Photo-induced structural dynamics of o-nitrophenol by ultrafast electron diffraction.

The photo-induced dynamics of o-nitrophenol, particularly its photolysis, has garnered significant scientific interest as a potential source of nitrous acid in the atmosphere. Although the photolysis products and preceding photo-induced electronic structure dynamics have been investigated extensively, the nuclear dynamics accompanying the non-radiative relaxation of o-nitrophenol on the ultrafast timescale, which include an intramolecular proton transfer step, have not been experimentally resolved. Herein, we present a direct observation of the ultrafast nuclear motions mediating photo-relaxation using ultrafast electron diffraction. This work spatiotemporally resolves the loss of planarity which enables access to a conical intersection between the first excited state and the ground state after the proton transfer step, on the femtosecond timescale and with sub-Angstrom resolution. Our observations, supported by ab initio multiple spawning simulations, provide new insights into the proton transfer mediated relaxation mechanism in o-nitrophenol.

Relativistic ultrafast electron diffraction at high repetition rates.

The ability to resolve the dynamics of matter on its native temporal and spatial scales constitutes a key challenge and convergent theme across chemistry, biology, and materials science. The last couple of decades have witnessed ultrafast electron diffraction (UED) emerge as one of the forefront techniques with the sensitivity to resolve atomic motions. Increasingly sophisticated UED instruments are being developed that are aimed at increasing the beam brightness in order to observe structural signatures, but so far they have been limited to low average current beams. Here, we present the technical design and capabilities of the HiRES (High Repetition-rate Electron Scattering) instrument, which blends relativistic electrons and high repetition rates to achieve orders of magnitude improvement in average beam current compared to the existing state-of-the-art instruments. The setup utilizes a novel electron source to deliver femtosecond duration electron pulses at up to MHz repetition rates for UED experiments. Instrument response function of sub-500 fs is demonstrated with < 100 fs time resolution targeted in future. We provide example cases of diffraction measurements on solid-state and gas-phase samples, including both micro- and nanodiffraction (featuring 100 nm beam size) modes, which showcase the potential of the instrument for novel UED experiments.

Rehybridization dynamics into the pericyclic minimum of an electrocyclic reaction imaged in real-time.

Electrocyclic reactions are characterized by the concerted formation and cleavage of both σ and π bonds through a cyclic structure. This structure is known as a pericyclic transition state for thermal reactions and a pericyclic minimum in the excited state for photochemical reactions. However, the structure of the pericyclic geometry has yet to be observed experimentally. We use a combination of ultrafast electron diffraction and excited state wavepacket simulations to image structural dynamics through the pericyclic minimum of a photochemical electrocyclic ring-opening reaction in the molecule α-terpinene. The structural motion into the pericyclic minimum is dominated by rehybridization of two carbon atoms, which is required for the transformation from two to three conjugated π bonds. The σ bond dissociation largely happens after internal conversion from the pericyclic minimum to the electronic ground state. These findings may be transferrable to electrocyclic reactions in general.

Conformer-specific photochemistry imaged in real space and time.

Conformational isomers (conformers) of molecules play a decisive role in biology and organic chemistry. However, experimental methods for investigating chemical reaction dynamics are typically not conformer-sensitive. We report on a gas-phase megaelectronvolt ultrafast electron diffraction investigation of α-phellandrene undergoing an electrocyclic ring-opening reaction. We directly imaged the evolution of a specific set of α-phellandrene conformers into the product isomer predicted by the Woodward-Hoffmann rules in real space and time. Our experimental results are in quantitative agreement with nonadiabatic quantum molecular dynamics simulations, which provide considerable detail of how conformation influences the time scale and quantum efficiency of photoinduced ring-opening reactions.

Liquid-phase mega-electron-volt ultrafast electron diffraction.

The conversion of light into usable chemical and mechanical energy is pivotal to several biological and chemical processes, many of which occur in solution. To understand the structure-function relationships mediating these processes, a technique with high spatial and temporal resolutions is required. Here, we report on the design and commissioning of a liquid-phase mega-electron-volt (MeV) ultrafast electron diffraction instrument for the study of structural dynamics in solution. Limitations posed by the shallow penetration depth of electrons and the resulting information loss due to multiple scattering and the technical challenge of delivering liquids to vacuum were overcome through the use of MeV electrons and a gas-accelerated thin liquid sheet jet. To demonstrate the capabilities of this instrument, the structure of water and its network were resolved up to the 3 rd hydration shell with a spatial resolution of 0.6 Å; preliminary time-resolved experiments demonstrated a temporal resolution of 200 fs.

Photodissociation of aqueous I 3 - observed with liquid-phase ultrafast mega-electron-volt electron diffraction.

