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Pathogenesis and Immunity
Enhancement or Attenuation of Disease by Deletion of Genes from Citrus Tristeza Virus
Satyanarayana Tatineni, William O. Dawson
Satyanarayana Tatineni
aCitrus Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, Lake Alfred, Florida, USA
bUnited States Department of Agriculture, Agricultural Research Service, and Department of Plant Pathology, University of Nebraska—Lincoln, Lincoln, Nebraska, USA
William O. Dawson
Stem pitting is a common virus-induced disease of perennial woody plants induced by a range of different viruses. The phenotype results from sporadic areas of the stem in which normal xylem and phloem development is prevented during growth of stems. These alterations interfere with carbohydrate transport, resulting in reduced plant growth and yield. Citrus tristeza virus (CTV), a phloem-limited closterovirus, induces economically important stem-pitting diseases of citrus. CTV has three nonconserved genes (p33, p18, and p13) that are not related to genes of other viruses and that are not required for systemic infection of some species of citrus, which allowed us to examine the effect of deletions of these genes on symptom phenotypes. In the most susceptible experimental host, Citrus macrophylla, the full-length virus causes only very mild stem-pitting symptoms. Surprisingly, we found that certain deletion combinations (p33 and p18 and/or p13) induced greatly increased stem-pitting symptoms, while other combinations (p13 or p13 plus p18) resulted in reduced stem pitting. These results suggest that the stem-pitting phenotype, which is one of more economically important disease phenotypes, can result not from a specific sequence or protein but from a balance between the expression of different viral genes. Unexpectedly, using green fluorescent protein-tagged full-length virus and deletion mutants (CTV9Δp33 and CTV9Δp33Δp18Δp13), the virus was found at pitted areas in abnormal locations outside the normal ring of phloem. Thus, increased stem pitting was associated not only with a prevention of xylem production but also with a proliferation of cells that supported viral replication, suggesting that at random areas of stems the virus can elicit changes in cellular differentiation and development.
Viruses are obligate parasites with relatively small genetic coding capacity and must exploit a portion of host genes for invasion, multiplication, and spread. Although viruses of plants have been focused upon because of the diseases that they cause, the ultimate interaction when a virus evolves with a host is likely no disease. Yet, as viruses interact with plant hosts, they do sometimes cause disease. When disease occurs in a plant, it is often accidental due to the virus moving to a new host presented to it by agricultural practices. Disease symptoms usually occur on portions of the plant that develop and grow subsequent to viral infection. Rarely do symptoms occur in areas of the plant that are fully developed at the time of infection. Disease often results from interference with differentiation or development. Yet, when diseases do occur, they can cause severe damage to plants, and in agricultural crops, diseases cause economic losses, sometimes even preventing some crops from being grown.
Interference with differentiation or development results in numerous phenotypes. Lack of chloroplast development resulting in chlorosis is probably the most common virus-induced symptom. The resulting reduced photosynthesis causes reduced growth. Stem pitting is a common virus-induced phenotype of perennial woody plants that results from interference with stem growth. A range of different viruses distributed throughout the plant virus taxon induces stem pitting in a range of plant species, including numerous Prunus species, apples, vinifera grapevines, citrus, and avocado, usually resulting in a slow decline of growth and poor yields. In healthy and in normally developed areas of infected trees, the cambium, which is between the phloem and xylem, divides and differentiates in opposite horizontal directions, producing new xylem on the inward side and new phloem on the bark side, resulting in increased girth of the tree trunk and branches. Stem pits develop in areas where development is disrupted. The surrounding areas grow normally, leaving the disrupted areas as indented areas or pits. Although this disease phenotype is common in virus-infected perennial woody plants, there is little understanding of the processes that cause the stem pits.
Citrus tristeza virus (CTV), a phloem-limited virus, is a member of the genus Closterovirus of the family Closteroviridae (2, 14). The 19.3-kb single-stranded positive-sense genomic RNA of CTV is organized into 12 open reading frames (ORFs) (12, 15). ORFs 1a and 1b are directly translated from the genomic RNA as two overlapping polyproteins that encode two papain-like proteinases and methyltransferase-, helicase-, and RNA-dependent RNA polymerase-like domains (12). The 10 3′ genes are dispensable for replication at the single-cell level and are expressed through a nested set of 3′-terminal subgenomic (sg) RNAs (10, 20). CTV encodes a signature gene block, conserved among the members of Closteroviridae, comprising a 6-kDa hydrophobic protein; HSP70h, a homologue of the ubiquitous cellular heat shock protein; a 61-kDa protein; and minor coat protein (CPm) and major coat protein (CP). The last four proteins are involved in the formation of flexuous filamentous bipolar virions of 2,000 nm by 10 to 12 nm (21, 24). The proteins CP, p20, and p23 were shown to be involved in suppression of host RNA silencing in Nicotiana spp. (13). Additionally, the members of Closteroviridae encode 1 to 5 unique species-specific nonconserved genes with no sequence identity with available sequences. CTV possesses three such genes (p33, p18, and p13), which are dispensable for systemic infection of certain citrus species (26). Recently, we reported that CTV was apparently able to extend its host range by acquiring these nonconserved genes (27). Acquisition of the p33 gene allowed systemic infection of sour orange and Eureka lemon trees, that of the p33 or the p18 ORF allowed infection of grapefruit trees, and that of the p33 or the p13 ORF allowed infection of calamondin trees (27).
The host range of CTV is limited to Citrus spp. and close relatives. Infections with almost all CTV isolates are symptomless in some citrus hosts; those isolates that do cause disease symptoms do so in only a small subset of their host range. Yet, some CTV isolates cause severe economic losses in citrus (2, 14). In the early 1900s, CTV destroyed entire citrus industries, particularly in South America. Currently, CTV continues to limit citrus production in much of the citrus-producing world. The virus causes a range of disease phenotypes in citrus, but the phenotype that is presently the most economically important is referred to as "stem pitting." Trees with severe stem pitting grow poorly, lack vigor, and yield small, unmarketable fruit. Sasaki et al. (19) found that isolates causing severe stem pitting had a tissue tropism slightly different from the meristematic cells at the interface between phloem and xylem. Brlansky et al. (4) examined stem pitting induced by CTV by light and electron microscopy. In areas of the stem pits, the cambium appeared to be missing and the lack of new xylem formation resulted in a depression or pit in the surface of the stem as the normal areas continued increasing the girth of the stem. Proportionally to the density of pits, the function of the phloem and plant growth and vigor are reduced.
Certain isolates of CTV cause stem pitting in specific citrus varieties (9, 11). The remarkable feature of the CTV-stem pitting association is the high degree of specificity. For example, some isolates cause stem pitting in sweet orange but not in grapefruit. Others cause stem pitting in grapefruit but not in sweet orange. Others cause the phenotype in both, and others cause it in neither. This level of specificity continues throughout a range of other citrus species and even among different cultivars within a species, such that there are literally hundreds of phenotypes. Thus, among its whole host range, a particular isolate will cause stem pitting in only a small subset, perhaps only one species or even a subset of the species. Why is the induction of stem pitting so specific?
We created an infectious cDNA clone of CTV (20) that has allowed us to examine its interaction with its hosts (7, 26) and recently showed that the virus acquired three genes to extend its host range (27). The ability to systemically infect some young citrus trees with CTV mutants with deletions of combinations of the p33, p18, and p13 genes provided an opportunity to examine the effect of the lack of these genes on the development of symptoms caused by CTV. In the most susceptible experimental host, Citrus macrophylla, the full-length virus causes only very mild stem-pitting symptoms. Surprisingly, we found that certain deletion combinations induced greatly increased stem-pitting symptoms, while other combinations resulted in reduced stem pitting. These results suggest that the stem-pitting phenotype, which is one of the more economically important disease phenotypes, can result from a balance between the expression of different viral genes. The general expectation was that stem pitting induced by CTV would be due to specific viral sequences that could be used to identify isolates causing severe stem pitting. However, the changes in stem-pitting phenotypes shown here were due to removal of sequences instead of different sequences. Unexpectedly, at the pitted areas, the virus was found in locations outside the normal ring of phloem. Increased stem pitting was associated not only with a prevention of xylem production but also with a proliferation of cells that supported viral replication, suggesting that the virus can elicit changes in cellular differentiation at random areas of stems.
Maintenance of CTV deletion mutants and inoculation of citrus plants.The following were produced from cDNA constructs previously described (20, 22, 23, 26): full-length cloned virus CTV9; deletion mutants CTV9Δp33, CTV9Δp18, CTV9Δp13, CTV9Δp33Δp18, CTVΔp33Δp13, CTV9Δp18Δp13, and CTV9Δp33Δp18Δp13; green fluorescent protein (GFP)-tagged wild-type virus CTV9-GFPC3; and deletion mutants CTV9Δp33-GFPC3 and CTV9Δp33Δp18Δp13-GFPC3. The virus-infected plants were maintained in C. macrophylla under greenhouse conditions. Bark pieces or buds from infected plants were used to graft transmit the virus. A minimum of 4 to 5 plants were inoculated and maintained in a temperature-controlled greenhouse. The grafted plants infected by these viruses were pruned at 1 week after inoculation, which was followed by growth of a new flush. The infectivity of test plants was determined by analyzing extracts from young stems by double-antibody sandwich indirect enzyme-linked immunosorbent assay (DAS-I-ELISA) using CTV-specific antiserum (8).
The main stems of plants infected with the full-length virus (CTV9) and deletion mutants at 3 months postinoculation (mpi) were examined with the bark removed, and pictures were taken under visible light.
Examination of GFP fluorescence in stems and stem pits.Cross sections of stems with bark tissue and vertical sections of wood with bark removed from plants infected with CTV9-GFPC3, CTV9Δp33-GFPC3, or CTV9Δp33Δp18Δp13-GFPC3 were examined for GFP fluorescence under a Zeiss Stemi SV 11 UV fluorescence dissecting microscope (Carl Zeiss Jena GmbH, Jena, Germany) with an attached Olympus Q-color 5 camera (Olympus America, Inc., Center Valley, PA).
Deletions of the p33, p18, and/or p13 ORF greatly affect stem-pitting symptoms.Previously, we reported that deletions in the p33, p18, or p13 ORFs in all possible combinations in CTV (T36) (Fig. 1A) still allowed the virus to systemically infect the following hosts: C. macrophylla, Mexican lime (C. aurantifolia), sweet orange (C. sinensis), C. indica, C. hystrix, C. micrantha, Persian lime (C. latifolia), and citron (C. medica) plants (27). These results allowed the examination of the effect of deletion of genes on symptom production. Except on C. macrophylla and Mexican lime plants, the full-length virus (CTV9) and all deletion mutants induced mild or symptomless infections with little or no stem pitting. However, some of the mutants induced severe stem pitting in both C. macrophylla and Mexican lime plants. The phenotypes of the mutants in both plants were similar, but for brevity, here we report the results only for C. macrophylla.
(A) Schematic diagram of the genomic organization of CTV (CTV9) with ORFs (open boxes) showing the following encoded proteins: PRO, papain-like proteinase domains; MT, methyltransferase-like domain; HEL, helicase-like domain; RdRp, RNA-dependent RNA polymerase-like domains, IDR, interdomain region; HSP70h, homolog of heat shock protein 70; CPm, minor coat protein; CP, major coat protein. The replication gene block, quintuple gene module (closterovirus hallmark genes), and virion assembly genes are indicated. The internal genes of CTV are expanded below the genomic organization of CTV9, with deletions in single, double, and triple ORFs indicated with black boxes. The location and nucleotide coordinates of deletions in CTV genome in constructs B to H are indicated. (B) Deletions in the p33 or p18 ORF and a deletion of the p33 ORF in combination with deletion of p18 and/or p13 ORFs induced severe stem-pitting symptoms in C. macrophylla at 3 months postinoculation. The bark tissue from the main stem of full-length virus (CTV9)- or deletion mutant-infected C. macrophylla plants was removed, and pictures were taken under normal lighting.
Because the full-length virus was known to induce mild stem pits on the mature trunk and limbs at 2 to 3 mpi, we examined the stem-pitting symptom phenotype of the deletion mutants. CTV with a deletion of the p33 or p18 ORF induced more severe stem pitting with a substantial increase in the number and depth of pits (Fig. 1B). The mutant with the deletion of the p18 ORF induced stem pits slightly milder than those of the mutant with the p33 deletion; nonetheless, the stem pits elicited by both deletion mutants were severe compared to those elicited by the full-length virus (Fig. 1B). In contrast, CTV with a deletion of the p13 ORF induced no visible stem pits, and it induced even milder symptoms than the full-length virus (Fig. 1B). These data demonstrated that deletion of the p33 ORF (presence of the p18 and p13 ORFs) or the p18 ORF (presence of the p33 and p13 genes) increased the stem pitting by CTV, but the deletion of the p13 ORF (presence of the p33 and p18 ORFs) resulted in reduced stem-pitting symptoms.
We next examined the effects of double or triple gene deletions on induction of stem pits. The mutants with deletions of the p18 and/or the p13 ORF plus the p33 ORF induced greatly increased numbers and sizes of stem pits compared to those induced by mutants with the p33 deletion alone (Fig. 1B). In contrast, the mutant with deletions of both the p18 and p13 ORFs induced no visible stem pits, which was similar to the finding for the mutant with the single deletion of the p13 ORF. Both of these mutants induced reduced stem pitting compared to the full-length virus. Even though the mutant with the deletion of the p18 ORF induced severe stem pits, the mutant with deletions of the p18 ORF plus the p13 ORF induced no visible stem pits. The triple-gene-deletion mutant, CTV9Δp33Δp18Δp13, induced stem pits that were the most severe compared to those induced by the other deletion mutants (Fig. 1B).
Association of virus-infected cells with stem pits.We developed the full-length virus and deletion mutants CTV9Δp33 and CTV9Δp33Δp18Δp13 with a GFP marker in order to monitor virus movement and distribution in citrus trees (6, 26). The GFP ORF was constructed as an extra gene between the CPm and CP genes, and free GFP accumulates in the cytoplasm of cells as CTV replicates (6). The location of CTV replication is limited to phloem-associated cells, companion cells, and phloem parenchyma cells. Since CTV does not replicate in sieve elements or xylem cells, GFP does not accumulate in these cells (7). In cross sections of stems, a ring of meristematic cambium cells is located between the phloem to the outside and the xylem on the inside. Normally, the virus is limited to the ring of phloem-associated cells (6, 26). In rapidly growing citrus, the bark will slip, such that when the bark is pulled away from the tree at the loose cambium, the phloem is attached to the inside of the excised bark, while the xylem remains with the stem. In cross sections of CTV9-GFPC3-infected mature stems, as expected, the fluorescence was restricted to the ring of phloem-associated cells (Fig. 2A).
