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1.
J Biophotonics ; 16(9): e202300148, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37280718

RESUMEN

A dynamically tunable metal clad planar waveguide having 0.62PMN-0.38PT material is simulated and optimized for detection of cancer cells. Angular interrogation of the TE0 mode of waveguide shows that critical angle increases greater than the resonance angle with increasing of cover refractive index, which limits the detection range of waveguide. To overcome this limitation, proposed waveguide applies a potential on the PMN-PT adlayer. Although a sensitivity of 105.42 degree/RIU was achieved at 70 Volts in testing the proposed waveguide, it was found that the optimal performance parameters were obtained at 60 Volts. At this voltage, the waveguide demonstrated detection range 1.3330-1.5030, a detection accuracy 2393.33, and a figure of merit 2243.59 RIU-1 , which enabled the detection of the entire range of the targeted cancer cells. Therefore, it is recommended to apply a potential of 60 Volts to achieve the best performance from the proposed waveguide.


Asunto(s)
Metales , Neoplasias , Vibración , Neoplasias/diagnóstico por imagen
2.
Appl Opt ; 61(28): 8540-8552, 2022 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-36256172

RESUMEN

We report here the first implementation of chemically specific imaging in the exhaust plume of a gas turbine typical of those used for propulsion in commercial aircraft. The method used is chemical species tomography (CST) and the target species is CO2, absorbing in the near-infrared at 1999.4 nm. A total of 126 beams propagate transverse to the plume axis, along 7 m paths in a coplanar geometry, to probe a central region of diameter ≈1.5m. The CO2 absorption spectrum is measured using tunable diode laser spectroscopy with wavelength modulation, using the second harmonic to first harmonic (2f/1f) ratio method. The engine is operated over the full range of thrust, while data are recorded in a quasi-simultaneous mode at frame rates of 1.25 and 0.3125 Hz. Various data inversion methodologies are considered and presented for image reconstruction. At all thrust levels a persistent ring structure of high CO2 concentration is observed in the central region of the measurement plane, with a raised region in the middle of the plume assumed to be due to the engine's boat tail. With its potential to target various exhaust species, the CST method outlined here offers a new approach to turbine combustion research, turbine engine development, and aviation fuel research and development.

3.
Sci Rep ; 12(1): 3114, 2022 02 24.
Artículo en Inglés | MEDLINE | ID: mdl-35210470

RESUMEN

On 11th March 2020, the UK government announced plans for the scaling of COVID-19 testing, and on 27th March 2020 it was announced that a new alliance of private sector and academic collaborative laboratories were being created to generate the testing capacity required. The Cambridge COVID-19 Testing Centre (CCTC) was established during April 2020 through collaboration between AstraZeneca, GlaxoSmithKline, and the University of Cambridge, with Charles River Laboratories joining the collaboration at the end of July 2020. The CCTC lab operation focussed on the optimised use of automation, introduction of novel technologies and process modelling to enable a testing capacity of 22,000 tests per day. Here we describe the optimisation of the laboratory process through the continued exploitation of internal performance metrics, while introducing new technologies including the Heat Inactivation of clinical samples upon receipt into the laboratory and a Direct to PCR protocol that removed the requirement for the RNA extraction step. We anticipate that these methods will have value in driving continued efficiency and effectiveness within all large scale viral diagnostic testing laboratories.


Asunto(s)
SARS-CoV-2
4.
Sci Rep ; 12(1): 2883, 2022 02 21.
Artículo en Inglés | MEDLINE | ID: mdl-35190592

RESUMEN

We report the development of a large scale process for heat inactivation of clinical COVID-19 samples prior to laboratory processing for detection of SARS-CoV-2 by RT-qPCR. With more than 266 million confirmed cases, over 5.26 million deaths already recorded at the time of writing, COVID-19 continues to spread in many parts of the world. Consequently, mass testing for SARS-CoV-2 will remain at the forefront of the COVID-19 response and prevention for the near future. Due to biosafety considerations the standard testing process requires a significant amount of manual handling of patient samples within calibrated microbiological safety cabinets. This makes the process expensive, effects operator ergonomics and restricts testing to higher containment level laboratories. We have successfully modified the process by using industrial catering ovens for bulk heat inactivation of oropharyngeal/nasopharyngeal swab samples within their secondary containment packaging before processing in the lab to enable all subsequent activities to be performed in the open laboratory. As part of a validation process, we tested greater than 1200 clinical COVID-19 samples and showed less than 1 Cq loss in RT-qPCR test sensitivity. We also demonstrate the bulk heat inactivation protocol inactivates a murine surrogate of human SARS-CoV-2. Using bulk heat inactivation, the assay is no longer reliant on containment level 2 facilities and practices, which reduces cost, improves operator safety and ergonomics and makes the process scalable. In addition, heating as the sole method of virus inactivation is ideally suited to streamlined and more rapid workflows such as 'direct to PCR' assays that do not involve RNA extraction or chemical neutralisation methods.


