The Plasmodium falciparum, Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation

<b>Background</b> Malaria parasites undergo, in the vertebrate host, a developmental switch from asexual replication to sexual differentiation leading to the formation of gametocytes, the only form able to survive in the mosquito vector. Regulation of the onset of the sexual phase remains largely unknown and represents an important gap in the understanding of the parasite's complex biology. <b>Methods:</b> The expression and function of the Nima-related kinase Pfnek-4 during the early sexual development of the human malaria parasite Plasmodium falciparum were investigated, using three types of transgenic Plasmodium falciparum 3D7 lines: (i) episomally expressing a Pfnek-4-GFP fusion protein under the control of its cognate pfnek-4 promoter; (ii) episomally expressing negative or positive selectable markers, yeast cytosine deaminase-uridyl phosphoribosyl transferase, or human dihydrofolate reductase, under the control of the pfnek-4 promoter; and (iii) lacking a functional pfnek-4 gene. Parasite transfectants were analysed by fluorescence microscopy and flow cytometry. In vitro growth rate and gametocyte formation were determined by Giemsa-stained blood smears. <b>Results:</b> The Pfnek-4-GFP protein was found to be expressed in stage II to V gametocytes and, unexpectedly, in a subset of asexual-stage parasites undergoing schizogony. Culture conditions stimulating gametocyte formation resulted in significant increase of this schizont subpopulation. Moreover, sorted asexual parasites expressing the Pfnek-4-GFP protein displayed elevated gametocyte formation when returned to in vitro culture in presence of fresh red blood cells, when compared to GFP- parasites from the same initial population. Negative selection of asexual parasites expressing pfnek-4 showed a marginal reduction in growth rate, whereas positive selection caused a marked reduction in parasitaemia, but was not sufficient to completely abolish proliferation. Pfnek-4- clones are not affected in their asexual growth and produced normal numbers of stage V gametocytes. <b>Conclusions:</b> The results indicate that Pfnek-4 is not strictly gametocyte-specific, and is expressed in a small subset of asexual parasites displaying high rate conversion to sexual development. Pfnek-4 is not required for erythrocytic schizogony and gametocytogenesis. This is the first study to report the use of a molecular marker for the sorting of sexually-committed schizont stage P. falciparum parasites, which opens the way to molecular characterization of this pre-differentiated subpopulation

Social vs. sensual: the struggle for identity within the characters of Clarín’s La Regenta.

This study approaches the novel La Regenta, by Leopoldo Alas Clarín, by exploring the three types of forces that act upon the characters of Fermín de Pas and Ana de Ozores as they construct their personal identities. The Catholic Church acts in the novel as it controls its female devotees and protects its corrupted clergy. Physical influences in the form of scientific discourse and corporal desires also motivate and control the characters. Societal expectations are expressed in the form of filial duties and behavioral pressures on women to conform to the ideal of the ángel del hogar. The conflicting influences of these forces create a world in which Fermín and Ana must struggle in order to understand themselves

Nanoimprinted superlattice metallic photonic crystal as ultraselective solar absorber

A two-dimensional superlattice metallic photonic crystal (PhC) and its fabrication by nanoimprint lithography on tantalum substrates are presented. The superior tailoring capacity of the superlattice PhC geometry is used to achieve spectrally selective solar absorption optimized for high-temperature and high-efficiency solar-energy-conversion applications. The scalable fabrication route by nanoimprint lithography allows for a high-throughput and high-resolution replication of this complex pattern over large areas. Despite the high fill factor, the pattern of polygonal cavities is accurately replicated into a resist that hardens under ultraviolet radiation over an area of 10  mm². In this way, cavities of 905 nm and 340 nm width are achieved with a period of 1 μm. After pattern transfer into tantalum via a deep reactive ion-etching process, the achieved cavities are 2.2 μm deep, separated by 85–95 nm wide ridges with vertical sidewalls. The room-temperature reflectance spectra of the fabricated samples show excellent agreement with simulated results, with a high spectral absorptance approaching blackbody absorption in the range from 300 to 1900 nm and a steep cutoff. The calculated solar absorptivity of this superlattice PhC is 96% and its thermal transfer efficiency is 82.8% at an operating temperature of 1500 K and an irradiance of 1000  kW/m².United States. Army Research Office (W911NF-13-D-0001)United States. Department of Energy (DE-SC0001299

