Diatom Proteomics Reveals Unique Acclimation Strategies to Mitigate Fe Limitation

Phytoplankton growth rates are limited by the supply of iron (Fe) in approximately one third of the open ocean, with major implications for carbon dioxide sequestration and carbon (C) biogeochemistry. To date, understanding how alteration of Fe supply changes phytoplankton physiology has focused on traditional metrics such as growth rate, elemental composition, and biophysical measurements such as photosynthetic competence (Fv/Fm). Researchers have subsequently employed transcriptomics to probe relationships between changes in Fe supply and phytoplankton physiology. Recently, studies have investigated longer-term (i.e. following acclimation) responses of phytoplankton to various Fe conditions. In the present study, the coastal diatom, Thalassiosira pseudonana, was acclimated (10 generations) to either low or high Fe conditions, i.e. Fe-limiting and Fe-replete. Quantitative proteomics and a newly developed proteomic profiling technique that identifies low abundance proteins were employed to examine the full complement of expressed proteins and consequently the metabolic pathways utilized by the diatom under the two Fe conditions. A total of 1850 proteins were confidently identified, nearly tripling previous identifications made from differential expression in diatoms. Given sufficient time to acclimate to Fe limitation, T. pseudonana up-regulates proteins involved in pathways associated with intracellular protein recycling, thereby decreasing dependence on extracellular nitrogen (N), C and Fe. The relative increase in the abundance of photorespiration and pentose phosphate pathway proteins reveal novel metabolic shifts, which create substrates that could support other well-established physiological responses, such as heavily silicified frustules observed for Fe-limited diatoms. Here, we discovered that proteins and hence pathways observed to be down-regulated in short-term Fe starvation studies are constitutively expressed when T. pseudonana is acclimated (i.e., nitrate and nitrite transporters, Photosystem II and Photosystem I complexes). Acclimation of the diatom to the desired Fe conditions and the comprehensive proteomic approach provides a more robust interpretation of this dynamic proteome than previous studies.This work was supported by National Science Foundation grants OCE1233014 (BLN) and the Office of Polar Programs Postdoctoral Fellowship grant 0444148 (BLN). DRG was supported by National Institutes of Health 5P30ES007033-10. AH and MTM were supported by Natural Sciences and Engineering Research Council of Canada. RFS and PWB were supported by the New Zealand Royal Society Marsden Fund and the Ministry of Science. This work is supported in part by the University of Washington's Proteomics Computer Resource Centre (UWPR95794). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Patterns of sexualised recreational drug use and its association with risk behaviours and sexual health outcomes in men who have sex with men in London, UK: a comparison of cross-sectional studies conducted in 2013 and 2016

Objective: London has one of the highest identified prevalence of chemsex (sexualised recreational drug use) among men who have sex with men (MSM) in Europe. We examine MSM’s patterns of chemsex and its association with HIV/STI risk behaviours, STI diagnoses, and sexual healthcare-seeking behaviours, including if HIV testing behaviour met UK national guidelines (3-monthly if engaging in chemsex). Methods: Cross-sectional survey data from 2013 (n=905) and 2016 (n=739) were collected using anonymous, self-administered questionnaires from MSM recruited in commercial gay venues in London, UK. Descriptive and multivariable analyses, stratified by self-reported HIV status, were conducted. Adjusted prevalence ratios (aPR) with 95% confidence intervals (CI) were calculated. Results: Comparing the 2013 and 2016 surveys; chemsex prevalence in the past year remained stable, in both HIV-negative/unknown-status MSM (20.9% in 2013 vs 18.7% in 2016, p=0.301) and HIV-positive MSM (41.6% in 2013 vs 41.7% in 2016, p=0.992). Combined 2013-2016 data showed that compared to other MSM, those reporting chemsex were more likely to report HIV/STI risk behaviours, including condomless anal intercourse with serodifferent HIV-status partners (HIV-negative/unknown-status men: aPR 2.36, 95% CI 1.68-3.30; HIV-positive men: aPR 4.19, 95% CI 1.85-9.50), and STI diagnoses in the past year (HIV-negative/unknown-status men: aPR 2.10, 95% CI 1.64-2.69; HIV-positive men: aPR 2.56, 95% CI 1.57-4.20). 69.0% of HIV-negative/unknown-status men reporting chemsex attended sexual health clinics and 47.6% had tested for HIV more than once in the past year. Conclusions: Chemsex in London MSM remained stable but high, particularly among HIV-positive men. Irrespective of HIV status, chemsex was associated with engagement in HIV/STI risk behaviours. Frequency of HIV testing in the past year among HIV-negative/unknown-status men was below national recommendations. Promoting combination prevention strategies, including 3-monthly HIV/STI testing, access to PrEP/ART, and behavioural interventions among MSM reporting chemsex, remain vital to address sexual health inequalities in MSM