Asunto(s)
Prueba de Ácido Nucleico para COVID-19/métodos , COVID-19/diagnóstico , Contención de Riesgos Biológicos/métodos , Calor , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , SARS-CoV-2/genética , Manejo de Especímenes/métodos , Inactivación de Virus , Animales , COVID-19/virología , Línea Celular , Humanos , Ratones , Virus de la Hepatitis Murina/genética , ARN Viral/genética , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad
5.
J Thromb Haemost ; 20(3): 729-741, 2022 03.
Artículo en Inglés | MEDLINE | ID: mdl-34846792

RESUMEN

AIMS: P-selectin is a key surface adhesion molecule for the interaction of platelets with leukocytes. We have shown previously that the N-terminal domain of Staphylococcus aureus extracellular fibrinogen-binding protein (Efb) binds to P-selectin and interferes with platelet-leukocyte aggregate formation. Here, we aimed to identify the minimal Efb motif required for binding platelets and to characterize its ability to interfering with the formation of platelet-leukocyte aggregates. METHODS AND RESULTS: Using a library of synthetic peptides, we mapped the platelet-binding site to a continuous 20 amino acid stretch. The peptide Efb68-87 was able to bind to resting and, to a greater extent, thrombin-stimulated platelets in the absence of fibrinogen. Dot blots, pull-down assays and P-selectin glycoprotein ligand-1 (PSGL-1) competitive binding experiments identified P-selectin as the cellular docking site mediating Efb68-87 platelet binding. Accordingly, Efb68-87 did not bind to other blood cells and captured platelets from human whole blood under low shear stress conditions. Efb68-87 did not affect platelet activation as tested by aggregometry, flow cytometry and immunoblotting, but inhibited the formation of platelet-leukocyte aggregates (PLAs). Efb68-87 also interfered with the platelet-dependent stimulation of neutrophil extracellular traps (NETs) formation in vitro. CONCLUSIONS: We have identified Efb68-87 as a novel selective platelet-binding peptide. Efb68-87 binds directly to P-selectin and inhibits interactions of platelets with leukocytes that lead to PLA and NET formation. As PLAs and NETs play a key role in thromboinflammation, Efb68-87 is an exciting candidate for the development of novel selective inhibitors of the proinflammatory activity of platelets.


Asunto(s)
Selectina-P , Trombosis , Plaquetas/metabolismo , Fibrinógeno/metabolismo , Humanos , Inflamación/metabolismo , Leucocitos/metabolismo , Selectina-P/metabolismo , Péptidos/metabolismo , Activación Plaquetaria , Trombosis/metabolismo
6.
iScience ; 24(11): 103370, 2021 Nov 19.
Artículo en Inglés | MEDLINE | ID: mdl-34816105

RESUMEN

Autonomously oscillating circadian clocks resonate with daily environmental (zeitgeber) rhythms to organize physiology around the solar day. Although entrainment properties and mechanisms have been studied widely and in great detail for light-dark cycles, entrainment to daily temperature rhythms remains poorly understood despite that they are potent zeitgebers. Here we investigate the entrainment of the chronobiological model organism Neurospora crassa, subject to thermocycles of different periods and fractions of warm versus cold phases, mimicking seasonal variations. Depending on the properties of these thermocycles, regularly entrained rhythms, period-doubling (frequency demultiplication) but also irregular aperiodic behavior occurs. We demonstrate that the complex nonlinear phenomena of experimentally observed entrainment dynamics can be understood by molecular mathematical modeling.

