncogenes using DNA from isolated minimal residual cancer cells. Significant correlations were found between genomic alterations of the DCC - and c-erbB-2 genes in disseminated breast cancer cells and actuarial relapse-free survival. Furthermore, increasing numbers of genomic imbalances measured in disseminated tumour cells were significantly associated with worse prognosis of recurrent disease. Logistic regression and Cox multivariate analysis led to the identification of genomic imbalances as an independent prognostic factor. Determination of disseminated tumour cells by genotyping of oncogenes and tumour suppressor genes seems not only to be a useful adjunct in follow up of carcinoma patients but provides also valuable additional individualized prognostic and predictive information in breast cancer patients beyond the TNM system.

Genetic analysis of immunomodulating factors in sporadic Parkinson's disease.

Immunomodulating factors have been shown to play a role in the pathogenesis of Parkinson's disease (PD) by biochemical methods. In order to investigate functionally important genes of the tumor necrosis factor alpha (TNFalpha) pathway we studied the frequency of DNA polymorphisms in the interleukin 6 (IL6), the TNFalpha, and the TNFalpha receptor 1 (TNFR1) genes in 264 sporadic German PD patients and in 183 age and sex matched German healthy controls. Analyzing the TNFalpha-308 polymorphism we found heterozygous individuals carrying alleles 1 and 2 more frequently in patients with a relative risk of 1.56 (p = 0.046, p(c) = 0.13, chi2 = 3.98). In contrast, the frequency of the B/2 haplotype described by the TNFR1-609 and TNFRI+36 polymorphisms was significantly decreased in our PD patients group (p = 0.0097, p(c) = 0.048, chi2 = 6.69) with a relative risk reduced to 0.52. Our results suggest an involvement of immunomodulating factors in the pathogenesis of sporadic PD as revealed by a molecular genetic approach.

Lack of association between the interferon regulatory factor-1 (IRF1) locus at 5q31.1 and multiple sclerosis in Germany, northern Italy, Sardinia and Sweden.

Interferon regulatory factor-1 (IRF-1) is a transcriptional inducer of the interferon-beta (IFN-beta) gene and other interferon-stimulated genes. A GT repeat polymorphism in the 7th intron of the IRF-1 gene was used as a marker to test for association with multiple sclerosis (MS) in a case-control study including individuals from Germany, Northern Italy and Sweden. In none of these populations, did we find any significant allelic association with disease. This lack of association was confirmed by testing transmission disequilibrium of individual IRF1 alleles in a representative sample of Sardinian simplex MS families. No deviation of the expected 50% transmission rates was seen. Therefore, our work does not provide evidence in favor of IRF1 being a candidate for conferring genetic susceptibility to, or protection against, MS in Europe.

Thermally boosted pumping of neodymium lasers.

Pumping at 885 nm from thermally excited ground-state levels directly to the Nd:YAG upper lasing level is experimentally demonstrated by use of a Ti:sapphire pump laser. This approach utilizes thermal energy contained within the laser medium to provide part of the pump energy required to achieve population inversion. Slope efficiency increased by 12% compared with traditional pump band excitation (lambda(pump) = 808 nm) and by 7% compared with ground-state direct pumping (lambda(pump) = 869 nm). The combined transition from the first and second thermally excited Stark components of the ground state (4I(9/2)) to the upper lasing level (4F(3/2)) has characteristics that make thermally boosted pumping a suitable candidate for use with diode lasers: reasonable absorption (1.8 cm(-1)) and bandwidth (2.7 nm FWHM). A model suggests that, compared with traditional 808-nm pumping, heat could be reduced by 40% by use of thermally boosted pumping.

Correction of birefringence and thermal lensing in nonreciprocal resonators by use of a dynamic imaging mirror.

Enhanced correction of thermally induced birefringence in the presence of strong single-pass, azimuthally dependent bipolar focusing was achieved in single-rod laser oscillators by use of an adaptive optic rear mirror with image relay and aberration correction capabilities. Together with a Faraday rotator, the imaging variable radius mirror was successfully tested in stable and unstable Nd:Cr:GSGG power oscillators under variable pump power conditions from 0 to 800 W. Birefringence correction in the absence of ray retracing was achieved.

Analysis of an IFN-gamma gene (IFNG) polymorphism in multiple sclerosis in Europe: effect of population structure on association with disease.

An intronic dinucleotide polymorphism in the IFN-gamma gene (IFNG) was used as a marker for testing association with multiple sclerosis (MS). Disease association was analyzed in case-control sets sampled from four geographically separate European populations (Germany, Northern Italy, Sardinia, and Sweden). Only in the Swedish was a weak disease association of the IFNG allele pattern found, mainly due to a higher frequency of IFNG allele I1 in MS patients. No evidence for association was found in the German or Northern Italian populations. These results contrast with the situation in Sardinia. We have recently reported transmission disequilibrium of IFNG allele I2 in Sardinian MS siblings not carrying the predisposing DRB1 *03 or *04 alleles (Ann. Neurol. 44, 841-842, 1998). Further analysis now shows that I2 is significantly more often transmitted to DRB1 *03-/*04- males, than to DRB1 *03-/*04- females. The odds ratio (OR) for IFNG-associated susceptibility to MS in the total Sardinian DRB1*03-/*04- group was 1.88 for I2 heterozygotes but amounted to 8.235 for I2 homozygotes, suggestive of a recessive mode of inheritance. Score test-based statistics pointed to an I2 allele dosage effect acting in susceptibility. Comparison of the IFNG allele frequencies in seven European populations (Northern Finnish, Southern Finnish, Swedish, Danish, German, Italian, and Sardinian) revealed a highly different distribution pattern. We introduced latitude as a score variable in order to test for trend in binomial proportions. This test statistic showed that for both most common alleles, I1 and I2 (compiled allele frequency about 85%), a significant opposite north-to-south trend is seen throughout Europe. This effect is primarily due to the extreme values found in the outlier populations of Finland and Sardinia. Our findings are discussed with respect to recent literature pertinent to the role of the IFNG chromosome region in autoimmune diseases.