GFP-tagged CTV deletion mutant-infected cells associated with pits in mature stems of C. macrophylla at 3 months postinoculation. (A) Cross sections of stems showing the presence of fluorescence from GFP-tagged viruses in the stems. Note that fluorescence was restricted to phloem-related tissue in CTV9-GFPC3-infected stems, whereas in deletion mutant-infected plants, a substantial amount of fluorescence was detected in areas normally composed of xylem cells. (B) CTV9-GFPC3-, CTV9Δp33-GFPC3-, and CTV9Δp33Δp18Δp13-GFPC3-infected C. macrophylla mature stems with the bark removed showing pitting symptoms under visible light (VIS) and UV light (UV) and stems split vertically shown under UV light (VS-UV).
Since CTV9Δp33 and CTV9Δp33Δp18Δp13 induced severe stem pitting compared to the relatively mild stem pitting induced by the full-length virus, we next examined how the stem pits were associated with the virus-infected cells. We examined the mature wood of trees infected with GFP-tagged full-length virus (CTV9) and deletion mutant CTV9Δp33 or CTV9Δp33Δp18Δp13 at 3 mpi. After the bark was removed, the new xylem on the outside of the woody stems was observed under a fluorescence microscope for the presence of GFP fluorescence associated with infected cells in the woody stems. As expected from previous observations (6, 7, 26), most of the fluorescence by GFP was restricted to the inside of the bark that was removed, except that a few fluorescent areas were associated with the few pitted areas induced by the full-length virus, CTV9 (Fig. 2B). In contrast, at 3 mpi the mature wood (bark removed) from trees infected with CTV9Δp33-GFPC3 or CTV9Δp33Δp18Δp13-GFPC3 had a large number of large fluorescent areas associated with the more confluent pitted areas (Fig. 2B). In those areas, some cells susceptible to CTV did not remain attached to the bark but instead remained in the pitted areas, with much more GFP fluorescence associated with the pits of the severely affected stems.
To further confirm the association of virus with stem pits, we examined the vertical sections of the stems with the bark removed from GFP-tagged full-length virus- and deletion mutant-infected plants at 3 mpi (Fig. 2B). We found a large amount of fluorescent patches deep in the wood extending close to the central (pith) region of the wood in CTV9Δp33-GFPC3- and CTV9Δp33Δp18Δp13-GFPC3-infected plants (Fig. 2B). In contrast, a few tiny fluorescent patches were observed at the periphery of wood from GFP-tagged full-length virus-infected plants (Fig. 2B). Taken together, our data suggest that a large amount of virus is associated with stem pits in C. macrophylla.
Cross sections of similar stems with the bark (including phloem and cambium) still attached were examined. In cross sections of CTV9-GFPC3-infected mature stems examined at 3 mpi, the fluorescence was restricted to the ring of phloem-associated cells, as described above (Fig. 2A). In contrast, examination of cross sections from CTV9Δp33-GFPC3- and CTV9Δp33Δp18Δp13-GFPC3-infected mature stems resulted in fluorescence visible deep into the stem instead of limited to the normal ring of phloem (Fig. 2A). Thus, the distribution of the deletion mutants in mature stems was substantially different from that of the full-length virus.
It should be kept in mind that stems that develop pitting grow in the presence of the virus. We next examined the progressive development of stem pits in infected plants relative to the amount of growth that occurred after infection by taking a series of cross sections from the top of young stems of side branches, where the virus had recently infected and begun replication, to the lower parts of the stems that were more mature and had been infected much longer (Fig. 3). These cross sections retained the bark and phloem. The fluorescence from CTV9-GFPC3-infected plants was mostly restricted to the rings of phloem-related tissues in young as well as mature stems (Fig. 3B, top row). In CTV9Δp33-GFPC3- and CTV9Δp33Δp18Δp13-GFPC3-infected plants, the fluorescence was also mostly restricted to the ring of phloem-related tissues in young stems (Fig. 3B, middle and bottom rows). However, as cross sections of more mature tissues were progressively obtained, the fluorescence at stem pits no longer was limited to the phloem ring but was shifted progressively toward the inner areas of the stem (Fig. 3B, middle and bottom rows).
CTV deletion mutants progressively induced increased stem-pitting symptoms with the maturity of the stem in C. macrophylla. (A) Schematic diagram of a C. macrophylla plant showing the positions of cross sections used for examination for the presence of GFP fluorescence. A series of cross sections from the young (top) toward the mature (bottom) side branches at 3 months postinoculation was observed under a fluorescence microscope. The position of grafted bud (inoculum) from infected plants onto healthy C. macrophylla is indicated. (B) Cross sections of C. macrophylla branches infected with CTV9-GFPC3 (top row), CTV9Δp33-GFPC3 (middle row), and CTV9Δp33Δp18Δp13-GFPC3 (bottom row). Note that cross sections from CTV9-GFPC3-infected plants showed the uniform localization of fluorescence mostly restricted to the phloem-related tissue in the bark tissue. In contrast, cross sections from GFP-tagged deletion mutant-infected stems showed progressively increased accumulation of fluorescence outside the phloem-related tissue as the result of severe stem pits elicited by the deletion mutants.
Previously, we reported that CTV contained nonconserved genes (p33, p18, and p13) and that different combinations of these genes allow extension of the CTV host range (27). Yet, the virus can systemically infect most of its hosts with one, two, or three of the genes deleted. A few other viruses have been shown to tolerate deletion of a gene, but these mutants generally cause attenuated symptoms in their hosts (e.g., see references 17, 18, and 25). Although the CTV deletion mutants induced mild symptoms in most other citrus species (26, 27), in this study, we were surprised to find that these deletion mutants affected stem-pitting symptoms in C. macrophylla.
Different isolates of CTV induce a myriad of phenotypes in different citrus varieties and species (9, 11, 14). Vein clearing, leaf cupping, and temporary yellowing and stunting of young seedlings are phenotypes used in greenhouse diagnosis, but these symptoms have little effect on the growth and yield of large trees. The major concern today around the world is stem pitting, which makes citrus production not economical. Production in areas where isolates that cause severe stem pitting are endemic can be economical only if susceptible varieties, normally oranges and grapefruit, are not grown or by preinoculation of trees with isolates inducing mild pitting to reduce losses. Areas where isolates causing severe pitting are not endemic invest in quarantine measures to prevent their introduction. A major limitation is identification of an isolate causing severe stem pitting. Isolates of CTV that cause severe stem pitting can be identified in greenhouse tests by inoculating indicator hosts, but this assay takes about a year and requires large amounts of greenhouse space and labor. A major goal has been to identify viral sequences specific to severe stem pitting so that detection can rely on rapid antibody-based or PCR assays. In fact, it has been proposed that sequencing of many isolates of the virus and comparison of the sequences of stem-pitting isolates with those of non-stem-pitting isolates would define the causal sequences. However, the increase in stem pitting by CTV in C. macrophylla did not result from a certain sequence that could be targeted for identification. Instead, severe disease resulted from removal of sequences. The full-length virus that caused minimal stem pitting contains all of the sequences of the severe stem-pitting deletion mutants.
The formation of pits was apparently due to the inhibition of production of new xylem in the localized affected area, as shown by Brlansky et al. (4). The normally developing surrounding areas continue to grow, leaving a depression or pit at the affected area. What was unexpected was that in severely pitted areas, GFP fluorescence as a marker of virus replication was observed in regions normally made up of mature xylem or wood. It has been shown that the result of deletions in other viruses is to allow the virus to lose restriction to the phloem and move into nonvascular mesophyll cells (16) or alter the intestinal tropism within its insect vector (3). However, the CTV deletion mutants did not move into nonvascular mesophyll cells. Instead, they were found in a group of cells that appeared to be on the woody side of the phloem. In normally developing trees, most of the cells in this area differentiate into tracheary elements, which essentially consist of dead cells with thick walls connected into vessels for water transport. Interspersed in this area are live ray cells that transport nutrients from the phloem. In the full-length virus-infected trees, the fluorescence of GFP was always limited to the phloem ring outside the wood. However, increased stem pitting was associated with virus-infected cells in areas not normally infected. Since CTV multiplies and produces GFP only in living cells and free GFP was not found in noninfected adjacent cells (7), it would not be expected that the virus could produce GFP in mature xylem cells without virus replication or that GFP made in other cells could accumulate in xylem. However, it should be kept in mind that this is a process that occurs over a period of time and the stem increases in girth as the plant grows in the presence of the viral infection. These results suggest that the process of forming a stem pit not only involves the lack of production of new xylem in the affected area, resulting in a depression in the wood, but also affects development and causes cells within the pitted area to continue living and to be susceptible to CTV invasion and replication. Brlansky et al. (4) found that phloem-associated ray cells in citrus stem pitted by CTV were abnormal and disorganized. Virus-induced stem pitting might parallel regeneration after bark girdling where xylem cells dedifferentiate and transdifferentiate to form new cambium and phloem cells (28). Since CTV is normally restricted to phloem-associated cells, these masses of infected cells might be phloem-related cells. However, these cells apparently do not develop into functional phloem sieve elements that move carbohydrates, first, because CTV does not replicate in those cells and, more importantly, because the phenotype of severely stem-pitted trees is reduced phloem function. Further study at the microscopic anatomical level will be necessary to determine how the differentiation of the stem cells is altered by the severe stem-pitting mutants to cause pitting and reduction of tree growth. However, the CTV deletion mutants provide defined mutants that induce defined stem-pitting phenotypes that should be useful tools for further analysis of this phenomenon.
How did deletions in CTV induce severe stem pitting? One possibility was that the deletions caused an increase in expression of other CTV genes, which in turn induced the stem pitting. A common rule of expression of genes of viruses that express multiple genes via subgenomic RNAs is that genes positioned nearer the 3′ terminus tend to be produced in larger amounts, and movement of the genes closer to the 3′ terminus by deletion of intervening genes increases levels of expression (5). CTV follows this pattern. When the lowly expressed p33 or p6 gene was moved to near the 3′ terminus by deletion of all of the intervening genes, their mRNAs were increased many fold (1, 20). The largest deletion that would be expected to increase production of the upstream genes (CP, CPm, p61, HSP70h, p33, and p6) would be the deletion of both the p13 and p18 genes. However, this deletion induced the least amount of stem pitting. Increased stem pitting was often associated with deletion of the p33 gene. However, the p33 gene is positioned most 5′ of the genes expressed by subgenomic mRNAs, and its deletion does not increase expression of any other genes. Thus, it seems unlikely that the increased stem pitting due to deletions in the CTV genome was due to changes in expression of other viral genes.
Deletion of the p13 ORF tended to be correlated with reduced stem pitting. Thus, deletion mutants that retained the p13 gene (deletion of p33, p18, or p33 plus p18) tended to have the most stem pitting, which might suggest that the p13 gene product was involved in induction of stem pits. However, the triple-deletion mutant, which did not have the p13 gene, induced severe stem pitting, demonstrating that interpretation is not so simple. In contrast, increased stem pitting was generally associated with deletion of the p33 ORF. Mutants with the absence of the p33 ORF (deletion of the p33 plus the p18 ORFs and the p33 plus the p18 and p13 ORFs) induced severe stem pitting. Thus, mutants retaining the p33 gene (deletions of p13, p18, or p13 plus p18) had the smallest amounts of stem pitting. These results suggest that the presence of the p33 protein could be correlated with reduced stem pitting (and its absence increases it). However, the mutant with the deletion of the p18 ORF (with p33 and p13 retained) induced moderate stem pitting. Overall, the production of stem pits or no stem pits appears to be related more to a balance between expression of the p33 and p13 genes and possibly p18 genes.
In general, deletions in CTV resulted in a substantial increase in the stem-pitting disease of citrus examined here. Yet, there are different phenotypes of stem pitting. Some trees have large stem pits that are readily visible in tree trunks and limbs without removing the bark. Other trees exhibit "cheesy bark" stem pitting, which is a high density of very small pits. There is a continuum of levels in between. Some cause a rapid decline of tree growth and yield, while others cause little damage to the tree. Additionally, there is the extreme specificity between virus isolates and different citrus species and varieties. It should be noted that most of the other hosts examined did not form stem pits when infected with these mutants (27; results presented here). There is no reason to think that all of the different stem-pitting phenotypes in different citrus hosts would be caused by the same virus-host interactions. Moreover, it should be kept in mind that within the host with stem pitting, much of the infected tissue is normal. It is only the occasional area that becomes a pit. This is a complex disease. Yet, here we have been able to associate viral alterations with one phenotype of stem pitting as a first step in understanding this process.
This research was supported in part by an endowment from the J. R. and Addie S. Graves family and grants from the Florida Citrus Production Research Advisory Board and the National Research Initiative for the USDA Cooperative State Research, Education and Extension Service.
We thank Cecile Robertson for excellent technical assistance and Stephen M. Garnsey for valuable advice.
Received 12 April 2012.
Accepted 8 May 2012.
Accepted manuscript posted online 16 May 2012.
Copyright © 2012, American Society for Microbiology. All Rights Reserved.
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Prokhnevsky AI,
Peremyslov VV,
Napuli AJ,
Dolja VV
. 2002. Interaction between long-distance transport factor and Hsp70-related movement protein of beet yellows virus. J. Virol. 76:11003–11011.
Sasaki A,
Tsuchizaki T,
Saito Y
. 1980. Distribution of Citrus tristeza virus antigen in citrus tissues, p 17–18. Proc. 8th Congr. Int. Org. Citrus Virologists.
. 1999. An engineered closterovirus RNA replicon and analysis of heterologous terminal sequences for replication. Proc. Natl. Acad. Sci. U. S. A. 96:7433–7438.
. 2000. Closterovirus encoded HSP70 homolog and p61 in addition to both coat proteins function in efficient virion assembly. Virology 278:253–265.
. 2001. Amplification of Citrus tristeza virus from a cDNA clone and infection of citrus trees. Virology 280:87–96.
. 2003. Frameshift mutations in infectious cDNA clones of Citrus tristeza virus: a strategy to minimize the toxicity of viral sequences to Escherichia coli. Virology 313:481–491.
. 2004. Closterovirus bipolar virion: evidence for initiation of assembly by minor coat protein and its restriction for the genomic RNA 5′ region. Proc. Natl. Acad. Sci. U. S. A. 101:799–804.
Stenger DC,
Gildow FE
. 2005. Complete deletion of wheat streak mosaic virus HC-Pro: a null mutant is viable for systemic infection. J. Virol. 79:12077–12080.
. 2008. Three genes of Citrus tristeza virus are dispensable for infection and movement throughout some varieties of citrus trees. Virology 366:297–307.
Robertson CJ,
. 2011. A plant virus evolved by acquiring multiple nonconserved genes to extend its host range. Proc. Natl. Acad. Sci. U. S. A. 108:17366–17371.
Zhang J,
. 2011. Molecular features of secondary vascular tissue regeneration after bark girdling in Populus. New Phytol. 192:869–884.
Journal of Virology Jul 2012, 86 (15) 7850-7857; DOI: 10.1128/JVI.00916-12
You are going to email the following Enhancement or Attenuation of Disease by Deletion of Genes from Citrus Tristeza Virus