A new compact soft x ray spectrometer for resonant inelastic x ray scattering studies at PETRA III

We present a newly designed compact grating spectrometer for the energy range from 210 eV to 1250 eV, which would include the K amp; 945;1,2 emission lines of vital elements like C, N, and O. The spectrometer is based on a grazing incidence spherical varied line spacing grating with 2400 l mm at its center and a radius of curvature of 58 542 mm. First, results show a resolving power of around 1000 at an energy of 550 eV and a working spectrometer for high vacuum 10 amp; 8722;4 mbar environment without losing photon intensit

MERLIN - A meV resolution beamline at the ALS

An ultra-high resolution beamline is being constructed at the Advanced Light Source (ALS) for the study of low energy excitations in strongly correlated systems with the use of high-resolution inelastic scattering and angle-resolved photoemission. This new beamline, given the acronym Merlin (for meV resolution line), will cover the energy range 10-150 eV. The monochromator has fixed entrance and exit slits and a plane mirror that can illuminate a spherical grating at the required angle of incidence (as in the SX-700 mechanism). The monochromator can be operated in two different modes. In the highest resolution mode, the energy scanning requires translating the monochromator chamber (total travel 1.1 m) as well as rotating the grating and the plane mirror in front of the grating. The resolution in this mode is practically determined by the slits width. In the second mode, the scanning requires rotating the grating and the plane mirror. This mode can be used to scan a few eV without a significant resolution loss. The source for the beamline is a 1.9 m long, 90 mm period quasi periodic EPU. The expected flux at the sample is higher than 10 photons/s at a resolving power of 5 × 10 in the energy range 16-130 eV. A second set of gratings can be used to obtain higher flux at the expense of resolution. © 2007 American Institute of Physics. 11

A new tool for the chemical genetic investigation of the Plasmodium falciparum Pfnek-2 NIMA-related kinase

Background: Examining essential biochemical pathways in Plasmodium falciparum presents serious challenges, as standard molecular techniques such as siRNA cannot be employed in this organism, and generating gene knock-outs of essential proteins requires specialized conditional approaches. In the study of protein kinases, pharmacological inhibition presents a feasible alternative option. However, as in mammalian systems, inhibitors often lack the desired selectivity. Described here is a chemical genetic approach to selectively inhibit Pfnek-2 in P. falciparum, a member of the NIMA-related kinase family that is essential for completion of the sexual development of the parasite. Results: Introduction of a valine to cysteine mutation at position 24 in the glycine rich loop of Pfnek-2 does not affect kinase activity but confers sensitivity to the protein kinase inhibitor 4-(6-ethynyl-9H-purin-2-ylamino) benzene sulfonamide (NCL-00016066). Using a combination of in vitro kinase assays and mass spectrometry, (including phosphoproteomics) the study shows that this compound acts as an irreversible inhibitor to the mutant Pfnek2 likely through a covalent link with the introduced cysteine residue. In particular, this was shown by analysis of total protein mass using mass spectrometry which showed a shift in molecular weight of the mutant kinase in the presence of the inhibitor to be precisely equivalent to the molecular weight of NCL-00016066. A similar molecular weight shift was not observed in the wild type kinase. Importantly, this inhibitor has little activity towards the wild type Pfnek-2 and, therefore, has all the properties of an effective chemical genetic tool that could be employed to determine the cellular targets for Pfnek-2. Conclusions: Allelic replacement of wild-type Pfnek-2 with the mutated kinase will allow for targeted inhibition of Pfnek-2 with NCL-00016066 and hence pave the way for comparative studies aimed at understanding the biological role and transmission-blocking potential of Pfnek-2. © 2016 The Author(s)

Missed opportunities for diagnosis and treatment of diabetes, hypertension, and hypercholesterolemia in a Mexican American population, Cameron County Hispanic Cohort, 2003-2008

