Prevalence of Hepatitis E Virus Antibodies, Israel, 2009–2010

We investigated prevalence of hepatitis E virus in a sample of the population of Israel. The overall seroprevalence of antibodies to the virus was 10.6% (95% CI 8.4%–13.0%); age-adjusted prevalence was 7.6%. Seropositivity was associated with age, Arab ethnicity, low socioeconomic status, and birth in Africa, Asia, or the former Soviet Union

Generation of ligand-receptor alliances by "SEA" module-mediated cleavage of membrane-associated mucin proteins

A mechanism is described whereby one and the same gene can encode both a receptor protein as well as its specific ligand. Generation of this receptor–ligand partnership is effected by proteolytic cleavage within a specific module located in a membrane resident protein. It is postulated here that the "SEA" module, found in a number of heavily O-linked glycosylated membrane-associated proteins, serves as a site for proteolytic cleavage. The subunits generated by proteolytic cleavage of the SEA module reassociate, and can subsequently elicit a signaling cascade. We hypothesize that all membrane resident proteins containing such a "SEA" module will undergo cleavage, thereby generating a receptor–ligand alliance. This requires that the protein subunits resulting from the proteolytic cleavage reassociate with each other in a highly specific fashion. The same SEA module that serves as the site for proteolytic cleavage, probably also contains the binding sites for reassociation of the resultant two subunits. More than one type of module can function as a site for proteolytic cleavage; this can occur not only in one-pass membrane proteins but also in 7-transmembrane proteins and other membrane-associated proteins. The proposal presented here is likely to have significant practical consequences. It could well lead to the rational design and identification of molecules that, by binding to one of the cleaved partners, will act either as agonists or antagonists, alter signal transduction and, hence, cellular behavior

Inferring Numbers of Wild Poliovirus Excretors Using Quantitative Environmental Surveillance

Response to and monitoring of viral outbreaks can be efficiently focused when rapid, quantitative, kinetic information provides the location and the number of infected individuals. Environmental surveillance traditionally provides information on location of populations with contagious, infected individuals since infectious poliovirus is excreted whether infections are asymptomatic or symptomatic. Here, we describe development of rapid (1 week turnaround time, TAT), quantitative RT-PCR of poliovirus RNA extracted directly from concentrated environmental surveillance samples to infer the number of infected individuals excreting poliovirus. The quantitation method was validated using data from vaccination with bivalent oral polio vaccine (bOPV). The method was then applied to infer the weekly number of excreters in a large, sustained, asymptomatic outbreak of wild type 1 poliovirus in Israel (2013) in a population where >90% of the individuals received three doses of inactivated polio vaccine (IPV). Evidence-based intervention strategies were based on the short TAT for direct quantitative detection. Furthermore, a TAT shorter than the duration of poliovirus excretion allowed resampling of infected individuals. Finally, the method documented absence of infections after successful intervention of the asymptomatic outbreak. The methodologies described here can be applied to outbreaks of other excreted viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), where there are (1) significant numbers of asymptomatic infections; (2) long incubation times during which infectious virus is excreted; and (3) limited resources, facilities, and manpower that restrict the number of individuals who can be tested and re-tested

Human cancer cells express MUC1-ARF in both the nucleus and cytoplasm.

<p>T47D breast cancer cells (Panels A and C) and COLO357 pancreatic cancer cells (Panels B) were immunostained with anti-MUC1-ARF mAb MPR2G10 followed by red-labeled secondary antibody. DAPI (blue, demonstrating nuclei: white stippled ovals) and green-labeled phalloidin (labeling actin filaments) are shown in the merged images (Panels A' and B'). Immunostaining of T47D cells with anti-MUC1-ARF mAb MPR2G10 is abrogated when done in the presence of competing MUC1-ARF peptide (compare Panels C with Panels D). Simultaneous immunostaining of ZR75 breast cancer cells with anti-MUC1-TM antibodies (DMB5F3 mAbs directly green labeled) and anti-MUC1-ARF antibodies (MPR2G10, red labeled) in the absence of MUC1-TM-junction peptide is shown in Panels E, while the effect of adding MUC1-TM-junction peptide to an identical immunostaining of ZR75 breast cancer cells is shown in Panels F. (Panel G) Lysates of human COLO357 cancer cells were resolved on SDS-PAGE, western blotted, and probed with anti-MUC1-ARF MPR2G10. MUC1-ARF protein is indicated by the filled red arrow head (left panel). Immunoreactivity is abrogated by addition of competing free MUC1-ARF peptide (right panel). (Panel H, left side, labeled MUC1-ARF) Equivalent amounts of protein from either cytoplasmic or nuclear (cyt or nuc) T47D cell extracts were analyzed by a sandwich ELISA that detects MUC1-ARF. Competing MUC1-ARF peptide (ARF pep) added to the detecting biotinylated anti-MUC1-ARF MPR4B3, abolished signal in both cytoplasmic and nuclear samples, whereas non-relevant peptide (non-rel. pep) had no effect. (Panel H, right side, labeled MUC1-TM)- analysis of nuclear and cytoplasmic T47D cell extracts with a sandwich ELISA detecting MUC1-TM protein. (Panel I) Mouse DA3 mammary tumor cells expressing human MUC1 cDNA were immunostained with anti MUC1-ARF antibody MPR2G10 and red-labeled secondary antibody followed by DAPI staining. High magnification images of orthogonal projections of confocal laser microscopy are shown for anti-MUC1-ARF (Panel I-i), DAPI (Panel I-ii) and merged images (Panel I-iii). (Panel J) Untransfected mouse DA3 mammary tumor cells (DA3-PAR, left panels) or transfected with human MUC1 cDNA (DA3-MUC1, J, right panels) were stained with DAPI and immunostained with anti-MUC1-ARF monoclonal followed by red-labeled secondary antibody. DAPI staining alone (blue) and the merged images of DAPI plus red anti-ARF immunostaining (DAPI + anti-ARF) are shown. A parallel set of cells (Panel J, lower panels, plus etop.) were treated with Etoposide, a DNA topoisomerase II inhibitor.</p

