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Artykuły w czasopismach na temat "Cst Genes"
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Calvo, Olga, Nathalie Grandin, Antonio Jordán-Pla, Esperanza Miñambres, Noelia González-Polo, José E. Pérez-Ortín i Michel Charbonneau. "The telomeric Cdc13–Stn1–Ten1 complex regulates RNA polymerase II transcription". Nucleic Acids Research 47, nr 12 (22.04.2019): 6250–68. http://dx.doi.org/10.1093/nar/gkz279.
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Abstract Specialized telomeric proteins have an essential role in maintaining genome stability through chromosome end protection and telomere length regulation. In the yeast Saccharomyces cerevisiae, the evolutionary conserved CST complex, composed of the Cdc13, Stn1 and Ten1 proteins, largely contributes to these functions. Here, we report genetic interactions between TEN1 and several genes coding for transcription regulators. Molecular assays confirmed this novel function of Ten1 and further established that it regulates the occupancies of RNA polymerase II and the Spt5 elongation factor within transcribed genes. Since Ten1, but also Cdc13 and Stn1, were found to physically associate with Spt5, we propose that Spt5 represents the target of CST in transcription regulation. Moreover, CST physically associates with Hmo1, previously shown to mediate the architecture of S-phase transcribed genes. The fact that, genome-wide, the promoters of genes down-regulated in the ten1-31 mutant are prefentially bound by Hmo1, leads us to propose a potential role for CST in synchronizing transcription with replication fork progression following head-on collisions.
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Cho, Il-Je, Doyeon Kim, Eun-Ok Kim, Kyung-Hwan Jegal, Jae-Kwang Kim, Sang-Mi Park, Rongjie Zhao, Sung-Hwan Ki, Sang-Chan Kim i Sae-Kwang Ku. "Cystine and Methionine Deficiency Promotes Ferroptosis by Inducing B-Cell Translocation Gene 1". Antioxidants 10, nr 10 (28.09.2021): 1543. http://dx.doi.org/10.3390/antiox10101543.
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Ferroptosis is a type of programmed necrosis triggered by iron-dependent lipid peroxidation. We investigated the role of B-cell translocation gene 1 (BTG1) in cystine and methionine deficiency (CST/Met (−))-mediated cell death. CST/Met (−) depleted reduced and oxidized glutathione in hepatocyte-derived cells, increased prostaglandin-endoperoxide synthase 2 expression, and promoted reactive oxygen species accumulation and lipid peroxidation, as well as necrotic cell death. CST/Met (−)-mediated cell death and lipid peroxidation was specifically inhibited by pretreatment with ferroptosis inhibitors. In parallel with cell death, CST/Met (−) blocked global protein translation and increased the expression of genes associated with the integrated stress response. Moreover, CST/Met (−) significantly induced BTG1 expression. Using a BTG1 promoter-harboring reporter gene and siRNA, activating transcription factor 4 (ATF4) was identified as an essential transcription factor for CST/Met (−)-mediated BTG1 induction. Although knockout of BTG1 in human HAP1 cells did not affect the accumulation of reactive oxygen species induced by CST/Met (−), BTG1 knockout significantly decreased the induction of genes associated with the integrated stress response, and reduced lipid peroxidation and cell death in response to CST/Met (−). The results demonstrate that CST/Met (−) induces ferroptosis by activating ATF4-dependent BTG1 induction.
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Brockmann, Kathrin, Stefanie Lerche, Sarah Selina Dilger, Johannes Georg Stirnkorb, Anja Apel, Ann-Kathrin Hauser, Inga Liepelt-Scarfone i in. "SNPs in Aβ clearance proteins". Neurology 89, nr 23 (8.11.2017): 2335–40. http://dx.doi.org/10.1212/wnl.0000000000004705.
