text:
Towing camper over septic fields?
Hey all...
I have a 1300 pound pop up camper. I have a 425 pound riding mower. If i wanted to move the camper around, think its safe to tow it over the septic fields? I have in ground sprinkler too.
Septic field pipes (NOT the tank, the fields) are typically at least 14-18 inches below grade. It is generally OK to run a light truck over them. Consider how they are built, first the field is excavated to grade, then the pipes are installed and covered with gravel, finally the soil cover is put on. This is all done using heavy equipment. The tank itself is usually reinforced concrete, and rated H-10 or H-20, depending on the manufacturer. An H-10 or H-20 rated tank can support a pickup truck easily. Of course, you may have an old tank that is not rated, so best to check your plans before driving on top of the tank.
Great info. The home was built in 1998. Do you think there would be anything old-school about the field, or would that age weaken it at all? I sense that I'd be fine towing this camper across that area...but just want to be sure. I won't be going anywhere near the tank.
|All times are GMT -5. The time now is 01:08 PM.
vBulletin Security provided by vBSecurity v2.2.2 (Pro) - vBulletin Mods & Addons Copyright © 2017 DragonByte Technologies Ltd.
User Alert System provided by Advanced User Tagging (Pro) - vBulletin Mods & Addons Copyright © 2017 DragonByte Technologies Ltd.


text:
Originally Posted by ChienAboyeur
Yes, it was told to narrativist gamers. Actually, anytime I GM a game, I tell that the session is how role playing, not looking for builds, not looking for the elaboration of a narrative etc
How people call things do not change what things are, when they are things like RPG.
Narrative game players can call their games RPGs. That wont make them RPGs.
They could call football RPG.

subset: commoncrawl


text:
Article Preview
State's High Court Upholds Reforms In Liquor Controls
ALBANY, Jan. 7 (AP) -- The state's highest court today upheld the constitutionality of two key segments of Governor Rockefeller's liquor reform program, which was adopted at a special session of the State Legislature last year.
January 08, 1965
Thank you for visiting The New York Times archive.
New York Times subscribers* enjoy full access to TimesMachine—view 129 years of New York Times journalism, as it originally appeared. 99¢ for your first 4 weeks.
Or, purchase this article individually for $3.95 and download a high-resolution PDF.
* Does not include NYT Now or Premium Crosswords subscribers.
Already a subscriber? Log in to view this article »


text:
Thank you for your vote! You have already voted for this video!
Report Download
Submited by: Miaford
Joined: 6 years ago
miaford secret movie on 1/28/15 05:01 from chaturbate
Duration: 120:39 Views: 218590 Added: 1 year ago
Categories: Solo Webcam Chaturbate Redhead Strip Close-up Big Tits Straight Masturbation
There are no comments for this video.