A Search for the Damped Ly-alpha Absorber at z=1.86 toward QSO 1244+3443 with NICMOS

We have carried out a high-resolution imaging search for the galaxy associated with the damped Lyman-alpha (DLA) absorber at z=1.859 toward the z_{em}=2.48 quasar QSO 1244+3443, using the HST and the NICMOS. Images were obtained in the broad filter F160W and the narrow filter F187N with camera 2 on NICMOS with the goal of detecting the rest-frame optical continuum and the H-alpha line emission from the DLA. After PSF subtraction, two weak features are seen at projected separations of 0.16-0.24" from the quasar. Parts of these features may be associated with the DLA absorber, although we cannot completely rule out that they could be artifacts of the point spread function (PSF). If associated with the DLA, the objects would be ~1-2 h_{70}^{-1} kpc in size with integrated flux densities of 2.5 and 3.3 mu Jy in the F160W filter, implying luminosities at lambda_{central}=5600 A in the DLA rest frame of 4.4-5.9 x 10^{9} h_{70}^{-2} L_{solar} at z=1.86, for q0=0.5. However, no significant H-alpha line emission is seen from these objects, suggesting low star formation rates (SFRs). Our 3 sigma upper limit on the SFR in the DLA is 1.3 h_{70}^{-2} M_{solar}/yr for q0 = 0.5 (2.4 h_{70}^{-2} M_{solar} yr^{-1} for q0 = 0.1). This together with our earlier result for LBQS 1210+1731 mark a significant improvement over previous constraints on the star formation rates of DLAs. A combination of low SFR and some dust extinction is likely to be responsible for the lack of H-alpha emission. Alternatively, the objects, may be associated with the quasar host galaxy. In any case, our observations suggest that the DLA is not a large bright proto-disk, but a compact object or a low-surface brightness galaxy. If the two features are PSF artifacts then the constraints on DLA properties are even more severe.Comment: 28 pages, 9 figures. Accepted for publication in The Astrophysical Journal. Figures are given at a slightly lower resolution here, to decrease file sizes. The higher resolution versions can be found in the Ap

Prevalence of significant liver disease in human immunodeficiency virus-infected patients exposed to Didanosine: A cross sectional study

AIM: To identify significant liver disease [including nodular regenerative hyperplasia (NRH)] in asymptomatic Didanosine (DDI) exposed human immunodeficiency virus (HIV) positive patients. METHODS: Patients without known liver disease and with > 6 mo previous DDI use had liver stiffness assessed by transient elastography (TE). Those with alanine transaminase (ALT) above upper limit normal and/or TE > 7.65 kPa underwent ultrasound scan (U/S). Patients with: (1) abnormal U/S; or (2) elevated ALT plus TE > 7.65 kPa; or (3) TE > 9.4 kPa were offered trans-jugular liver biopsy (TJLB) with hepatic venous pressure gradient (HVPG) assessment. RESULTS: Ninety-nine patients were recruited, median age 50 years (range 31-70), 81% male and 70% men who have sex with men. Ninety-five percent with VL 9.4 kPa. Seventeen (17%) met criteria for TJLB, of whom 12 accepted. All had HVPG < 6 mmHg. Commonest histological findings were steatosis (n = 6), normal architecture (n = 4) and NRH (n = 2), giving a prevalence of previously undiagnosed NRH of 2% (95%CI: 0.55%, 7.0%). CONCLUSION: A screening strategy based on TE, liver enzymes and U/S scan found a low prevalence of previously undiagnosed NRH in DDI exposed, asymptomatic HIV positive patients. Patients were more likely to have steatosis highlighting the increased risk of multifactorial liver disease in this population

The reliability of two visual motor integration tests used with children

Occupational therapists often assess the visual motor integration (VMI) skills of children and young people. It is important that therapists use tools with strong psychometric properties. This study aims to examine the reliability of 2 VMI tests. Ninety-two children between the ages of 5 and 17 years (response rate of 31%) completed 2 VMI tests: the Developmental Test of Visual Motor Integration (DTVMI) and the Full Range Test of Visual Motor Integration (FRTVMI). Cronbach\u27s alpha coefficient was used to examine the internal consistency of the 2 VMI tests whereas Spearman\u27s rho correlation was used to evaluate the test&ndash;retest reliability, intrarater reliability, and interrater reliability of the 2 VMI tests. The Cronbach\u27s alpha coefficient for the DTVMI was .82 and .72 for the FRTVMI. The test&ndash;retest reliability coefficient was .73 (p = .000) for the DTVMI and .49 (p = .05) for the FRTVMI. The interrater correlation was significant for both the DTVMI at .94 (p = .000) and FRTVMI at .68 (p = .001). The DTVMI intrarater reliability correlation result was .90 (p = .000) and the FRTVMI at .85 (p = .000). Overall, the DTVMI exhibited a higher level of reliability than the FRTVMI. Both VMI tests appear to exhibit reasonable levels of reliability and are recommended for use with children and young people.<br /

