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MINI REVIEW article
Sec. Crop Biology and Sustainability
Volume 4 - 2020 | https://doi.org/10.3389/fsufs.2020.00021
Cauliflower mosaic virus (CaMV) Biology, Management, and Relevance to GM Plant Detection for Sustainable Organic Agriculture
- Department of Plant Pathology, University of California, Davis, Davis, CA, United States
In today's global market, some organic farmers must meet regulatory requirements to demonstrate that their plants and feedstocks are genetically modified organism (GMO)-free. Many GM plants are engineered to contain a promoter from the plant virus, Cauliflower mosaic virus (CaMV), in order to facilitate expression of an engineered target gene. The relative ubiquity of this CaMV 35S promoter (P35S) in GM constructs means that assays designed to detect GM plants often target the P35S DNA sequence, but these detection assays can yield false-positives from plants that are infected by naturally-occurring CaMV or its relatives within the Caulimoviridae. This review places CaMV infection and these ambiguous GM plant detection assays in context, serving as a resource for industry professionals, regulatory bodies, and researchers at the nexus of organic farming and global commerce. We first briefly introduce GM plants from a regulatory perspective, and then we describe CaMV biology, transmission, and management practices, highlighting the relatively widespread nature of CaMV infection in both GM and non-GM crops within the Brassicaceae and Solanaceae families. Finally, we discuss current knowledge of public food safety related to the consumption of CaMV-infected produce.
Organic agriculture, an ecological production management system that serves to promote and enhance biodiversity, biogeochemical cycles, and soil biological activity, has become increasingly popular, with global retail sales reaching more than $80 billion (Brantsæter et al., 2017; Mie et al., 2017). Organic agriculture relies on fertilizers of organic origin, such as compost, and encourages the use of biological pest control. It allows for the use of natural substances, such as pyrethrin and rotenone, while prohibiting most synthetic fertilizers and pesticides (some synthetic substances, such as copper sulfate and elemental sulfur, can be allowed). However, genetically modified organisms (GMOs) and plant growth regulators are prohibited in organic farming in regulated markets (Santhoshkumar et al., 2017).
A genetically modified organism (GMO) is any organism with genetic material that has been altered using genetic engineering techniques, and in the case of GM plants, a new trait is typically introduced to the plant. GM plants can confer advantages such as herbicide resistance, disease resistance, stress (e.g., drought) resistance, and/or insect tolerance (Oliver, 2014), yet GMO use in commercial crops and feedstocks is becoming regulated in some countries (Sharples, 1982; Maghari and Ardekani, 2011; Zhang et al., 2016; Tsatsakis et al., 2017). For example, many countries in the European Union (EU) have banned or imposed legislation to regulate GMOs (Davison, 2010; Lau, 2015; Wong and Chan, 2016). GMO regulation has impacts on international trade (Krueger and Le Buanec, 2008; Ramessar et al., 2008), and the popularity of organic foods, along with the tight regulations surrounding GMOs in some markets, have made GMO detection technologies essential for product labeling and traceability. Indeed, many countries have instituted labeling laws stating that products must be labeled when they contain GMOs above a certain threshold concentration (Wong and Chan, 2016). These thresholds can vary—for example, the threshold is 0.9% in the EU and 5% in Japan, whereas the U.S. has voluntary labeling, and China has “yes-or-no” labeling (Fu et al., 2015). In the U.S., a new law has just passed with mandatory labeling for some GMO products that will be effective in 2020.1
More than 80% of engineered genetic constructs in GM plants are built with the 35S promoter (P35S) from Cauliflower mosaic virus (CaMV) and/or the NOS terminator (TNOS) derived from the soil-borne bacterium, Agrobacterium tumefaciens (Figure 1A). Specifically, as of 2015, P35S and TNOS were used in 65.7 and 53.49%, respectively, of commercialized GMOs, with at least one of the two used in 81.4% of these constructs (Chaouachi et al., 2013; Wu et al., 2014; Fu et al., 2015). Thus, most GMO detection methods are based on marker sequences for P35S and TNOS detected through polymerase chain reaction (PCR) or quantitative PCR (qPCR) (Holden et al., 2010; Wu et al., 2014; Fraiture et al., 2015). However, it is known that CaMV infection of non-GM plants can yield false-positive results in some GMO detection assays, due to the presence of the P35S region in both the CaMV genome and many GMO constructs (Figures 1A,B; Wolf et al., 2000; Chaouachi et al., 2008; Becker and Ulrich, 2018). Although a detailed consideration of A. tumefaciens infections is beyond the scope of this mini-review, A. tumefaciens is naturally found in the soil and can infect plants, so assays designed to detect TNOS can yield false-positive GMO results too, as described elsewhere (Wolf et al., 2000; Escobar and Dandekar, 2003; Yang et al., 2013; Nabi et al., 2016; Becker and Ulrich, 2018).
Figure 1. Schematic representation of (A) a genetically modified (GM) plant construct and (B) the Cauliflower mosaic virus (CaMV) genome, highlighting the shared P35S promoter sequence. (A) In a GM plant, a gene conferring a specific trait (in blue) is inserted, and this transgene is expressed via the 35S promoter, P35S (in yellow) and the Nos terminator, TNOS (in orange). (B) The coding sequences of the seven CaMV genes (I-VII) encoding the seven proteins (Pl-P7) are represented with blue arrows. The CaMV 35S promoter (P35S) is shown in yellow and the 19S promoter (P19S) is shown in orange. P35S overlaps with the end of gene VI. Gene IV overlaps with the end of gene III and the beginning of gene V.
Here, we synthesize relevant information for navigating issues related to CaMV infection and false-positive GM plant detection due to CaMV infection, with particular relevance to sustainable organic farming and international trade of organic products in GMO-regulated markets. We outline CaMV biology and the impacts of CaMV infection on plant health and yield, recommended management practices for reducing CaMV infection of crop plants, available detection assays for GM plants with relevance to CaMV infection, and current knowledge of the safety of human consumption of CaMV in whole or in part.
Cauliflower mosaic virus (CaMV) Biology, Pathology, and Transmission
Cauliflower mosaic virus (CaMV) belongs to the Caulimoviridae family of circular, double-stranded DNA viruses. It predominantly infects members of the Brassicaceae family, including radish, turnip, canola, mustard, cauliflower, broccoli, and cabbage. Some CaMV strains (D4 and W260) are also able to infect Solanaceae species, such as devil's trumpets (genus Datura) and tobacco plants (genus Nicotiana) (Scholelz and Shepherd, 1988). CaMV genetic variants have been described in different host species with different symptoms, virulence, and transmission rates (Covey et al., 1990; Al-Kaff and Covey, 1994; Doumayrou et al., 2013; Yasaka et al., 2014), and recent studies have identified a high diversity of CaMV genomic sequences (Farzadfar et al., 2014; Gong and Han, 2017; Becker and Ulrich, 2018; Sukal et al., 2018).
The CaMV genome consists of approximately 8,000 base-pairs of circular, double-stranded DNA. The genome encodes seven genes (gene I to gene VII), also called P1 to P7 for encoded proteins 1-7. CaMV replicates by reverse transcription (Haas et al., 2002), and its genes are transcribed from two promoters, the 19S and 35S promoters (Figure 1B), which are DNA sequences that define where transcription of a gene (or group of genes) begins. After entry into a plant host cell, the CaMV virus particle migrates to the nucleus, where the viral genome is separated from the viral particle (called decapsidation). The viral DNA is transcribed via the two promoters (P19S and P35S) into two long messenger RNAs (mRNAs), the 19S mRNA that encodes protein P6 and the 35S mRNA that encodes the other six proteins. Translation of the 19S mRNA results in the production of protein P6, which aggregates in numerous cytoplasmic virus factories (Harries et al., 2009; Angel et al., 2013; Rodriguez et al., 2014; Schoelz et al., 2016; Schoelz and Leisner, 2017), where the translation of other viral proteins will take place: the movement protein P1 (required for cell-to-cell movement), the helper component P2 (required for aphid transmission), the virus-associated protein P3, the coat protein P4, and the reverse transcriptase P5. Protein P7 has never been detected in planta and can be deleted by mutagenesis without any effect on virus infection or transmission (Dixon et al., 1986; Wurch et al., 1990).