text:
Mammalian microRNAs: a small world for fine-tuning gene expression
- First Online:
- Received:
- Accepted:
DOI: 10.1007/s00335-005-0066-3
- Cite this article as:
- Sevignani, C., Calin, G.A., Siracusa, L.D. et al. Mamm Genome (2006) 17: 189. doi:10.1007/s00335-005-0066-3
- Citations
- 1.1k Downloads
The basis of eukaryotic complexity is an intricate genetic architecture where parallel systems are involved in tuning gene expression, via RNA-DNA, RNA-RNA, RNA-protein, and DNA-protein interactions. In higher organisms, about 97% of the transcriptional output is represented by noncoding RNA (ncRNA) encompassing not only rRNA, tRNA, introns, 5′ and 3′ untranslated regions, transposable elements, and intergenic regions, but also a large, rapidly emerging family named microRNAs. MicroRNAs are short 20-22-nucleotide RNA molecules that have been shown to regulate the expression of other genes in a variety of eukaryotic systems. MicroRNAs are formed from larger transcripts that fold to produce hairpin structures and serve as substrates for the cytoplasmic Dicer, a member of the RNase III enzyme family. A recent analysis of the genomic location of human microRNA genes suggested that 50% of microRNA genes are located in cancer-associated genomic regions or in fragile sites. This review focuses on the possible implications of microRNAs in post-transcriptional gene regulation in mammalian diseases, with particular focus on cancer. We argue that developing mouse models for deleted and/or overexpressed microRNAs will be of invaluable interest to decipher the regulatory networks where microRNAs are involved.
What are microRNAs?
The central dogma of classical biology is that genetic information flows from DNA to RNA to proteins. Therefore, "genes" are synonymous with proteins and a gene is defined as a protein-coding region with associated regulatory signals (Mattick 2003). MicroRNAs represent new stars in the gene regulation galaxy, and there is a strong interest among researchers in different fields to understand their mechanism of action and identify their targets. The definition of a gene in the genomics era should be expanded to be a "transcription unit" or "a complete chromosomal segment responsible for making a functional product" (Okazaki et al. 2002). Because of the complex genetic architecture of eukaryotic cells, this definition is still not complete, and additional issues such as overlap, alternative splicing, or transcribed pseudogenes have to be considered (Snyder and Gerstein 2003). Thus, the term "gene" could be expanded to include microRNAs (and other functional ncRNAs).
MicroRNA websites for "in silico" cloning, target identification, and the Registry
"In silico" cloning
C. elegans, Human
Dr. Pavel Tomancak (personal communication)
Target identification
Diana MicroT
miRNA–target prediction
RNA effective
MicroRNAs database
The MicroRNAs Registry
How are microRNAs produced?
More than a decade ago, Ambros and colleagues discovered that lin-4, a gene known to control the timing of C. elegans larval development, does not code for any protein but instead codes for a pair of small RNAs (Lee et al. 1993). One RNA is longer (∼70 nt) and can fold in a stem-loop structure, as the precursor of the shorter RNA (∼22 nt). The Ambros and Ruvkun laboratories later discovered that the small lin-4 RNAs had an antisense complementarity to multiple sites in the 3′ UTR of the lin-14 protein-coding gene (Lee et al. 1993; Wightman et al. 1993). The sequence complementarity between the noncoding lin-4 RNA and the 3′ UTR of the protein-coding lin-14 gene suggested that lin-4 regulated translation of lin-14 by binding to the 3′ UTR. It was later shown that lin-4 regulated the lin-14 gene during early larval development as well as the lin-28 gene during late larval development by a mechanism involving RNA-RNA binding that led to suppression of translation (Moss et al. 1997). The shorter lin-4 RNA is now known as the founding member of the class of microRNA genes (Lagos-Quintana et al. 2001; Lau et al. 2001; Lee and Ambros 2001).
The majority of microRNAs (70%) are located in introns and/or exons, and approximately 30% are located in intergenic regions (Rodriguez et al. 2004) (Fig. 1). The first group of microRNAs from introns and/or exons is oriented in sense with the exon-coding "host" gene and, therefore, may be transcribed as part of annotated genes. The second group of microRNAs are transcribed from intergenic regions or gene deserts (Lagos–Quintana et al. 2001; Lau et al. 2001; Lee and Ambros 2001), indicating that they form independent transcription units (Lee et al. 2002). The third group of microRNAs are derived from introns and/or exons of annotated genes but are transcribed in the antisense orientation, suggesting that they too form their own transcription units. Tightly linked microRNAs may be transcribed as polycistronic messengers; however, microRNAs separated by more than 50 kb tend to represent independent transcription units (Baskerville and Bartel 2005). RNA polymerases II and III are candidates for pri-microRNAs (primary) transcription (Fig. 1).
In animals (Fig. 1), the first step in microRNA maturation is the nuclear cleavage of the several-kb-long pri-microRNA, which releases an approximately 70-nt, cropped, hairpin-shaped intermediate known as pre-microRNA (precursor). The Drosha RNase III endonuclease is responsible for this nuclear processing (Bartel 2004; Lee et al. 2003; Zeng et al. 2005). Drosha cannot cleave without a partner, forming a complex with DGCR8 (a product of the DiGeorge syndrome critical region gene 8), which contains two double-stranded RNA-binding domains (Han et al. 2004; Tomari and Zamore 2005). The hairpin precursor is actively transported from the nucleus to the cytoplasm by exportin 5 (Yi et al. 2003; Lund et al. 2004). A second enzyme, an RNase III endonuclease called Dicer, is responsible for generating an approximately 21-nt, short, single-stranded RNA that is the mature microRNA (Lee et al. 2003). Dicer was first recognized for its role in generating siRNAs that mediate RNA interference (RNAi) (Bernstein et al. 2001). After cleavage in the cytoplasm, the microRNA pathways of plants and animals share some other steps with RNA silencing, and the cleavage products become incorporated as single-stranded RNAs into the ribonucleoprotein RNA-induced silencing complex RISC (Hammond et al. 2000). The RISC has been purified from fruit fly and human cells and in both cases contains a member of the Argonaute protein family (essential for gene silencing in Caenorhabditis elegans, Neurospora, and Arabidopsis), which is thought to be a core component of the complex (Hammond et al. 2001; Mourelatos et al. 2002). Once incorporated in the cytoplasmic RISC complex, the microRNA will specify cleavage if the mRNA has sufficient complementarity to the microRNA, or it will block translation if the microRNA does not have sufficient complementarity, resulting in reduced expression of the corresponding protein. MicroRNA regulation of the microRNA:mRNA duplex is mediated mainly through multiple complementary sites in the 3′ UTRs, but there are many exceptions. MicroRNAs may also bind the 5′ UTR and/or the coding region of mRNAs, resulting in a similar outcome (Fig. 1).
How are microRNAs working?
The functions of microRNA are various, such as the control of leaf and flower development in plants (Aukerman and Sakai 2003) or the modulation of hematopoietic lineage differentiation in mammals (Chen et al. 2004). Several groups have uncovered roles for microRNAs in the coordination of cell proliferation and cell death during development and in stress resistance and fat metabolism (Ambros 2003). For example, the Drosophila microRNA gene (mir-14) suppresses cell death and is required for normal fat metabolism (Xu et al. 2003), while the bantam locus encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila (Brennecke et al. 2003). It was recently shown that, in addition to their regulatory functions, cellular microRNAs mediate antiviral defenses in human cells (Lecellier et al. 2005).
However, the function of most microRNAs is not known. The antisense single-stranded microRNAs can bind specific mRNA transcripts through sequences that are significantly, though not completely, complementary to the target mRNA. This process is also known as post-transcriptional gene regulation (PTGS). Some microRNAs can downregulate large numbers of target mRNAs (Lim et al. 2005), and it has been speculated that microRNAs could regulate approximately 30% of the human genome (Bartel 2004). MicroRNAs seem to be responsible for fine regulation of gene expression, "tuning" the cellular phenotype during delicate processes like development and differentiation in all organisms, from plants to mammals. Many microRNAs are conserved in sequence between distantly related organisms (see The MicroRNA Registry in Table 1), suggesting that these molecules participate in essential processes (Lagos-Quintana et al. 2003; Pasquinelli et al. 2000). One example is the cluster of mir-16-1 and mir-15a, which is highly conserved in nine of ten primate species (Berezikov et al. 2005).
Target identification has been hampered by the fact that in animals, in contrast to plants, microRNAs do not bind perfectly to their targets. A few nucleotides typically remain unbound, yielding complex secondary structures. Mammalian genes can have more than one microRNA target site in their 3′ UTRs and one microRNA can target more than one mRNA. Bioinformatics approaches have been developed to search for the most thermodynamically favorable microRNA:mRNA duplex interactions (Table 1). Several computational procedures such as DianaMicroT, TargetScan, and miRanda (Lewis et al. 2003; Kiriakidou et al. 2004; Enright et al. 2003) are available to predict microRNA targets. Another set of programs developed for target identification includes RNAhybrid, RNAcalibrate, and RNAeffective (Rehmsmeier et al. 2004), which work by searching for the most energetically favorable hybridization sites for the smaller microRNA within the larger mRNA (Table 1). Only a few target mRNAs have been experimentally proven and studied in vitro, but the numbers of such confirmed interactions are expected to sharply increase as prediction tools become more sophisticated. Generally, the proven mRNA targets are produced from different chromosomes as the corresponding microRNAs (such as mir-19a on chromosome 13q31.3 having as a target the mRNA of the PTEN tumor suppressor gene located on chromosome 10q23.3), but examples of microRNAs targeting "neighbor" exon-coding genes are also known. An important example is that of mir-196, which leads to the cleavage and degradation of mRNA from the homeobox gene HOXB8 (Yekta et al. 2004).
Functional noncoding RNAsa
ncRNA symbol
ncRNA name
Size (kb)
Antisense Igfar RNA
Silencing autosomal imprinted genes
H19 fetal liver RNA
Human, mouse
Imprinted embryonic transcript, tumor supppressor gene, and/or associated with tumor progression in breast
Brannan et al. (1990); Lottin et al. (2002); Kanduri et al. (2005)
Imprinted in Prader-Willi syndrome
Human, mouse
Prader-Willi syndrome, imprinting process
Noncoding transcript in T cells
Transcript present only in activated CD4+ T-cell clones, Regulation of neighboring
RNA on the X1
Fruit fly
Dosage compensation
Antisense orientation of Xist
Regulation in the early steps of X inactivation
Lee et al. (1999)
X-inactivation Specific transcript
X-chromosome inactivation
Brockdorff et al. (1992); Avner and Heard (2001)
MicroRNAs and human cancer
MicroRNAs and human cancer
Cancer type
MicroRNA alterations
Deletions and down regulation of mir-15 andmir-16 (∼68% of cases)
Distinct microRNA profiling signatures in leukemias
MicroRNA signatures associated with prognostic factors and disease progression
B-cell lymphoma
Accumulation of mir-155 and BIC gene
Primary or mature levels of microRNAs derived from the mir-17-92 locus are often increased
Colorectal cancer
Reduced mir-143 and mir-145 (∼80% cases)
Mir-21 is overexpressed in human malignant cell lines and tumor tissues
Human cancer
50% of microRNAs located in minimal LOH/Amplified regions
Lung cancer
Reduced expression of Let-7 associated with short postoperative survival (∼40% cases)
Let-7 microRNAs regulates expression of RAS
Malignant lymphoma
Genomic amplification of mir-17-91-18- 19a-20-19b-92 cluster
Pediatric Burkitt's lymphoma
High expression of precursor mir-155/BIC
Profiles of human tumors
Distinct microRNA expression signatures
The first report linking microRNAs and cancer (Calin et al. 2002) involved chronic lymphocytic leukemia (CLL), the most common form of adult leukemia in the Western world. Hemizygous and/or homozygous loss at chromosome 13q14 occurs in more than half of CLL cases. Loss of chromosome 13q14 is also found in more than 50% of mantle cell lymphomas (Stilgenbauer et al. 1998), in approximately 30% of multiple myeloma (Elnenaei et al. 2003), and in approximately 70% of prostate cancers (Dong et al. 2001), suggesting that one or more tumor suppressor genes located at chromosome 13q14 are involved in the pathogenesis of human tumors (Calin et al. 2002). However, detailed genetic analysis, including extensive loss of heterozygosity (LOH), mutation, and expression studies failed to demonstrate the consistent involvement of any of the 12 protein-coding genes located in or close to the deleted region. A cluster of two microRNAs, mir-15a and mir-16-1, was found within the minimal region of deletion (∼30 kb) at 13q14 and to be deleted or downregulated in approximately 70% of CLL samples. Similarly, a t(12;13) chromosome translocation in a patient with CLL was found to decrease the mir-15a and mir-16-1 precursors (Calin et al. 2002). A similar cluster of microRNAs, named mir-15b and mir-16-2, is located on human Chromosome 3 but is expressed at very low levels in lymphoid cells.
The first genome-wide systematic search for correlations between the genomic positions of microRNAs and cancer-associated genomic regions (CAGRs) provided a catalog of microRNAs possibly involved in cancer (Calin et al. 2004a). One hundred eighty-six microRNAs were mapped to the human genome and compared with the location of previously reported nonrandom genetic alterations identified in human tumors and with cloned fragile sites (FRAs). Minimal regions of LOH are suggestive of the presence of tumor suppressor genes, minimal regions of amplification for the presence of oncogenes, while common breakpoint regions are found close to both types of cancer genes. Some common fragile sites predispose to DNA instability in cancer cells; indeed they are preferential sites of sister chromatid exchange, translocation, deletion, amplification, or integration of plasmid DNA and tumor-associated viruses. This study concluded that 19% (35 of 186) of microRNAs are located inside or near FRAs and the relative incidence of microRNAs inside FRAs occurred at a rate nine times higher than in non-FRAs. A significant proportion, 52.5% (98 of 186), of microRNAs are in CAGRs described in a variety of tumors such as lung, breast, ovarian, colon, gastric, and hepatocellular carcinoma, as well as leukemias and lymphomas.
Little was known about the expression levels of microRNA genes in normal and neoplastic cells until 2002. To assess cancer-specific expression levels for hundreds of microRNAs is time-consuming and requires a large amount of total RNA and the use of radioactive isotopes. cDNA microarrays are useful tools for identifying different patterns of expression in a large number of samples. The first developed oligonucleotide microRNA microarray chips (Liu et al. 2004a), containing hundreds of human precursor and mature microRNA probes, identified distinct patterns of microRNA expression in human and mouse tissues (tissue-specific microRNA expression signatures). Another method to determine microRNA expression levels involves the use of a bead-based flow cytometric technique (Lu et al. 2005a). Since the development of these two methods of assessing global microRNA expression, several commercially available platforms have been developed for microRNA gene expression profiling.
Global analysis of microRNA gene expression has shown that these profiles can be used to classify the developmental lineage and differentiation state of tumors; in addition, microRNA profiles appear to provide a distinct signature for tumors which is more precise than that provided by protein-coding genes (Lu et al. 2005a). These findings lead to a unique opportunity to use these microRNA profiles for tumor diagnostics.
Specific differences in microRNA expression between CLL samples and normal CD5+ B cells, the latter representing normal cells corresponding to CLL malignant cells, were found (Calin et al. 2002). Two clusters of CLL samples were identified that were associated with the presence or absence of ZAP-70 expression, a predictor of early disease progression. Low levels of ZAP-70 (identified by both Western blots and flow cytometry) are associated with good prognosis. Two microRNA signatures were associated with the presence or absence of mutations in the expressed immunoglobulin variable-region genes or with deletions at chromosome 13q14, respectively. Different microRNA expression profiles suggested that specific microRNAs can have prognostic significance in CLL, thus expanding the spectrum of prognostic markers (Calin et al. 2004b). A unique microRNA expression signature composed of 13 genes can differentiate cases with low or high ZAP-70 expression and cases with unmutated or mutated IgVH. Furthermore, both mir-15a and mir-16-1 are members of this signature, suggesting an important functional role in the pathogenesis of human CLL (Calin et al. 2005).
The list of microRNAs reported to be involved in cancer is growing continuously (Table 3). One report showed that mir-143 and mir-145 exhibited reduced levels of the mature microRNA in colon cancer patients, colon cancer cell lines, and precancerous adenomatous polyps when compared to normal tissues by Northern blot (Michael et al. 2003). On the contrary, levels of unprocessed hairpin precursors were not altered. It is possible that the reduced accumulation of the mature microRNA reflects early changes in neoplastic cells or that this depletion is related to their activity in binding and inactivating target mRNAs in the cytoplasm. Putative targets (human mRNAs that display imperfect complementarity of 70%–100%) for mir-143 and mir-145 include several genes that encode components of signal transduction pathways (such as RAF, RHO, and G-proteins) and chromatin-mediated control of gene expression were found by computer search (Table 3).
In an independent study, it was shown that mir-143 regulates adipocyte differentiation (Esau et al. 2004). When antisense oligonucleotides (ASOs) targeting human microRNAs were transfected into cultured human primary subcutaneous preadipocytes, mir-143 was upregulated as adipocytes differentiated. Bioinformatics approaches predicted several putative microRNA targets. ERK5/BMK1 (which promotes cell growth and proliferation in response to tyrosine kinase signaling) was predicted to be a mir-143 target and, in fact, ERK5 protein levels were upregulated in mir-143 ASO-treated adipocytes. However, because each microRNA can regulate multiple target genes, mir-143 could directly inhibit ERK5 by binding the 3′ UTR of ERK5, or indirectly by binding to another target gene (Esau et al. 2004).
Another study reported reduced expression of the let-7 microRNA in human lung cancer and the correlation with clinicopathologic features (Takamizawa et al. 2004). Reduction in the expression (>80%) levels of let-7 was observed in 60% (12 of 20) of cancer cell lines and 44% (7 of 16) of lung tumors when compared with normal lung tissues by Northern blots. The reduced expression was significantly associated with shortened survival of patients after potentially curative resection. One hundred forty-three cases had been followed for more than five years after surgery and were used to study the prognostic significance of let-7. One hundred forty-three non-small-cell lung carcinoma (NSCLC) cases were classified into clusters 1 and 2 (cluster 1 with low let-7 expression and higher disease stages; cluster 2 with high let-7 expression and lower disease stages). The difference in postoperative survival between clusters 1 and 2 was highly significant. In addition, overexpression of let-7 in the A549 lung adenocarcinoma cell line inhibited lung cancer cell growth in vitro (Takamizawa et al. 2004). Several observations led to the conclusion that let-7 acts as a tumor suppressor gene in lung tissue. Several let-7 family members, including let-7a-2, let-7c, and let-7g, have been mapped to minimally deleted regions in lung cancers (Calin et al. 2004a). Let-7 expression is lower in lung tumors than in normal lung tissue, while RAS protein is significantly higher in lung tumors (Johnson et al. 2005). Finally, expression of the RAS oncogene is regulated by let-7, and overexpression of let-7 has been shown to inhibit lung tumor cell line growth (Johnson et al. 2005).
High expression of human BIC RNA has been found in Hodgkin's lymphoma (van den Berg 2003). In contrast, an analysis of non-Hodgkin's lymphoma (NHL) did not reveal any remarkable upregulation of BIC expression. The BIC locus was originally identified as a common retroviral integration site in avian leukosis virus-induced B-cell lymphomas (Tam et al. 1997). The human BIC locus also encodes a microRNA, mir-155 (Tam 2001; Lagos-Quintana et al. 2002). The conditio sine qua non for the development of Burkitt's lymphoma is activation of the MYC oncogene, mostly by chromosomal translocations in which MYC is juxtaposed next to an immunoglobulin enhancer. But activation of MYC alone is not sufficient for full malignancy; in fact, MYC cooperates with other oncogenes. Further studies involving a human B-cell line that contains a tetracycline-regulated c-MYC gene have shown that overexpression of the c-MYC oncogene leads to increased expression of six microRNAs, two of which are mir-17-5p and mir-20a (O'Donnell et al. 2005). These microRNAs were found to negatively regulate expression of the E2F1 transcription factor. Since E2F1 is also a target of c-MYC, these results allude to an intricate network that exquisitely regulates gene expression, ultimately controlling cell proliferation.
In childhood Burkitt's lymphoma, there has been reported a 100-fold upregulation of the hairpin precursor mir-155 (Metzler et al. 2004). It could be speculated that mir-155 directly downregulates one of the MYC antagonists such as MAD1, MXI1, or ROX/MNT. Accumulation of mir-155 and BIC RNA was also reported in other B-cell lymphomas (Eis et al. 2005).
It was recently reported that mir-21 is upregulated in both human glioblastomas as well as cultured glioblastoma multiforma cells and cell lines compared to primary astrocyte cultures derived from normal tissue; inhibition of mir-21 expression led to the activation of caspases and cell death in vitro (Chan et al. 2005). These data suggest a role for mir-21 in malignant glioblastoma, most likely by influencing expression of genes involved in apoptosis (Chan et al. 2005).
Overall, the studies to date clearly implicate a causative role for microRNAs in human cancer. The intricate network of signals regulating gene expression requires a delicate balance between protein-coding genes and microRNA genes. Disruption of this delicate relationship by mutation can tip the balance in favor of the development of cancer.
MicroRNAs and other human diseases
MicroRNAs and other human diseases
Protein alteration
Fragile X syndrome
Loss of expression of the fragile X mental retardation protein (FMRP)
Peng et al. (2004)
Spinal muscular atrophy (SMA)
Reduction of expression in Survival of Motor Neurons protein (SMN)
DiGeorge syndrome
22q11.21 microdeletion with loss of expression of DGCR8
Spinal muscular atrophy (SMA) is an inherited neurodegenerative disease with neuromuscular symptoms. Survival of Motor Neurons protein (SMN) is a component of the SMN complex that has critical functions in the assembly and restructuring of diverse ribonucleoprotein (RNP) complexes. Components of the SMN complex (GEMIN3 and GEMIN4) are also in a separate complex that contains eIF2C2, a member of the Argonaute protein family. This novel complex contains numerous microRNAs that form novel RNPs named miRNPs (Mourelatos et al. 2002). This link between microRNAs and a devastating neurodegenerative disease is intriguing and will be of great interest to determine what effect these microRNAs have on miRNPs in SMA patients.
DiGeorge syndrome is characterized by a few specific cardiac malformations, facial deformity, and certain endocrine and immune anomalies. The DGCR8 (DiGeorge syndrome critical region gene 8) protein and Drosha interact in human cells. Knockdown of the Drosophila dmDGCR8 resulted in a 5–23-fold accumulation of some pri-microRNAs, confirming a role in mediating the genesis of miRNAs from the pri-microRNA transcript (Landthaler et al. 2004; Gregory et al. 2004).
MicroRNAs have also been shown to regulate insulin secretion (Poy et al. 2004). Overexpression of mir-375 (a pancreatic islet-specific microRNA) suppresses glucose-induced insulin secretion, and, conversely, inhibition of endogenous mir-375 by ASOs enhances insulin secretion in murine pancreatic α- and β-cell lines. Bioinformatics approaches predicted several putative targets with a potential role in insulin secretion such as MAPK4, MKI1, VTI1A, and MTPN; the last two proteins are known to be involved in vesicle transport of neurons and in neurotransmitter release.
These studies represent the beginning links between microRNAs and human disease. Further investigations are likely to reveal the involvement of additional microRNAs and their targets in simple and complex genetic diseases.
Mouse models for microRNA studies
Almost all mammalian microRNAs are highly conserved and more than 220 mouse microRNAs were cloned by conventional methods (Lagos–Quintana et al. 2002; Seitz et al. 2004) (see The MicroRNA Registry in Table 1). It has been shown that a tissue-specific microRNA dominates the population of expressed microRNAs, suggesting important roles in tissue differentiation. For example, in mouse liver, mir-122a and mir-122b represent 72% of all cloned microRNAs; these microRNAs were not detected in the other tissues analyzed. In spleen, mir-143 was the most abundant microRNA. In colon, mir-142 was cloned several times; however, because of the strong RNase activity in small intestine and pancreas, too few microRNA sequences were obtained from these tissues to reach statistical significance.
Using a microRNA microarray approach, microRNA expression was analyzed in 17 mouse organs and tissues (Babak et al. 2004). More than half of the 78 microRNAs detected were expressed in specific adult tissues. The results were confirmed by Northern blotting and were consistent with microRNA expression data reported in the literature. Furthermore, this study is consistent with microRNA expression profiling in human tissues (Liu et al. 2004). For example, seven of eight microRNAs were brain-specific in both mice and humans.
Another report gave new light on the world of microRNA regulation in humans and mice (Seitz et al. 2004). By bioinformatics approaches, 46 potential microRNAs located in the human imprinted 14q32 domain have been identified. Forty of these microRNAs are clustered and most of them are arranged in tandem arrays. The majority of these genes map within an approximately 40-kb region, making it the largest microRNA cluster that has been described. In mouse, this microRNA cluster is conserved at the homologous region on Chromosome 12. The detected microRNAs are expressed in embryos and placenta, whereas in the adult their expression pattern is limited to the brain. Expression of these microRNAs comes solely from the maternally inherited chromosome, which is imprinted. Regulation of their expression is dictated by a region located approximately 200 kb upstream from the microRNA cluster (Seitz et al. 2004).
Research has shown how microRNAs have a role in fine-tuning specific Hox mRNA expression patterns during mouse development (Mansfield et al. 2004). A novel approach was used to monitor microRNA tissue-specific expression patterns during embryogenesis. Until now, microRNA expression studies were done by Northern blots, tissue-specific RNA cloning, and microarrays, but all of these methods are not useful for displaying spatiotemporal expression patterns. In situ hybridization methods are still difficult to adapt in vertebrate embryos because of the small size of microRNAs. An alternative approach, previously used in Drosophila melanogaster, was used where several reporter transgenes ("sensors") were made to detect microRNAs in the embryo. The sensor was a constitutively expressed reporter gene (lacZ) containing microRNA complementary sequences in the 3′ UTR. In cells lacking the corresponding microRNAs, the reporter transgene is stable and will express b-galactosidase. In contrast, cells expressing the corresponding microRNA degrade the lacZ message by an RNA interfering pathway (RNAi), thus resulting in absence of b-galactosidase activity.
To prove the biological significance of microRNAs in human biology and in cancer in particular, the development of mouse models for deleted and/or overexpressed microRNAs will be of invaluable interest for deciphering the regulatory networks where microRNAs are involved. Several groups have started this task by disrupting Dicer1 in mice. Loss of Dicer1 by replacement of exon 21 with a neomycin-resistance cassette leads to lethality early in embryogenesis at day 7.5 of development; Dicer1-null embryos were found to be depleted of pluripotent stem cells (Bernstein et al. 2003). Another group generated Dicerex1/2 mice that have a deletion of the amino acid sequences from the first and second exons of the dicer gene (Yang et al. 2005). Dicerex1/2 homozygous embryos died between days 12.5 and 14.5 of gestation, again demonstrating that Dicer is necessary for normal mouse development. In addition, blood vessel formation and maintenance in Dicerex1/2 embryos and yolk sacs were severely compromised, with altered expression of Vegf, Flt1, Kdr, and Tie1 being detected in the mutant embryos. This study suggests that Dicer has a role in embryonic angiogenesis, probably through processing of microRNAs that regulate expression levels of key angiogenic regulators (Yang et al. 2005).
Although Dicer-null embryonic stem (ES) cells are viable, they are defective in RNA interference (RNAi) and the generation of microRNAs (Kanellopoulou et al. 2005). These mutant ES cells displayed severe defects in differentiation both in vitro and in vivo. An increase in centromeric repeat transcripts was detected, whereas the expression of homologous small double-stranded RNAs was markedly reduced in Dicer-deficient ES cells (Kanellopoulou et al. 2005; Murchison et al. 2005). Re-expression of Dicer in the knockout cells rescued these phenotypes. These Dicer-deficient cells were able to undergo gene silencing when provided with synthetic siRNAs (Kanellopoulou et al. 2005; Murchison et al. 2005), indicating that Dicer function is required for processing of microRNAs but not required for the action of microRNAs once they are produced. However, the status of cytosine methylation of centromeric satellite sequences remains in question. Taken together, these studies suggest that the essential role of Dicer in the production of mature microRNAs is central for biological processes in mammalian organisms, ranging from stem cell differentiation to the integrity and function of centromeric heterochromatin (Kanellopoulou et al. 2005).
Critical insights into the roles of individual proteins of the microRNA processing machinery have been provided by in situ hybridization studies of Argonaute (Ago1-Ago4) family members in mouse embryos (Liu et al. 2004b; Lu et al. 2005b). Studies in embryos at days 9.5–14.5 of development have shown that Argonaute family members are restricted in both their temporal and spatial patterns of expression. However, Ago2 shows the broadest expression in mouse embryos and is consistent with the fact that Ago2-deficient mice die early in development (Liu et al. 2004b). Thus, the differential expression of Ago family members suggests specific roles for these proteins during organogenesis, most likely involving the production of microRNAs that are controlled at different times and in specific tissues by the presence of Ago proteins.
One example of the use of mouse models to study the roles of specific microRNAs in cancer was provided by He et al. (2005). Their initial observation was that the human mir 17-92 cluster was overexpressed by more than 5-fold in a majority of human B-cell lymphomas. To determine whether overexpression of this cluster could drive B-cell lymphomagenesis in mouse, the authors derived hematopoeitic stem cells from a susceptible Eμ-myc transgenic mouse and transduced them with a retrovirus carrying the mir 17-92 cluster. Injection of transduced cells into normal mice resulted in acceleration of the development of B-cell lymphomas; in contrast, other subsets of microRNAs expressed in these cells did not have similar effects (He et al. 2005).
The ability to manipulate the genome of mice coupled with the multitude of inbred strain backgrounds provides invaluable tools to genetically dissect the biological functions of individual microRNAs. Further studies of mouse models will provide significant insights into the roles of different microRNAs in development and disease processes.
A link between microRNAs and human diseases, in particular, with cancer as focused on in this review, has been recently shown. To better understand the biological significance of microRNAs and of alterations found in human diseases (such as the effect of microRNA mutations or variations in expression) the development of mouse models is mandatory. Regulation of gene expression in the mammalian genome, during development, differentiation, and disease, is a complex and multitasked system. To make transcripts instead of proteins is energetically "less expensive" for cells and could be the reason why regulation at the RNA level is "cheaper" and more efficient than at the protein level. Penetrance in gene expression requires a fine regulation and microRNAs could have a role in different phenotypic expressions of the same gene. The ability to simultaneously regulate large sets of genes by a single microRNA appears to be at the heart of control of multiple pathways that include morphogenesis and cell fate decisions, response to infectious organisms, and centromeric heterochromatin structure. MicroRNAs appear to be at the center of the balance between apoptosis and proliferation in cancer cells and thus could provide novel approaches for therapeutics. MicroRNAs are small stars in the genome galaxy with many surprises still in store.
The authors thank Dr. Marina Markova for reviewing the manuscript. This work was supported by Program Project Grants P01CA76259, P01CA81534, and P30CA56036 from the National Cancer Institute to CMC. GAC was the recipient of a Kimmel Scholar award. LDS was supported by NCI grants PO1 CA72027 and RO1 CA89560.