7.
Sci Rep ; 10(1): 22224, 2020 12 17.
Artículo en Inglés | MEDLINE | ID: mdl-33335302

RESUMEN

Theory predicts that self-sustained oscillations require robust delays and nonlinearities (ultrasensitivity). Delayed negative feedback loops with switch-like inhibition of transcription constitute the core of eukaryotic circadian clocks. The kinetics of core clock proteins such as PER2 in mammals and FRQ in Neurospora crassa is governed by multiple phosphorylations. We investigate how multiple, slow and random phosphorylations control delay and molecular switches. We model phosphorylations of intrinsically disordered clock proteins (IDPs) using conceptual models of sequential and distributive phosphorylations. Our models help to understand the underlying mechanisms leading to delays and ultrasensitivity. The model shows temporal and steady state switches for the free kinase and the phosphoprotein. We show that random phosphorylations and sequestration mechanisms allow high Hill coefficients required for self-sustained oscillations.


Asunto(s)
Proteínas CLOCK/metabolismo , Animales , Proteínas CLOCK/genética , Relojes Circadianos/fisiología , Ritmo Circadiano/fisiología , Biología Computacional , Retroalimentación Fisiológica , Mamíferos , Modelos Biológicos , Neurospora crassa/fisiología , Fosforilación , Biosíntesis de Proteínas , Transcripción Genética
8.
Sci Rep ; 10(1): 14796, 2020 09 09.
Artículo en Inglés | MEDLINE | ID: mdl-32908156

RESUMEN

To curb the staggering health burden attributed to air pollution, the sustainable solution for India would be to reduce emissions in future. Here we project ambient fine particulate matter (PM2.5) exposure in India for the year 2030 under two contrasting air pollution emission pathways for two different climate scenarios based on Representative Concentration Pathways (RCP4.5 and RCP8.5). All-India average PM2.5 is expected to increase from 41.4 ± 26.5 µg m-3 in 2010 to 61.1 ± 40.8 and 58.2 ± 37.5 µg m-3 in 2030 under RCP8.5 and RCP4.5 scenarios, respectively if India follows the current legislation (baseline) emission pathway. In contrast, ambient PM2.5 in 2030 would be 40.2 ± 27.5 (for RCP8.5) and 39.2 ± 25.4 (for RCP4.5) µg m-3 following the short-lived climate pollutant (SLCP) mitigation emission pathway. We find that the lower PM2.5 in the mitigation pathway (34.2% and 32.6%, respectively for RCP8.5 and RCP4.5 relative to the baseline emission pathway) would come at a cost of 0.3-0.5 °C additional warming due to the direct impact of aerosols. The premature mortality burden attributable to ambient PM2.5 exposure is expected to rise from 2010 to 2030, but 381,790 (5-95% confidence interval, CI 275,620-514,600) deaths can be averted following the mitigation emission pathway relative to the baseline emission pathway. Therefore, we conclude that given the expected large health benefit, the mitigation emission pathway is a reasonable tradeoff for India despite the meteorological response. However, India needs to act more aggressively as the World Health Organization (WHO) annual air quality guideline (10 µg m-3) would remain far off.

9.
Int J Mol Sci ; 20(12)2019 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-31248072

RESUMEN

Autonomous endogenous time-keeping is ubiquitous across many living organisms, known as the circadian clock when it has a period of about 24 h. Interestingly, the fundamental design principle with a network of interconnected negative and positive feedback loops is conserved through evolution, although the molecular components differ. Filamentous fungus Neurospora crassa is a well-established chrono-genetics model organism to investigate the underlying mechanisms. The core negative feedback loop of the clock of Neurospora is composed of the transcription activator White Collar Complex (WCC) (heterodimer of WC1 and WC2) and the inhibitory element called FFC complex, which is made of FRQ (Frequency protein), FRH (Frequency interacting RNA Helicase) and CK1a (Casein kinase 1a). While exploring their temporal dynamics, we investigate how limit cycle oscillations arise and how molecular switches support self-sustained rhythms. We develop a mathematical model of 10 variables with 26 parameters to understand the interactions and feedback among WC1 and FFC elements in nuclear and cytoplasmic compartments. We performed control and bifurcation analysis to show that our novel model produces robust oscillations with a wild-type period of 22.5 h. Our model reveals a switch between WC1-induced transcription and FFC-assisted inactivation of WC1. Using the new model, we also study the possible mechanisms of glucose compensation. A fairly simple model with just three nonlinearities helps to elucidate clock dynamics, revealing a mechanism of rhythms' production. The model can further be utilized to study entrainment and temperature compensation.