CaMV is transmitted between host plants by more than 27 aphid species in a non-persistent and non-circulative manner (Kennedy et al., 1962), meaning that after an aphid acquires the virus from an infected plant, the virus does not circulate or replicate within the insect. The virus is retained for a short period (a few hours) in the aphid stylets (mouthparts), where CaMV receptor candidates have recently been identified (Bak et al., 2012, 2013a,b; Martinière et al., 2013; Webster et al., 2018). The virus can then be released to initiate a new infection during aphid feeding on healthy plants. There are no known cases of CaMV transmission via seeds.
CaMV in Agriculture and management practices to reduce CaMV infection
CaMV is a widespread virus in temperate regions and can cause significant loss in Brassicaceae crops, especially in cases of co-infection with other viruses (Shepherd, 1981; Sutic et al., 1999; Spence et al., 2007; Li et al., 2019). CaMV incidence can easily exceed 70%, and subsequent yields may be reduced up to 20–50% (Shepherd, 1981; Sutic et al., 1999). CaMV can affect plant development, especially in early infections, and the production of flowers can be blocked. Low seed yields have also been reported from plants with CaMV infection (Sutic et al., 1999). The virus can induce a range of systemic symptoms, such as chlorosis (loss of green leaf color), mosaic (patches of light and dark green on leaves), vein clearing (abnormal clear or translucent color of veins), and/or stunting (Figure 2). CaMV survives in Brassicaceae crop and weed hosts, including wild radish, turnip weed, canola, mustard, cauliflower, broccoli and cabbage, and weed hosts are known reservoirs for the virus outside the growing season (Farzadfar et al., 2005). Although global GM crop regulations vary widely (see discussion above), canola is a good example of a common GM crop that can also be infected by CaMV. It has recently been shown that water stress can influence CaMV virulence and transmission: under well-watered conditions, viral load, virulence, and transmission rate increased, whereas under water deficit, transmission rate, and virulence decreased (Bergès et al., 2018).
Figure 2. CaMV symptoms on leaves. For each plant type, the left panel shows an uninfected leaf and the right panel shows a leaf from a CaMV-infected plant with typical symptoms, such as mosaic (mottling, e.g., lighter and/or darker green patches, puckered or curled leaves) or vein clearing (yellow or white veins). Top left: turnip (Brassica rapa), bottom left: watercress (Nasturtium officinale), right: canola (Brassica napus).
The best way to minimize CaMV infection is to inhibit aphid contact with seedlings, which are very susceptible to virus infection (Jenkinson and Glynne Jones, 1951; Farzadfar et al., 2005; Shah et al., 2015). Seedbeds can be isolated from aphids with a barrier of cereals or by growing the seedlings under insect-proof mesh (Jenkinson and Glynne Jones, 1951; Broadbent, 1957; Shah et al., 2015). A barrier of cereals may act as a sink for the viruses and/or as a physical barrier, such that aphids will be more likely to land on the tall cereals first and lose their virus contents while probing (Simons, 1957; Toba et al., 1977; Alegbejo and Uvah, 1986; Difonzo et al., 1996; Fereres, 2000). Still, non-persistently transmitted viruses are difficult to control, since the insects only need to feed on the plant briefly to release viruses (Fereres, 2000; Bak et al., 2019), and pesticides are usually not an effective solution because aphids can transmit viruses before the pesticide has an effect (Simons, 1957; Jayasena and Randles, 1985).
CaMV can also be transmitted mechanically by sap inoculation using contaminated hands and pruning tools (Yasaka et al., 2014). It has been shown that CaMV can stay for hours on surfaces such as doors, phones, and gloves, and can be exchanged by hand-shaking (Jiang et al., 1998; Dancer, 2014), so disinfecting tools, equipment, and anything that contacts plants will reduce infection. Though it did not include CaMV, a study that tested how to prevent propagation of plant viruses, such as Pepino mosaic virus (PepMV), Potato spindle tuber viroid (PSTVd), Tomato mosaic virus (ToMV), and Tobacco mosaic virus (TMV), examined sixteen commercially available disinfectants and found that 10% bleach is the most effective solution for preventing viral infection of plants in greenhouse facilities (Li et al., 2015).
GM Plant Detection Methods
Many GM plant detection assays target the 35S promoter (P35S) and/or the terminator NOS (TNOS). Polymerase chain reaction (PCR) was the first technique applied to GM plant detection, and it was mainly used as a fast (~2 h) and low-cost method (Gachet et al., 1998; Iloh et al., 2018; Grohmann et al., 2019). Real-time PCR, also known as qPCR, was developed later and became the preferred technique, providing both qualitative and quantitative results by measuring both the presence and concentration of gene sequences in a given sample (Akiyama et al., 2009; Holden et al., 2010; Wu et al., 2014). Both PCR and qPCR have been reviewed extensively elsewhere (e.g., VanGuilder et al., 2008; Emerson et al., 2017). In the context of GM plant detection, both are used to identify specific genetic regions of interest, based on the hybridization of primers to conserved DNA (or, in some cases, RNA) regions flanking the genetic sequence to be identified, followed by the amplification of the sequence of interest by a polymerase enzyme. In the case of PCR, the result is typically binary (detection or non-detection) and can be visualized by gel electrophoresis. More quantitative results can be attained by qPCR, including the concentration and/or number of copies of a specific genetic region, which can be useful if a threshold allowable concentration has been set by a regulatory agency.
Multiplex qPCR can allow for amplification of several DNA targets at the same time, e.g., for the simultaneous detection of multiple GMO markers in a single reaction (Akiyama et al., 2009; Singh et al., 2016; Cottenet et al., 2019). For example, in Cottenet et al. (2019), the authors developed a new GMO screening method based on two multiplex real-time PCR reactions, targeting six major GM markers in one reaction and six other GM events in another reaction. The method showed a broad screening capacity, due to the large number of targets, and the limit of detection ranged from 0.005 and 0.02% (Cottenet et al., 2019). However, the same fluorophore was used for all markers, so detection was based on the presence or absence of signal with no ability to distinguish among the markers. To distinguish among markers in the same reaction, a high level of multiplexing can be achieved using qPCR with different fluorophores for each target; depending on the instrument, up to five distinguishable targets have been successfully amplified in a single multiplexed qPCR reaction (Reller et al., 2013; Datukishvili et al., 2015). For example, we recently developed a multiplex qPCR assay that can distinguish CaMV infection from GM plants containing P35S in a single reaction, based on detection of four different targets: P35S, CaMV gene III, TNOS, and actin (a universal plant gene used as a positive control for the assay) (Bak and Emerson, 2019).
Given the increasing number and diversity of GMOs developed and the number of molecular biology technologies available in addition to PCR and qPCR, many different GMO detection methods have been developed (Broeders et al., 2012; Fraiture et al., 2015; Demeke and Dobnik, 2018) (for more information, see Supplementary Table 1 and references therein). Databases of GMO sequences and detection assays have also been compiled, facilitating the identification of an appropriate detection method or the development of a new technique for a given need (Dong et al., 2008; Petrillo et al., 2015; Wilkes et al., 2017). To date, there is no international standardized GMO or CaMV detection method, but PCR and qPCR (most commonly targeting P35S and/or TNOS) are still the most commonly used (Grohmann et al., 2019). From a regulatory standpoint, detection thresholds are typically evoked for various requirements. For example, in the European Union, detection of >0.9% GMO content requires a GMO label, but for specific GMOs that are not allowed in the EU under any circumstances, GMO contents must be 0%, which, practically speaking, means below the limit of detection of the assay (Davison, 2010). Thus, for regulatory purposes, the chosen GMO detection method must be sensitive enough to detect a GMO at the required threshold concentration(s), and qPCR is the most widely used method that provides this quantitative information.