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Objective:To evaluate whether genetic variants in β-amyloid (Aβ) clearance proteins are associated with CSF levels of Aβ1-42 on a biological level and the onset of dementia on a clinical level in Parkinson disease (PD).Methods:We analyzed genetic variants known to be involved in Aβ clearance in a PD group comprising 456 patients, 103 of them with dementia. Single nucleotide polymorphisms in the genes APOE, cystatin C (CST), and membrane metalloendopeptidase (MME) were evaluated in relation to demographic variables, clinical phenotypes, and CSF Aβ1-42 levels using a cross-sectional approach.Results:Risk variants in the genes APOE and CST were associated with lower CSF Aβ1-42 levels. Clinically, patients with 2 risk alleles in CST tended to show a shorter interval from age at onset of PD to age at onset of dementia.Conclusions:This study suggests that genetic variants associated with Aβ clearance are involved in the pathogenesis of dementia in PD and possibly influence the onset of dementia.
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Dickinson, D. P., M. Thiesse, L. D. Dempsey i S. J. Millar. "Genomic Cloning, Physical Mapping, and Expression of Human Type 2 Cystatin Genes". Critical Reviews in Oral Biology & Medicine 4, nr 3 (kwiecień 1993): 573–80. http://dx.doi.org/10.1177/10454411930040034401.
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Humans carry one gene encoding cystatin C and six to eight genes with homology to an S-like cystatin hybridization probe. However, the precise composition and organization of the cystatin gene family remains to be established. Further, the pattern of tissue-specific expression has not been fully defined. We have previously shown that the type 2 cystatin genes are clustered together in a ca. 270 kb region (the CST locus). To determine the structure of this region, we have sought to clone the entire CST locus. Our approach has been to isolate cosmid and lambda genomic clones carrying cystatin genes and then to use "walk" probes derived from the end regions of these clones to identify other clones, which extend them. To date, we have obtained over 320 kb of distinct sequences. Based on restriction maps, sequencing, and hybridization analyses, we have identified eight apparently nonallelic copies of cystatin genes. These include one gene for cystatin C, four closely related genes encoding S-like cystatins, and three genes encoding relatively divergent sequences. Complete assembly of these clones into an unambiguous contiguous sequence is hampered by the presence of flanking locus-specific repetitive-like sequences. RNase protection assays used to characterize the tissue-specific patterns of expression showed that cystatin C is expressed at modest, comparable levels in all tissues examined, whereas expression of the CST 1 gene, encoding cystatin SA-I, was found to be restricted to a small subset of tissues, with the highest level in the submandibular gland. The cystatin gene family, therefore, appears to have evolved by tandem gene duplication, followed by the acquisition of control elements influencing the location and level of expression. The cystatin gene family is, thus, a potentially powerful system for the future study of mechanisms of gene regulation in human salivary glands.
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Boltz, Kara A., Katherine Leehy, Xiangyu Song, Andrew D. Nelson i Dorothy E. Shippen. "ATR cooperates with CTC1 and STN1 to maintain telomeres and genome integrity in Arabidopsis". Molecular Biology of the Cell 23, nr 8 (15.04.2012): 1558–68. http://dx.doi.org/10.1091/mbc.e11-12-1002.
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The CTC1/STN1/TEN1 (CST) complex is an essential constituent of plant and vertebrate telomeres. Here we show that CST and ATR (ataxia telangiectasia mutated [ATM] and Rad3-related) act synergistically to maintain telomere length and genome stability in Arabidopsis. Inactivation of ATR, but not ATM, temporarily rescued severe morphological phenotypes associated with ctc1 or stn1. Unexpectedly, telomere shortening accelerated in plants lacking CST and ATR. In first-generation (G1) ctc1 atr mutants, enhanced telomere attrition was modest, but in G2 ctc1 atr, telomeres shortened precipitously, and this loss coincided with a dramatic decrease in telomerase activity in G2 atr mutants. Zeocin treatment also triggered a reduction in telomerase activity, suggesting that the prolonged absence of ATR leads to a hitherto-unrecognized DNA damage response (DDR). Finally, our data indicate that ATR modulates DDR in CST mutants by limiting chromosome fusions and transcription of DNA repair genes and also by promoting programmed cell death in stem cells. We conclude that the absence of CST in Arabidopsis triggers a multifaceted ATR-dependent response to facilitate maintenance of critically shortened telomeres and eliminate cells with severe telomere dysfunction.