Asunto(s)
Relojes Circadianos/fisiología , Ritmo Circadiano/fisiología , Modelos Biológicos , Neurospora/fisiología
10.
Environ Pollut ; 242(Pt B): 1817-1826, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30078683

RESUMEN

Exposure to fine particulate matter (PM2.5) is one of the leading risk factors for the mortality and morbidity burden in India. Health benefit expected from mitigation of emissions from individual sectors is the key policy information to address this issue. Here we quantify the relative shares of four major year-round anthropogenic sources to ambient PM2.5 in India using a chemical transport model and estimate premature deaths that could have been avoided due to complete mitigation of emissions from these sources at state level. Population-weighted all-India averaged (±1σ) annual ambient PM2.5 exposures due to residential, transport, industrial and energy sectors in 2010 are estimated to be 26.2 ±â€¯12.5, 3.8 ±â€¯4.3, 5.5 ±â€¯2.7 and 2.2 ±â€¯2.3 µg m-3, respectively. Complete mitigation of emissions from the transport, industrial and energy sectors combined would avoid 92,380 (95% uncertainty interval (UI), 40,918-140,741) premature deaths annually, primarily at the urban hotspots. For the residential sector, this would result in avoiding 378,295 (95% UI, 175,002-575,293) premature deaths due to a reduction in ambient PM2.5 exposure in addition to the benefit of avoiding all premature deaths from household exposure. Bihar and Goa are expected to have the largest (289) and smallest (48) premature mortality burden per 100,000 population due to anthropogenic PM2.5 exposure. From policy perspective, controlling residential sources should be prioritized in view of the effectiveness of implementing mitigation measures and the expected larger health benefit at a regional scale. However, additional mitigation measures are advised at the urban hotspots to curb emissions from the other sectors to get maximum possible health benefit.


Asunto(s)
Contaminantes Atmosféricos/análisis , Contaminación del Aire/estadística & datos numéricos , Exposición a Riesgos Ambientales/estadística & datos numéricos , Material Particulado/análisis , Vivienda , Humanos , India , Mortalidad Prematura
11.
J Biol Chem ; 293(24): 9335-9344, 2018 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-29724824

RESUMEN

Kidney- and brain-expressed protein (KIBRA), a multifunctional scaffold protein with around 20 known binding partners, is involved in memory and cognition, organ size control via the Hippo pathway, cell polarity, and membrane trafficking. KIBRA includes tandem N-terminal WW domains, a C2 domain, and motifs for binding atypical PKC and PDZ domains. A naturally occurring human KIBRA variant involving residue changes at positions 734 (Met-to-Ile) and 735 (Ser-to-Ala) within the C2 domain affects cognitive performance. We have elucidated 3D structures and calcium- and phosphoinositide-binding properties of human KIBRA C2 domain. Both WT and variant C2 adopt a canonical type I topology C2 domain fold. Neither Ca2+ nor any other metal ion was bound to WT or variant KIBRA C2 in crystal structures, and Ca2+ titration produced no significant reproducible changes in NMR spectra. NMR and X-ray diffraction data indicate that KIBRA C2 binds phosphoinositides via an atypical site involving ß-strands 5, 2, 1, and 8. Molecular dynamics simulations indicate that KIBRA C2 interacts with membranes via primary and secondary sites on the same domain face as the experimentally identified phosphoinositide-binding site. Our results indicate that KIBRA C2 domain association with membranes is calcium-independent and involves distinctive C2 domain-membrane relative orientations.


Asunto(s)
Calcio/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Fosfatidilinositoles/metabolismo , Fosfoproteínas/metabolismo , Dominios C2 , Membrana Celular/metabolismo , Cristalografía por Rayos X , Humanos , Péptidos y Proteínas de Señalización Intracelular/química , Péptidos y Proteínas de Señalización Intracelular/genética , Modelos Moleculares , Fosfoproteínas/química , Fosfoproteínas/genética , Polimorfismo de Nucleótido Simple , Unión Proteica , Conformación Proteica
12.
Appl Opt ; 56(31): H57-H66, 2017 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-29091667