Hydra Islet- (Day)- Ajira Airlines 316 is flying towards Guam with Kate, Sayid, Hugo, Sun, Jack, and approximately twenty others passengers. Frank Lapidus is piloting the airliner. Lightning appears in the sky. The plane hits turbulence. Frank says "Oh no!" as the turbulence gets much worse. Night turns to day. The flight loses power. Hydra Island comes into view. The altimeter reading is 650' as the plane negotiates around a steep mountain on the main island. Power is restored. As the co-pilot is calling for a mayday, the numbers are being broadcast from the radio tower. The pilots see a runway on Hydra islet and attempt to land on it, over shooting it slightly and crashing the plane into the jungle. During the crash, a branch breaks through the windshield and impales/kills the co-pilot. The co-pilots blood covers Frank. Sun and Ben are onboard with about 20 passengers, but Jack, Kate, Hurley, and Sayid are somehow missing.
Hydra Islet- (Day)- Lapidus gathers the approximately 20 survivors and tells them the radio is dead, but he is sure that a rescue will be forthcoming. Ceaser tells the group that a building and animal cages are close and they should check it out for a radio. Ben slips away from the group. Sun and Lapidus follow Ben.
Hydra Islet- (Day)- Lapidus, Ben, and Sun assemble at the main island side of Hydra. There are three boats there. Ben tells Lapidus that he has people that he needs to take care of, also. Ben tells Lapidus that there is a small dock about ½ mile due South across the water that leads directly to a town where he used to live. Sun knocks out Ben with an oar before Ben can finish.
Hydra Islet- (Day)- (Off Camera)- Sun and Frank carry Ben to the makeshift infirmary and Franks takes care of the passengers immediate needs.
Hydra Islet- (Night)- Caesar is insides of Ben's office on Hydra islet. He is searching the office. He finds maps of the island and a shotgun. Ilana interrupts him. Ilana tells Caesar that we found a man, just south of here. He was just standing in the water. He's wearing a suit. He was not on the plane. Ceaser and Ilana go to the location where flight 316 landed (the plane did not break apart). A group of survivors are camped around the plane.
The strange man introduces himself as John Locke.
Submarine Dock- (Night)- Sun and Frank arrive at the submarine dock via the outrigger. As they approach, there is a sound of something moving in the foliage. It is the monster (clicking sounds accompany the movement). Sun, realizing that it is the monster, lies to Frank and tells him it is probably an animal. The two of them travel onward and reach the processing center. The center is in disarray and appears not to have been in use for many years. Whispers are heard. A light turns on in one of the building and Christian Shepard appears at the door. Sun asks Christian if he knows where her husband, Jin, is. Christian tells her to follow him. They enter the Processing center building. Christian looks at the photos on the wall, which depict the annual recruits every year. The first one he sees is from 1972 (this is the earliest reference to the Dharma initiative). Christian tells Sun Jin is with their friends and shows her the photo from 1977, which has Jack, Kate, and Hurley in it. Christian tells Sun she has a bit of a journey ahead. to enter.
ӏ am reаlly loving the theme/ԁеsign of уour webѕіtе.
Do you eveг run into аny bгowѕеr cοmpatibility problems?
Α handful of my blοg гeаԁers have comρlained about my sіtе not operatіng сoгrectlу in Explorer but looks great in Opera.