text:
Combating email spam
The topic for today is spam, and not the tasty tinned kind. Agent Matthew Stephens is here to help you spot, prevent and remove unwanted marketing or phishing messages from your email inbox, to protect you and your info from prying eyes and a mountain of digital paperwork.
Almost everyone with an email account gets spam messages on a regular basis. Especially if you sign up to company newsletters or tick the 'yes, please contact me' box when completing an online purchase. But don't worry, I'm going to help you spot and deal with email spam like an Agent.
Agent tip
Check the settings area of your email account to see if there's a 'spam filter' section which will allow for customisation and help with protection.
How do I identify a spam message?
Some spam messages contain product information from companies, while others have links to infected sites or requests for funding from fake foreign dignitaries. If you ever receive an email with a dodgy subject line, strange looking link or a request for your personal details, even if it's from a contact, don't reply, open any attachments or links.
How do I prevent spam?
- Turn your spam filter on. A spam filter scans incoming emails for common spam keywords and phrases, like 'money back guarantee', for example. Most free and business email accounts have spam filters built-in, including Gmail, Yahoo! And Hotmail. Make sure yours is turned on in email settings.
- Read every sign up form. Never sign up for anything on a website, application or social network before reading the form. Some companies will get paid to provide databases full of consenting people to third parties, who will then send out spam or marketing emails on a regular basis. Look out for the box that says "yes you can share my information" and make sure it is de-selected.
- Get a good internet security suite. While business email providers generally have antivirus suites to scan email attachments and prevent spam, some free email providers don't. Getting a good internet security suite for your home PC or laptop is beneficial. Kaspersky, Bullguard and Trend all do great internet security packages that have anti spam and email attachment scanners built-in.
What do I do if email spam gets into my inbox?
If something that looks like spam gets through your email filter, for instance an email entitled 'I can haz your moneys and your house?' don't be afraid of it. Simply select the offending message and
Agent tip
If you receive an SMS text message you're not sure about or don't recognise the person who sent it, don't reply or click on any links in the message.
click the 'Report Spam' button. This will then add the sender to your block list and send all emails coming from that address straight to your Spam Folder. Unfortunately, most spam doesn't come from the same sender, so the filter also stores key phrases, words and other information to help it to increase its awareness.
In Conclusion
Email spam can be effectively dealt with and prevented. Be vigilant with every email you open, every attachment you download and every link you follow, and keep a good internet security suite installed on your PC.