CaMV and Food Safety
Public concerns exist regarding whether plant viruses can infect humans, and a number of studies have attempted to answer this question (Bawa and Anilakumar, 2012; Rastogi Verma, 2013; Wunderlich and Gatto, 2015). Although a definitive, universal answer is not possible, here we point to existing literature for insights. Numerous viruses infect plants and are consumed through various types of fresh food and food products (Balique et al., 2015), and two widespread plant viruses (Pepper mild mottle virus, PMMoV, and tobacco mosaic virus, TMV) are the most comprehensively studied, in terms of their effects on human consumption. To the extent that human consumption of CaMV has been studied, we will first present results specific to CaMV, and then we will consider human consumption of PMMoV and TMV.
CaMV commonly and widely infects crucifers, also known as cole crops (cabbage, kale, cauliflower, broccoli, and mustard), and it has been reported that 10% of the cabbages and cauliflowers on sale in supermarkets are infected with CaMV (Raafat El–Gewely, 2001). In an epidemiological study of cauliflower in England, it was shown that 50% of the plants were infected with CaMV (Gong and Han, 2017), yet no ill effects or evidence of human pathogenicity have been found resulting from CaMV-infected plants (Hull et al., 2000). In terms of consumption of parts of CaMV, e.g., the P35S promoter in GM plants, some concerns have been raised, but to our knowledge, there are no scientific publications that have demonstrated a safety risk of eating CaMV, P35S, or any CaMV genetic sequences. In fact, many plants already contain sequences from members of the Caulimoviridae in their genomes as endogenous viral elements (EVEs), including commonly consumed crops, such as Vitis vinifera (grape), Oryza sativa (rice), and a variety of Citrus species (e.g., Citrus clementina, clementines) (Bertsch et al., 2009; Geering et al., 2014), meaning that we have been consuming parts of viruses in our food for centuries.
Still, we review some of the CaMV- and 35S promoter-specific concerns here for context. Sequence overlap exists between P35S and the coding sequences of one CaMV gene, gene VI, also known as P6 (Figure 1), indicating the potential for virus protein expression in humans (Podevin and Du Jardin, 2012). Another concern is the horizontal gene transfer risk with DNA recombination that can potentially occur between P35S and human genes, along with the potential for P35S to facilitate expression of human genes (Chiter et al., 2000; Morel and Tepfer, 2000; Nielsen and Townsend, 2004; Paparini and Romano-Spica, 2006; Bawa and Anilakumar, 2012). In two studies that have attempted to address these concerns, no evidence was observed for the activity of the P35S promoter in mammalian cells (Vlasák et al., 2003; Paparini and Romano-Spica, 2006). In terms of the potential for viruses to recombine with and acquire genes from transgenic plants (i.e., horizontal gene transfer from transgenic plants to a virus), earlier studies of CaMV have demonstrated that this is possible (Schoelz and Wintermantel, 1993; Wintermantel and Schoelz, 1996), but only under very specific conditions, such as strong selection pressure in the laboratory (Wintermantel and Schoelz, 1996).
PMMoV is a widespread plant virus that infects pepper and is found in numerous products containing chilis and peppers (sauce, spicy powder, etc.). For example, PMMoV was found in 57% of 28 pepper-based foods found in supermarkets (Colson et al., 2010). Colson et al. found that humans who consumed PMMoV presented a specific immune response to the virus, including fever and abdominal pain symptoms (Colson et al., 2010). This observation may not be the infection of human cells by the virus itself, but rather viral RNA interfering with the function of human RNA. Tobacco mosaic virus (TMV) is often present in smoked tobacco and, therefore, it is resistant to manufacturing processes (Smith, 1957; Wetter, 1975; Wahyuni et al., 2008). A study found that TMV was viable in 53% of the cigarettes of six different brands (Balique et al., 2012). In addition, 45% of the saliva from 12 smokers, compared to 0% of the saliva from 15 non-smokers, tested positive for TMV RNA (Liu et al., 2013). Another study found that exposure to TMV-infected tobacco products can induce an immune response to TMV in humans; specifically, using an ELISA assay, the authors found anti-TMV antibodies in tobacco smokers (Kamthan et al., 2016). Nevertheless, there was no direct evidence of viral infection in any of these studies, and the symptoms described seem to be an indirect response to the viruses. The extent to which these results for PMMoV and TMV are relevant to CaMV is largely unknown (for example, both PMMoV and TMV have RNA genomes, whereas CaMV is a DNA virus), but these cases demonstrate the potential for some minor symptoms associated with an autoimmune response to plant viruses in some cases. In general, plant viruses are not considered to present pathogenicity to humans (Balique et al., 2015).
In parallel with the development of new GMOs in agriculture, organic farming is increasing (Shi-ming and Sauerborn, 2006; Kamthan et al., 2016). As a result of these farming trends and, in some countries, regulation of GMOs, international trade can necessitate quantification of the GMO content of a given product. Thus, numerous GMO detection methods are emerging (Kamle et al., 2017; Salisu et al., 2017; Umesha and Manukumar, 2018), but the potential for crop infection with CaMV can lead to false-positive results in the most commonly used GMO detection assays, due to the use of a CaMV-derived P35S promoter in many GM constructs (Lipp et al., 1999; Becker and Ulrich, 2018; Lübeck, 2019). This potential for false-positive GMO detection is particularly relevant for organic farmers trying to meet regulatory requirements for non-GM plants in a sustainable international market. Here we have provided information about GMO detection assays and how to disambiguate GMO detection from CaMV infection (e.g., by considering detection targets in the CaMV genome outside the P35S region, such as gene III), management practices to minimize CaMV infection (e.g., planting tall cereals around Brassicaceae crops and adding protective aphid-proof netting around seedlings), and our current understanding of the food safety risk associated with CaMV infection (CaMV is not currently known to pose a risk to humans, but specific studies related to CaMV impacts on human health are limited).
JE conceived of the article. AB performed the literature research and prepared the figures. AB and JE wrote and edited the article. All authors have approved of the final version of the manuscript.
We gratefully acknowledge Amway for funding this work in the form of salary support for AB from a grant awarded to JE.
Conflict of Interest
We have no conflicts of interest to report. In the interest of full transparency, this work was funded by Nutrilite by Amway from a grant awarded to JE. This synthesis article was developed solely by JE and AB, who provided periodic progress reports to Nutrilite by Amway via conference calls. The funding body played no part in directing the research.
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fsufs.2020.00021/full#supplementary-material
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Keywords: Cauliflower mosaic virus, CaMV, organic agriculture, qPCR, GMO
Citation: Bak A and Emerson JB (2020) Cauliflower mosaic virus (CaMV) Biology, Management, and Relevance to GM Plant Detection for Sustainable Organic Agriculture. Front. Sustain. Food Syst. 4:21. doi: 10.3389/fsufs.2020.00021
Received: 29 November 2019; Accepted: 17 February 2020;
Published: 31 March 2020.
Edited by:Avtar Krishan Handa, Purdue University, United States
Reviewed by:James Schoelz, University of Missouri, United States
Tahira Fatima, Purdue University, United States
Copyright © 2020 Bak and Emerson. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Joanne B. Emerson, firstname.lastname@example.org