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Büdel, Thomas, Esther Kuenzli, Mathieu Clément, Odette J. Bernasconi, Jan Fehr, Ali Haji Mohammed, Nadir Khatib Hassan, Jakob Zinsstag, Christoph Hatz i Andrea Endimiani. "Polyclonal gut colonization with extended-spectrum cephalosporin- and/or colistin-resistant Enterobacteriaceae: a normal status for hotel employees on the island of Zanzibar, Tanzania". Journal of Antimicrobial Chemotherapy 74, nr 10 (30.07.2019): 2880–90. http://dx.doi.org/10.1093/jac/dkz296.
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Abstract Objectives For low-income countries, data regarding the intestinal colonization with extended-spectrum cephalosporin-resistant (ESC-R) and colistin-resistant (CST-R) Enterobacteriaceae in the community are still scarce. Here, we investigated this phenomenon by analysing hotel employees in Zanzibar. Methods During June to July 2018, rectal swabs from 59 volunteers were screened implementing selective enrichments and agar plates. Species identification was achieved using MALDI-TOF MS. Strains were characterized using microdilution panels (MICs), microarray, PCRs for mcr-1/-8, repetitive extragenic palindromic-PCR (rep-PCR) and WGS. Results Colonization prevalence with ESC-R-, CST-R- and mcr-1-positive Enterobacteriaceae were 91.5%, 66.1% and 18.6%, respectively (average: 2.2 strains per volunteer). Overall, 55 ESC-R Escherichia coli (3 also CST-R), 33 ESC-R Klebsiella pneumoniae (1 also CST-R), 17 CST-R E. coli and 21 CST-R K. pneumoniae were collected. The following main resistance genes were found: ESC-R E. coli (blaCTX-M-15-like, 51.0%), ESC-R K. pneumoniae (blaCTX-M-9-like, 42.9%), CST-R E. coli (mcr-1, 55%) and CST-R K. pneumoniae (D150G substitution in PhoQ). ESBL-producing E. coli mainly belonged to ST361, ST636 and ST131, whereas all those that were mcr-1 positive belonged to ST46 that carried mcr-1 in a 33 kb IncX4 plasmid. ESBL-producing K. pneumoniae mainly belonged to ST17, ST1741 and ST101, whereas CST-R strains belonged to ST11. Conclusions We recorded remarkably high colonization prevalence with ESC-R and/or CST-R Enterobacteriaceae in hotel staff. Further research in the local environment, livestock and food chain is warranted to understand this phenomenon. Moreover, as Zanzibar is a frequent holiday destination, attention should be paid to the risk of international travellers becoming colonized and thereby importing life-threatening pathogens into their low-prevalence countries.
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Takahashi, Tadanobu, Kouki Murakami, Momoe Nagakura, Hideyuki Kishita, Shinya Watanabe, Koichi Honke, Kiyoshi Ogura i in. "Sulfatide Is Required for Efficient Replication of Influenza A Virus". Journal of Virology 82, nr 12 (16.04.2008): 5940–50. http://dx.doi.org/10.1128/jvi.02496-07.
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ABSTRACT Sulfatide is abundantly expressed in various mammalian organs, including the intestines and trachea, in which influenza A viruses (IAVs) replicate. However, the function of sulfatide in IAV infection remains unknown. Sulfatide is synthesized by two transferases, ceramide galactosyltransferase (CGT) and cerebroside sulfotransferase (CST), and is degraded by arylsulfatase A (ASA). In this study, we demonstrated that sulfatide enhanced IAV replication through efficient translocation of the newly synthesized IAV nucleoprotein (NP) from the nucleus to the cytoplasm, by using genetically produced cells in which sulfatide expression was down-regulated by RNA interference against CST mRNA or overexpression of the ASA gene and in which sulfatide expression was up-regulated by overexpression of both the CST and CGT genes. Treatment of IAV-infected cells with an antisulfatide monoclonal antibody (MAb) or an anti-hemagglutinin (HA) MAb, which blocks the binding of IAV and sulfatide, resulted in a significant reduction in IAV replication and accumulation of the viral NP in the nucleus. Furthermore, antisulfatide MAb protected mice against lethal challenge with pathogenic influenza A/WSN/33 (H1N1) virus. These results indicate that association of sulfatide with HA delivered to the cell surface induces translocation of the newly synthesized IAV ribonucleoprotein complexes from the nucleus to the cytoplasm. Our findings provide new insights into IAV replication and suggest new therapeutic strategies.