text:
The Seram mountain pigeon (Gymnophaps stalkeri) is a species of bird in the pigeon family Columbidae that is endemic to the island of Seram in Indonesia, where it inhabits hill forest. It was long considered to be a subspecies of the Buru mountain pigeon, but was split on the basis of differences in appearance. It is a medium-sized pigeon with a buff-pink face and breast, wine-pink underparts, a grey nape, crown, back of neck, and thighs, and dark chestnut belly and underside of the tail.

The Seram mountain pigeon feeds on fruit and feeds in flocks of up to 50 birds. The only known nest was observed in September in old-growth montane forest and contained one egg. It is listed as being of least concern by the International Union for Conservation of Nature (IUCN) on the IUCN Red List due to its stable population and sufficiently large range.

Taxonomy and systematics

The Seram mountain pigeon was described as Columba stalkeri by the Scottish ornithologist William Ogilvie-Grant in 1911 on the basis of specimens from the Kanobi Mountains on Seram, Indonesia. It was subsequently merged with the Buru mountain pigeon in the genus Gymnophaps by the Australian amateur ornithologist Gregory Mathews in 1927. It was again split as a distinct species on basis of differences in appearance in 2007 by Frank Rheindt and Robert Hutchinson.

The generic name Gymnophaps is derived from the Ancient Greek words γυμνος (gumnos), meaning bare, and φαψ (phaps), meaning pigeon. The specific name stalkeri is in honour of Wilfred Stalker, an Australian natural history collector who collected the type specimens of the species on a British Ornithologists' Union expedition. Seram mountain pigeon is the official common name designated by the International Ornithologists' Union. Other common names for the species include Seram long-tailed mountain-pigeon.

The Seram mountain pigeon is one of four species in the mountain pigeon genus Gymnophaps, which is found in Melanesia and the Maluku Islands. It forms a superspecies with the other species in its genus. Within its family, the genus Gymnophaps is sister to Lopholaimus, and these two together form a clade sister to Hemiphaga. The species has no subspecies.

Description
The Seram mountain pigeon is a medium-sized pigeon that is  long and weighs . It has a deep buff-pink face and breast, paler on the chin, changing to wine-pink on the underparts. The thighs are grey and the nape, crown, and back of neck are pale, pure blue-grey, while the belly and underside of the tail are dark chestnut with wine-grey fringes to the feathers. The eyes are scarlet to yellow with purplish red to scarlet orbital skin, the bill is yellow with a purple cere, and the legs are purple. Juveniles have browner upperparts, have browner underparts, and are darker below the tail. They also have dull red orbital skin, dull red legs, and a white-tipped grey bill with a dull red cere.

Distribution and habitat
The Seram mountain pigeon is endemic to Seram in the Maluku Islands. It mainly inhabits hill forest at elevations of , but is typically more common above . One record of the species found seven individuals at an elevation of .

Behaviour and ecology
The species is very social, frequently being seen in flocks of over 20 birds. It feeds on fruit. Foraging flocks can have over 20 birds, with some being as large as 50 individuals.

Breeding
The Seram mountain pigeon has been observed making a display flight in February. The display is similar to that of Papuan mountain pigeons, males of which launch themselves upwards from a perch before plummeting down and returning. However, the display of the Seram mountain pigeon is lower and less steep, and it continues flying forward after descending instead of returning to the perch immediately.

Its only known nest was observed in September in primary montane forest at an elevation of . The nest was placed in a crevice on a dead, mossy branch  above the ground. It was lined loosely with twigs and moss, and contained a single white egg.

Status
The Seram mountain pigeon is listed as being of least concern by the International Union for Conservation of Nature (IUCN) on the IUCN Red List due to its stable population and sufficiently large range. Its population has not been estimated, but it is reported as being more common than the Buru mountain pigeon is on Buru, and is especially common at altitudes of .

References

Seram mountain pigeon
Birds of Seram
Endemic fauna of Seram Island
Taxa named by William Robert Ogilvie-Grant
Seram mountain pigeon

redpajama_set_name: RedPajamaWikipedia


This piano is designed by Morganic. Open 08:00-21:30.
Oh, we dont know what we are doingk with music, we are just having fun with it.. and then these expert musos turn up and show us the real skillz of it.
Very lovely gardens to play piano in!
Beautiful sunny park, with commuters scurrying home from work at 5pm. This girl stopped and said 'I'd love to play you!' She sat down and started to play a classical piece softly and beautifully - a fine antidote to the peak hour traffic.


text:
The Resource !UNKNOWN LABEL
!UNKNOWN LABEL
Evans, Edward Steptoe, 1879-1945
Letter, dated 11 April 1907, from Edward Steptoe Evans (1879-1945) assistant librarian at the Virginia State Library to United States Senator John Warwick Daniel (1842-1910) concerning the investigation and report on the purchase of books for traveling school libraries by head librarian John Pendleton Kennedy (1871-1946). Kennedy was accused of having paid higher prices for books and of receiving kick-backs. After an investigation by the Library Board 11-14 March 1907, an inconclusive report was given to Governor Claude A. Swanson (1862-1939). Kennedy resigned 6 July 1907 and was succeeded by H. R. McIlwaine (1864-1934). Evans, who sided against Kennedy, resigned 12 July 1907 and was succeeded by Earl G. Swem (1870-1965)
1907 Apr. 11
Virginia State Library
Personal papers
Swanson, Claude Augustus, 1862-1939
Kennedy, John Pendleton, 1871-1946
Daniel, John W., (John Warwick), 1842-1910
Personal papers collection;, 23309
Biographical or historical data
Edward Steptoe Evans was born 24 May 1879 in Bedford County, Virginia. He graduated from George Washington University. Evans began working as assistant librarian at the Virginia State Library in 1904. Evans resigned from the library 12 July 1907 and moved to Florida. In Florida, he became involved in a company that produced block tackles for loading automobiles on frieght cars. After helping the company expand, Evans moved to Detroit, Michigan, where he founded Evans Products Company, which produced items for cars and trains. His company became a success. Evans served as a captain in the Transportation Corps during World War I. He became an advocate of aviation and exploration. Cape Evans in the Antarctic Ocean and Mount Edward Evans in Greenland are named for him. Evans expanded his business in the years before World War II, and during the war, produced weapons and supplies for the United States Army. Evans and his wife Virginia Epes McCormick Evans had two sons. Evans died in Detroit 6 September 1945.
Form designation
Photostats (negative).

 Data from !UNKNOWN LABEL - Library of Virginia

Data Citation of the Item !UNKNOWN LABEL
http://link.virginiamemory.com/portal/UNKNOWN-LABEL/WGYC6YMPUI8/
http://library.link/portal/UNKNOWN-LABEL/WGYC6YMPUI8/

redpajama_set_name: RedPajamaCommonCrawl


Make a card or projects using animals you find on a farm.
I found a great tip in some of the tutorials for the Cricut Explore-when you are looking for an image, if you click on the round circle with the italic "i" in it-it will tell you what cartridge the image came from. Handy to know, huh?
Cricut: barn (Create a Critter), Bull (Noah's ABC Animals) Cloud (Bits & Pieces). All images were cut from scrap stash.
Embossing folders: Clouds (Darice) Swiss Dots (Cuttlebug).
Ink: Versafine Onyx, Colorbox White Frost.
Enter a project for a chance to win Verve stamps!