RESUMEN

This paper reports open-path in situ measurements of atmospheric carbon dioxide at Gandhinagar (23.2156°N, 72.6369°E) and Ahmedabad (23.0225°N, 72.5714°E) in the heavily industrialized state of Gujarat in western India. Calibration-free second harmonic wavelength modulation spectroscopy (2f WMS) is used to carry out accurate and fully automated measurements. The mean values of the mole fraction of carbon dioxide at four locations were 438 ppm, 495 ppm, 550 ppm, and 740 ppm, respectively. These values are much higher than the current global average of 406.67 ppm. A 1 mW, 2004-nm vertical cavity surface-emitting laser is used to selectively interrogate the R16 transition of carbon dioxide at 2003.5 nm (4991.2585 cm-1). The 2f WMS signal corresponding to the gas absorption line shape is simulated using spectroscopic parameters available in the HITRAN database and relevant laser parameters that are extracted in situ from non-absorbing spectral wings of the harmonic signals. The mole fraction of carbon dioxide is extracted in real-time by a MATLAB program from least-squares fit of the simulated 2f WMS signal to the corresponding experimentally obtained signal. A 10-mW, 1392.54-nm distributed feedback laser is used at two of the locations to carry out water vapor measurements using direct absorption spectroscopy. This is the first instance of a portable tunable diode laser spectroscopy system being deployed in an urban location in India to measure atmospheric carbon dioxide and water vapor under varying traffic conditions. The measurements clearly demonstrate the need to adopt tunable diode laser spectroscopy for precise long-term monitoring of greenhouse gases in the Indian subcontinent.

13.
Opt Lett ; 42(11): 2138-2141, 2017 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-28569866

RESUMEN

We report the first demonstration, to the best of our knowledge, of accurate real-time noninvasive measurement of the absolute cumulative mole fraction of metabolic carbon dioxide emitted by Escherichia coli and Staphylococcus aureus over a period of several hours of their life cycles using a recently developed calibration-free wavelength modulation spectroscopy technique. A 1 mW vertical-cavity surface-emitting laser is used to interrogate a single rotational vibrational absorption line of carbon dioxide at 2003.5 nm. The measurements are immune to laser intensity fluctuations and variable optical coupling that is inevitable in such free-space coupled experiments that run over 10-18 h. The cumulative carbon dioxide mole fraction follows the characteristic modified Gompertz model that is typical of bacterial growth in batch cultures. The characteristic growth parameters are extracted from this curve. The technique can be readily extended to study multiple volatile organic compounds that bacteria are known to emit.

14.
ACS Appl Mater Interfaces ; 8(32): 21077-88, 2016 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-27447357

RESUMEN

Developing generic strategies for building adaptable or multifunctional bioplatforms is challenging, in particular because protein immobilization onto surfaces often causes loss of protein function and because multifunctionality usually necessitates specific combinations of heterogeneous elements. Here, we introduce a generic, modular bioplatform construction strategy that uses cage-like supramolecular multienzyme complexes as highly adaptable building blocks immobilized directly and noncovalently on graphene. Thermoplasma acidophilum dihydrolipoyl acyltransferase (E2) supramolecular complexes organize as a monolayer or can be controllably transferred onto graphene, preserving their supramolecular form with specific molecular recognition capability and capacity for engineering multifunctionality. This E2-graphene platform can bind enzymes (here, E1, E2's physiological partner) without loss of enzyme function; in this test case, E1 catalytic activity was detected on E2-graphene over 6 orders of magnitude in substrate concentration. The E2-graphene platform can be multiplexed via patterned cotransfer of differently modified E2 complexes. As the E2 complexes are robust and highly customizable, E2-graphene is a platform onto which multiple functionalities can be built.


Asunto(s)
Grafito/química , Complejos Multienzimáticos , Proteínas , Thermoplasma
15.
J Biol Chem ; 291(6): 2764-76, 2016 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-26627825