domain: agronomy


Patient-derived cell lines and animal models have proven invaluable for the understanding of human intestinal diseases and for drug development although both inherently comprise disadvantages and caveats. genes of intestinal diseases. Therefore, here we discuss how patient-derived organoids should be grown and how advanced genome-editing tools may be applied to research on modeling of cancer and infectious diseases. We also highlight practical applications of organoids ranging from basic studies to drug screening and precision medicine. screens in order to predict which drugs will perform as intended (Mokry et al., 2014; van de Wetering et al., 2015). Two-dimensional (2D) monocultures of cell lines lose cell-matrix interactions that are necessary to maintain phenotypes and thus fail to sustain cellular functions that exist in tissues (Jabaji et al., 2013; Gould et al., 2015). Although animal models recapitulate physiology closely, the most obvious problem is the fundamental difference between animal and human being biology (Huch et al., 2013a,b; Karthaus et al., 2014; Nanduri et al., 2014; Gould et al., 2015). The wide-spread execution of organoid systems provides a even more physiologically Azacitidine inhibitor relevant system for high-throughput testing during drug finding (Eglen and Randle, 2015; Walsh et al., 2016). An organoid means three-dimensional (3D) cells originating from body organ stem cells, embryonic stem cells (ESCs), or induced pluripotent stem cells (iPSCs), with framework and function just like those of the initial body organ to some extent (McCracken et al., 2011; Fuller et al., 2012; Knoblich and Lancaster, 2014; Ordonez-Moran et al., 2015; Tamminen et al., 2015; Wieck et al., 2015). Up to now, different organoid systems have already been successfully founded from a particular body organ and could become extended infinitely (Sato et al., 2011; Hisha et al., 2013; Mahe et al., 2013; Takebe et al., 2013; Clevers and Gehart, 2015; Rookmaaker et al., 2015; Xinaris et al., 2015; Yin et al., 2016). For intestinal epithelial organoids, termed the gastric also, intestinal, or colonic epithelial constructions (Stange et al., 2013; DeWard et al., 2014; Watson et al., 2014; Rookmaaker et al., 2015; Kuo and Salahudeen, 2015). organoids possess different advantages over traditional pet versions and cell tradition systems in human being physiological study and disease modeling because (i) an organoid in 3D tradition can form and expand everywhere, thus simulating body organ advancement and morphological features (Mahe et al., 2013; Lancaster and Knoblich, 2014); (ii) an organoid produced from a human being body organ can maintain steadily its features stably and solely after passaging for most decades without significant hereditary or Mouse monoclonal to IgG1 Isotype Control.This can be used as a mouse IgG1 isotype control in flow cytometry and other applications physiological adjustments (DiMarco et al., 2014; Grabinger et al., 2014); (iii) GI organoid versions can be quickly founded by isolating epithelial crypts through the mouse GI system or a human GI biopsy, and crypts can grow into crypt-villus structures in less than 7 days; (iv) the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 (CRISPR/Cas9) system has made it feasible to correct or change Azacitidine inhibitor the human genome (Barker et al., 2010); (ii) bone morphogenetic protein (BMP) inhibitor Noggin diffuses throughout the organoid from the culture medium; this situation leads to the lack of a BMP signaling gradient in intestinal organoids (Shroyer and Wong, 2007); (iii) in human colon and intestinal organoid culture, Wnt and other factors (such as p38 inhibitor, transforming growth factor beta (TGF-) inhibitor, and nicotinamide) prevent stem cell differentiation and reduce cell diversity (Miyoshi et al., 2012; de Lau et al., 2014; Germann et al., 2014; Krausova and Korinek, 2014); (iv) intestinal epithelial organoids consist mainly of epithelial cells without interaction with mesenchymal cells (Wilson et al., 2015). This problem can be overcome with embryonic or iPSCs that can form organoids including subepithelial myofibroblasts, immune cells, and enteric nerves (Spence et al., 2011; Takebe et al., 2013). In addition, modeling of intestinal diseases can be improved by introducing additional cell types from lineages other than the epithelium (Lindemans et al., 2015; Vetizou et al., 2015; Pastula et al., 2016). For instance, co-culturing of an intestinal organoid with Paneth cells significantly increases the plating efficiency (Shamir and Ewald, 2014). A microvascular niche has also been successfully generated through co-culturing Azacitidine inhibitor of tumor cells with stromal cells and endothelial cells (Shamir and Ewald, 2014). To study the complex interaction of intestinal epithelial cells (IECs) and immune cells or microbiota, a model has been optimized to have features similar to those of the system (Table ?Table11). An organoid is cultured together with cytokines produced by immune cells or directly with CD8+ T cells (Lindemans et al., 2015; Vetizou et al., Azacitidine inhibitor 2015). This system allows researchers to examine the proliferation, activation,.


