Membrane-associated proteins of ejaculated sperm from Morada Nova rams

AbstractThe objective was to describe the profile of membrane proteins from sperm of tropically adapted Morada Nova rams (N = 5). Samples from protein-enriched fractions of ejaculated sperm (containing 400 μg of protein) were separated by two-dimensional electrophoresis and respective maps analyzed using PDQuest software (version 7.3.0; Bio-Rad). Proteins were identified using tandem mass spectrometry. Also, membrane proteins were incubated with antibodies against binder of sperm protein (BSP) 1 and bodhesin 2 (Bdh-2), components of vesicular gland secretion. For membrane proteins of ejaculated sperm, an average of 133 ± 4.6 spots were detected per gel, of which, 107 spots were consistently present on all gels. Sixty-eight spots and 37 proteins were identified using mass spectrometry, corresponding to 71.6% of the intensity of all spots detected. Three major spots identified as ram seminal vesicle protein (RSVP) 14 represented approximately 30% of the intensity of all spots. Two of the most intense spots in the gel reacted against anti-BSP1, at 14 kDa. In addition, four low molecular weight spots reacted with anti-Bdh-2 antibodies. Proteins RSVP and Bdh-2 belong to the BSP and spermadhesin families, respectively, and were previously reported as major components of ram seminal proteins. Additional proteins identified in the sperm membrane two-dimensional maps included alpha-2-heparan sulfate-glycoprotein, plasma glutamate carboxypeptidase, arylsulfatase A, cathelicidin, heat shock protein 70 kDa, angiotensin-converting enzyme, leucine aminopeptidase, and clusterin. Some proteins were present as multiple isoforms, such as tubulin (12), alpha-2-heparan sulfate-glycoprotein (5), ATP synthase (5), Bdh-2 (4) and RSVP14 (3). Based on gene ontology analysis, the most common biological processes associated with the membrane proteins were cellular processes (34%), response to stimulus (14%), and metabolic processes (11%). Binding (37%) and catalytic activity (32%) corresponded to the most frequent molecular functions for those proteins. In conclusion, we identified a diverse cohort of components of membrane proteins in ram sperm. Major proteins previously reported in seminal plasma, such as RSVP14 and Bdh-2, were also extracted from sperm membranes. Knowledge of sperm proteins is crucial for elucidating mechanisms underlying their association with sperm function

The Complex Topology of Chemical Plants

We show that flowsheets of oil refineries can be associated to complex network topologies that are scale-free, display small-world effect and possess hierarchical organization. The emergence of these properties from such man-made networks is explained as a consequence of the currently used principles for process design, which include heuristics as well as algorithmic techniques. We expect these results to be valid for chemical plants of different types and capacities.Comment: 7 pages, 5 figures and 1 tabl

Simple heuristics for the assembly line worker assignment and balancing problem

We propose simple heuristics for the assembly line worker assignment and balancing problem. This problem typically occurs in assembly lines in sheltered work centers for the disabled. Different from the classical simple assembly line balancing problem, the task execution times vary according to the assigned worker. We develop a constructive heuristic framework based on task and worker priority rules defining the order in which the tasks and workers should be assigned to the workstations. We present a number of such rules and compare their performance across three possible uses: as a stand-alone method, as an initial solution generator for meta-heuristics, and as a decoder for a hybrid genetic algorithm. Our results show that the heuristics are fast, they obtain good results as a stand-alone method and are efficient when used as a initial solution generator or as a solution decoder within more elaborate approaches.Comment: 18 pages, 1 figur

Comparison of blood neoangiogenesis and lymphatic vascularization in colorectal adenomas from patients with and without concomitant colorectal cancer