NICMOS Imaging of a Damped Lyman-alpha Absorber at z=1.89 toward LBQS 1210+1731 : Constraints on Size and Star Formation Rate

We report results of a high-resolution imaging search (in rest frame H-$\alpha$ and optical continuum) for the galaxy associated with the damped Lyman-$\alpha$ (DLA) absorber at $z=1.892$ toward the $z_{em}=2.543$ quasar LBQS 1210+1731, using HST/NICMOS. After PSF subtraction, a feature is seen in both the broad-band and narrow-band images, at a projected separation of 0.25\arcsec from the quasar. If associated with the DLA, the object would be $\approx 2-3$ $h_{70}^{-1}$ kpc in size with a flux of $9.8 \pm 2.4$ $\mu$Jy in the F160W filter, implying a luminosity at $\lambda_{central}=5500$ {\AA} in the rest frame of $1.5 \times 10^{10}$ $h_{70}^{-2}$ L$_{\odot}$ at $z=1.89$, for $q_{0}=0.5$. However, no significant H-$\alpha$ emission is seen, suggesting a low star formation rate (SFR) (3 $\sigma$ upper limit of 4.0 $h_{70}^{-2}$ M$_{\odot}$ yr$^{-1}$), or very high dust obscuration. Alternatively, the object may be associated with the host galaxy of the quasar. H-band images obtained with the NICMOS camera 2 coronagraph show a much fainter structure $\approx 4-5$ $h_{70}^{-1}$ kpc in size and containing four knots of continuum emission, located 0.7\arcsec away from the quasar. We have probed regions far closer to the quasar sight-line than in most previous studies of high-redshift intervening DLAs. The two objects we report mark the closest detected high-redshift DLA candidates yet to any quasar sight line. If the features in our images are associated with the DLA, they suggest faint, compact, somewhat clumpy objects rather than large, well-formed proto-galactic disks or spheroids.Comment: 52 pages of text, 19 figures, To be published in Astrophysical Journal (accepted Dec. 8, 1999

Diatom Proteomics Reveals Unique Acclimation Strategies to Mitigate Fe Limitation

Phytoplankton growth rates are limited by the supply of iron (Fe) in approximately one third of the open ocean, with major implications for carbon dioxide sequestration and carbon (C) biogeochemistry. To date, understanding how alteration of Fe supply changes phytoplankton physiology has focused on traditional metrics such as growth rate, elemental composition, and biophysical measurements such as photosynthetic competence (Fv/Fm). Researchers have subsequently employed transcriptomics to probe relationships between changes in Fe supply and phytoplankton physiology. Recently, studies have investigated longer-term (i.e. following acclimation) responses of phytoplankton to various Fe conditions. In the present study, the coastal diatom, Thalassiosira pseudonana, was acclimated (10 generations) to either low or high Fe conditions, i.e. Fe-limiting and Fe-replete. Quantitative proteomics and a newly developed proteomic profiling technique that identifies low abundance proteins were employed to examine the full complement of expressed proteins and consequently the metabolic pathways utilized by the diatom under the two Fe conditions. A total of 1850 proteins were confidently identified, nearly tripling previous identifications made from differential expression in diatoms. Given sufficient time to acclimate to Fe limitation, T. pseudonana up-regulates proteins involved in pathways associated with intracellular protein recycling, thereby decreasing dependence on extracellular nitrogen (N), C and Fe. The relative increase in the abundance of photorespiration and pentose phosphate pathway proteins reveal novel metabolic shifts, which create substrates that could support other well-established physiological responses, such as heavily silicified frustules observed for Fe-limited diatoms. Here, we discovered that proteins and hence pathways observed to be down-regulated in short-term Fe starvation studies are constitutively expressed when T. pseudonana is acclimated (i.e., nitrate and nitrite transporters, Photosystem II and Photosystem I complexes). Acclimation of the diatom to the desired Fe conditions and the comprehensive proteomic approach provides a more robust interpretation of this dynamic proteome than previous studies