Efficient pumping scheme for neodymium-doped materials by direct excitation of the upper lasing level.

An efficient pumping scheme that involves direct excitation of the upper lasing level of the Nd(3+) ion is demonstrated experimentally. The results obtained for direct upper laser level pumping of Nd:YAG R2 (869 nm) and Nd:YVO(4) (880 nm) were compared with traditional approximately 808-nm pump band excitation. A tunable cw Ti:sapphire laser was used as the pump source. In Nd:YAG, the oscillator slope efficiency increased by 10% and the threshold decreased by 11%. In Nd:YVO(4), the slope efficiency increased by 5% and the threshold decreased by 11%. These results agree with theory. The increase in optical efficiency indicates that laser material thermal loading can be substantially reduced.

Working beyond the static limits of laser stability by use of adaptive and polarization-conjugation optics.

Strong thermo-optical aberrations in flash-lamp-pumped Nd:Cr:GSGG rods were corrected to yield TEM(00) output at twice the efficiency of Nd:YAG. A hemispherical resonator operating at the limit of stability was employed. As much as 3 W of average power in a Gaussian beam (M(2) approximately 1) was generated. Unique features were zero warm-up time and the ability to vary the repetition rate without varying energy, near- and far-field profiles, or polarization purity. Thermal focusing and astigmatism were corrected with a microprocessor-controlled adaptive-optics backmirror composed of discrete elements (variable-radius mirror). A reentrant resonator coupled polarizer losses back into the laser rod and corrected depolarization.

Dynamic Correction of Thermal Focusing in Nd:YAG Confocal Unstable Resonators by Use of a Variable Radius Mirror.

Excellent beam quality and divergence stability over a wide pump power range was demonstrated in a Q-switched, Nd:YAG, positive branch confocal unstable resonator by using a one degree-of-freedom, adaptive optic. Unlike single-element flexible-membrane mirrors, the variable radius mirror (VRM) consisted of a lens and mirror, whose separation determined the VRM's effective radius of curvature. This simple method enabled low cost and efficient thermal focusing compensation. The VRM was demonstrated by producing a 300-mJ Q-switch or 1-J free-running at a beam quality factor M(2) that varied between 1.2 and 1.8 as the average output power varied between 0 and 33 W.

Genetic predisposition to multiple sclerosis as revealed by immunoprinting.

This study was designed to examine the immunogenetic background predisposing to multiple sclerosis (MS). Three hundred fifty-eight clinically well-characterized MS patients from Germany were investigated and compared to 395 healthy control subjects. Each individual was genotyped for 22 polymorphic markers located within or close to immunorelevant candidate genes including HLA-DRB1*, T-cell receptor (TCR), cell interaction molecules, cytokines, and cytokine receptor genes. Altogether, approximately 17,000 genetic analyses were performed. Patients were grouped according to the course of MS-relapsing-remitting or chronic progressive. Most of the genetic markers were not associated with increased risk or their exact contribution was not clear (e.g., tumor necrosis factor). The relative risks for HLA-DRB1*15+ and DRB1*03+ individuals were 3.64 and 1.42, respectively. In both groups of patients, certain TCRB gene polymorphisms were risk factors. In DRB1*03+ individuals the relative risk was increased (> 22) when a specific TCRBV6S3 allele was also inherited. Furthermore, distinct linkage disequilibria of TCRBV6S1/TCRBV6S3 elements in patients and control subjects strongly suggested an additional risk factor in the TCRBV region for DRB1*15+ individuals. These findings are discussed with respect to the pathogenesis and rational approaches to the therapy of MS.

Dose response study of enrofloxacin against Riemerella anatipestifer septicaemia in Muscovy and Pekin ducklings.

The efficacy of enrofloxacin against Riemerella anatipestifer (formerly Pasteurella anatipestifer) septicaemia of Muscovy and Pekin ducklings was assessed in an artificial challenge model which reproduced typical duck riemerellosis with very high mortality. Mortality, clinical signs, gross lesions, microbiological clearance, feed intake and body weight gain were the efficacy criteria studied. A pulsing medication in drinking water for 4 h at 25, 50 or 100 ppm enrofloxacin on the first day, followed by 12.5, 25 or 50 ppm, respectively, on the following 4 days, provided clinical cure of R. anatipestifer septicaemia at all dosages tested. An optimum dosage of 50 ppm enrofloxacin, followed by 25 ppm was established.

Channeling of terawatt laser pulses by use of hollow waveguides.

Subpicosecond laser pulses at power levels in excess of 1 TW were channeled through hollow microcapillary tubes by use of a combination of grazing-incidence dielectric and plasma-wall reflection mechanisms. Maximum input and output intensities were 10(17) and 10(16) W/cm(2) through 50-microm radius by 3-cm-long glass microcapillary tubes with as few as two waveguide modes being excited. 133-microm radius tubes as long as 13 cm resulted in successful channeling with an extinction coefficient of 0.2 cm(-1) and a plasma-wall reflectivity of 80%.