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Sign up for our newsletter. Please visit my Marijuana lightbox. A survey found that 55 percent of men reported ever smoking marijuana in their lifetimes, and 11 percent said that they currently smoked marijuana. Sign Up Please enter a valid email address and try again. Chan School associate professor of nutrition and epidemiology Jorge Chavarro, Ph. Men concerned about their fertility are often warned away from marijuana use, which has been linked to lower sperm counts in previous research. Previous related studies suggest that adverse effects on male reproductive health correlate to cannabis use.
Study Links Marijuana Use With Higher Sperm Counts
PA The men included in the research - mostly white, educated, with an average age of 36 - were all part of couples seeking help with conception from the Massachusetts General Hospital Fertility Centre between and , and most of the men had normal sperm counts. The researchers also linked greater marijuana use to higher levels of testosterone based on the blood samples collected. Is marijuana smoking associated with semen quality, sperm DNA integrity or serum concentrations of reproductive hormones among subfertile men? The sperm of marijuana smokers, it seems, is alright. The study was published in Human Reproduction. Another interpretation, she said, was 'that men with higher testosterone levels are more likely to engage in risk-seeking behaviours, including smoking marijuana'. Fifty-five per cent reported having used cannabis at some point in the past, and 11 per cent said they currently used the drug.
The result came as a big surprise to scientists measuring the sperm counts of more than men from couples attending a fertility clinic. Life on Earth is not as plentiful and may soon be extinct. Meghan Collie GlobalNews. A4M Redefining Medicine: We know a lot less than we think we know," according to EurekAlert. The survey asked the men about past and present drug use, as well as other lifestyle choices.
Sign up for our Health IQ newsletter. Results should be interpreted with caution as the researchers note that further study is required on the health effects of cannabis. These include a higher risk of addiction and developing psychiatric disorders such as schizophrenia. For permissions, please e-mail: Voted one of this year's top 10 Cannabis Strains. Share this article via facebook Share this article via twitter Share this article via messenger Share this with Share this article via email Share this article via flipboard Copy link. Chan School of Public Health researchers wrote in their paper that they expected marijuana use to be associated with lower sperm count; previous research has shown this, but such studies have typically focused on men with drug abuse history.

domain: health


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SILVER AND GOLD SOCIETY TRIP PLANS
Thursday, May 12, 2017 - Legally Blonde The Musical - 2 pm Matinee, Temple Theatre, 120 Carthage St., Sanford, NC, 27330, 7:30 pm, $21 per person, group rate for over 20 guests. A fabulously fun international award-winning musical based on the adored movie, Legally Blonde The Musical, follows the transformation of Elle Woods as she tackles stereotypes, snobbery, and scandal in pursuit of her dreams. This action-packed musical explodes on the stage with memorable songs and dynamic dances. Equal parts hilarious and heart-warming, this musical is so much fun it should be illegal! Sign up in January.
Sunday - June 4, 2017 - Avenue Q - 3 pm - Raleigh Little Theatre, 301 Pogue Street, Raleigh NC 27607, $15 pp; no group rate because of the special First Sunday price. A musical; a recent college grad, Princeton, moves into a shabby New York apartment all the way out on Avenue Q. Along with his new friends, Princeton struggles to find a job, date, and discover an ever-elusive purpose in life. Filled with gut-busting humor and a delightfully catchy score, not to mention puppets, this truly unique show has quickly become a favorite for audiences everywhere. Sign-up in February.
Advisory: Avenue Q may not be appropriate for kids under 13 due to adult situations, language, and nude puppets. Parental guidance is suggested. Be sure to check with your parents!

domain: artistic


Denver (AP) Despite withering criticism from Governor Owens,
the University of Colorado's Board of Regents approved a 20%
tuition hike for in–state students, Thursday. Regents approved the plan 8–to–1, saying it was their only choice in the face of declining state funding.
The decision will cut spending on personnel, maintenance and
other areas by more than $5 million dollars. Regents had initially proposed a 28% tuition hike, which prompted strong criticism from the governor and state education officials.
Owens is also criticizing the decision and is threatening to
bring the issue up when the legislature reconvenes in January.
C.U. Faculty Council President Rod Muth says the governor's
criticism is unfounded, because state funding for the university has plummeted under Owens' administration.
Copyright 2005 by the Associated Press. All Rights Reserved.
© KKCO NBC 11 News - All Rights Reserved


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The Murray River is the greatest of Australia's rivers. The source of its water is the snowy Australian Alps. Salt is a natural feature in many Australian landscapes. The Murray-Darling Basin’s low rainfall and high evaporation have combined to concentrate salt in the groundwater. This salt is produced naturally from the underground brine. A red pigment, carotene, is secreted from the salt tolerant algae. The underground saline waters have been laying dormant for thousands of years. Australian Murray River salt crystals are light & delicate peach colored flakes. They have a wonderfully mild flavor. The texture is ideal for use as a finishing salt. The crystals melt quickly and evenly making Murray River flake salt ideal for finishing, roasting, and baking. Why is it Pink? This pure ancient sea salt contains no additives or preservatives, but is loaded with natural minerals and elements such as magnesium and calcium. These natural mineral brine's give the salt a lovely pink color, as well as a unique and delicious flavor. Our Gourmet Pink Salt are enjoyed by food connoisseurs the world over. Place in a small finger bowl and sprinkle over meals prior to eating. Taste the difference!
|Common Uses||Finishing, Table & Cooking|

domain: chemistry


Latest Hindi Movie Songs Lyrics
Switch to the dark mode that's kinder on your eyes at night time.
Switch to the light mode that's kinder on your eyes at day time.
You are here:
Starring: Dev Anand, Dharmendra, Ashok Kumar, Jackie ShroffDirector: Ashok TyagiProducer(s): T.P. AggarwalMusic Director(s): Jatin Pandit, Lalit PanditLyricist(s): Anand Bakshi, Mehboob Alam KotwalComposer(s): Jatin Pandit, Lalit Pandit
All lyrics are property and copyright of their owners. All the lyrics are provided for educational purposes only. © 2020 Latest Hindi Songs Lyrics