The Initial Mass Function of Low-Mass Stars and Brown Dwarfs in Young Clusters

We have obtained images of the Trapezium Cluster (140" x 140"; 0.3 pc x 0.3 pc) with the Hubble Space Telescope Near-Infrared Camera and Multi-Object Spectrometer (NICMOS). Combining these data with new ground-based K-band spectra (R=800) and existing spectral types and photometry and the models of D'Antona & Mazzitelli, we find that the distributions of ages of comparable samples of stars in the Trapezium, rho Oph, and IC 348 indicate median ages of \~0.4 Myr for the first two regions and ~1-2 Myr for the latter. The low-mass IMFs in these sites of clustered star formation are similar over a wide range of stellar densities and other environmental conditions. With current data, we cannot rule out modest variations in the substellar mass functions among these clusters. We then make the best estimate of the true form of the IMF in the Trapezium by using the evolutionary models of Baraffe et al. and an empirically adjusted temperature scale and compare this mass function to recent results for the Pleiades and the field. All of these data are consistent with an IMF that is flat or rises slowly from the substellar regime to about 0.6 Msun, and then rolls over into a power law that continues from about 1 Msun to higher masses with a slope similar to or somewhat larger than the Salpeter value of 1.35. For the Trapezium, this behavior holds from our completeness limit of ~0.02 Msun and probably, after a modest completeness correction, even from 0.01-0.02 Msun. These data include ~50 likely brown dwarfs. We test the predictions of theories of the IMF against various properties of the observed IMF.Comment: 34 pages, 13 figures, for color image see http://cfa-www.harvard.edu/~kluhman/trap/colorimage.jp

Self-reported difficulties with everyday function, cognitive symptoms, and cognitive function in people with HIV

BACKGROUND: We determined factors associated with self-reported decline in activities of daily living (ADLs) and symptoms of cognitive impairment in HIV positive (HIV+) adults in five European clinics. METHODS: HIV+ adults underwent computerized and pen-and-paper neuropsychological tests and questionnaires of cognitive symptoms and ADLs. We considered cognitive function in five domains, psychosocial factors and clinical parameters as potentially associated with symptoms. Separate regression analyses were used to determine factors associated with decline in ADL (defined as self-reported decline affecting ≥2 ADLs and attributed to cognitive difficulties) and self-reported frequency of symptoms of cognitive impairment. We also estimated the diagnostic accuracy of both questionnaires as tests for cognitive impairment. RESULTS: 448 patients completed the assessments (mean age 45.8 years, 84% male, 87% white, median CD4 count 550 cells/mm, median time since HIV diagnosis 9.9 years, 81% virologically suppressed [HIV-1 plasma RNA <50 copies/mL]). Ninety-six (21.4%) reported decline in ADLs and attributed this to cognitive difficulties. Self-reported decline in ADLs and increased symptoms of cognitive impairment were both associated with worse performance on some cognitive tests. There were also strong associations with financial difficulties, depressive and anxiety symptoms, unemployment, and longer time since HIV diagnosis. Both questionnaires performed poorly as diagnostic tests for cognitive impairment. CONCLUSION: Patients' own assessments of everyday function and symptoms were associated with objectively-measured cognitive function. However, there were strong associations with other psychosocial issues including mood and anxiety disorders and socioeconomic hardship. This should be considered when assessing HIV-associated cognitive impairment in clinical care or research studies

Platelet Glycoprotein lib: Chromosomal Localization and Tissue Expression

The GPIIb-IIIa complex functions as a receptor for cytoadhesive proteins on the platelet surface. Both GPIIb and GPIIIa are synthesized by a human erythroleukemia (HEL) cell line. We isolated several cDNA clones by screening a HEL cell cDNA library with an oligonucleotide derived from amino acid sequence of GPIIb. Nucleotide and amino acid sequences were determined from 703 bp of one of these clones. Amino acid sequence of purified platelet GPIIb peptides confirmed the identity of the clone. The cDNA encodes the carboxyl terminus of the large (a) subunit of GPIIb and all of the smaller (f6) subunit of GPIIb. By hybridizing the cDNA directly to chromosomes separated by dual laser chromosome sorting, the gene for GPIIb was mapped to chromosome 17. Northern blot analysis showed a - 3.4-kb GPIIb mRNA in HEL cells. We also compared the amino acid sequences determined from eight additional platelet GPIIb peptides with the derived amino acids from a published HEL cell GPIIb cDNA, and the platelet and HEL cell proteins appear to be the same. Despite previous reports that vascular endothelial cells and monocytes contain GPIIb, no GPIIb mRNA was observed in either type of cell. Thus, GPIIb appears to be specific for the platelet-megakaryocyte membrane and is distinct from the a subunits of the adhesion receptors in other normal tissues