Blood and lymphatic vessel proliferation is essential for tumor growth and progression. Most colorectal carcinomas develop from adenomas (adenoma-carcinoma sequence) in a process due to accumulation of molecular genetic alterations. About 5% of adenomatous polyps are expected to become malignant, but data on the differential angiogenic patterns of these lesions in patients with and without concomitant cancer are missing. The aim of the present study is to compare the angiogenic and lymphatic patterns of adenomatous polyps from patients with and without sporadic cancer. Thirty adenomatous polyps (15 from patients with another principal malignant lesion, and 15 from patients without cancer) were submitted to immunohistochemical staining for CD105 (marker for neoangiogenesis) and D2-40 (marker for lymphatic endothelium). Microvessel density and total vascular area were determined by computer image analysis to quantify the immunostained and total areas, and to assess the number of microvessels. Adenomas from patients with carcinoma showed significantly higher values of total vascular area determined by immunostaining for CD105 (cutoff value = 4386 µm²; P = 0.019) and of lymphatic microvessel density determined by immunostaining with D2-40 (cutoff value = 11.5; P = 0.041) when compared with those from patients without cancer. The present data indicate a significant increase in blood microvascular area and in lymphatic microvascular counts in adenomas removed from patients with cancer.59359

Optical Tweezers 3d Photonic Force Spectroscopy

Since optical tweezers trapped microspheres can be used as an ultrasensitive force measurements technique, the knowledge of its theoretical description is of utmost importance. However, even the description of the incident electromagnetic fields under very tight focusing, typical of the optical trap, is not yet a closed problem. Therefore it is important to experimentally obtain whole accurate curves of the force as a function of wavelength, polarization and incident beam 3D position with respect to the center of the microsphere. Theoretical models for optical forces such as the Generalized Lorenz-Mie theory, can then be applied to the precisely evaluated experimental results. Using a dual trap in an upright standard optical microscope, one to keep the particle at the equilibrium position and the other to disturb it we have been able to obtain these force curves as a function of x, y and z position, incident beam polarization and also wavelength. Further investigation of optical forces was conducted for wavelengths in and out Mie resonances of the dielectric microspherical cavities for both TM and TE modes.6131Ashkin, A., Dziedzic, J.M., Bjorkholm, J.E., Chu, S., Observation of a single-beam gradient force trap for dielectric particles (1986) Opt. Lett., 11, pp. 288-290Ashkin, A., Dziedzic, J.M., Optical trapping and manipulation of viruses and bacteria (1987) Science, 235, pp. 1517-1520Grier, D.G., A revolution in optical manipulation (2003) Nature, 424, pp. 810-816Neuman, K.C., Block, S., Optical trapping (2004) Rev. Sci. Instrum., 75, pp. 2787-2809Lock, J.A., Calculation of the radiation trapping force for laser tweezers by use of generalized Lorenz-Mie theory. I. Localized model description of an on-axis tightly focused laser beam with spherical aberration (2004) Appl. Opt., 43, pp. 2532-2544Lock, J.A., Calculation of the radiation trapping force for laser tweezers by use of generalized Lorenz-Mie theory. II. On-axis trapping force (2004) Appl. Opt., 43, pp. 2545-2554Mazolli, A., Neto, P.A.M., Nussenzveig, H.M., Theory of trapping forces in optical tweezers (2003) Proc. Royal Soc. London Ser. A Math. Phys. Eng. Sci., 459, pp. 3021-3041Fontes, A., Neves, A.A.R., Moreira, W.L., De Thomaz, A.A., Barbosa, L.C., De Paula, A.M., Cesar, C.L., Double optical tweezers for ultrasensitive force spectroscopy in microsphere Mie scattering (2005) Appl. Phys. Lett., 87. , Art. No. 221109Ren, K.F., Gouesbet, G., Gréhan, G., Integral localized approximation in generalized Lorenz-Mie theory (1998) Appl. Opt., 37, pp. 4218-4225Lock, J.A., Excitation efficiency of a morphology-dependent resonance by a focused Gaussian beam (1998) J. Opt. Soc. Am. A, 15, pp. 2986-2994Davis, L.W., Theory of electromagnetic beams (1979) Phys. Rev. A, 19, pp. 1177-1779Ren, K.F., Gréhan, G., Gouesbet, G., Radiation pressure forces exerted on a particle arbitrarily located in a gaussian beam by using the generalized Lorenz-Mie theory and associated resonance effects (1994) Opt. Commun., 108, pp. 343-354Ren, K.F., Gréhan, G., Gouesbet, G., Symmetry relations in generalized Lorenz-Mie theory (1994) J. Opt. Soc. Am. A, 11, pp. 1812-181

Force Spectroscopy And Two Photon Excited Luminescence In An Optical Tweezers System

