A human embryonic kidney 293T cell line mutated at the Golgi -mannosidase II locus

Disruption of Golgi -mannosidase II activity can result in type II congenital dyserythropoietic anemia and can induce lupus-like autoimmunity in mice. Here, we isolate a mutant human embryonic kidney (HEK) 293T cell line, called Lec36, that displays sensitivity to ricin that lies between the parental HEK 293T cells, whose secreted and membrane-expressed proteins are dominated by complex-type glycosylation, and 293S Lec1 cells, which only produce oligomannose-type N-linked glycans. The stem cell marker, 19A, was transiently expressed in the HEK 293T Lec36 cells, and in parental HEK 293T cells with and without the potent Golgi -mannosidase II inhibitor, swainsonine. Negative-ion nano-electrospray ionization mass spectra of the 19A N-linked glycans from HEK 293T Lec36 and swainsonine-treated HEK 293T cells were qualitatively indistinguishable and, as shown by collision-induced dissociation spectra, dominated by hybrid-type glycosylation. Nucleotide sequencing revealed mutations in each allele of MAN2A1, the gene encoding Golgi -mannosidase II: a point mutation in one allele mapping to the active site and an in-frame deletion of twelve-nucleotides in the other. Expression of wild-type but not the mutant MAN2A1 alleles in Lec36 cells restored processing of the 19A reporter glycoprotein to complex-type glycosylation. The Lec36 cell line will be useful for expressing therapeutic glycoproteins with hybrid-type glycans and provides a sensitive host for detecting mutations in human MAN2A1 causing type II congenital dyserythropoietic anemia

Violence and affective states in contemporary Latin America

This special issue brings together scholars interested in the analysis of the social, cultural and affective dimensions of violence. The contributions explore the connections between situated experiences of violence and shifting affective states, relations, sensations and contingencies in contemporary Latin America. The articles consider how violence might constitute a nexus for the production of subjectivities and forms of identification, relationality and community, alterity and belonging, in a range of Latin American contexts including Argentina, Brazil, Guatemala, Mexico and in the Mexican diaspora in Spain

Effect of co-trimoxazole on mortality in HIV-exposed but uninfected children in Botswana (the Mpepu Study): a double-blind, randomised, placebo-controlled trial

Background Co-trimoxazole prophylaxis reduces mortality among HIV-infected children, but efficacy in HIV-exposed but uninfected (HEU) children in a non-malarial, low-breastfeeding setting with a low risk of mother-to-child transmission of HIV is unclear. Methods HEU children in Botswana were randomly assigned to receive co-trimoxazole (100 mg/20 mg once daily until age 6 months and 200 mg/40 mg once daily thereafter) or placebo from age 14–34 days to age 15 months. Mothers chose whether to breastfeed or formula feed their children. Breastfed children were randomly assigned to breastfeeding for 6 months (Botswana guidelines) or 12 months (WHO guidelines). The primary outcome, analysed by a modified intention-to-treat approach, was cumulative child mortality from treatment assignment to age 18 months. We also assessed HIV-free survival by duration of breastfeeding. This trial is registered with ClinicalTrials.gov, number NCT01229761. Findings From June 7, 2011, to April 2, 2015, 2848 HEU children were randomly assigned to receive co-trimoxazole (n=1423) or placebo (n=1425). The data and safety monitoring board stopped the study early because of a low likelihood of benefit with co-trimoxazole. Only 153 (5%) children were lost to follow-up (76 in the co-trimoxazole group and 77 in the placebo group), and 2053 (72%) received treatment continuously to age 15 months, death, or study closure. Mortality after the start of study treatment was similar in the two study groups: 30 children died in the co-trimoxazole group, compared with 34 in the placebo group (estimated mortality at 18 months 2·4% vs 2·6%; difference –0·2%, 95% CI –1·5 to 1·0, p=0·70). We saw no difference in hospital admissions between groups (12·5% in the cotrimoxazole group vs 17·4% in the placebo group, p=0·19) or grade 3–4 clinical adverse events (16·5% vs 18·4%, p=0·18). Grade 3–4 anaemia did not differ between groups (8·1% vs 8·3%, p=0·93), but grade 3–4 neutropenia was more frequent in the co-trimoxazole group than in the placebo group (8·1% vs 5·8%, p=0·03). More co-trimoxazole resistance in commensal Escherichia coli isolated from stool samples was seen in children aged 3 or 6 months in the co-trimoxazole group than in the placebo group (p=0·001 and p=0·01, respectively). 572 (20%) children were breastfed. HIV infection and mortality did not differ significantly by duration of breastfeeding (3·9% for 6 months vs 1·9% for 12 months, p=0·21). Interpretation Prophylactic co-trimoxazole seems to offer no survival benefit among HEU children in non-malarial, low-breastfeeding areas with a low risk of mother-to-child transmission of HIV

Cotrimoxazole Prophylaxis and Risk of Severe Anemia or Severe Neutropenia in HAART-Exposed, HIV-Uninfected Infants