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Kimura, Takefumi, Takero Nakajima, Yuji Kamijo, Naoki Tanaka, Lixuan Wang, Atsushi Hara, Eiko Sugiyama, Eiji Tanaka, Frank J. Gonzalez i Toshifumi Aoyama. "Hepatic Cerebroside Sulfotransferase Is Induced by PPARα Activation in Mice". PPAR Research 2012 (2012): 1–10. http://dx.doi.org/10.1155/2012/174932.
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Sulfatides are one of the major sphingoglycolipids in mammalian serum and are synthesized and secreted mainly from the liver as a component of lipoproteins. Recent studies revealed a protective role for serum sulfatides against arteriosclerosis and hypercoagulation. Although peroxisome proliferator-activated receptor (PPAR)αhas important functions in hepatic lipoprotein metabolism, its association with sulfatides has not been investigated. In this study, sulfatide levels and the expression of enzymes related to sulfatide metabolism were examined using wild-type (+/+),Ppara-heterozygous (+/−), andPpara-null (−/−) mice given a control diet or one containing 0.1% fenofibrate, a clinically used hypolipidemic drug and PPARαactivator. Fenofibrate treatment increased serum and hepatic sulfatides inPpara(+/+) and (+/−) mice through a marked induction of hepatic cerebroside sulfotransferase (CST), a key enzyme in sulfatide synthesis, in a PPARα-dependent manner. Furthermore, increases in CST mRNA levels were correlated with mRNA elevations of several known PPARαtarget genes, and such changes were not observed for other sulfatide-metabolism enzymes in the liver. These results suggest that PPARαactivation enhances hepatic sulfatide synthesis via CST induction and implicate CST as a novel PPARαtarget gene.
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Zia, Saadiya, Netasha Khan, Komal Tehreem, Nazia Rehman, Rokayya Sami, Roua S. Baty, Faris J. Tayeb i in. "Transcriptomic Analysis of Conserved Telomere Maintenance Component 1 (CTC1) and Its Association with Leukemia". Journal of Clinical Medicine 11, nr 19 (29.09.2022): 5780. http://dx.doi.org/10.3390/jcm11195780.
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Telomere length (TEL) regulation is important for genome stability and is governed by the coordinated role of shelterin proteins, telomerase (TERT), and CST (CTC1/OBFC1/TEN1) complex. Previous studies have shown the association of telomerase expression with the risk of acute lymphoblastic leukemia (ALL). However, no data are available for CST association with the ALL. The current pilot study was designed to evaluate the CST expression levels in ALL. In total, 350 subjects were recruited, including 250 ALL cases and 100 controls. The subjects were stratified by age and categorized into pediatrics (1–18 years) and adults (19–54 years). TEL and expression patterns of CTC1, OBFC1, and TERT genes were determined by qPCR. The univariable logistic regression analysis was performed to determine the association of gene expression with ALL, and the results were adjusted for age and sex in multivariable analyses. Pediatric and adult cases did not reflect any change in telomere lengths relative to controls. However, expression of CTC1, OBFC1, and TERT genes were induced among ALL cases. Multivariable logistic regression analyses showed association of CTC1 with ALL in pediatric [β estimate (standard error (SE)= −0.013 (0.007), p = 0.049, and adults [0.053 (0.023), p = 0.025]. The association of CTC1 remained significant when taken together with OBFC1 and TERT in a multivariable model. Furthermore, CTC1 showed significant association with B-cell ALL [−0.057(0.017), p = 0.002) and T-cell ALL [−0.050 (0.018), p = 0.008] in pediatric group while no such association was noted in adults. Together, our findings demonstrated that telomere modulating genes, particularly CTC1, are strongly associated with ALL. Therefore, CTC1 can potentially be used as a risk biomarker for the identification of ALL in both pediatrics and adults.