text:
Tips about Saving Energy Dollars in Your House
An average U.S. If you have an opinion about video, you will seemingly require to read about wholesale building energy efficiency. family spends more than $1,600 a year on home bills, yet making some basic changes around the home could save money and make heating and cooling systems more effective, according to World Energy Solutions, a publicly traded energy services company situated in St. Petersburg, Fla. By assessing equipment and facilities, World Energy Solutions (symbol: WEGY) helps firms reduce maintenance costs and their utility usage and extend the life of these equipment. "Many of-the methods we use for the commercial customers may also be placed on the home," states Benjamin Croxton, ceo of World Energy Solutions. Learn further on this affiliated article - Click here: the infographic. "There are many common-sense, low-cost and no-cost approaches to lower your home energy use in addition to many new technologies which can be put on your home's energy-consuming systems." Here are a few guidelines from the American Council for an Efficient Economy on things homeowners can do to make their homes more energy efficient: * Ignore the temperature of the water heater to the environment. To get different interpretations, we understand people check out: study energy management system. * Use energy-saving controls on refrigerators, dishwashers, washers and clothes dryers. * Use compact fluorescent lamps, that may save three-quarters of the energy used by incandescents. First to become changed should be any 60-watt to 100-watt lamps which can be used several hours per day. * Have your heating and cooling systems maintained in the spring and fall. Duct sealing also can improve overall performance and the energy efficiency of your furnace or central ac. Visit energy efficiency talk to study how to study this enterprise. * Clean or replace heater, air conditioner and heat-pump filters. * Assess your heating and cooling systems to find out in the event that you must replace or retrofit them to create them work more proficiently to provide exactly the same comfort, or better, with less energy. "If your home's central air-conditioning system is finished a decade old, a new state-of-the-art system will save you 30 percent or more of your home's air-conditioning expense," states George Walker, air-conditioning expert with World Energy Solutions..