Background: Prophylactic cotrimoxazole is recommended for infants born to HIV-infected mothers. However, cotrimoxazole may increase the risk of severe anemia or neutropenia. Methods: We compared the proportion of HIV-exposed uninfected (HIV-EU) infants experiencing incident severe anemia (and separately, severe neutropenia) between a prospective cohort receiving prophylactic cotrimoxazole from 1 to 6 months vs. infants from two prior trials who did not receive cotrimoxazole. Infants were from rural and urban communities in southern Botswana. Results: A total of 1705 HIV-EU infants were included. Among these 645 (37.8%) were fed with iron-supplemented formula from birth. Severe anemia developed in 87 (5.1%) infants, and severe neutropenia in 164 (9.6%) infants. In an analysis stratified by infant feeding method, there were no significant differences in the risk of severe anemia by prophylactic cotrimoxazole exposure–risk difference, −0.69% (95% confidence interval [CI] −2.1 to 0.76%). Findings were similar in multivariable analysis, adjusted odds ratio (aOR) 0.35 (95% CI 0.07 to 1.65). There were also no significant differences observed for severe neutropenia by cotrimoxazole exposure, risk difference 2.0% (95% CI −1.3 to 5.2%) and aOR 0.80 (95% CI 0.33 to 1.93). Conclusions: Severe anemia and severe neutropenia were infrequent among HIV-exposed uninfected infants receiving cotrimoxazole from 1–6 months of age. Concerns regarding hematologic toxicity should not limit the use of prophylactic cotrimoxazole in HIV-exposed uninfected infants. ClinicalTrials.gov Registration Numbers NCT01086878 (http://clinicaltrials.gov/show/NCT01086878), NCT00197587 (http://clinicaltrials.gov/show/NCT00197587), and NCT00270296 (http://clinicaltrials.gov/show/NCT00270296)

First Study of Combined Blazar Light Curves with FACT and HAWC

For studying variable sources like blazars, it is crucial to achieve unbiased monitoring, either with dedicated telescopes in pointing mode or survey instruments. At TeV energies, the High Altitude Water Cherenkov (HAWC) observatory monitors approximately two thirds of the sky every day. It uses the water Cherenkov technique, which provides an excellent duty cycle independent of weather and season. The First G-APD Cherenkov Telescope (FACT) monitors a small sample of sources with better sensitivity, using the imaging air Cherenkov technique. Thanks to its camera with silicon-based photosensors, FACT features an excellent detector performance and stability and extends its observations to times with strong moonlight, increasing the duty cycle compared to other imaging air Cherenkov telescopes. As FACT and HAWC have overlapping energy ranges, a joint study can exploit the longer daily coverage given that the observatories' locations are offset by 5.3 hours. Furthermore, the better sensitivity of FACT adds a finer resolution of features on hour-long time scales, while the continuous duty cycle of HAWC ensures evenly sampled long-term coverage. Thus, the two instruments complement each other to provide a more complete picture of blazar variability. In this presentation, the first joint study of light curves from the two instruments will be shown, correlating long-term measurements with daily sampling between air and water Cherenkov telescopes. The presented results focus on the study of the variability of the bright blazars Mrk 421 and Mrk 501 during the last two years featuring various flaring activities.Comment: 6 pages, 2 figures. Contribution to the 6th International Symposium on High Energy Gamma-Ray Astronomy (Gamma2016), Heidelberg, Germany. To be published in the AIP Conference Proceeding

High-frequency climate variability in the Holocene from a coastal-dome ice core in east-central Greenland

An ice core drilled on the Renland ice cap in east-central Greenland contains a continuous climate record dating through the last glacial period. The Renland record is valuable because the coastal environment is more likely to reflect regional sea surface conditions compared to inland Greenland ice cores that capture synoptic variability. Here we present the δ¹⁸O water isotope record for the Holocene, in which decadal-scale climate information is retained for the last 8 kyr, while the annual water isotope signal is preserved throughout the last 2.6 kyr. To investigate regional climate information preserved in the water isotope record, we apply spectral analysis techniques to a 300-year moving window to determine the mean strength of varying frequency bands through time. We find that the strength of 15–20-year δ¹⁸O variability exhibits a millennial-scale signal in line with the well-known Bond events. Comparison to other North Atlantic proxy records suggests that the 15–20-year variability may reflect fluctuating sea surface conditions throughout the Holocene, driven by changes in the strength of the Atlantic Meridional Overturning Circulation. Additional analysis of the seasonal signal over the last 2.6 kyr reveals that the winter δ¹⁸O signal has experienced a decreasing trend, while the summer signal has predominantly remained stable. The winter trend may correspond to an increase in Arctic sea ice cover, which is driven by a decrease in total annual insolation, and is also likely influenced by regional climate variables such as atmospheric and oceanic circulation. In the context of anthropogenic climate change, the winter trend may have important implications for feedback processes as sea ice retreats in the Arctic