RESUMEN

Extracellular fibrinogen-binding protein (Efb) from Staphylococcus aureus inhibits platelet activation, although its mechanism of action has not been established. In this study, we discovered that the N-terminal region of Efb (Efb-N) promotes platelet binding of fibrinogen and that Efb-N binding to platelets proceeds via two independent mechanisms: fibrinogen-mediated and fibrinogen-independent. By proteomic analysis of Efb-interacting proteins within platelets and confirmation by pulldown assays followed by immunoblotting, we identified P-selectin and multimerin-1 as novel Efb interaction partners. The interaction of both P-selectin and multimerin-1 with Efb is independent of fibrinogen. We focused on Efb interaction with P-selectin. Excess of P-selectin extracellular domain significantly impaired Efb binding by activated platelets, suggesting that P-selectin is the main receptor for Efb on the surface of activated platelets. Efb-N interaction with P-selectin inhibited P-selectin binding to its physiological ligand, P-selectin glycoprotein ligand-1 (PSGL-1), both in cell lysates and in cell-free assays. Because of the importance of P-selectin-PSGL-1 binding in the interaction between platelets and leukocytes, we tested human whole blood and found that Efb abolishes the formation of platelet-monocyte and platelet-granulocyte complexes. In summary, we present evidence that in addition to its documented antithrombotic activity, Efb can play an immunoregulatory role via inhibition of P-selectin-PSGL-1-dependent formation of platelet-leukocyte complexes.


Asunto(s)
Proteínas Bacterianas/metabolismo , Plaquetas/metabolismo , Glicoproteínas de Membrana/metabolismo , Monocitos/metabolismo , Selectina-P/metabolismo , Staphylococcus aureus/metabolismo , Plaquetas/patología , Proteínas Sanguíneas/metabolismo , Humanos , Monocitos/patología
16.
Bioinformation ; 12(5): 266-278, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-28246460

RESUMEN

Oysters are economically and ecologically important bivalves, with its calcareous shell and delicious meat. The shell composition is a blend of inorganic crystals and shell proteins that form an organic matrix which protects the soft inner tissue of the oyster. The objective of the study was to compare the composition of organic matrix proteins (OMP) of two phylogenetically related species: the Hong Kong oyster (Crassostrea hongkongensis) and the Portuguese oyster (Crassostrea angulata) which differ in their shell hardness and mechanical properties. C. hongkongensis shells are comparatively stronger than C. angulata. Modern shotgun proteomics has been used to understand the nature of the OMP and the variations observed in the mechanical properties of these two species of oyster shells. After visualizing proteins on the one (1DE) and two-dimensional electrophoresis (2DE) gels, the protein spots and their intensities were compared using PDQuest software and 14 proteins of C. hongkongensis were found to be significantly different (student׳s t-test; p<0.05) when compared to the C. angulata. Furthermore, shell OMP separated on 1DE gels were processed using Triple TOF5600 mass spectrometry and 42 proteins of C. hongkongensis and 37 of C. angulata identified. A Circos based comparative analysis of the shell proteins of both oyster species were prepared against the shell proteome of other shell forming gastropods and molluscs to study the evolutionary conservation of OMP and their function. This comparative proteomics expanded our understating of the molecular mechanism behind the shells having different hardness and mechanical properties.

17.
Opt Lett ; 40(17): 4086-9, 2015 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-26368718

RESUMEN

This Letter demonstrates a new calibration-free 2f wavelength modulation spectroscopy (WMS) technique to measure gas concentration and pressure without the need for laser precharacterization. A 1650-nm laser diode is used for methane concentration and pressure measurements for pressures up to 4 bar and for a modulation index (m) of 2.2. All laser parameters such as the intensity, linear and nonlinear intensity modulation (IM), frequency modulation (FM) characteristics, the phase difference ψ1 between the FM and the linear IM, and the phase difference ψ2 between the FM and the nonlinear IM are accurately estimated in situ and in real time. This technique accounts for variations in these parameters that arise due to scanning of the laser's center wavelength, laser temperature variations, and aging of the laser. The laser is modulated at its phase quadrature frequency at which the linear IM and the FM are orthogonal to each other (ψ1=90°). This ensures that the two linear IM-dependent distorting Fourier components are orthogonal to the detection axis, and the undistorted 2f signal is recovered. This simplifies the simulation and gas parameter-extraction process. Finally, 2f RAM nulling is implemented to remove the significant absorption-independent 2f residual amplitude-modulation (RAM) signal that is seen to cause significant distortion of the 2f signal and detector saturation.


