13 research outputs found

    Differential effects of human and plant N-acetylglucosaminyltransferase I (GnTI) in plants

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    In plants and animals, the first step in complex type N-glycan formation on glycoproteins is catalyzed by N-acetylglucosaminyltransferase I (GnTI). We show that the cgl1-1 mutant of Arabidopsis, which lacks GnTI activity, is fully complemented by YFP-labeled plant AtGnTI, but only partially complemented by YFP-labeled human HuGnTI and that this is due to post-transcriptional events. In contrast to AtGnTI-YFP, only low levels of HuGnTI-YFP protein was detected in transgenic plants. In protoplast co-transfection experiments all GnTI-YFP fusion proteins co-localized with a Golgi marker protein, but only limited co-localization of AtGnTI and HuGnTI in the same plant protoplast. The partial alternative targeting of HuGnTI in plant protoplasts was alleviated by exchanging the membrane-anchor domain with that of AtGnTI, but in stably transformed cgl1-1 plants this chimeric GnTI still did not lead to full complementation of the cgl1-1 phenotype. Combined, the results indicate that activity of HuGnTI in plants is limited by a combination of reduced protein stability, alternative protein targeting and possibly to some extend to lower enzymatic performance of the catalytic domain in the plant biochemical environment

    First evidence of the Hepatitis E virus in environmental waters in Colombia

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    RESUMEN: Hepatitis E virus (HEV) is one of the main causes of acute viral hepatitis of enteric transmission. HEV has been detected in environmental samples in several countries from Europe and Asia, constituting a risk factor for waterborne infection. In Colombia, HEV has been identified in samples obtained from patients as well as from swine, but no environmental studies have been carried out. To determine if HEV is present in environmental waters, samples from the main source of drinking water plant and of wastewater system of eight municipalities and two villages of Antioquia state (North West Colombia), were collected between December 2012 and April 2014. The HEV genome was detected by RT-PCR in 23.3% (7/30) of the samples from the main source of drinking water plants and in 16.7% (5/30) from sewage. Viral concentrates obtained from three positive sewage samples were used to inoculate HepG2 cell cultures that were followed for one month; however, the viral genome was not detected in any cell culture. This study demonstrates the circulation of HEV in both source of drinking water plants and wastewater in Antioquia state, Colombia. The presence of HEV in environmental waters could be a risk for waterborne transmission in this population. The findings of the present study, together with the evidence of HEV circulation in human and swine in Colombia, should be consider by public health authorities for the development of surveillance programs and the inclusion of HEV infection diagnosis in the guidelines of viral hepatitis in the country. This is the first report of HEV in environmental samples in Colombia and the second one in Latin America

    Vacuole-Targeted Proteins: Ins and Outs of Subcellular Localization Studies

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    International audienceAccurate and efficient demonstrations of protein localizations to the vacuole or tonoplast remain strict prerequisites to decipher the role of vacuoles in the whole plant cell biology and notably in defence processes. In this chapter, we describe a reliable procedure of protein subcellular localization study through transient transformations of Catharanthus roseus or onion cells and expression of fusions with fluorescent proteins allowing minimizing artefacts of targeting

    Hepatitis E Viruses : Emerging and Transboundary Animal Viruses

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    Hepatitis E virus (HEV), member of genus Orthohepevirus in the family Hepeviridae, is the leading cause of hepatitis E. More than 20 million cases of hepatitis E virus (HEV) infection have been reported globally, out of which 3.3 million suffered from symptomatic illness and 44,000 deaths were recorded. Hepatitis E is considered highly significant on account of its predominance in both developed and developing nations due to poor sanitation and low-grade drinking water. In India and Africa, the major cause of hepatitis E was water contaminated with faecal matter. Reports from East and Southeast Asia signified the role of animal reservoir, especially pigs, for HEV. In Western Asia, the major cases of hepatitis E were associated with blood transfusion. However, in Europe and South and North America, the significant causes of hepatitis E were associated with the consumption of pork products. Till date, major rely is on the enzyme immunoassays involving antibodies for detection and diagnosis purpose. Still, there is a need for the improvement and development of new approaches and methods for diagnosing hepatitis E virus as well as developing HEV vaccines
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