Almost 2ft tall, this large colorful birdhouse is sure to get noticed indoors or out in the garden. Features a tall curvy roof, large front porch, and ornately framed opening.
Made exclusively by aGardenPlace, our handcrafted birdhouses are are built of solid wood, and handpainted with a love of whimsy.
Built to endure the outdoors, but sure to bring a a smile displayed indoors, too.
Overall Size: 9.5" W x 11" D x 23.5" H.


text:
Accusations fly as Colombian hostage mission fails
A Venezuelan-led mission to rescue three hostages from leftist rebels in Colombia's jungles fell apart Monday. The guerrillas accused Colombia's military of sabotaging the handoff.
Colombian police stand guard near a Venezuelan helicopter bearing Red Cross insignias at the airport in Villavicencio, Colombia, on Monday. Rebels said the military sabotaged a hostage hand-off.Fernando Vergara / AP
A Venezuelan-led mission to rescue three hostages, including a young boy, from leftist rebels in Colombia's jungles fell apart Monday, with the guerrillas accusing Colombia's military of sabotaging the promised handoff.
Colombian President Alvaro Uribe dismissed the claim as a lie by the Revolutionary Armed Forces of Colombia, or FARC, instead suggesting the guerrillas could be backing out of the deal brokered by President Hugo Chavez because they don't have the boy hostage.
"The FARC terrorist group doesn't have any excuse. They've fooled Colombia and now they want to fool the international community," Uribe said from the central Colombian city where Venezuela helicopters have been waiting since Friday for word from the guerrillas on where the hostages could be picked up.
He made the shocking suggestion that the guerrillas "don't dare to keep their promises because they don't have the boy, Emmanuel" — who the FARC announced two weeks ago they'd free along with his mother, Clara Rojas, and former congresswoman Consuelo Gonzalez.
Uribe said his government had given Venezuela and the international Red Cross coordinating the mission every guarantee that its military would not obstruct the handover, even promising to create a cease-fire corridor to allow the rebels to escort their hostages through the jungles to the pickup point.
Observers leave town
Former Argentine President Nestor Kirchner and observers from France, Switzerland and four other Latin American nations abandoned Villavicencio on Monday.
"Shame on Colombia, shame on Uribe," Oliver Stone, the American filmmaker, said shortly before boarding one of three Venezuelan jets carrying the observers back to Caracas. Stone, who was invited by Chavez to document the handover, added "the FARC have no motive not to release these hostages."
Uribe said a 3-year old child named Juan David Gomez, matching the description of Emmanuel provided by escaped hostages and suffering from malnutrition, malaria and jungle-born leishmaniasis, may have been living for the past two and a half years with at a foster home in Bogota.
The child was turned over in the eastern city of San Jose del Guaviare, a FARC stronghold, in 2005 by a man who said he was the boy's great uncle and who authorities now believe was his father. The boy's mother was reported as disappeared, according to the child welfare agency case file read to journalists by peace commissioner Luis Carlos Restrepo.
The Colombian leader said only DNA tests were required to prove or disprove "this hypothesis" — which he said could be done as soon as the boy's grandmother returns from Caracas, where she was awaiting the handover of her daughter and grandson.
Ivan Rojas, brother of Clara Rojas, said his family was having a meeting to digest Uribe's revelation.
Chavez accuses Colombia's president
Chavez, even while holding out the possibility the boy may not be in the FARC's hands, accused Uribe of traveling to Villavicencio "to dynamite the hostage rescue mission with a smoke screen."
"Hopefully the hypothesis of Uribe is correct. The FARC would look very bad to the entire world in light of such a lie," he added.
Speaking earlier on Venezuelan state television, Chavez said the rebels wrote in a letter that "the military operational attempts in the zone impede us for now from turning over" the three hostages.
In the letter, the FARC said that "insisting on (a handover) in these conditions would be putting at risk" the lives of hostages and guerrillas sent to turn them over.
Uribe last month abruptly ended Chavez's efforts to broker a wider swap of 44 high-profile hostages — including three American defense contractors and former Colombian presidential candidate Ingrid Betancourt — for hundreds of jailed rebels.
The U.S.-allied Uribe has used some $600 million in annual military and intelligence aid from Washington to push the half-century-old insurgency deeper into the jungle.


text:
Q: Rails associations and passing another model :id as an argument I have two associated models: Job and Candidate. The Candidate model is updated when a user applies to a job listing through the candidates 'new' view. Since I've associated Candidate with the Job model, I can't save any new instances of Candidate. I know that I have to pass job_id as a parameter so that any new instance of Candidate references a particular Job, but I just cant figure out what I'm doing wrong. Also, the Job model is associated with the Devise User model, but Candidate is not associated with the User model. Could that be the issue? Thanks in advance.
I've tried a ton of things in my candidates controller, but now I've put it back to how it was before I associated the Candidate model with Job model, so I can start over again as if I just ran the migration to AddJobToCandidate
candidates_controller.rb
class CandidatesController < ApplicationController
  def index
    @candidates = Candidate.all
  end

  def new
    @candidate = Candidate.new
    render layout: false
  end

  def single
    @candidate = Candidate.find(params[:id])
  end

  def create
    @candidate = Candidate.new(candidate_params)
    if @candidate.save
      redirect_to root_path
    else
      render "new"
    end
  end

  private
  def candidate_params
    params.require(:candidate)
          .permit(:first_name, :last_name, :email, :cover_letter, :ph, :job_id)
  end

end

views/candidates/new.html.erb

Work at <%= current_user.company %>

<%= simple_form_for @candidate do |form| %>
  <%= form.input :first_name, label: "First Name" %>
  <%= form.input :last_name, label: "Last Name" %>
  <%= form.input :email, label: "Email" %>
  <%= form.input :cover_letter, label: "Cover Letter" %>
  <%= form.input :ph, label: "Phone Number" %>

  <%= form.button :submit %>
<% end %>

routes.rb
Rails.application.routes.draw do
  get 'candidates/index'
  get 'candidates/new/:id' => 'candidates#new', :as => 'apply'
  get 'candidates/single/:id' => 'candidates#single', :as => 'viewcandidate'
  get 'jobs/index'
  get 'jobs/new'
  get 'jobs/listing/:id' => 'jobs#listing', as: 'listing'
  get 'jobs/listings', :as => 'careerpage'
  get 'jobs/dashboard'
  get 'jobs/single/:id' => 'jobs#single', :as => 'viewjob'
  devise_for :users, controllers: { sessions: "users/sessions" }
  resources :candidates
  resources :jobs
  root 'jobs#dashboard'
end

views/jobs/listing.html.erb the only view that links to candidates/new.html/erb

<%= @job.title %>

<%= @job.description %>

<%= link_to "Apply", apply_path(@job.id) %>

Candidate/Job association migration
class AddJobToCandidate < ActiveRecord::Migration[5.2]
  def change
    add_reference :candidates, :job, foreign_key: true
  end
end

jobs_controller.rb (don't think this is necessary but I'll include anyway)
class JobsController < ApplicationController
   before_action :authenticate_user!

  def index
    @jobs = current_user.jobs.all
  end

  def dashboard

  end

  def new
    @job = Job.new
  end

  def listing
    @job = Job.find(params[:id])
    render layout: false
  end

  def listings
    @jobs = current_user.jobs.all
    render layout: false
  end

  def single
    @job = Job.find(params[:id])
    @candidates = Candidate.all
  end

  def create
    @job = current_user.jobs.new(job_params)
    if @job.save
      redirect_to jobs_index_path
    else
      render "new"
    end
  end

  private
  def job_params
    params.require(:job)
          .permit(:title, :description, :url, :user_id)
  end

end

No error is shown, but model is not updated.
One of the things I tried in candidate_controller #create action was:
def create
    @candidate = @job.candidates.create(candidate_params)
    if @candidate.save
      redirect_to root_path
    else
      render "new"
    end
  end

and the error I get is 'undefined method `candidates' for nil:NilClass'
Another thing I tried was adding this line in the #create action:
@job = Job.find(params[:id])

and the error I get is 'Couldn't find Job without an ID'

A: When you arrive at candidate#new you're just discarding the Job ID. You're not making it part of the data submitted by the form to be picked up in the create controller.
Given you have completely abandoned RESTful routes, the quickest solution would be to add this to the form:
 <%= form.hidden_field :job_id, value: params[:id] %>

However, I highly recommend you look at RESTful routes and nested resources. You have a mess where the ID passed to Candidate#new is the ID of a different model. While you're at it you may want to reconsider having a candidate belong to a job, what happens when a candidate is interested in multiple jobs? They create a new profile each time? In job sites one would normally have a Job, a Candidate (or User / Profile) and then a JobApplication, which joins the two.

redpajama_set_name: RedPajamaStackExchange


Safeguarding is at the forefront of everyday life at Lindens Primary School. We pride ourselves on keeping every child happy, safe and secure. We acknowledge that e-safety is becoming evermore vital in our role to protect children.
We are now a member of e-safety support, who will work with us to offer support to staff, parents and your children on how to keep themselves safe when using the Internet.
The documents below offer advice on specific e-safety issues and include key facts to support children to enjoy and explore the Internet safely.


text:
Q: The specified user does not have a valid profile. Unable to load XXX.dll In a website, I get this error:
The specified user does not have a valid profile.  Unable to load XXX.dll

This happens when any page load is attempted.

A: Set to Load User Profile true in the apppool advanced settings in IIS:

More info is here: http://www.iis.net/learn/manage/configuring-security/application-pool-identities


J Am Geriatr Soc. 2017 Nov;65(11):2516-2521. doi: 10.1111/jgs.15061. Epub 2017 Oct 12.
Readily Identifiable Risk Factors of Nursing Home Residents' Oral Hygiene: Dementia, Hospice, and Length of Stay.
Zimmerman S1,2, Austin S3, Cohen L4, Reed D1, Poole P1, Ward K1, Sloane PD1,5.
School of Social Work, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
East Carolina University, Brody School of Medicine, Greenville, North Carolina.
Duke Clinical Research Institute, School of Medicine, Duke University, Durham, North Carolina.
Department of Family Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
The poor oral hygiene of nursing home (NH) residents is a matter of increasing concern, especially because of its relationship with pneumonia and other health events. Because details and related risk factors in this area are scant and providers need to be able to easily identify those residents at most risk, this study comprehensively examined the plaque, gingival, and denture status of NH residents, as well as readily available correlates of those indicators of oral hygiene, including items from the Minimum Data Set (MDS).
Oral hygiene assessment and chart abstract conducted on a cross-section of NH residents.
NHs in North Carolina (N = 14).
NH residents (N = 506).
Descriptive data from the MDS and assessments using three standardized measures: the Plaque Index for Long-Term Care (PI-LTC), the Gingival Index for Long-Term Care (GI-LTC), and the Denture Plaque Index (DPI).
Oral hygiene scores averaged 1.7 (of 3) for the PI-LTC, 1.5 (of 4) for the GI-LTC, and 2.2 (of 4) for the DPI. Factors most strongly associated with poor oral hygiene scores included having dementia, being on hospice care, and longer stay. MDS ratings of gingivitis differed significantly from oral hygiene assessments.
The findings identify resident subgroups at especially high risk of poor oral health who can be targeted in quality improvement efforts related to oral hygiene; they also indicate need to improve the accuracy of how MDS items are completed.