Asunto(s)
Láseres de Semiconductores , Calibración , Metano , Factores de Tiempo
18.
Sci Total Environ ; 533: 495-505, 2015 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-26186464

RESUMEN

Fuzzy-analytical hierarchical process (F-AHP) can be extended to determine fuzzy air quality health index (FAQHI) for deducing health risk associated with local air pollution levels, and subjective parameters. The present work aims at determining FAQHI by considering five air pollutant parameters (SO2, NO2, O3, CO, and PM10) and three subjective parameters (population sensitivity, population density and location sensitivity). Each of the individual pollutants has varying impacts. Hence the combined health effects associated with the pollutants were estimated by aggregating the pollutants with different weights. Global weights for each evaluation alternatives were determined using fuzzy-AHP method. The developed model was applied to determine FAQHI in Howrah City, India from daily-observed concentrations of air pollutants over the three-year period between 2009 and 2011. The FAQHI values obtained through this method in Howrah City range from 1 to 3. Since the permissible value of FAQHI (as calculated for NAAQS) for residential areas is 1.78, higher index values are of public health concern to the exposed individuals. During the period of study, the observed FAQHI values were found to be higher than 1.78 in most of the day in the months of January to March, and October to December. However, the index values were below the recommended limit during rest of the months. In conclusion, FAQHI in Howrah city was above permissible limit in winter months and within acceptable values in summer and rainy months. Diurnal variations of FAQHI showed a similar trend during the three-year period of assessment.


Asunto(s)
Contaminantes Atmosféricos/normas , Contaminación del Aire/estadística & datos numéricos , Exposición a Riesgos Ambientales/estadística & datos numéricos , Exposición a Riesgos Ambientales/normas , Humanos , India , Estaciones del Año
19.
Methods Mol Biol ; 1258: 209-22, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25447866

RESUMEN

Transient transfection of human embryonic kidney cells (HEK 293) enables the rapid and affordable lab-scale production of recombinant proteins. In this chapter protocols for the expression and purification of both secreted and intracellular proteins using transient expression in HEK 293 cells are described.


Asunto(s)
Escherichia coli/genética , Células HEK293/metabolismo , Mamíferos/genética , Proteínas Recombinantes/genética , Animales , Escherichia coli/metabolismo , Humanos , Mamíferos/metabolismo , Proteínas Recombinantes/metabolismo , Transfección/métodos
20.
PLoS One ; 9(1): e87063, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24489835

RESUMEN

Metagenomic analyses have advanced our understanding of ecological microbial diversity, but to what extent can metagenomic data be used to predict the metabolic capacity of difficult-to-study organisms and their abiotic environmental interactions? We tackle this question, using a comparative genomic approach, by considering the molecular basis of aerobiosis within archaea. Lipoylation, the covalent attachment of lipoic acid to 2-oxoacid dehydrogenase multienzyme complexes (OADHCs), is essential for metabolism in aerobic bacteria and eukarya. Lipoylation is catalysed either by lipoate protein ligase (LplA), which in archaea is typically encoded by two genes (LplA-N and LplA-C), or by a lipoyl(octanoyl) transferase (LipB or LipM) plus a lipoic acid synthetase (LipA). Does the genomic presence of lipoylation and OADHC genes across archaea from diverse habitats correlate with aerobiosis? First, analyses of 11,826 biotin protein ligase (BPL)-LplA-LipB transferase family members and 147 archaeal genomes identified 85 species with lipoylation capabilities and provided support for multiple ancestral acquisitions of lipoylation pathways during archaeal evolution. Second, with the exception of the Sulfolobales order, the majority of species possessing lipoylation systems exclusively retain LplA, or either LipB or LipM, consistent with archaeal genome streamlining. Third, obligate anaerobic archaea display widespread loss of lipoylation and OADHC genes. Conversely, a high level of correspondence is observed between aerobiosis and the presence of LplA/LipB/LipM, LipA and OADHC E2, consistent with the role of lipoylation in aerobic metabolism. This correspondence between OADHC lipoylation capacity and aerobiosis indicates that genomic pathway profiling in archaea is informative and that well characterized pathways may be predictive in relation to abiotic conditions in difficult-to-study extremophiles. Given the highly variable retention of gene repertoires across the archaea, the extension of comparative genomic pathway profiling to broader metabolic and homeostasis networks should be useful in revealing characteristics from metagenomic datasets related to adaptations to diverse environments.


Asunto(s)
Oxidorreductasas de Alcohol/genética , Archaea/enzimología , Archaea/genética , Genómica/métodos , Lipoilación , Familia de Multigenes , Aerobiosis , Oxidorreductasas de Alcohol/metabolismo , Anaerobiosis , Biotina/metabolismo , Genes Arqueales , Ligasas/genética , Filogenia , Especificidad por Sustrato , Ácido Tióctico/metabolismo
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