Developing femtosecond resolution methods for directly observing structural dynamics is critical to understanding complex photochemical reaction mechanisms in solution. We have used two recent developments, ultrafast mega-electron-volt electron sources and vacuum compatible sub-micron thick liquid sheet jets, to enable liquid-phase ultrafast electron diffraction (LUED). We have demonstrated the viability of LUED by investigating the photodissociation of tri-iodide initiated with a 400 nm laser pulse. This has enabled the average speed of the bond expansion to be measured during the first 750 fs of dissociation and the geminate recombination to be directly captured on the picosecond time scale.

The photochemical ring-opening of 1,3-cyclohexadiene imaged by ultrafast electron diffraction.

The ultrafast photoinduced ring-opening of 1,3-cyclohexadiene constitutes a textbook example of electrocyclic reactions in organic chemistry and a model for photobiological reactions in vitamin D synthesis. Although the relaxation from the photoexcited electronic state during the ring-opening has been investigated in numerous studies, the accompanying changes in atomic distance have not been resolved. Here we present a direct and unambiguous observation of the ring-opening reaction path on the femtosecond timescale and subångström length scale using megaelectronvolt ultrafast electron diffraction. We followed the carbon-carbon bond dissociation and the structural opening of the 1,3-cyclohexadiene ring by the direct measurement of time-dependent changes in the distribution of interatomic distances. We observed a substantial acceleration of the ring-opening motion after internal conversion to the ground state due to a steepening of the electronic potential gradient towards the product minima. The ring-opening motion transforms into rotation of the terminal ethylene groups in the photoproduct 1,3,5-hexatriene on the subpicosecond timescale.

Implementation and modeling of a femtosecond laser-activated streak camera.

A laser-activated streak camera was built to measure the duration of femtosecond electron pulses. The streak velocity of the device is 1.89 mrad/ps, which corresponds to a sensitivity of 34.9 fs/pixels. The streak camera also measures changes in the relative time of arrival between the laser and electron pulses with a resolution of 70 fs RMS. A full circuit analysis of the structure is presented to describe the streaking field and the general behavior of the device. We have developed a general mathematical model to analyze the streaked images. The model provides an accurate method to extract the pulse duration based on the changes of the electron beam profile when the streaking field is applied.

Mega-electron-volt ultrafast electron diffraction at SLAC National Accelerator Laboratory.

Ultrafast electron probes are powerful tools, complementary to x-ray free-electron lasers, used to study structural dynamics in material, chemical, and biological sciences. High brightness, relativistic electron beams with femtosecond pulse duration can resolve details of the dynamic processes on atomic time and length scales. SLAC National Accelerator Laboratory recently launched the Ultrafast Electron Diffraction (UED) and microscopy Initiative aiming at developing the next generation ultrafast electron scattering instruments. As the first stage of the Initiative, a mega-electron-volt (MeV) UED system has been constructed and commissioned to serve ultrafast science experiments and instrumentation development. The system operates at 120-Hz repetition rate with outstanding performance. In this paper, we report on the SLAC MeV UED system and its performance, including the reciprocal space resolution, temporal resolution, and machine stability.

Expression and functionality of Toll-like receptor 3 in the megakaryocytic lineage.

BACKGROUND: In addition to their key role in hemostasis, platelets and megakaryocytes regulate immune and inflammatory responses, in part through their expression of Toll-like receptors (TLRs). Among the TLRs, TLR3 recognizes dsRNA associated with viral infection. Thrombocytopenia is a frequent complication of viral infection. However, the expression and functionality of TLR3 in megakaryocytes and platelets is not yet well understood. OBJECTIVE: To study the expression and functionality of TLR3 in the megakaryocytic lineage. METHODS AND RESULTS: RT-PCR, flow cytometric and immunofluorescence assays showed that TLR3 is expressed in CD34(+) cells, megakaryocytes, and platelets. Immunoblotting assays showed that stimulation of megakaryocytes with two synthetic agonists of TLR3, Poly(I:C) and Poly(A:U), activated the nuclear factor-κB (NF-κB), phosphoinositide 3-kinase (PI3K)/Akt, extracellular signal-related kinase (ERK)1/2 and p38 pathways. TLR3-megakaryocyte activation resulted in reduced platelet production in vitro and interferon-ß release through the PI3K-Akt and NF-κB signaling pathways. TLR3 ligands potentiated the aggregation mediated by classic platelet agonists. This effect was also observed for ATP release, but not for P-selectin or CD40L membrane exposure, indicating that TLR3 activation was not involved in α-granule release. In addition, TLR3 agonists induced activation of the NF-κB, PI3K-Akt and ERK1/2 pathways in platelets. Reductions in platelet production and platelet fibrinogen binding mediated by Poly(I:C) or Poly(A:U) were prevented by the presence of an inhibitor of the TLR3-dsRNA complex. CONCLUSIONS: Our findings indicate that functional TLR3 is expressed in CD34(+) cells, megakaryocytes, and platelets, and suggest a potential role for this receptor in the megakaryopoiesis/thrombopoiesis alterations that occur in viral infections.