text:
Apr 18, 2011 #072
Contact: John Austin
UMKC Health and Life Sciences Schools team up for 2011 Health Sciences Research Summit
Public invited to see UMKC Faculty and Student research on display
KANSAS CITY, Mo. -- The University of Missouri-Kansas City (UMKC) Health and Life Sciences Schools will co-host the UMKC Health Sciences Research Summit on Wednesday, April 20, from 11 a.m. - 3 p.m. in the UMKC Health Sciences Building on the Hospital Hill Campus. The purpose of Research Day is to make the UMKC community aware of the health sciences related research being conducted in the Schools of Biological Sciences, Dentistry, Medicine, Nursing and Pharmacy and to foster research collaborations across disciplines and schools.
During the event, students and faculty of the UMKC Health and Life Sciences Schools will showcase their research in the form of poster presentations, with more than 115 posters on display throughout the Health Sciences Building. Judy Baker, Regional Director, Region VII, U.S. Department of Health and Human Services, will be the Keynote Speaker at the Research Summit Opening Ceremony at 11 a.m. in room 3301 of the Health Sciences Building.
A panel of judges will select one poster presented by students from each of four categories as the best in terms of scientific merit and impact on health sciences. The student winners will each receive a cash honorarium as a token of appreciation for their research efforts. Posters will be presented in the following categories:
- Clinical, Behavioral and Practice Research
- Medicinal and Biological Sciences
- Pharmaceutical Science
- Pharmacology & Toxicology
About the UMKC Health & Life Sciences Schools
The University of Missouri-Kansas City Schools of Dentistry, Medicine, Nursing and Pharmacy are located on the UMKC Health Sciences Campus at Hospital Hill and the School of Biological Sciences is located on the UMKC Volker Campus. UMKC is one of fewer than 30 universities in the nation to have four major health sciences schools centrally located on one campus. The next generation of health practitioners and scientists educated at UMKC benefit from the innovative learning and research environment provided by the University and its clinical and research partners.
About the University of Missouri-Kansas City
The University of Missouri-Kansas City (UMKC), one of four University of Missouri campuses, is a public university serving more than 15,000 undergraduate, graduate and professional students. UMKC engages with the community and economy based on a four-part mission: life and health sciences; visual and performing arts; urban issues and education; and a vibrant learning and campus life experience. For more information about UMKC, visit www.umkc.edu. You can also find us on Facebook, follow us on Twitter and watch us on YouTube.
This information is available to people with speech or hearing impairments by calling Relay Missouri at (800) 735-2966 (TT) or (800) 735-2466 (voice).


text:
Leave a Reply
You must be logged in to post a comment.
Conversations shaping the future of emerging media
Film News: Par's wild card tries to overcome tracking with word of mouth — Paramount's "Super 8" secured first last night with $12.2 million from 3,379 locations.
See the article here:
$12 mil wins Friday for 'Super 8′
You must be logged in to post a comment.
Talk NYC/WW is your daily download of the tech, marketing and advertising news you need to know. It's smartly curated to keep you up to speed on the innovators and innovations that are shaking up the digital world today.


text:
Zurich chocolate show treats nation of chocoholics
By Caroline Copley and Nathalie Olof-Ors
ZURICH (Reuters) - Exhibitors at Zurich's Salon du Chocolat are celebrating "Swissness" with chocolate watches and Easter egg clocks as the travelling confectionary fair opens its doors for the first time in a country renowned for its love of chocolate.
Founded in Paris in 1994, the 'Salon du Chocolat' gala has toured the world to destinations such as Tokyo, New York and Cairo, and offers smaller, independent artisans the chance to exhibit their latest wares.
One such company is Swiss-based "Chocowatch", which launched its novelty chocolate timepieces sculpted into a rock from the Swiss Alps in February this year.
Designed by Francis Gerber, who has worked as a graphic designer for Richemont's watch brand Jaeger lecoultre, the 70 percent cocoa watches come in two styles: milk chocolate and dark chocolate and retail at 14.50 Swiss francs ($15.98).
His Renault car laden with the edible watches, distributor Bernard Matthey toured Swiss tourist hotspots, such as Verbier, Zermatt and Interlaken - popular with Asian visitors who are snapping up the real Swiss watches.
"We're not in Lucerne yet, because we didn't have time to stop," said Matthey's wife Mary Josee. "But we hope to get there soon."
Home to chocolate companies like Nestle and Lindt & Spruengli, famed for its gold foil-wrapped bunnies, the Swiss invented milk chocolate in 1875 and have been a nation of chocoholics ever since.
The Swiss consumed 94,008 tonnes of chocolate products in 2011, according to statistics from the Association of Swiss Chocolate Manufacturers. Continued...