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The ins and outs of Hemp Fabric
The two components of a garment production cycle that create the most waste and exploit the most people is in the fabric production and the garment sewing (also known as CMT).
Our fabric is all produced sustainably and ethically.
About hemp fibres:
Hemp is an incredibly old fibre which can be traced back 8000 BC in the Middle East and China where it was used for textiles, ropes, fishnets, the oil for cosmetic purposes and the seeds for food. It is a fast growing plant (no wonder its called weed!), requires very little water, and uses minimal nutrients from the soil. Hemp plant roots aerate the soil, leaving it rich for future crops. It has a huge yield of 1500 pounds of fibre per acre (compared to 500 pounds from organic cotton) and doesn't require any pesticides or herbicides. The umbrella shape of the plant means it doesn't require herbicides as it out-competes any weeds for sun.
Hemp fabrics are strong, absorbent, durable, and insulating which mean they often take time to wear in, but last a very long time once worn and washed a few times. They don’t stretch out of shape which makes them a perfect upholstery fabric, because it can be pulled tight and retain its shape. Cotton fabric is softer and more comfortable against the skin than hemp fabric, which is why the two are often combined for fabrics, you get the best of both!
Our manufacturer grows over 600,000 square yards of organic hemp, meaning their yield is one of the highest and most sustainable in the world. They use of rainwater for irrigation which reduces water consumption, while free-grazing cow manure is used to fertilize. Hemp is naturally antibacterial and there is no use of pesticides used in their fields. The hemp plant naturally grows very densely (hence a high yield!) which also means there is no need for pesticides as the plant naturally out-competes any weeds for sunlight.
Hemp grows roots that penetrate deep into the soil, which increases nutrients in the soil and eliminate weeds and diseases. Planting hemp contributes greatly to the balance of CO2 content in the atmosphere. One hectare of hemp can sequester and store 20 tons of CO2 in the cellulose carbon and another 500kg of CO2 in the soil within one hundred days growth cycle.
In addition, hemp plant cells have the ability to avoid damage from heavy metal ions, which selectively concentrate in the roots of plants. This means that hemp is an excellent natural heavy metal adsorbent, which has an effective cleaning effect on water and soil in the area.
Our fabric is made from the ‘bast’ hemp plant variety, cultivated in the mountains of Northern China; comfortable, breathable, and cool to the touch. It is a luxurious textile that can be woven to be soft and fine or thick and durable. It is one of the strongest, natural fibres and its antibacterial properties as well as its resistance to UV rays make it ideal for athletic wear.
Hemp in short...
- improves land
- takes time to wear in
- often combined with organic cotton
- doesn't use much water to grow
- no pesticides
- mechanically processed
THE BENEFITS OF HEMP CLOTHING:
Breathable fabric,The cracks distributed on the surface of hemp fiber are connected with small holes, which makes it excellent in moisture absorption and sweat discharge. It is estimated that wearing hemp fabric, compared with cotton fabric, can reduce the body temperature by about 5 ℃.
Antibacterial mould suppression, Hemp fiber is hollow, usually rich in oxygen, so that anaerobic bacteria can not survive. According to the analysis and determination of high frequency plasma emission spectrometer, the hemp fiber also contains phenolic substances, which can resist a variety of microorganisms and damage the formation of mold microorganisms. Therefore, hemp fiber has excellent anti-mildew and bacteriostatic effect.
Antistatic, The molecular structure of hemp fiber is stable and the capacity of generating static electricity is very low. Therefore, the hemp textile will not cause discharge, dust adsorption and pilling due to the clothing swing friction.
WANT TO READ MORE?:


ABSTRACT:Hydrographic data and current meter records from the East Greenland Current, collected during theperiod 1987 to 1989 around the latitude of the central Greenland Sea, reveal the formation of a watermass having the characteristics of Denmark Strait Overflow Water (DSOW). The downstream(southwestward) transport of this newly formed water mass can be substantial, amounting to roughly halfof the outflow of DSOW through Denmark Strait as reported in the literature. The formation of DSOW inthe East Greenland Current results from isopycnic mixing of recirculated Atlantic Water and Upper ArcticIntermediate Water. Substantial formation occurs when the neccessary conditions for baroclinic instabilityare satisfied and the current variability due to mesoscale eddies is high; this is subject to significantchanges from year to year.
AWI Organizations > Climate Sciences > Climate Dynamics
AWI Organizations > Climate Sciences > Sea Ice Physics


Chevrolet Equinox Headlight & Parts
About Headlight & Parts
Modern composite headlights are integrated into the body design for better aesthetics and aerodynamics. These give manufacturers the ability to provide running lights, turn signal lights, and parking lights in the headlight housing, if desired, and to use a variety of lighting like LEDs, projector beams, halos, etc. However, the plastic lenses on these lights degrade over time, becoming abraded from road debris, and discolored, dull and blurry from the sun and external conditions. Many of the replacement headlights appear, install and work much like the factory models, and comply with all acceptable highway use standards. Custom lights are often bolt-on replacements, but some wiring changes can be needed. Projector headlights create a clearer, more concentrated light beam with less scattering loss. Whereas the conventional parabolic reflector reflects light outward in a scattered pattern, the elliptical reflector of the projector beam focuses light into a single focal point in front of the bulb. LED light are highly durable, with an average lifetime of more than 50,000 hours, and the light they emit is much clearer and cleaner than incandescent light.
The key role of the headlights on cars is to illuminate the roadway and facilitate fatigue-free and safe driving. Headlights and their light sources are vital vehicle components to ensure safety. The nature and the location of the light on an automobile and its design, light sources, colors, and photometric values are all regulated by law. We use them to notify other drivers of our presence at night and in foggy situations or a mountain pass where visibility might be very low. Earlier, the headlights were acetylene based flames which had to be manually lit by the driver. Now, there are both high and low beam functions for a lot of driving conditions, and we can toggle the car headlights on or off from inside the passenger compartment as per the requirement. The switch is connected to the 12-volt power supply which runs through the vehicle, and then when it is activated, it sends electricity to the headlights, which powers them up.
Where does Headlight & Parts fit?
The headlights are mounted on the front end of the car, usually below the car hood, on both right and left sides, encased by the bumper panel and the grille, for maximum illumination and safety.
Chevrolet Equinox Headlight & Parts At PartsAvatar
There are many parts to every vehicle, in reality, thousands, but some systems that regulate what your vehicle is doing and how it is doing can be more important than others. The headlight & parts are one of the most significant, and if there are some faults in it, then it should be replaced with a brand new one. At PartsAvatar, we offer the largest array of vehicle headlight & parts, so you will never have to wait for quality parts to fix your vehicle. We offer the best prices on auto parts and deliver them in the shortest time possible from our warehouses spread all across Canada.
How can you find your Chevrolet Equinox Headlight & Parts
Need headlight & parts at a good price? No matter what the headlight & parts requirements are, we’re ready to help you out. We have a wide range of headlight & parts in stock that are specific to the design of each car. If you need one for your car, then what you need to do is choose your car year, make and model from the menu at the top left. If you have a preference for a specific vehicle headlight & parts manufacturer, you can also make a selection for it. You may also find the correct part in the search bar. Once you are sure that you have made the selection for the right fitment then you can simply choose the part you need and make a final purchase.
Top Headlight & Parts Brands
PartsAvatar brings you economical vehicle parts without compromising on quality brands. Our parts are sourced from famous and leading performance parts brands, for example, Blue Streak (Hygrade Motor), Philips, Anzo USA, and many more. We offer handpicked quality headlight & parts to ensure that we have only the top manufacturers in each line of auto items. Now there is no need to choose between an inexpensive and a high-quality part, as you can have them both. A portion of our replacement parts come with lifetime warranties that are superior to OEM parts!