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Cheng, Li-Wu, Omkar Vijay Byadgi, Chin-En Tsai, Pei-Chi Wang i Shih-Chu Chen. "Pathogenicity and Genomic Characterization of a Novel Genospecies, Bacillus shihchuchen, of the Bacillus cereus Group Isolated from Chinese Softshell Turtle (Pelodiscus sinensis)". International Journal of Molecular Sciences 24, nr 11 (1.06.2023): 9636. http://dx.doi.org/10.3390/ijms24119636.
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The Chinese softshell turtle (CST; Pelodiscus sinensis) is a freshwater aquaculture species of substantial economic importance that is commercially farmed across Asia, particularly in Taiwan. Although diseases caused by the Bacillus cereus group (Bcg) pose a major threat to commercial CST farming systems, information regarding its pathogenicity and genome remains limited. Here, we investigated the pathogenicity of Bcg strains isolated in a previous study and performed whole-genome sequencing. Pathogenicity analysis indicated that QF108-045 isolated from CSTs caused the highest mortality rate, and whole-genome sequencing revealed that it was an independent group distinct from other known Bcg genospecies. The average nucleotide identity compared to other known Bcg genospecies was below 95%, suggesting that QF108-045 belongs to a new genospecies, which we named Bacillus shihchuchen. Furthermore, genes annotation revealed the presence of anthrax toxins, such as edema factor and protective antigen, in QF108-045. Therefore, the biovar anthracis was assigned, and the full name of QF108-045 was Bacillus shihchuchen biovar anthracis. In addition to possessing multiple drug-resistant genes, QF108-045 demonstrated resistance to various types of antibiotics, including penicillins (amoxicillin and ampicillin), cephalosporins (ceftifour, cephalexin, and cephazolin), and polypeptides, such as vancomycin.
Rozprawy doktorskie na temat "Cst Genes"
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Charles, Ian George. "Proteus mirabilis and cat". Thesis, University of Leicester, 1986. http://hdl.handle.net/2381/35192.
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Proteus mirabilis PM13 is a well characterized chloramphenicol-sensitive isolate which spontaneously gives rise to resistant colonies on solid media containing chloramphenicol (50ug/ml) at a plating efficiency of between 10-4 and 10-5 per cell per generation. When a chloramphenicol resistant colony is grown in liquid medium in the absence of the antibiotic for I50 generations a population of predominantly sensitive cells arises. The cat gene responsible for the phenomenon is chromosomal, and has been cloned from P.mirabilis PMI3 with DNA prepared from cells grown in the absence or the presence of chloramphenicol. Recombinant plasmids which confer resistance to chloramphenicol carry an 8.5-kb PstI fragment irrespective of the source of host DNA. The location of The cat gene within the PstI fragment was determined by Southern blotting with a cat consensus 'active - site' oligonucleotide (5'-CCATCACAGACGGCATGATG-3') corresponding to the expected amino acid sequence of the active site region of chloramphenicol acetyltransferase. DNA sequence analysis has revealed a high degree of homology between the P. mirabllls cat -gene and the type I ca-t variant (Tn9), 76% at the amino acid level and 73% when nucleotides in the coding sequence are compared. The mechanism for the appearance and disappearance of chloramphenicol resistance in P. mirabilis appears to be associated with a host-specific trans-acting element which controls cat gene expression. A precedent for such a control network is given by phase variation in Salmonella typhimurium, where an invertible DNA segment controls the transcription of a trans-acting regulatory element. A comparison of the 5' regions of the S.typhimurium flagellin genes in and H2, which are alternately expressed by a flip-flop control mechanism with the 5' region of P.mirabilis cat show blocks of homology. Whether or not this homology is significant in the regulation of cat gene expression has not been determined.
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Day, Christopher. "Genes Involved in Osteoclastogenesis". Thesis, Griffith University, 2005. http://hdl.handle.net/10072/367814.
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Osteoclast formation is a complex process requiring the temporal activation of a yet unknown number of transcription factors. Osteoclast differentiation is dependent on two cytokines: macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL). These agents induce gene expression changes during the differentiation process, presumably by inducing transcription factors. A search for genes that are regulated in the developing osteoclast was performed using both differential display and gene arrays. Differential display revealed a novel member of the krüppel-associated box (KRAB) containing transcription repressor, KROCS, which was shown to be down regulated during osteoclast formation. The potential targets for this gene remain unknown, as do the targets of the majority of the other members of the krüppel associated box (KRAB) containing family of transcription repressors. In addition to KROCS, three other genes were also identified including ADO21, PRO1859 and an endogenous retrovirus like gene and the regulation of vitamin D up-regulated protein (VDUP) was also confirmed.