text:
Anna Serquina
My goal is to help raise the seed money to fund ALS research in UMass headed by Dr. Robert H. Brown. I plan to do this by crowd-sourcing donations through family, friends, and networks. In conjunction with this, I will be running the 2012 Boston Marathon with the UMass ALS Champion Fund Team (and hopefully finishing it before they close the course on me, right?).
Please click on the link below on the left to learn more about the UMass ALS Champion Fund.
Online donations are processed via Amazon Payments.
If you prefer to pay via personal check, please make it out to "UMass ALS Champion Fund" and in the memo field write "Anna Serquina" so that they know which team member you are supporting. Please mail your donation to:
Katie Friend
UMass Medicine Development Office
333 South St.
Shrewsbury, Massachusetts 01545
"I have fought the good fight. I have run the race. I have kept the faith."
CR Impact Points
Click the VOTE button to give Anna 100 CrowdRise Impact Points (CIPs). Vote for the Volunteers and Fundraisers that are answering the call to service, raising money for charity, and making an impact for their causes. Come back and Vote every months. CIPs can earn you the highly coveted, highly respected CrowdRise Royalty Status.
CR Impact Points
If you're seeing the grey VOTE button, you should feel great about yourself. It means you already voted for Anna this month. Come back next month and vote again if you still love Anna. Thanks to your vote, Anna earned 100 CrowdRise Impact Points (CIPs). CIPs can earn you the highly coveted, highly respected CrowdRise Royalty Status.
Total Impact Points
Anna Serquina's Fundraisers
I'm working on creating my first super compelling Fundraiser so please keep hitting refresh over and over until my first Fundraiser shows up. Or just wait for me to email you about it.


text:
2 Gas related Questions
View Single Post
JIm Sam
Registered User
Join Date: Nov 2006
Location: Boston Area
Posts: 10
Edits: 0
Thanks: 0
Thanked 1 Time in 1 Post
JIm Sam's Photo Album
Hello to All,
First I don't know how to reply to individual posts as I would like yet. (New to Boards)....So I will give a general reply.
1. - Thanks to all who took the time to respond.
2. - I have gas (old) in the carb..checked with site plug on side of bowl.
At first the needle and seat were stuck so there was too much gas. I pulled the needle and seat and cleaned and seems OK now.
3. - The fuel pump is brand new.
4. - I put an in-line filter before and one after the pump.
5. - I think the car may idle if I can get it to smoothen out a bit. I think I need to try and keep it running for about 5 minutes to see if it works better when warm.....I believe that the idle circuit of the carb should allow the car to run. I don't know much about the idle circuit in detail. This is a Holley 4160/50 - 600cfm.
6. - I am running with a hand choke..(no cable hooked up yet).This could be a slight problem as I think I soaked the plugs the other day with the choke closed to long. I couldn't get out of the car to open the choke by hand. Now I am trying to start with the choke almost fully open. This is how the car rn years ago...(No winter driving)
7. - So to dmorris 1200, sir Joab, One MOre Time, johnsongrass1....I say THANKS....nas ask an additional question....
8. - I took the carb apart and sprayed with carb cleaner quite a bit. I sprayed through all the passages I could see etc. Do you guys think this will work or do I need to disassemble the carb and let it soak in some sort of cleaner solution. Could use advice on brand..
I am somewhat stumped at this point....can I save my pretty new carb????.....Thanks......Jim S
JIm Sam
View Public Profile
Find all posts by JIm Sam


text:
I feel as if TJ is the same person who played Clementine on The Walking Dead. However, I could find absolutely nothing to confirm it. It wouldn't surprise me if Telltale used the same voice actors.
I just want it to be confirmed.
I feel as if TJ is the same person who played Clementine on The Walking Dead. However, I could find absolutely nothing to confirm it. It wouldn't surprise me if Telltale used the same voice actors.
I just want it to be confirmed.
Yeah, Clem's voice actress also plays Toad Jr. Here's a tweet confirming it:
Just about every voice actor in The Wolf Among Us so far has played a character in the Walking Dead.
The only new ones listed were Cia Court who plays Faith and Janet Lipsey who plays Holly.
You know what? I'm calling it right now. Dave Fennoy (Lee) plays Bluebeard.
Andy St John is Bigby Wolf. "You ate a lot of people back in your days", Colin said. True, true.
I thought Mark Barbolak (unnamed cop) is Colin. Same voice.
Nope. Straight from the horse's (err...pig's) mouth: http://www.telltalegames.com/profile/35651/brian-sommer (Second paragraph of the latest post)
Some more ironic banter between the two of them:
Colin: "I'm hungry too. You don't see me tearing the flesh of your bones."
Bigby: "But you would if you could."
Colin: "...Probably."
I knew I recognized some of the voices! TJ, Snow, Beast and Gren I could totally tell and I knew Bigby had a familar voice but I was thinking Hershel.
Wow, I only knew a small handful of those returning actors. Nice list.
Chuck's actor being Bufkin especially humors me.
You're psichic!! I'm sure you asked the mirror who would play Bluebeard in the Wolf Among Us!
God, I hope the red-haired guy played by Sam Joan ends up being the killer.
Bigby: Why'd you do it, man? What do have against young, attractive women?
Cabbie: 75% of players chose to save one over me. Just wanted to even the score.
Until I looked it up, I thought for sure the voice actress who did Faith was the same one who did Carly.
interesting... seems tellltale sticks to their voice actors a lot, cause im pretty sure at least crane's voice can be heard in bttf (soup kitchen guy) and the sam & max games too afair. at least.
some of the voice actors are doing a really good job, mr toad, bufkin and hershel are the same guy? wtf....
the other voice actors are good too, of course. well... okay, i guess i remembered cranes voice so well, cause i dont like it that much, but thats the only one
so during the bar scene bigby and woodsman are the same person, I.E. adam harrington is having a conversation with himself. lol
Riot and Telltale seem to have a lot of mutual voice actors. Adam Harrington and Brian Sommer both play a handful of League characters. I think a few of the others do, as well.
-Melissa Hutchison: Ashe
-Gavin Hammon: Pantheon/Varus/Bloodlord Vlad
-Chuck Kourouklis: Darius
-Cia Court: VI
Bound to be more


text:
Souped-Up Utility Knife
Utility knives can be used for everything from opening a box of door hardware to cutting drywall and trimming insulation. Olfa, the company that invented the snap-off blade, has just released a new knife designed with the contractor in mind. The LA-X has an automatic blade lock, a rubberized fiberglass handle, and a metal fin at the rear end that can be used to open a can of paint or as a quick and dirty screwdriver. The knife uses 18mm heavy-duty snap-off blades.
¦ Price: $7. 800-962-6532. www.olfa.com.


text:
FAU Teaching "Stand Your Ground" Course
Florida Atlantic University is offering a course that concentrates on the state's ``stand your ground'' self-defense law.
The course is being offered during fall semester by FAU's School of Public Administration at the Boca Raton campus.
Program coordinator Richard Mangan says national publicity surrounding the recent George Zimmerman trial in central Florida helped spur the creation of the course. It's being offered as an elective.
Florida's self-defense law generally removed a person's duty to retreat if possible in the face of danger. It was the first of its kind in the nation when passed in 2005. Now, about two dozen states have similar laws.
Zimmerman was acquitted of all charges last month in the shooting of Miami teenager Trayvon Martin. While "stand your ground" was not invoked by Zimmerman's attorneys at the trial, those wishing to repeal the law used the attention garnered by the trial to push for such a repeal.
Just this week, Florida's legislature announced that they would not be gathering for a special session to deal with the self defense law.
Copyright: AP/Photo: Getty Images


text:
The Foothill College Respiratory Therapy Program will present an application workshop to help clarify and answer questions you may have about the 2013 application. The workshop is Wednesday, April 16, at 5 p.m. in Room 5 p.m. Admission is free; parking is $3.
What to Bring:
• Download and print the 2013 Respiratory Therapy application at http://www.foothill.edu/bio/programs/respther/app.php;
• Pencil/pen, highlighter, and paper for note taking;
• All of your college transcripts (unofficial copies are acceptable).
Be aware that the completed 2013 application and all support documentation is due by May 30, 2013.
Parking Lots 5 and 6 provide the closest access to Room 5307. Visitors must purchase the required campus parking permit for $3. Purchase permit from dispensers in any student lot. Dispensers accept one-dollar bills and quarters. Foothill College is located off I-280 on El Monte Road in Los Altos Hills. For additional information, call or e-mail Kerry West at (650) 949-7538 or firstname.lastname@example.com.