Up to now optical spectroscopies have analyzed the scattered light or the heat generated by absorption as a function of the wavelength to get information about the samples. Among the light matter interaction phenomena one that has almost never been used for spectroscopy is the direct photon momenta transfer. Probably because the forces involved are very small, varying from hundreds of femto to tens of pico Newtons. However, the nowadays very popular Optical Tweezers can easily accomplish the task to measure the photon momenta transfer and may be the basis for the Optical Force Spectroscopy. We demonstrate its potential as such a tool by observing more than eight Mie resonance peaks of a single polystyrene microsphere, and showed the capability to selective couple the light to either the TE, TM or both microsphere modes depending of the beam size, the light polarization and the beam positioning. The Mie resonances can change the optical force values by 30-50%. Our results also clearly show how the beam polarization breaks the usually assumed azimuthal symmetry by Optical Tweezers theories. We also obtained the spectrum from the two photon excited luminescence using the Optical Tweezers to hold a single bead suspended and a femtosecond Ti:sapphire laser for the non-linear excitation. This spectrum shows the pair of peaks due to both TE and TM spherical cavity modes. We have been able to observe more than 14 Mie resonance peaks in the TPE luminescence. Our results are in good agreement with optical force calculations using Maxwell stress tensor and partial wave decomposition of the incident beam approximated to a 3th order gaussian beam.593017Ashkin, Dziedzic, J.M., Bjorkholm, J.E., Chu, S., Observation of a single-beam gradient force trap for dielectric particles (1986) Opt. Lett., 11, pp. 288-290Ashkin, A., Dziedzic, J.M., Optical trapping and manipulation of viruses and bacteria (1987) Science, 235, pp. 1517-1520Grier, D.G., A revolution in optical manipulation (2003) Nature, 424, pp. 810-816Felgner, H., Frank, R., Schliwa, M., Flexural rigidity of microtubules measured with the use of optical tweezers (1996) J. Cell Sci., 109, pp. 509-516Sakata-Sogawa, K., Kurachi, M., Sogawa, K., Fujii-Kuriyama, Y., Tashiro, H., Direct measurement of DNA molecular length in solution using optical tweezers: Detection of looping due to binding protein interactions (1998) Eur. Biophys. J., 27, pp. 55-61Berg, H.C., Berry, R.M., Absence of a barrier to backwards rotation of the bacterial flagellar motor demonstrated with optical tweezers (1997) Proc. Natl. Acad. Sci. U.S.A., 94, pp. 14433-14437Huruta, R.R., Mechanical properties of stored red blood cells using optical tweezers (1998) Blood, 92, pp. 2975-2977Brandão, M.M., Optical tweezers for measuring red blood cell elasticity: Application to the study of drug response in sickle cell disease (2003) Eur. J. Haematol., 70, pp. 207-211Ashkin, A., Dziedzic, J.M., (1977) Phys. Rev. Lett., 38, pp. 1351-1355Barber, P.W., Chang, R.K., (1988) Optical Effects Associated with Small Particles, , Word Scientific, SingaporeVan De Hulst, H.C., (1981) Light Scattering by Small Particles, , Dover, New YorkAshkin, A., Dziedzic, J.M., Observation of optical of dieletric particles by light-scattering (1981) Appl. Optics, 20, pp. 1803-1814Benner, R.E., Barber, P.W., Owe, J.F., Chang, R.K., Observation of structure resonances in the fluorescence spectra from microspheres (1980) Physical Review Letters, 44, pp. 475-478Leung, C.H., She, T.C., Lee, W.K., Positions of low order morphology dependent resonances determined by elastic light scattering (1995) J. Opt. Soc. Am. B, 12, pp. 1259-1266Schaschek, K., Popp, J., Kiefer, W., Observation of morphology dependent in- And output-resonances in time dependent Raman spectra of optically levitated microdroplets (1993) J. Raman Spectrosc., 24, pp. 69-75Ashkin, A., Dziedzic, J.M., Observation of resonances in radiation pressure on dieletric particles (1977) Phys. Rev. Lett., 38, pp. 1351-1354Bisht, P.B., Fukuda, K., Hirayama, S., Steady-state and time-resolved fluorescence study of some dyes in polymer microspheres showing morphology dependent resonances (1996) J. Chem. Phys., 105, pp. 9349-9361Arnold, S., Shift of whispering gallery modes in microspheres by protein adsorption (2003) Opt. Lett., 28, pp. 272-274Vollmer, F., Multiplexed DNA quantification by spectroscopic shift of two microsphere cavities (2003) Biophys. J., 85, pp. 1974-1979Ren, K.F., Gréhan, G., Gouesbet, G., Radiation pressure forces exerted on a particle arbitrarily located in a gaussian beam by using the generalized Lorenz-Mie theory and associated resonance effects (1994) Opt. Commun., 108, pp. 343-354Ren, K.F., Gréhan, G., Gouesbet, G., Symmetry relations in generalized Lorenz-Mie theory (1994) J. Opt. Soc. Am. A, 11, pp. 1812-1817Bohren, C.F., Huffman, D.R., (1983) Absorption and Scattering of Light by Small Particles, , Wiley, New YorkVan De Hulst, H.C., (1981) Light Scattering by Small Particles, , Dover, New YorkDavis, L.W., Theory of electromagnetic beams (1979) Phys. Rev. A, 19, pp. 1177-1779Richards, B., Wolf, E., Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system (1959) Proc. R. Soc. London A, 253, pp. 358-379Ren, K.F., Gouesbet, G., Gréhan, G., Integral localized approximation in generalized Lorenz-Mie theory (1998) Appl. Opt., 37, pp. 4218-4225Lock, J.A., Excitation efficiency of a morphology-dependent resonance by a focused Gaussian beam (1998) J. Opt. Soc. Am. A, 15, pp. 2986-2994Bateman, J.B., Weneck, E.J., Eshler, D.C., Determination of Particle Size and Concentration from Spectrophotometric Transmission (1959) J. Colloid Sci., 14, pp. 308-32