For the second day running parliament and press have been giving the greatest attention to the social-media scribblings of a rookie parliamentarian Naz Shah, moreover scribblings that were scribbled when she can have had no idea she'd be an MP, let alone parliamentary private secretary to the shadow chancellor of the exchequer less than a year later.
Ignoring the frankly astounding news from the Hillsborough Inquiry the Daily Mail ( formerly associated with the British Union of Fascists, its founder openly fraternised with Hitler) for example gave over its whole front page to a fake media storm around Naz Shah and her uneducated views on the Israel-Palestine conflict.
Shah has buckled, thrice issued ever more grovelling public apologies, resigned as a PPS, said she no longer believes the things she believed last year, knows better now etc. The psychobabble is in full flow "healing" "deep empathy" "sorry I hurt my family, my friends, parliament" etc.
It is said, and seems to have been accepted in some unlikely places, that Shah is guilty of "anti-Semitism". She is not, at least on the evidence which has emerged to date. Her comments are half-witted to be sure, and their appearance at this time will prove a mighty headache for Labour leader Jeremy Corbyn (who is actually what this is all about) and may prove very significant in British politics over the next few months. But they are not anti-Semitic.
I have no reason to defend Naz Shah except that it is necessary to stand up for the truth always. Not just morally but practically, for the Naz Shah affair is just another straw being laid on the back of Corbyn, and it may prove the last one. It is part of a bigger, wider political operation being mounted by supporters of Israel to redefine not just the political narrative but the language itself.
But before all that, this must be said. Anti-Semitism exists. It is a foul poison which subjected Jews for centuries to discrimination, bigotry, pogrom followed by the Holocaust, the greatest crime in human history in which millions of Jews were exterminated by Hitler-fascism and their "Christian" European allies while the rest of the world barred their gates against those fleeing the early stages of the industrialised attempt at genocide.
Anti-Semitism is a right-wing "nationalist" curse which deems, for example British Jews as not British at all but aliens who should be encouraged by all means necessary to leave their own countries and leave to wherever, anywhere but here.
It was the standard narrative in British fascism until the 1950s when black non-Jewish immigrants began to take the place of the Jews as the scapegoats and whipping boys for societal ills. Jewish "control" of banks and many businesses, landlords and mountebanks of all kinds were presumed by default to be Jews. This British fascism crystalised around Sir Oswald Mosely, a former Tory and Labour grandee whose attempt to ape Hitler and Mussolini had the full support of the family which still owns the Daily Mail (including straight financial support) It could if Mosely had been successful have led to a very British Holocaust, which would I am sure have been supported and facilitated by sections of the ruling class.
Such anti-Semites still exist today, a very grand now deceased Tory MP regularly described to me both former Foreign Secretary Sir Malcolm Rifkind and former Home Secretary Leon Brittain as "Jew Boys". Another leading Tory personally said to me that Mrs Thatcher's cabinet contained "more Estonians than Etonians", and he didn't mean it as a compliment.
There is no doubt either that amongst some opponents of the state of Israel, anti-Semitism exists. I have had to ban, from both my radio and TV shows, and on social media platforms callers who supported the Palestinians and in doing so exhibited clear anti-Semitic views. Many others I have had to chastise for the potentially deadly sin of conflating Jews, Judaism, Zionism and Israel. These are all to be found on YouTube by the way.
While, I think, none of those I have banned were Muslims, there is no doubt that anti-Semitism exists amongst some poorly educated Muslims in Britain also.
So I am certainly not here to argue that anti-Jewish hatred, like other forms of nascent hate crimes does not exist, It does..
Since the late stages of Jeremy Corbyn's leadership campaign, when the establishment began to panic over his potential victory, an offensive began to paint him and his principle ally John McDonnell MP the now shadow Chancellor as somehow anti-Semitic. Nothing could be further from the truth, literally.
McDonnell is a former Trotskyite. Trotsky was a Jew. Both men are influenced by Karl Marx. Marx was a Jew. Both men are anti-racist as an article of faith. They have spent their lives fighting racism in all its forms. Given their politics, how could it possibly be otherwise?
Both, and here's the rub, have also been life-long campaigners for the Palestinians. Every platform they shared, march they attended, donation they made was made in good faith in support of the just cause of the Palestinians. They may have made mistakes along the way, may have chosen words poorly from time to time, but only the malignant could maintain that any mistakes they made were as a result of some deeply concealed hatred of Jews. For those who know them, as I have for forty years, such a suggestion would be laughable if it were not so serious.
Time then to turn to Naz Shah MP and her no doubt late night social media scribbles.
There are three (at least so far) posts in the eye of this storm. I shall enumerate them for ease of reference.
1. She commented that it would be a good idea to relocate ISRAEL (my emphasis) to the United States. This was accompanied by a map showing the hugeness of the USA and the smallness of Israel. After all she said, the US was spending plenty of tax-dollars maintaining the Zionist state where it currently is, it would be cheaper to relocate it. This a foolish idea of course, but contains a serious point.
If the maintenance of the Zionist state is so important to the US, why does it have to be maintained on the land that clearly belongs to someone else, the Palestinians? Palestine was small, and was obliterated to make way for Israel. Why?
It isn't as if the founders of the state of Israel were always committed to building their state in Palestine. There was no Biblical reason for doing so, the Zionist leaders were overwhelmingly atheist. Zionism is a nationalist idea not a religious one (which is why some of the most religious Jews continue to oppose it to this day.) The founders of Zionism negotiated seriously with the then dominant British Empire over proposals to locate their European settler state in all manner of places, Uganda, the Seychelles and Argentina being just three of them.
And it isn't that there has been no "transportation" or "relocation" of populations in this story already. The transportation of 800,000 Palestinians from their homes and their lands is no Facebook post, it actually happened! This 800,000 driven from their land by mass murder and terror in the lifetime of some people alive today, have become millions of Palestinians "transported" to the four corners of the earth as usually stateless refugees.
Neither is the idea of a country ceasing to exist alien to this conflict. Look at your map and see if you can find the country formerly known as Palestine.
As it happens I believe the only way this conflict can now be resolved rules out the transportation of any more people from this tiny, Holy Land. I am against any further relocation of anybody, except the right of the Palestinians to return to their homes. I fight for one democratic secular state from the Jordan River to the sea in which all the Jews in Israel, together with all the Christians and Muslims as equal citizens live - one person one vote - in a new country called Israel-Palestine.
But those who reject this idea and who endorse the wiping of Palestine off the map and the permanent disinheritance of its people have no right to a fit of the vapours over Naz Shah.
Of course a more experienced politician, with a better grasp of history, would not have placed the word "transportation" in the same sentence as one which was dealing with the even fictional fate of millions of Jews. That politician would have had in mind the "transportation" to the death camps of millions of Jews in the Holocaust. Naz Shah isn't that politician. But she was not supporting the transportation of anybody to their deaths, but to America, where most of the Jews in the world already live overwhelmingly successful lives. The idea is foolish, a joke even, but to pretend a political activist as she was at the time is somehow the re-embodiment of Eichmann over it is self-serving rubbish.
2. By writing, again on Facebook, in trying to get people voting in an internet poll on Palestine the words "the Jews are rallying" Shah is supposedly guilty of some anti-Semitic outrage. This is even more risible than the first charge. If I wrote the words, say, "the Muslims are rallying" for Sadiq Khan in an internet post it would be (I hope) completely untrue, but it would be no kind of Islamophobic slur. And then there is the small matter of Israel's PM Netanyahu going on TV on Israel's last election night to warn his supporters that, er, "the Arab voters are...rallying"
3. Over a picture of Martin Luther King under arrest, Shah wrote that one should remember that everything Hitler did was legal. This point is obvious. Just because something is legal doesn't mean it is right. Do I really need to waste more of your time with that?
You may well ask why I have taken this many words to oppose the witch-hunt against Naz Shah. She, after all smeared me relentlessly during our election campaign in 2015 - including on these very matters - with the support of the very forces seeking to devour her now.
It is not because I am some St Francis of Assisi or uniquely forgiving by nature. It is not because when she wrote these things she was a member of my party Respect and hoping to be our candidate in Bradford. It is because if we allow opposition to Israel to become virtually a criminal offence, and on the eve of the centenary of the Balfour Declaration which first gave Palestine away, this would be the ultimate betrayal not just of the Palestinians but our own rights to speak out.
And if we allow false charges of anti-Semitism to be the rope to hang Corbyn, everything will be lost. Which is, why, of course the Israel lobby is trying so very very hard.
George Galloway is a British politician, broadcaster, and writer. He is the leader of the UK Respect Party. George is the producer, writer and presenter of the soon to be released feature film The Killing$ of Tony Blair (Bulldog Films Ltd). He is currently campaigning for election as Mayor of London, to be held in May.


Great for new students or new employees to get acquainted with the campus or a perfect introduction on orientation day. For large corporate campuses, employees can easily familiarize themselves with their new working environment. Choose between printed handouts or digital versions for internet or emailing. You simply provide us with the size, a list of elements or buildings that you would like to include, addresses (if they are located on city streets) and we'll do the rest.


text:
Q: Converting DD-MM char to dates Having a lot of trouble converting my data to date format. I'm performing analysis on weather data for a class and haven't even gotten to the analysis part. This part isn't required for the class but is bugging the hell out of me.
In general I'm trying compare weather patterns across years. The dates are formatted as dd-mmm in the rows, with years in each column.
First off, I appreciate any insight in handling weather data like this in general.
I'm trying to convert to date because it seems like good practice and for ease in plotting the temperature over time. Example code below:
# sample weather data
df <- data.frame("DAY" = c("1-Jul", "2-Jul", "3-Jul"),
                 "1996" = c(98,97,97),
                 "1997" = c(86,90,93))
df

# convert column to date format
dates <- as.Date(df$DAY, '%d-%b')

# new df with as.Date column
newdf <- cbind(dates, df)

# new column is class "Date"
class(newdf$dates)

# want date in format dd-mmm but converting it changes to class(character)
dates_changed <- format(dates, "%d-%b")
class(dates_changed)

I've managed to convert the original dd-mmm format to yyyy-mm-dd format, but this isn't desired as years are in the columns. I might just end up keeping this format as it enables me to graph in the dates in order but it doesn't seem ideal.
Appreciate any insight.