text:
There's just something about three-part harmony that draws me in and even more so if the three-part harmonies are blood related. The other night (Tuesday, October 23), a new family trio, The Henningsens, stopped by the WYRK studios to play for me and some YRK staff members.
This family trio is made up of Brian (dad), Aaron (son) and (daughter) Clara. These singer/songwriters originate from Illinois and now have planted roots in Nashville looking to bring their family harmonies to country radio. You may not have heard their name before, but I'm sure you've some of the songs they've written, like The Band Perry hit "You Lie."
The Henningsens seem to have the whole package, and even better, they're down to earth, grounded and seem very happy as a family. Those three things alone give them a leg up in my opinion. Their first single is called "American Beautiful" — check it out!
Subscribe to 106.5 WYRK on
Subscribe to 106.5 WYRK on


text:
Wake up!
View Single Post
Alfa Seltzer
Registered User
Location: Marietta, GA
Join Date: Jun 2012
Posts: 18
Mein Auto: 1986 Alfa Romeo Spider
View My Garage
I loosened the mounts and jacked the engine by the a/c compressor. Space is very tight and the pan has to be wiggled/angled every few inches in just the right direction to get it free. I'd suggest the cherry picker/balance bar method.
Removing/installing the pan bolts is a giant pain even with the right extensions/u-joints. You are going to lose Your knuckles doing this and it wound up taking me more time than it would have to just pull the tranny or front crossmember.
Alfa Seltzer
View Public Profile
Find all threads started by Alfa Seltzer
Find all posts by Alfa Seltzer
View My Garage
Add Alfa Seltzer to Your Ignore List


text:
Bag it up in our nice bags and wallets. Discover our backpacks, tote bags, coin purses, wallets, make-up cases, zip cases, laptop cases, over the shoulder bags, card cases and all things needed to carry your best belongings with you. Whatever adventure you've planned, you'll be totes ready.
Find new arrivals in New In
For the latest news, get on our list


text:
RITTMAN Metal bats can be hazardous to one's health when held during a thunderstorm, and can also be a dangerous threat to a pitcher's earned run average. The aluminum whooping sticks used by both Orrville and Buckeye in the Rittman Div. II Sectional final had no ill effects on either side, as the Red Riders squeaked out a 3-1 victory no more than five minutes before the heavens unleashed lighting and rain on the area. Orrville (10-13) had only three hits, but played solid defense and received a strong pitching effort to advance to the district semifinals.
"We don't have any kids in the individual leaders' column in (the newspapers)," Red Riders coach Rick Smith said. "But when we play good fundamental baseball, we are a good team. We did the basics like throw strikes and make the plays in the field. It came down to us making more plays than they did."
The success of any defense begins on the mound, a place where Orrville stood strong with the combined performances of Josh Raber and Adam Samson. Raber, a sophomore, started and pitched into the fifth inning and allowed one unearned run on three hits. He retired the first eight hitters he faced, and would have left with a zero on the scoreboard if not for an error that allowed a run to score in the fourth. Four of the first nine hitters to face Raber swung at the first pitch.
"He's been solid all year on the mound," said Smith of his pitcher who had a 2.21 ERA before Thursday's game. "He throws strikes, and I'm not surprised he was getting so many people to swing early because of the way he pitches."
Samson shut down the Bucks (12-10) for the final three innings to earn the save. The visitors had a runner reach base in each of the three innings Samson pitched, but the Red Riders' southpaw used the good movement on his fastball and the help from the seven players behind him to avoid any damage.
Buckeye had the tying run stroll to the plate once in the sixth and twice in the seventh, but neither time could get the hit it needed off of Samson.
"I didn't think about any of that out there," said Samson of the potential trouble he faced in his outing. "My job is just to go out and throw strikes. I didn't throw more than three curveballs all day because they weren't hitting my fastball. My fastball was working, so I kept using it."
The Bucks did connect with a fastball twice with runners on, but the Red Riders' defense kept the ball in the infield each time. On one occasion, Samson fielded a ball between the mound and first base and made the throw to an outstretched Cory Bishop, who later made another Gumby-like stretch to complete a double play in the seventh.
"Playing good defense is the biggest thing you can do to keep a game from getting any closer," said Bishop, who also had one of Orrville's three hits. "Both of our pitchers did a great job and kept us in the game."
Buckeye starter Dan Sekerak did everything he could to keep his club close. Sekerak gave up only one earned run in six innings of work while striking out four. The Bucks made two errors in the third inning that opened the door for two of the Red Riders' runs.
"Dan had tremendous control tonight ... he deserved better," Buckeye coach Scott Sorrell said. "If the team didn't play hard that would be one thing, but our guys hustled and gave a great effort. Holding them to three runs isn't bad."
The only earned run came thanks to Samson's RBI double in the third frame. Samson later singled for Orrville's third and final hit in the fifth.
Joe Vardon can be reached at (330) 287-1649 or email@example.com.


text:
The first ever United Nations-sanctioned International Day of Yoga will take place on June 21, 2015. Widely recognized as the father of yoga in the West, Paramahansa Yogananda introduced millions of seekers to India's ancient Yoga science.
DaijiWorld, an online news portal, wrote in an article dated March 5: "If India's idea of an International Yoga Day has found easy acceptance in the UN, much of the credit must go to Paramahansa Yogananda, India's first yoga guru in the U.S....He played a huge role in laying the foundation for yoga in the United States a century ago.
Training the Conscious and Subconscious Minds for Success—Part 1
Paramahansa Yogananda
A previously unpublished talk in which the Master teaches how to fully harness the unlimited hidden powers of our mind for accomplishing material and spiritual goals.
See full content list, read samples of articles, and listen to audio excerpt.
As we announced last month, in the coming year Self-Realization Fellowship will be introducing a variety of new spiritual programs for teens and children. Plans for many of the programs are still in the development phase, but we are happy to share here the following information about one of the new offerings.


text:
A blog by Southern writer and historian Dale Cox, Civil War Florida shares information on and discusses the events of the Civil War in Florida. Topics of interest include troops, battles, skirmishes, campaigns, raids, forts, naval actions, ships, soldiers, officers, books and historic sites.
Sunday, October 5, 2008
The Raid on Marianna - Conclusion
2 comments:
Nice set of posts on Marianna. Though you might be interested... I just posted some a biographical sketch about one of the USCT troops (from the 82nd USCT) involved in the action there. http://southernunionistschronicles.wordpress.com/2008/10/09/noah-thorntons-story/
- Robert Moore
- October 10, 2008 at 7:27 AM
- Dale said...
Thank you for letting me know about this. I took a look and found it absolutely fascinating.
I would love to hear from you with any other information you have on Union troops that were active in Florida!
- October 10, 2008 at 4:55 PM