Stationary phase corrections in the process of bosonization of multi-quark interactions

The functional integration over the auxiliary bosonic variables of cubic order related with the effective action of the Nambu -- Jona-Lasinio model with 't Hooft term has recently been obtained in the form of a loop expansion. Even numbers of loops contribute to the action, while odd numbers of loops are assigned to the measure. We consider the two-loop corrections and analyse their effect on the low-lying pseudoscalar and scalar mass spectra, quark condensates and weak decay constants. The results are compared to the leading order calculations and other approaches.Comment: 22 pages, LaTeX, to appear in European Physics Journal

Adsortividade do herbicida "Zeta" em solos de variadas texturas

To estimate the herbicide rate which inhibits 50% (I50) of the growth of bioindicator plants in soils of different textures and organic matter levels, which could interfere on the bioactivity of the tested herbicide, a completely randomized experiment was carried out with seven treatments (0, 50, 100, 150, 200, 250 and 300 g a.i./ ha) and four replications, in five kinds of soil textures. The analyzed parameters were: total fresh biomass, dry shoot biomass and dry root biomass of the sorghum plants. According to the results, it can be concluded that in sandy soils the herbicide shows a great vertical mobility, and that high levels of AP+ in the soil can minimize the herbicide effect on plants. The experiment also has shown that the higher the level of active clays and of organic matter in the soil, the higher the rate of herbicide required for a given weed control level.Com o objetivo de estimar a dose do herbicida "Zeta" que inibe 50% I50) do crescimento de plantas bioindicadoras em solos com diferentes texturas e teores de matéria orgânica, podendo estes ter uma influência marcante sobre a bioatividade do herbicida testado, utilizou-se o delineamento experimental inteiramente casualizado com 7 tratamentos (0, 50, 100, 150, 200, 250 e 300 g i.a./ha) e 4 repetições, sendo os tratamentos dispostos em 5 tipos de texturas de solo. Os parâmetros avaliados foram: biomassa fresca total, biomassa seca da parte aérea e biomassa seca do sistema radicular das plantas de sorgo. De acordo com os resultados obtidos pode-se concluir que em solos com textura tendendo à arenosa o herbicida apresenta uma grande mobilidade vertical, e que altos teores de AP+ no solo podem mascarar, de certa forma, a atuação do herbicida na planta. Em linhas gerais, o experimento permitiu inferir que quanto maior os teores de argilas ativas e de matéria orgânica presentes no solo, maior a dose do herbicida requerida para um dado controle percentual de plantas daninhas