The Promise of Eminent Revival Bundle arrived in Wednesday's main client update to Dota 2. It is available only to Battle Pass owners who have reached level 615 and includes items from five previous Immortal Treasures to commemorate the history of The International.
The Dota 2 items include the Cauldron of Xahryx of Eminent Revival, the Leviathan Whale Blade of Eminent Revival, the Lightning Orchid of Eminent Revival, the Fin King's Charm of Eminent Revival, and the Tormented Staff of Eminent Revival.
Each of the items in the bundle has been redesigned from their original incarnations to match the theme of TI8, and players can unlock a further upgraded style through play. The items will not be available for purchase or trade on the marketplace.
Also arriving in the update are new lore-based tips, hero-specific tips, and updates to existing tips. These tips will now display when the game is paused.
The past two main client updates have focused on Dota Plus and Battle Pass changes and spectating bugs, respectively.


The Physician-Focused Payment Model Technical Advisory Committee (PTAC) requests stakeholder feedback on: (1) a new process PTAC established for providing initial feedback to individuals and stakeholder entities who submit models to PTAC; (2) the requirements that PTAC has established for model submission; and (3) PTAC's processes for reviewing and making recommendations on those proposals. PTAC posted a list of specific issues on which it is seeking feedback.
Stakeholders can submit feedback by email to PTAC@hhs.gov. The deadline for comment submission is September 7, 2018.
For more information on the PTAC visit the McDermottPlus MACRA Resource Center or contact Sheila Madhani at 202-204-1459 or smadhani@mcdermottplus.com or Mara McDermott at 202-204-1462 or mmcdermott@mcdermottplus.com.


This piece is featured in this month's issue of The BeZine. To go to the interactive table of contents, click HERE.
Addressing this topic is a tricky proposition for me. How do I write about "Music" after a lifetime of being in its company, serious collegiate study, professional and semi-professional music-making and now coming to an ever-changing place of informal interaction with it? It is as daunting as writing about "Being Female".
The music of the soul, singing, is not without dukkha, the intrinsic suffering of human life. Aside from Art or Artifice, singing is a conduit for emotion as vulnerable and raw as any primal utterance. Those who have guessed this often try to manipulate it or manufacture it for their own uses. Or they try to lose their egos and get as close to being on the edge as they can. Who are the great "emotive" singers you can name? Judy Garland is our favorite. Her story and her relationship with her music is a painful one, but we love to hear her inimitable voice and styling. I used to play my Wizard of Oz record over and over again and try to sound just like her…before I was 10. Before I knew much about suffering at all. She was all of 16 on the recording. She kept singing that song throughout her career. She knew exactly how to wring all the pathos of her life from that melody by the time she died. She did it repeatedly, convincingly each time.
Song pouring forth like tears for the relief of an aching heart. Music is a channel for all the emotion that lives within us, be it deep sorrow, longing, suffering, yearning, passion, joy or triumph. Have you never brought an embryonic ache to maturity by playing the right music? Have you not fed a wild impulse by stomping out an insistent rhythm and letting your voice, your body move along with it? Music is my companion, my teacher, my soul mate. It accompanies me as I discover myself, like my breath, my heartbeat. It is biological and intellectual, a genius of Life like an inalienable right. I could not endure existence without it; I can not imagine Freedom without it.
My late husband was a singer, a gifted tenor. When he died, 300 people came to his memorial service to sing their good-byes – solos, congregational hymns and choir pieces. They sang as the living and imagined that there must be Music after death. They could not bear it to be otherwise. Though Death is entirely unknown and a very different (and luckier, as Walt Whitman would say) experience, I would not be surprised if there was music in it. Perhaps it is the very essence of all experience, conscious or not.
imagine what might happen if you simply followed your bliss and did what you love.
A life may emerge that is not what you dreamed or expected or even what you may have been promised. Still, it is actual and dynamic…and there you are, being yourself and still doing what you love. That's not a bad outcome, is it?
Today is Day 3 of my mother's Birthday Project. (Happy St. Stephen's Day as well. "Come on over; we'll celebrate getting stoned"…one of my mother's quips.) On the docket are 10 musical memories. My mother began her formal music training at the age of 5 when she started piano lessons. To this day, she plays and sings for the residents of her senior community quite regularly.
Her musicianship far exceeds mine, even though I have a B.A. in Music/Voice Performance. She has an M.A. in Church Music. She can improvise at the piano in various keys as well as play the organ: pedal keyboard and two manuals at once. I am "keyboard proficient" and can play the pump organ at the museum…meaning, essentially, I can read piano music and ride a bicycle at the same time. NOT the same skill set. My favorite arrangement is her at the piano bench and me singing alongside.
1) She is a young girl, her mother calls out proudly, "Anne Louise, play Clair de lune." She rolls her eyes. Not again! Consequently, I don't think I've ever heard her play it.
2) I am a young girl, a wee little kindergartener. Mom tucks me into the bottom bunk bed at night and sings, "Now the day is over/ night is drawing nigh/ shadows of the evening/ steal across the sky. Jesus gives the weary/calm and sweet repose/ with his tend'rest blessings/ may thine eyelids close." She kisses my forehead. "Ni' – night, d'good girl!" All is well. I hear her voice complete in my memory. Every note.
3) Mom is studying at Concordia Teacher's College. She needs to do some organ practice, and I'm not in school. Perhaps I'm sick? So she takes me with her. The organ is enormous. The room is large and institutional. I sit beside her and watch. I am fascinated by the pedal keyboard. Mom lets me crawl around on it, picking out tunes. I play "Peter Peter Pumpkin Eater" on the black keys, like I do on the piano. It booms out all over the room. This is great!!
4) I am about seven years old. I am the youngest member of our church choir which consists of my parents, my 3 older sisters, and a few others. I sit in the front row of the loft with the sopranos, leaning out to see the candles in the Christmas Eve procession. I am singing Midnight Mass with the adult choir, and I am going to stay awake through the whole service! How exciting to be allowed to sing out like an angel from up here instead of being stuck in the basement in the church nursery! Anthem's over. That sure was fun! There's a pile of coats in the corner of the pew….my, I'm feeling sleepy. I'll just rest a bit before the next hymn…oh! What? We're going home now? Did I miss the recessional? Drat!
5) I am about eleven years old. I've been taking piano lessons for 3 years. I practice before school every morning, while Mom washes the breakfast dishes in the kitchen. I am out in the living room, struggling away with a new piece. I hear Mom calling out from behind the swinging kitchen door, "It's F-sharp, Priscilla! Look at your key signature." I look. She's right. How did she know that from the other room?! I am trying to play a piano reduction of Dance of the Hours by Amilcare Ponchielli. I can't get the stresses right to make the piece dance. It comes out stiff. I'm playing what's written on the page, aren't I? Mom comes in, "Think of it this way…try singing in your head… cot-tage cheee-eese, cot-tage cheee-eese…" Suddenly, it clicks! Oh, this is that piece from Fantasia with the hippos in tutus!! I've got it now! But cottage cheese? What made her think of that?!
7) I am in High School. I am dating a guy whose mother was a concert pianist. He sings in the community college choir and has a great voice. My mother highly approves. She invites him over for dinner. Afterwards, she sits at the piano and pulls out some sheet music: Jerome Kern's "All The Things You Are". I've never heard it before, but it's a great fit – the style, the sentiment, the voice. I am in romantic heaven. Months later, he invites me to one of his Jazz Choir concerts. "I have a surprise for you," he says and puts a piece of paper in my hands. On the program, I see he has a solo. Yes, you can guess the song. He tells me he's dedicating it to me. Yes, that was Jim, my husband for 24 years, until his death.
8) So I go off to college to study, um, music. I'm 400 miles away from home. Meanwhile, back at the ranch, my mother and Jim have been singing in church together and have formed, along with some other church music colleagues, a group called Renascense (or some archaic spelling pronounced ren-NAY-sense). I've done an entire blog post on this memory in the past, titled "Christmas 1982" and you can read it here.
9) Finally, I am a junior in college. I have just been inducted into the Phi Beta Kappa society. My grandmother purchased the gold PhiBet key as a gift to me, and I went to the awards banquet, alone. I'm told a bit later that I should come to the annual Senior Awards Ceremony in May, even though I'm not really a senior. Well, maybe I am. I have enough credits to be. I figure it's related to the Phi Beta Kappa thing and tell my mother about it over the phone. "Just a bit of news, Mom. I know you have a 9-year-old at home to take care of, but there's going to be this other ceremony…" That sunny morning in Southern California, I am seated in Balch Hall with the choir and all the senior women of Scripps, glowing with promise. It's a beautifully festive day. I scan the crowd…and there's my mother! What?! She came all the way down here for this little ceremony? The awards are given out. The next one sounds interesting: The Gladys Pattison Music award, given to "the most deserving student in the field of music for the purpose of enriching her music library". Drum roll, please….yes! It's me! I am surprised; I beam. Afterwards, I find my mother. She hands me a little gift. It's a music box, wrapped in keyboard paper. I turn the handle and hear the opening notes of Beethoven's Moonlight Sonata. Professor Lamkin, the choir master, joins us and suggests that we all go out to lunch…off campus. What a treat! I stare in amazement at Mom, who is not the spontaneous type. "How did you just up and take off to be here?" I ask. "Oh, honey. I've been planning this for weeks." Oh. Well, that explains it.
10) And in closing, every medley eventually ends up with "My Buddy". This is mom's signature when she's been at the piano a while. No matter what key she's in, no matter where she's been dabbling, she always figures out how to incorporate this theme. "Miss your voice, the touch of your hand, just long to know that you understand, my Buddy….my Buddy, your Buddy misses you." I miss you, Mom. Thanks for all the music! I look forward to much more in the New Year!
Recently (well, almost 2 months ago now), we had a marvelous nighttime adventure in Chicago with my youngest, Emily. We went to Ravinia, the outdoor musical festival, for a Brahms concert. We bought only lawn seats, not the more expensive Pavilion seats. It rained all day, not too fiercely, but fairly steadily. We found that we were among the few diehard music fans that did not let that deter us from setting up a picnic on the wet lawn and dining happily under our umbrellas. When the music started officially (after a brief practice during our picnic), we packed up the food and huddled beneath the umbrellas. The rain was falling in earnest by then. At intermission, an invitation came over the loudspeaker for anyone on the lawn to move into the pavilion, as most seats were empty. A kind man handed us tickets to a box seat well under the shelter, and we moved in to warm up and dry off. It was a thrilling evening, being out in the elements, listening to live music played by real, live, dedicated musicians from Germany…and the occasional roll of thunder.
BTW – on the menu: 5 different kinds of cheeses, 3 salads, handmade chocolate-dipped strawberries (thank you, Emily!), a light Chardonnay and the best beer on the planet (from Belgium – Maredsous).