text:
From Proteopedia
Structural highlights
Warning: this is a large structure, and loading might take a long time or not happen at all.
[FAS2_YEAST] Fatty acid synthetase catalyzes the formation of long-chain fatty acids from acetyl-CoA, malonyl-CoA and NADPH. The alpha subunit contains domains for: acyl carrier protein, 3-oxoacyl-[acyl-carrier-protein] reductase, and 3-oxoacyl-[acyl-carrier-protein] synthase. This subunit coordinates the binding of the six beta subunits to the enzyme complex.[HAMAP-Rule:MF_00101] [FAS1_YEAST] Fatty acid synthetase catalyzes the formation of long-chain fatty acids from acetyl-CoA, malonyl-CoA and NADPH. The beta subunit contains domains for: [acyl-carrier-protein] acetyltransferase and malonyltransferase, S-acyl fatty acid synthase thioesterase, enoyl-[acyl-carrier-protein] reductase, and 3-hydroxypalmitoyl-[acyl-carrier-protein] dehydratase.
Evolutionary Conservation
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
Fatty acids are among the major building blocks of living cells, making lipid biosynthesis a potent target for compounds with antibiotic or antineoplastic properties. We present the crystal structure of the 2.6-MDa Saccharomyces cerevisiae fatty acid synthase (FAS) multienzyme in complex with the antibiotic cerulenin, representing, to our knowledge, the first structure of an inhibited fatty acid megasynthase. Cerulenin attacks the FAS ketoacyl synthase (KS) domain, forming a covalent bond to the active site cysteine C1305. The inhibitor binding causes two significant conformational changes of the enzyme. First, phenylalanine F1646, shielding the active site, flips and allows access to the nucleophilic cysteine. Second, methionine M1251, placed in the center of the acyl-binding tunnel, rotates and unlocks the inner part of the fatty acid binding cavity. The importance of the rotational movement of the gatekeeping M1251 side chain is reflected by the cerulenin resistance and the changed product spectrum reported for S. cerevisiae strains mutated in the adjacent glycine G1250. Platensimycin and thiolactomycin are two other potent inhibitors of KSs. However, in contrast to cerulenin, they show selectivity toward the prokaryotic FAS system. Because the flipped F1646 characterizes the catalytic state accessible for platensimycin and thiolactomycin binding, we superimposed structures of inhibited bacterial enzymes onto the S. cerevisiae FAS model. Although almost all side chains involved in inhibitor binding are conserved in the FAS multienzyme, a different conformation of the loop K1413-K1423 of the KS domain might explain the observed low antifungal properties of platensimycin and thiolactomycin.
Inhibition of the fungal fatty acid synthase type I multienzyme complex.,Johansson P, Wiltschi B, Kumari P, Kessler B, Vonrhein C, Vonck J, Oesterhelt D, Grininger M Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):12803-8. Epub 2008 Aug 25. PMID:18725634[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
Proteopedia Page Contributors and Editors (what is this?)
Categories: Saccharomyces cerevisiae | Grininger, M | Johansson, P | Kessler, B | Kumari, P | Oesterhelt, D | Vonck, J | Vonrhein, C | Wiltschi, B | Cerulenin | Fa | Fatty acid biosynthesis | Fatty acid synthase | Hydrolase | Lipid synthesis | Lyase | Multifunctional enzyme | Nad | Nadp | Oxidoreductase | Phosphopantetheine | Phosphorylation | Transferase


text:
Summit Salute at the base of the Mt. Washington Auto Road
Saturday - August 5, 2017
Join other service members, veterans, & military families for this special FREE event at the base of Mt. Washington. Share stories, play outdoors, eat great food and enjoy a Mt. Washington sunset.
Registration now open: Click HERE to Register Today!
|2:30pm - Outdoor Fun
|Hiking, biking, games, children's activities and adventures - there's something for everyone
|4:00pm - Stories of Service: Stronger Together
|Child care is available during the program
|5:00pm - Grilled BBQ Dinner
|Fill your belly with a bounty of delicious food topped off with scrumptious dessert!
|6:30pm - 8:30pm - Sunset Summit Trips
|Discounted rides to the summit of Mt. Washington exclusively for Summit Salute attendees.
Contact info: email@example.com (603) 568-5780
This is an initiative of the NH Department of Health and Human Services and presented by NAMI New Hampshire (www.naminh.org) in partnership with NH Care Path (www.nhcarepath.org) and the Mt. Washington Auto Road / Great Glen Trails.


text:
Items in Cart:
New Mini Laser Effects Projector. Featuring two unique functions (sound activation and stroboflash), this laser effects projector proves that small size and economic price can still provide great fun.
Move with the music: With its sound activation feature, this Portable Laser Effects Projector can intelligently change its color/pattern to match the beat of the music! How? Just set your stereo system nearby and the projector's mic will pick up nearby music vibrations, so if you turn up the noise the laser will respond in style! That's not all...
Colour - Blue colour
Manufacturer Specifications
Product Notes
Package Contents


text:
Published: August 21, 2013 at 3:43 PMHaving been to Hong Kong Disneyland last December, I will complain that it is the Anaheim clone with a completely different feel. Main Street no longer has that Victorian charm. It is a mere facade. It is the same idea as Grizzly Gulch where the American West feeling, that I know, is completely lost in China. They should have done a tropical Adventureland version of Big Thunder Mountain instead.
Disneyland in China is not working in that people do not necessarily love Disneyland's penchant for themed environments that features American and European concepts and ideas. They love Mickey Mouse and the various characters. They also love expensive and elaborate merchandise and luxurious surroundings. The small scaled and modest Sleeping Beauty Castle seems crude in comparison and completely out of place in China. Perhaps that is why knock-offs don't work, and the originals don't work either.
Americans live in much more modest homes. Going to Disneyland seems like a luxury. Chinese live in high rises. Going to Hong Kong Disneyland with a modest castle and a quaint Main Street is like visiting a doll house.
I can see that the approach in Shanghai is much better than what was initially done in Hong Kong. This is a plus.
Lastly, I will welcome seeing the Monkey King theme park, and Mystic Manor.
Published: August 22, 2013 at 7:55 AMOkay, this was the absolute best article I have ever read anywhere on Chinese theme parks. I am going to use it as a reference for a paper I am writing on intellectual property issues in China. I already cleared it with my professor and she agreed it's an excellent primary source on what's happening in China.
I really have never understood why they built Hong Kong Disneyland or why they build DCA 1.0 in California. Both those projects seem like they were really done on the cheap and were half-baked. I don't know why they built them so cheaply and in such a rush...was it some kind of legal issue where they had to push the trigger and build something, even if they knew it would not be any good? Maybe they thought if they built anything they could just remodel it and make it good later. Seems stupid and wasteful to me but maybe there's some business related thing that I don't understand.
I really want to learn more about the copyright infringements and knockoffs in China. I hope someone adds in comments more information. I want to know why the Chinese government allows this...and if the government will ever shut down the knockoffs. Like, when Shanghai Disney opens will they shut down the creepy knockoff parks? A lot of Chinese are very poor and can't afford to go to Disney, so will those people then make do with the knockoff? I heard that Shanghai Disney is being geared just to the rich Chinese...so I wonder if it's just part of the plan that the poor will go to the knockoffs instead and nobody minds.
Please try to do more articles on this subject. This was just brilliant and I know I am going to get an A when I use it in my paper!
Published: August 22, 2013 at 2:22 PMHey "Okay". Your questions are so basic that I wish you had done a minimum of research before you asked your questions. Most people that follow the news from China, which isn't hard, will already know the answers to your questions.
Why HKDL? Eisner era. Pressler cut a lot out of the park. It is a badly done clone of Disneyland. DCA is a worse clone of Disney/MGM theme park.
Why Chinese government allows knockoffs? Many reasons. (1) Get ahead. They need money, quick money. They need jobs. They need expertise so the fastest why to get it is copy it first. Copies are best for the export market where they can sell plenty to foreigners.
(2) How can they shut it down? The government has shut it down for political reasons and to appease some manufacturers who are able to sue and win, but the companies close and quickly reopen. The environment of cheap labor allows this, AND if the manufacturer was once an OEM for a foreign company, it is quite easy to extend a product run and sell it as the original. Mostly, the government turns a blind eye to it.
(3) No, the creepy knock-off parks will close on their own incompetence. People will find Disney theme parks superior.
(4) Yes, many Chinese are poor and probably can't afford to go to Disneyland, but many Chinese has reached middle class and CAN afford it. Do you realize how many Chinese come to the US as tourists? PLENTY. The middle class is millions of people. I didn't think so myself until I was confronted with reality. I seen the Chinese tourists at home in Southern California and in Taiwan and Hong Kong. Many have money to burn. The poor will do what the Americans did 50 years ago. Save up for a once in a lifetime trip to Shanghai Disneyland.
Next time, ask researched questions.
Published: August 22, 2013 at 3:45 PMHere's a good article.
"Counterfeit cop: How a local attorney spots fake Rolexes and knockoff Louis Vuittons"
"Chinese companies frequently knock off American products, Austin says. He's convinced that if more American companies knew how much revenue is at stake and how relatively easy the process of bringing a claim against a counterfeiter is, more would take that step. Most companies assume it's a long, drawn-out legal process when in fact it typically takes only a few days."


text:
Items in Cart:
10 Days: If you are not satisfied with your purchase and you have a substantiated reason (the subject not as described or damaged), as will ask you to show pictures, please contact me within 48 hours of delivery. Refund within 10 days item back. Conversely shipping is obligation of the buyer!