Automatic flow-batch system for the sample treatment and determination of hydroxyproline in sausages.

In this study an automatic method for sample treatment and spectrophotometric determination of hydroxyproline in commercial sausages was developed. A flow-batch system that includes the steps of sample hydrolysis and determination of the analyte was designed. The method presents a linear range between 0.60 and 3.60 µg mL(-1) of hydroxyproline. A relative standard deviation of 1.68% was obtained from hydroxyproline standard solution (n=6, 1.20 µg mL(-1)) and the detection limit was 0.12 µg mL(-1). The sample throughput was 1 sample h(-1) while the reference method (AOAC) was carried out in about 17 h. This method employs 16 h of hydrolysis while in the proposed method the hydrolysis time was 15 min. For this purpose a pressure hydrolysis chamber with a low cost halogen lamp was used. The flow-batch system is simple and allows the use of both chambers (hydrolysis and detection) simultaneously. The obtained results with the flow-batch method are in good agreement with that obtained with the reference method. Moreover, it is good alternative to the quality control of meat products.

Picosecond imaging of low-density plasmas by electron deflectometry.

We have imaged optical-field ionized plasmas with electron densities as low as 10(13) cm(-3) on a picosecond timescale using ultrashort electron pulses. Electric fields generated by the separation of charges are imprinted on a 20 keV probe electron pulse and reveal a cloud of electrons expanding away from a positively charged plasma core. Our method allows for a direct measurement of the electron energy required to escape the plasma and the total charge. Simulations reproduce the main features of the experiment and allow determination of the energy of the electrons.

Perfil de resistencia de Staphylococcus spp aislados de hemocultivos en el Hospital Central del Instituto de Previsión Social / Resistance profile of Staphylococcus spp isolated from hemocultures in the Hospital Central of the Instituto de Prevision Social

Las infecciones nosocomiales por Staphylococcus spp constituyen uno de los problemas de mayor preocupación en salud pública en todo el mundo. Debido a que existen diferencias según centro hospitalario, paciente, área y tiempo de internación es esencial conocer el perfil de resistencia a los antibióticos más utilizados de las cepas de Staphylococcus involucradas en estas infecciones. El objetivo del presente estudio fue determinar el perfil de resistencia de los aislados de Staphylococcus aureus (Sau) y Estafilococo coagulasa negativo (ECN) de muestras de hemocultivos de pacientes adultos, pediátricos y recién nacidos internados entre el 1 de junio de 2005 al 30 de junio de 2006, en varios servicios del Hospital Central del Instituto de Previsión Social (IPS.).Se realizó un análisis retrospectivo de los resultados de antibiograma realizados en el Servicio de Microbiología del IPS. Los antibióticos evaluados fueron oxacilina (OXA), penicilina (PEN), eritromicina (ERY), clindamicina (CLI), ciprofloxacina (CIP), gentamicina (GEN), trimetoprim-sulfametoxazol (TMS), tetraciclina (TET) y vancomicina (VAN). De 5698 hemocultivos realizados en este periodo, 1706 (30%) fueron positivos. De estos, en 755 (44%) se aisló Staphylococcus spp; correspondiendo 327 (43,3%) a Sau y 428 (56,7%) a ECN. El perfil de resistencia para Sau y ECN fue respectivamente como sigue: OXA 64% y 87%, PEN 96% y 95%, ERY 48% y 52%, CIP 42% y 40%, GEN 48% y 48% y no presentaron R a VAN. El 58% de los Sau y el 56% de los ECN fueron resistentes a más de 4 antibióticos. La alta frecuencia de resistencia a la meticilina y la poliresistencia hallada en este trabajo deben ser tenidos en cuenta a la hora de indicar esquemas de antibióticos empíricos y para realizar ajustes oportunos de los ya iniciados. No se encontró ninguna cepa resistente a la vancomicina, que es considerada el mejor antimicrobiano disponible para el tratamiento de infecciones por estafilococos resistentes a las penicilinas que no son inhibidas por las penicilinasas.