Introduction Diabetes, hypertension, and hypercholesterolemia are common chronic diseases among Hispanics, a group projected to comprise 30% of the US population by 2050. Mexican Americans are the largest ethnically distinct subgroup among Hispanics. We assessed the prevalence of and risk factors for undiagnosed and untreated diabetes, hypertension, and hypercholesterolemia among Mexican Americans in Cameron County, Texas. Methods We analyzed cross-sectional baseline data collected from 2003 to 2008 in the Cameron County Hispanic Cohort, a randomly selected, community-recruited cohort of 2,000 Mexican American adults aged 18 or older, to assess prevalence of diabetes, hypertension, and hypercholesterolemia; to assess the extent to which these diseases had been previously diagnosed based on self-report; and to determine whether participants who self-reported having these diseases were receiving treatment. We also assessed social and economic factors associated with prevalence, diagnosis, and treatment. Results Approximately 70% of participants had 1 or more of the 3 chronic diseases studied. Of these, at least half had had 1 of these 3 diagnosed, and at least half of those who had had a disease diagnosed were not being treated. Having insurance coverage was positively associated with having the 3 diseases diagnosed and treated, as were higher income and education level. Conclusions Although having insurance coverage is associated with receiving treatment, important social and cultural barriers remain. Failure to provide widespread preventive medicine at the primary care level will have costly consequences

High Temperature Electron Localization in dense He Gas

We report new accurate mesasurements of the mobility of excess electrons in high density Helium gas in extended ranges of temperature $[(26\leq T\leq 77) K ]$ and density $[ (0.05\leq N\leq 12.0) {atoms} \cdot {nm}^{-3}]$ to ascertain the effect of temperature on the formation and dynamics of localized electron states. The main result of the experiment is that the formation of localized states essentially depends on the relative balance of fluid dilation energy, repulsive electron-atom interaction energy, and thermal energy. As a consequence, the onset of localization depends on the medium disorder through gas temperature and density. It appears that the transition from delocalized to localized states shifts to larger densities as the temperature is increased. This behavior can be understood in terms of a simple model of electron self-trapping in a spherically symmetric square well.Comment: 23 pages, 13 figure

Measurement of single electron emission in two-phase xenon

We present the first measurements of the electroluminescence response to the emission of single electrons in a two-phase noble gas detector. Single ionization electrons generated in liquid xenon are detected in a thin gas layer during the 31-day background run of the ZEPLIN-II experiment, a two-phase xenon detector for WIMP dark matter searches. Both the pressure dependence and magnitude of the single-electron response are in agreement with previous measurements of electroluminescence yield in xenon. We discuss different photoionization processes as possible cause for the sample of single electrons studied in this work. This observation may have implications for the design and operation of future large-scale two-phase systems.Comment: 11 pages, 6 figure

SAS6-like protein in Plasmodium indicates that conoid-associated apical complex proteins persist in invasive stages within the mosquito vector

The SAS6-like (SAS6L) protein, a truncated paralogue of the ubiquitous basal body/centriole protein SAS6, has been characterised recently as a flagellum protein in trypanosomatids, but associated with the conoid in apicomplexan Toxoplasma. The conoid has been suggested to derive from flagella parts, but is thought to have been lost from some apicomplexans including the malaria-causing genus Plasmodium. Presence of SAS6L in Plasmodium, therefore, suggested a possible role in flagella assembly in male gametes, the only flagellated stage. Here, we have studied the expression and role of SAS6L throughout the Plasmodium life cycle using the rodent malaria model P. berghei. Contrary to a hypothesised role in flagella, SAS6L was absent during gamete flagellum formation. Instead, SAS6L was restricted to the apical complex in ookinetes and sporozoites, the extracellular invasive stages that develop within the mosquito vector. In these stages SAS6L forms an apical ring, as we show is also the case in Toxoplasma tachyzoites. The SAS6L ring was not apparent in blood-stage invasive merozoites, indicating that the apical complex is differentiated between the different invasive forms. Overall this study indicates that a conoid-associated apical complex protein and ring structure is persistent in Plasmodium in a stage-specific manner