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Swiss Airlines Reservations process
All You Need To Know About Swiss Airlines Reservations Process
Swiss Airlines is one of the finest airlines and also known as the flag carrier of Switzerland. They are also counted among the top members of Star Alliance and has been operating its facilities among 102 destinations. Its business class services are considered top-notch all around the world especially in the European countries.
Steps for Swiss Airlines Reservations
Head over to the Swiss Airlines official homepage in your web browser
Select the Book option from the top header and then select your trip type from round-trip, One-way, Multi-destination
Select your departure & destination airport and then pick your travel dates as per the selected travel type
Now, enter the number of travelers and then choose your desired class from the drop-down menu
Hit the Search flights button to select your flights and then make your seat selection
Next, you can include more services to the itinerary and then enter the required passengers’ details
- At last, make the payments for your itinerary and confirm it
Hence, you should be able to fetch tickets for your selected Swiss Airlines flights. However, if you have any other doubts or queries then you can contact the reservation center to know more about Swiss Airlines Reservations.
An Over View On Swiss Airlines Business Class
Business Class by Swiss Airlines is well known for its luxurious in-flight services and ground services. If you have a membership in the Swiss Choice frequent flyer program then you can use your miles to make your Swiss Airlines Reservations in its Business Class. On the contrary, you can use the online booking portal to fetch your Swiss Airlines Business class flight tickets by paying some good bucks.
Top-Notch Services Offered In Swiss Airlines Business Class
Have a look at the best in class premium services offered in the Swiss Airlines Business class
You will get to enjoy 2-meter long flatbed convertible seats on the touch of a button
The seats will be integrated with massage functionality
Delicious Switzerland based cuisines along with best wines in the flights departing from Zurich
Swiss Universe – an in-flight entertainment program available on both Business & First class
- Better Luggage Allowances
Hand Baggage: 2X8 kg
Checked Baggage: 2X32 kg
- Exclusive access to all the Swiss and Star Alliance partner airline lounges
- Priority Check-ins and much more
if you have any other doubts or queries then you can contact the reservation center to know more about Swiss Airlines.

domain: travel


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Don Jazzy, D’Banj & Wande Coal Reunite (Video)
Don Jazzy, D’Banj & Wande Coal reunite and preform together at the birthday party of Aremo Segun Oniru at the Eko Hotel and Suites.
The fallout between Don Jazzy and D’Banj has been the talked about by their fans.
Nice to see them working together again.
Don Jazzy shared the clip on his IG..

domain: relationship


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This season of Yom HaShoah, three Holocaust survivors recently shared their stories at a“Prejudice & Memory: A Holocaust Exhibit” housed at the Wright Patterson Air Force Base’s U.S. Air Force Museum in Dayton, Ohio.
“Prejudice & Memory: A Holocaust Exhibit” is comprised of the photographs, artifacts, and recorded memories of concentration camp survivors, their families, liberators, and “righteous Gentiles” (non-Jews who helped Jews during the Holocaust), who now live in the Dayton, Ohio, area. The Holocaust Exhibit in Dayton is one of several in the U.S. constructed to honor and remember community members’ connections with the Nazis’ institutionalized genocide of Jews.
This is Larry’s story.
Trending: Science is Settled
Earl Lawrence “Larry” Reynolds found himself in Germany as a medic in the 5th Infantry Rifle Company, 71st Infantry Division, U.S. Army, at the end of World War II. While he saw combat, he got his fill of inhumanity off the battlefield.
One day, two of his fellow soldiers were traveling along a road in a jeep. One, an avid amateur photographer, had two cameras strapped across his chest, scouring the landscape for an interesting picture. They noticed something in the road. As they approached, they observed what Larry described, as a “’bag of bones’ trying to crawl down the road.”
The soldiers immediately contacted their commander for help. After the Red Cross arrived, the battalion began to investigate. What they discovered was Gunskirchen Lager, one of the many hidden starvation camps the Nazis constructed to irradiate the Jews.
Located midway between Lambach and Wels, Austria, and camouflaged by a beautiful forest, the Nazis had fenced off a section to use as a starvation camp.
When the 71st Infantry arrived, they discovered an incomplete barracks, which allowed the cold air to infiltrate the building while prisoners huddled inside. There was one cot and one blanket for each already barely clothed prisoner.
Larry remarked that while Dachau and other camps were originally built to hold political prisoners, the Nazis needed more space to hold more Jews, and more efficient, less expensive ways to kill them.
Many stories have been told about the Nazis digging ditches, lining up Jews, and shooting them. They would throw lime on the dead bodies, before setting up the next group to be executed. Once the war started, however, Hitler found this method to be too expensive, costing ammunition and valuable manpower needed on the Russian front.
To save ammunition, the Nazis next lined up their targets five deep, using one bullet to pierce through them all. Next, they would be pushed into the ditch, often while some were still alive, only to have lime and more bodies thrown on top of them while they gasped for air.
Architect Albert Speer offered an even more economic resolution to accelerate the Holocaust: the gas chamber. It was Speer who designed the gas chamber and the furnace used in concentration camps to quickly and easily dispose of Jews and other undesirables. These more efficient killing methods enabled Hitler to free up the resources he needed on the battlefield.
However, Larry pointed out that not all concentration camps used these methods. Gunskirchen Lager chose starvation.
The only food available to Jewish prisoners was what they could forage on the forest floor. At night, with only one blanket apiece, the prisoners would huddle up, layering the blankets and using body heat to keep warm. Inevitably, starvation, dehydration, and cold would claim a few casualties overnight.
On May 4, 1945, the 71st Infantry Division liberated Gunskirchen, one of the many sub-camps of the Mauthausen concentration camp in Austria. Some of the victims became honorary members of the unit.
Over the years, Larry visited with survivors of the camp. He recounted a visit he had with a survivor who in desperation had once swallowed whole snails, as that was all he could find to eat. Even though he could feel the snails moving in his stomach, he was thankful for the nourishment.
Larry relayed another man’s experience, which rendered the audience silent.
Upon arriving at the camp with his parents, sister, and her newborn baby, the sister pleaded with the guards not to harm her baby. One guard promptly grabbed the infant by its legs, smashed its head against a rock, and then casually threw it into the bushes. He then directed each family member where they should stand, acting as if his actions were nothing more than standard operating procedure.
Larry has also reunited with fellow veterans. The amateur photographer who discovered the escaped prisoner had a warming story about his hobby. As a traveling salesman, he often had spare time with nothing to do. While in Chicago, he noticed a business across the street from his hotel that advertised book binding. Having his box of war photos in the car trunk, he wondered if they could do something with them.
After proposing his idea to the woman at the front desk, she started going through the box of pictures to see what he had. She stopped on one photo, screamed, and then ran to the back office, frantically talking to her associate. A moment later, a man emerged and began inquiring about the photo.
“Where did you get this?” he asked.
“I took it,” Larry’s friend said.
“Where?” the man asked.
The soldier turned the photo over to find the date and location scribbled on the back. After relaying the information to the business owner, the man replied,
“This is my sister. We thought she was dead.”
His soldier friend never found out if the couple was able to locate his sister, but he was happy that he could give them hope.
Americans were not fully made aware of the atrocities of the Holocaust until LIFE Magazine found and published many photographs taken by the U.S. military.
Roosevelt knew. High ranking officials knew. But the average American citizen and active soldier during the war did not know. It was not until units like the 71st found and liberated the camps during the occupation after the war, that the true savage nature of the Nazi regime became exposed.
Many of those liberated from Gunskirchen Lager were fortunate to go on to have families and lead happy lives. One woman’s granddaughter even married Larry’s nephew. In fact, Earl Lawrence “Larry” Reynolds is my uncle.
It is imperative that Americans never forget what happened, and continue to discover and tell these stories from survivors and camp liberators. The fastest way for these atrocities to happen again is for us to live as if they never happened.
A partial or full audio tour of the museum can be heard on the Museum’s podcasts.
Hear another eye witness testimony of a 71st Division veteran who helped liberate Gunskirchen Lager:

domain: history


Since the MSM is starting to make note of the fact that the IRS won’t be processing itemized tax returns until next Monday (!#@#$##@ it!!!), I thought I’d go ahead and post my thoughts on a better tax system.
First, let me make it clear that I have not really looked at the “Flat Tax” or “Fair Tax” proposals that are already out there. It’s entirely possible my idea duplicates part or even all of it. At most, it was the idea of a flat tax rate that started this idea that’s been kicking around in my head for a while now.
So, here’s my idea for a Reasonable Tax.
- Only the states may levy taxes on citizens of that state or corporations based in that state, and ONLY on citizens of that state or corporations based in that state.
- Only income may be taxed. Income is defined as salary or wages for employment, interest and dividends from investments, earnings from rental property, or similar earned monies. Sales tax, property taxes, and “inheritance” or “estate” taxes will be eliminated.
- The maximum tax rate allowable is 10% of total income, annually. Period. Local taxes are deducted from this 10% maximum (e.g., if your county tax amounts to 2% of your income, the state can only tax you up to 8% of your income). States may restrict the maximum percentage localities may set for taxes.
- States may tax businesses or their owners/stockholders, but not both (no “double-dipping” by taxing a business’s income 10% and then the owners’/stockholders’ incomes 10%).
- The federal government may levy taxes on each state, and only on the states, and no more than 10% of the total taxes received by that state.
- The 10% rate is a statutory/Constitutional maximum. Each state government may elect to tax less than that 10% maximum. The federal government may elect to tax the states less than the 10% maximum.
- Tax rates must be flat. If a state sets the rate at 8%, it is 8% for all citizens regardless of income level. However, states may set a “poverty level” below which all taxes are waived (all or nothing). The same rules apply to the federal government in taxing states.
- Tax rates must be set by legislative act, which cannot be delegated, and are effective for a minimum of 3 years. The legislature may raise the tax rate within that 3 year period only with a 3/4 majority vote. Such override shall be effective for no longer than one year. At the end of that year, a new bill must be passed, again with a 3/4 majority vote of the legislature, or the tax rate automatically resets to the lower previous rate. The rate may be lowered at any time with a simple majority vote.
- Any bill changing the tax rate must specifically address only the tax rate, in the form of “Effective [date], the state income tax shall be X% calculated annually. Those with an income of $Y per year or less shall be exempt from any income tax.” Any bill changing the tax rate can only take effect at the beginning of the next tax year.
There are probably all sorts of specifics that would need to be worked out to reduce potential abuse and loopholes, but this is really more of a base to start from than a complete plan, anyway.
The intent here is to limit the power of both the federal and state governments by limiting the power to tax, to prevent the ruinous taxation of the people by setting an absolute maximum rate, and to motivate the states to limit the federal government’s power by placing the burden of funding of the federal government directly on the state governments. Additionally, it makes income tax more fair by taxing everyone the same percentage.


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Students who turn in research papers four months late are likely to be rewarded with a big fat zero; the Department of Justice, on the other hand, has to face the wrath of Sen. Patrick Leahy (D-VT), who chairs the Senate Judiciary Committee. Leahy was unhappy after the DoJ turned in a late report on the FBI's use of data mining, but he was unhappier still about the report's conclusions than its tardiness.
"This report raises more questions than it answers and demonstrates just how dramatically the Bush Administration has expanded the use of this technology, often in secret, to collect and sift through Americans' most sensitive personal information," Leahy said in a statement. "Unfortunately, the Congress and the American public know very little about these and other data mining programs, making them ripe for abuse."
The American public does know a bit more about the data mining efforts now that the DoJ report has been submitted to Congress. According to those who have seen a copy of the document, it spells out six different ways that the FBI uses data mining in an attempt to find criminals. Sure, there's a component that attempts to "score" potential terrorists, but there are also programs in place to detect identity theft rings, health insurance fraud, problems at Internet pharmacies, fraudulent home purchases, and false insurance claims from car accidents that never happened (BusinessWeek has a nice rundown on the categories).
The most controversial system is likely to be the newest one, a terrorist ranking system known as the System to Assess Risk (STAR). The system won't pop up bright red screens that say "Terrorist!" or anything, but government officials do admit that it will score those considered to be a potential threat. A higher score could earn you a visit from the Feds, though no one's saying exactly how the algorithms work or what information is included in the databases. What is known is that STAR will focus on foreigners and that such things as country of origin will be taken into account by the ranking system. Here on a visit from Pakistan? +25 right there.
Several of these projects sound relatively innocuous. Does anyone object, for instance, to the FBI using public real estate transaction information to nail people making fraudulent home purchases? But the government has really set itself up for suspicion when it rolls out such programs, especially when they relate to national security. That's because of the government's repeated interest in things like the now-defunct Total Information Awareness program—a program that was shot down by Congress.
Despite Congress' obvious uneasiness with massive data mining projects that target American citizens, the government has plowed ahead with things like ADVISE and CAPPS II and ATS, screening programs whose details are largely secret and whose information cannot be checked for accuracy by those that it most affects. Several Senators and Congressmen have even ended up on TSA watch lists, further fueling fears that non-targeted data mining is likely to finger plenty of innocent people. The TSA, which runs several of these programs, didn't help the government's position when it lied about how it was using private data to test CAPPS II.
The FBI itself has come under plenty of recent criticism for its (mis)use of National Security Letters sent to ISPs and its apparently illegal use of "exigent letters" to demand information without a warrant or an NSL. NSLs have been controversial for some time, but they entered the news again this week as the EFF made public a massive trove of data on them gleaned from a Freedom of Information Act request to the FBI. The concern over NSL use at the Bureau is likely to take away any "benefit of the doubt" that Congressional oversight committees might have been tempted to grant the FBI with respect to its data mining programs.
The STAR program appears to take past criticisms to heart by targeting a narrow subset of people in the US, but it's clear that Congress wants to know more about what's being done and what sort of safeguards are in place. The Senate Judiciary Committee held a hearing on data mining back in January that focused on the privacy implications of such programs; at that time, Leahy claimed that "52 different federal agencies are currently using data mining technology, and there are at least 199 different government data mining programs operating or planned throughout the federal government." Now that he knows about six more programs, another hearing may be in the works.

domain: law


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posted by stat .
ok there was a question that my teacher told me i got wrong
the question asked me to compute the mean which I did...
then the question went on to tell me compute the mean with another observation equal to zero
then the part that he drew the line through was my response to this question
Write a sentence about the effect of the zero on Joey\'s quiz average that mentions this property...
The property that this illustrate of the mean is the observations. Sense the number of observations increased but the sum of the observations stayed the same this brough down the mean by defintion...
ok he drew a line threw it and told me it was wrong so my question is how is this wrong?
What would a correct answer include?
Thanks for any Help!
It depends on what he taught in class, but I am guessing that what the problem was trying to illustrate is that arithmetic means are very sensitive to "wild" data, or erroneous data that are out of range. This is probably in preparation to the next topic, medians and mode.
What you have stated are mathematical facts and perhaps not considered as illuminating observations.

domain: mathematics