Nosocomial infections due to Staphylococcus spp areone of the major concerns in public health all over the world. Because there are differences according to health center, patient, area and days of hospitalization, the knowledge of the resistance profile of Staphylococcus spp to the most common antibiotics is essential. The objective of the present study was to determine the resistance pattern of Staphylococcus aureus (Sau) and coagulase-negative staphylococci strains isolated from hemoculture of adult, pediatric and neonatal patients, hospitalized in the Hospital Central del Instituto de Previsión Social from June 1, 2005 to June 30, 2006. Antibiogram results carried out at the Laboratory of Microbiology of the IPS were retrospectively reviewed. The antibiotics evaluated were oxacillin (OXA), penicillin (PEN), erythromycin (ERY), clindamycin (CLI), ciprofloxacin (CIP), gentamicin (GEN), trimethoprim-sulphametoxazol (TMS), tetracycline (TET) and vancomycin (VAN). Of the 5,698 hemocultures carried out in this period, 1706 (30%) were positive. Staphylococcus spp was isolated in 755 (44%) samples; 327 (43.3%) of them corresponded to Sau and 428 (56.7%) to ECN. Resistance profile for Sau and ECN were as follows: OXA 64% and 87%, PEN 96% and 95%, ERY 48% and 52%, CIP 42% and 40%, GEN 48% and 48% and VAN did not present resistance to any of the antibiotics tested. Fifty eight percent of all Sau and 56% of ECN isolates were resistant to more than 4 antibiotics. The high frequency of penicillin resistance and poly resistance found in this study have to be taken into account at the time of choosing an empirical antibiotic scheme and for the adjustment of those already implemented. No isolates were resistant to vancomycin which is still the best antibiotic available for the treatment of infections due to penicillin resistant Staphylococcus that are inhibited by penicillinase.

Glomerulopatías en el Paraguay. Reporte del registro de biopsias renales en 1072 casos / Glomerulopathies in Paraguay. Report of registry of renal biopsies in 1072 cases

La biopsia renal es el principal método de diagnóstico de las glomerulopatías. El Registro de biopsias renales es importante para conocer la situación epidemiológica y clínico­patológica de las enfermedades renales de un país y las variaciones geográficas entre los países o continentes. Para conocer la situación actual de las glomerulopatías del Paraguay se ha realizado el presente estudio en 1.365 biopsias renales registradas entre agosto de 1989 a junio de 2005 en el Departamento de Patología del IICS, que es el Centro de Referencia y Registro de Biopsias Renales del país, analizando sus aspectos histológicos, demográficos y las diferencias entre periodos quinquenales de estudio. Del total de casos, 1072 (75.5%) fueron glomerulopatías y, a diferencia de la mayoría de los países, se observó un predominio de glomerulonefritis secundarias (GNS) (57.3%) de las cuales 85.9% fueron nefritis lúpica (NL). En las glomerulonefritis primarias (GNP) la GN proliferativa mesangial (GNPM) y la membranoproliferativa (GNMP) fueron las mas frecuentes (39.8% y 28.2%), mientras que la Nefropatía por IgA (NIgA) y la Enfermedad de cambios mínimos (ECM) fueron raras (1.9% y 1.3%). No se observaron cambios significativos de incremento o disminución de tipos histológicos a través de los periodos de estudio tanto en las GNP como en las GNS. La existencia de este registro de biopsias renales fue la base para la creación del Registro de Glomerulopatías del Paraguay en el año 2004, cuyos datos son necesarios para establecer programas de tratamiento y prevención de las enfermedades glomerulares en nuestro país a fin de disminuir su progresión al estadío terminal.

Renal biopsy is the main diagnostic method of glomerulopathies. The registry of renal biopsies is important to know the epidemiological and clinical and pathological situation of renal diseases of a country and the geographic variations between countries or continents. In order to know the current situation of glomerulopathies in our country, we studied 1,365 renal biopsies registered between August 1989 to June 2005 at the Pathology Department of the IICS, the reference center and the Renal Biopsy Registry of the country, analyzing histological and demographic aspects as well as the differences between five­year periods of study. On the total cases, 1,072 (75.5%) were glomerulopathies while secondary glomerulonephritis (SGN) were more frequent (57.3 %) with 85.9% of lupus nephritis (LN) in contrast to most countries. Among primary glomerulonephritis (PGN), mesangioproliferative GN (MPGN) and membranoproliferative GN (MPGN) were predominant (39.8% and 28.2%) while IgA Nephropathy (IgAN) and Minimal Change Disease (ECM) were rare (1.9% and 1.3%). We did not observe significant changes of increases or decreases of histological types through the study periods in PGN as well as in SGN. The existence of this registry of renal biopsy was the base for the creation of the Paraguayan Registry of Glomerulopathies in 2004, which data are necessary to establish treatment and prevention programs of glomerular diseases in our country in order to decrease their progression to end­stage renal disease.