Nike golf balls have grown to become one of the most popular brands in golf. These imprinted and logo Nike golf balls make a great handout, tournament gift or sales premium. Their quality and playability are sure to please both the newcomer to the game and the die-hard player. Your company's logo or personal imprint can lie alongside one of the biggest names in golf -- or any sport for that matter.
Brand/Model 12-23 dz. 24-47 dz. 120+ dz.
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Ideas to use Numeracy in and around the home.
Ideas for using numeracy with your child.
Count on from any number you can see in the environment.
Look for number patterns around the home and the environment. Can you find house numbers for odd and even sequences?
Point out examples of numbers written as words in books, on signs, in papers etc.
How many people will we walk past on the way to and from school? Did we see more or fewer people on the way home? How many more / fewer people did we see?
Shopping receipts to find amounts less than £1. Make these amounts using 10p and 1p coins. Later make with the fewest amounts of coins.
Counting real money including 2p, 5p and 10p. Sort money into piles of 2p, 5p and 10p and encourage counting largest value first.


text:
First Pastor for Grace RP!
Home > Giving Thanks > First Pastor for Grace RP!
On Friday, January 22 of this year, Grace Reformed Church, a small mission church of the RPCNA in Columbia, Missouri (USA), welcomed her first pastor. The installation of Pastor Gary McNamee took place 11 years after this congregation first began. This is the story of God's faithful provision.
Congregation Formed
In October 2009, the Independent Reformed Church was formed with a steering committee consisting of Rev. Don Jefferson (PCA), Mr. Dave McLane, Rev. Jeff Yelton (RPCNA), and Dr. Noah Manring. The goal: to build a church committed to the Reformed Faith, with distinctive practices of exclusive psalm singing, a-cappella singing, and the use of the Authorized Version of the Bible for preaching. Rev. Jeff Yelton agreed to serve as pulpit supply.
Gary & Cynthia McNamee with the members of Grace Reformed Church
In June 2010, the Free Church Continuing (FCC) received the Independent Reformed Church as a preaching station under the new name of Grace Reformed Church. From January 2012 (when Rev. Yelton moved to Kansas) onward, Grace Reformed Church has sat under a variety of pastors providing pulpit supply. These include Rev. Jeff Yelton (RPCNA), Rev. Jonathan Mattull (FCC), Rev. Don Jefferson (PCA), Rev. Bob McFarland (RPCNA), Rev. John Keddie (FCC), Rev. Brent Evans (OPC), and Dr. Noah Manring (FCC).
Joining the RPCNA
In May of 2018, Rev. Bob Hemphill and Rev. Jonathan Haney of the RPCNA visited Columbia to preach and discuss the possibility of being taken into the RPCNA. On June 26, 2018, a petition was made at the RPCNA Synod meeting in Marion, Indiana to receive Grace Reformed Church as a mission work. In October 2018, the Ad Interim Commission of the Midwest Presbytery established a Temporary Governing Body to oversee Grace Reformed Church as a new mission work under the umbrella of the RPCNA.
The Temporary Governing Body consisted of Rev. Bob Hemphill (chair), Rev. Joe Allyn, Rev. Bob McFarland, Elder Romesh Prakashpalan, and Rev. Greg Stiner. Among their responsibilities were the reception of members, pulpit supply, oversight of the congregation's spiritual and financial welfare, and guidance toward full organization with officers and a pastor.
Installation of Gary McNamee as First Pastor
On December 4, 2019, Grace Reformed Church was formally welcomed into the Midwest Presbytery as a mission work of the RPCNA and the congregation began the process of searching for a pastor. On August 28, 2020, they issued a call to Gary McNamee.
On Friday, January 22, 2021, Gary McNamee was installed as the first pastor of Grace Reformed Church. Great is God's faithfulness!
RP Global Alliance thanks the session of Grace Reformed Church for this account. Stay tuned for part two, where Pastor McNamee will share a little of his journey to the pastorate.
February 5, 2021 at 1:40 pm Katrina van der Meer Bob Hemphill, Bob McFarland, Columbia, Columbia Missouri, Dr Noah Manring, Gary McNamee, Grace-Reformed-Church-Missouri-RPC, Jeff Yelton, Jonathan Haney, Midwest Presbytery, Mission Church, Missouri, Noah Manring, Reformed church Columbia, Reformed Church Missouri, RPCNA Giving Thanks, Latest News, Ordination / Installation, Slider Share via:


The Annual Independence Ministerial Alliance Beacon Heights concert is a tremendous group of local churches coming together to perform their best songs. There will be choirs, soloist, instrumentals and the like. You don't want to miss it! All free will proceeds from the concert benefit Truman Heritage Habitat for Humanity's mission.


The TOTO Promenade II 1G 1.0 GPF Toilet Tank with Right-Hand Trip Lever is designed for use with the Promenade II bowl. The Promenade II tank includes a tank to bowl gasket, tank to bowl hardware, and toilet bolt caps. The tank features TOTO''s industry leading 1G technology, offering a powerful flush that flushes completely and fully with only one gallon of water. This version of the tank comes equipped with a decorative right-hand chrome trip lever. The Promenade II meets the standards for EPA WaterSense, and California's CEC and CALGreen requirements. Additional items needed for installation and use must be purchased separately: toilet bowl, wax ring, toilet mounting bolts, water supply lines, and toilet seat. The Promenade II ST403U tank can be used on the toilet C404CUFG bowl.
Shipping: L:18"x W:9"x H:16" 31.5 lbs.
Mountainland Kitchen & Bath is proud to present this cotton finished toilet tank, by Toto. The ST403UR#01 is made from premium materials, this Toilet Tank offers great function and value for your home. This fixture is part of Toto's decorative Collection, so make sure to check out other styles of fixtures to accessorize your room.


text:
Q: What is the better way of writing multiple if statements in Ruby Rails? I'm pretty new to Ruby and have this scenario of writing multiple if statements. I want to optimize it and trying to find out if there are any other alternatives or better ways of writing the following conditions -
              if region == "uk"
                value = 1
              end
              if region == "us"
                value = 2
              end
              if region == "ind"
                value = 3
              end
              if region == "cn"
                 value = 4
              end

Problem statement - I can't be going on and on if other regions add up to my scenario. Any optimized way of writing the above code ?
Any teachings and solution would be really appreciated!

A: One way is to use a case statement:
value = case region
        when 'uk'
          1
        when 'us'
          2
        # etc.
        end

But when you've got a simple list like this with no logic beyond the lookup, you can build a hash with the value to lookup as the key, and the returned value as the value:
region_values = {
  'uk' => 1,
  'us' => 2,
  # etc.
}

value = region_values['uk']

A: The following are three options I would consider. Initially, I assume that the value of region is known to be one of the values in the array
REGIONS = ["uk", "us", "ind", "cn"]

Suppose
region = "ind"

#1
value =
case region
when "uk"  then 1
when "us"  then 2
when "ind" then 3
when "cn"  then 4
end
  #=> 3

#2
REGION_TO_ID = { "uk"=>1, "us"=>2, "ind"=>3, "cn"=>4 }

value = REGION_TO_ID[region]
  #=> 3

#3
value = REGIONS.index(region) + 1
  #=> 3

where REGIONS is as defined above.

If the value of region is not known to be one of the values in the array REGIONS one might first execute the following:
region = "moon"

raise ArgumentError, "'#{region}' is not an element of REGIONS" unless
  REGIONS.include?(region)
  #=> ArgumentError: 'moon' is not an element of REGIONS

Alternatively, in #1 at the end of the case statement one could insert
else raise ArgumentError, "'#{region}' is not an element of REGIONS"

and in #2 one could add
REGION_TO_ID.default_proc = proc { |_,region|
 raise ArgumentError, "'#{region}' is not an element of REGIONS" }

Then,
REGION_TO_ID['moon']
  #=> ArgumentError: 'moon' is not an element of REGIONS

See Hash#default_proc=.

A: There are few ways.
You can also wrap-up if statement in one liner instead of the block
value = 1 if region == "uk"
value = 2 if region == "us"
value = 3 if region == "ind"
value = 4 if region == "cn"

Can also do something like this
value = if region == "uk"
         1
        elsif region == "us"
         2
        elsif region == "ind"
         3
        elsif region == "cn"
         4
        end

But when comparing 2 values, better to use switch statement ie
value = case region
        when "uk"
         1
        when "us"
         2
        when "ind"
         3
        when "cn"
         4
        end


Hanoi has 16 museums in different districts which are opened in the regulated time every day. To visit in museum, tourists have to buy ticket.
Address: No. 1 Phạm Ngũ Lão, Tràng Tiền street, Hoàn Kiếm District, Hanoi.
Opening time: 8h00- 12h00 and 13h30- 17h.
Address: Nguyễn Văn Huyên street, Dịch Vọng, Cầu Giấy District, Hanoi.
Temple of Literature is famous as the most various complex in Hanoi, it is ranked on list of 23 special national relics. Hanoians always proud of Temple of Literature and it is a symbol of Hanoi travel.
Hoa Lo Prison is located in the heart of Hanoi. It is built in 1896 by French on the land of Phu Khanh village. The French occupied Hanoi in order to detain Vietnamese patriots who opposed their rule. They built one prison in 1896 in the middle of Hanoi and occupy all area of Phu Khanh Village to build prisons.
Cyclo is used in the past century, cyclo is becoming traditional culture beauty of Hanoi now. This is a suitable mean for tourists to visit in Hanoi.
Walking in Hanoi old quarters, you can easily see images of lacquer stores with unique and skilful works. Foreign tourists can buy it as souvenir for friend and family.
6 favourite experiences when travelling in Hanoi and the posts related.


text:
Atkinson named to 11th District House of Delegates seat
CHARLESTON, W.Va. – Martin "Rick" Atkinson III will represent residents of the 11th District in the West Virginia House of Delegates.
Gov. Earl Ray Tomblin today announced the appointment Sunday. Atkinson fills the vacancy created when former Delegate Bob Ashley was appointed to the West Virginia Senate on Sept. 15. The governor's appointment is effective immediately.
Atkinson, a West Virginia University graduate, retired from BPB-Stella Jones in 2013 and currently serves as director of sales for Mountain State Log Homes. He is an active volunteer with the 4-H Club in Roane County and has helped replace the cabins at Camp Sheppard. Atkinson is a member of First Baptist Church in Spencer.
He and his wife, Jeanette, live in Reedy.