MUC1-TM and MUC1-ARF expression in normal human kidney and pancreas.

<p>Serial sections of paraffin-embedded human pancreatic and renal tissues were immunohistochemically stained with anti-MUC1-TM SEA module antibodies (anti-MUC1-TM [DMB5F3]) and anti-MUC1-ARF antibodies (anti-MUC1-ARF [MPR2G10]) as indicated. Normal kidney is shown in Panels A-i (MUC1-TM) and A-ii (MUC1-ARF); larger fields and higher magnifications are shown in Panels C-i, C-i’ (MUC1-TM), and D-i, D-i’ (MUC1-ARF)]. Glomerulus, proximal tubule, and distal tubule are designated by <u><b>G</b></u>, <u><b>PT</b></u> and <u><b>DT</b></u> respectively. Filled green arrows designate sites of MUC1-TM-SEA protein at the cell surface whereas filled red arrows designate MUC1-ARF protein; absence of anti-MUC1-ARF immunoreactivity is shown by filled red and white arrows. Normal pancreatic tissue reacted with anti-MUC1-TM in the presence of ARF peptide (plus ARF pep) or in the presence of MUC1-TM SEA peptide (plus SEA pep), are shown in Panels B-i and B-i", respectively. MUC1-TM protein on the cell surface of ductal epithelial cells is competed out by MUC1-TM SEA peptide (B-i") but not by MUC1-ARF peptide (B-i). Conversely MUC1-ARF protein in the nuclei of pancreatic epithelial cells is competed out by MUC1-ARF peptide (B-ii") but not by MUC1-SEA peptide (B-ii). Larger fields and higher magnifications of pancreatic tissue is shown in E-i, E-i’ (MUC1-TM), and E-ii, E-ii’ (MUC1-ARF).</p

Detection of nuclear MUC1-ARF protein with polyclonal anti-MUC1-ARF antibodies and with three distinct anti-MUC1-ARF monoclonal antibodies, MPR2G10, MPR4B3 and MPR5C9.

<p>(A) Polyclonal anti-MUC1-ARF antibodies and (B) three independently isolated anti-MUC1-ARF monoclonal antibodies, MPR2G10, MPR4B3 and MPR5C9, were reacted in the presence of competing ARF peptide (B, panels 4), in its absence (B, panels 2), or with a non-relevant peptide (B, panels 3) with mouse DA3 cells stably transfected with and expressing human MUC1 DNA (DA3-MUC1) and with T47D human breast cancer cells that endogenously express MUC1. Parental DA3 cells (DA3-PAR) which do not express human MUC1 are shown in B, panels 1. Immunofluorescence of secondary antibody (red), DAPI staining of nuclei (blue) are shown in the merged images. (C) Mouse DA3 cells expressing human MUC1 reacted with anti-MUC1-ARF repeat monoclonal antibody MPR2G10 (left panel), compared with anti-MUC1-TM tandem repeat monoclonal antibody H23 (right panel), both followed by red-labelled secondary antibody.</p

Immunohistochemical analyses of MUC1-TM and MUC1-ARF expression in breast and pancreatic cancer.

<p>(A) Serial sections of breast cancer tissues from three distinct individuals were immunohistochemically stained with anti-MUC1-TM antibodies (anti-MUC1-TM [DMB5F3]), anti-MUC1-ARF antibodies (anti-MUC1-ARF [MPR2G10]) and with hematoxylin/eosin (H and E). Green arrows indicate sites of MUC1-TM reactivity at the cell surface, and red arrows designate MUC1-ARF reactivity in the nuclei. Absence of anti-MUC1-ARF immunoreactivity is shown by filled red and white arrows. (B) Binding specificity of anti-MUC1-ARF [MPR2G10] antibody is demonstrated by addition of either MUC1-ARF peptide or a non-relevant peptide, as indicated. Only MUC1-ARF peptide abrogates immunoreactivity. Serial sections of normal pancreas (C) or pancreatic cancer tissue (D), were immunohistochemically stained with anti-MUC1-TM [DMB5F3] or anti-MUC1-ARF [MPR2G10]. Both MUC1-TM and MUC1-ARF are expressed in the normal pancreatic tissue (C). In contrast, cancer tissue expresses only MUC1-TM. MUC1-ARF was not detected (red and white arrows).</p