Array analysis identified a number of other transcription factors regulated during osteoclast formation including the up-regulated NFATc1, GABP?, FBP, EGR1 and the repressed RelB and KOX31, a KRAB containing transcription repressor. The array also identified calmodulin 1 a member of the NFAT activation pathways as up-regulated by RANKL. The expression of NFATc1 to 4 in human osteoclasts was investigated showing NFATc1 to be the most expressed NFATc in osteoclasts. The transcription variants of NFATc1 were tested for expression differences showing that the mRNAs encoding the protein isoforms B and C were most expressed. The involvement of calmodulin, calcineurin and NFATc1 involvement in osteoclast formation was further studied by the use of inhibitors. BAPTA-AM is an intracellular chelator of calcium that prevents changes in calcium concentration. Phenoxybenzamine irreversibly binds calmodulin in the presence of calcium, inhibiting the action of calmodulin. Cyclosporin A (CsA) is an inhibitor of calcineurin. Use of both BAPTA-AM and phenoxybenzamine resulted in inhibition of osteoclast formation, decreasing the percentage of multinucleated cells from 54% in control cultures to 7.9% and 7.1% respectively. Both BAPTA-AM and phenoxybenzamine treated cells showed a marked reduction in TRAP activity with only 14.5% and 16.8% respectively staining positive for TRAP. This represents an approximate 60% reduction in TRAP positive cells compared to control osteoclasts. Both BAPTA-AM treated and phenoxybenzamine treated cells were negative for bone resorption. Addition of increasing doses of cyclosporin A (CsA) to M-CSF and RANKL treated cells resulted in the inhibition of multinucleated osteoclast formation. At 1000ng/mL CsA the formation of TRAP positive cells with more than one nucleus had reduced to less than 5% from 54% without the presence of CsA. Cells treated with 1000ng/mL CsA were unable to resorb bone, however the percentage of cells that were TRAP positive remained unchanged with CsA treatment. No significant decrease in expression of cathepsin K or TRAP transcripts were observed by real-time quantitative PCR (Q-PCR) in cells treated with CsA. Although all three agents inhibited the formation of multinuclear giant cells, both BAPTA-AM and phenoxybenzamine resulted in TRAP negative cells, whereas CsA resulted in TRAP positive cells. These results implicate the intracellular calcium increase caused by RANKL and calmodulin activation as a regulator of TRAP but place the calcineurin activation of NFATc1 downstream of TRAP induction. The regulation of a series of other genes was tested to determine if some RANKL mediated regulation of osteoclast genes were 'sensitive to CsA while others were 'resistant'. Of 28 genes tested, 13 were significantly affected by CsA and were considered 'sensitive' while the RANKL mediated regulation of 15 genes was unaffected by CsA and these were considered 'resistant'. This is strong evidence for two pathways of gene activation in osteoclasts, a CsA 'sensitive' pathway involving calcineurin, NFAT and possibly other transcription factors and a CsA 'resistant' pathway of gene activation, not dependent on calcineurin. Surprisingly, the RANKL mediated induction of NFATc1 was not inhibited by CsA, suggesting that NFATc1 induction is dependent on the resistant pathway. The identity of the second pathway (or pathways) is yet to be established, however the data indicate that this pathway mediate the RANKL sensitive regulation of at least one half of genes in human osteoclasts. The corollary if that only one half of osteoclast genes are dependent on calcineurin and presumably NFATc1 activation.
There was no unifying principle that separated the CsA resistant from sensitive pathways of RANKL regulation. Cell surface markers, chemokines and transcription factors were among those affected by CsA. Even classical osteoclast markers fell neatly into two categories. The RANKL mediated induction of calcitonin receptor (CalcR) was inhibited by more than 100 fold in the presence of CsA implicating NFAT/calcineurin in the regulation of CalcR expression in osteoclasts. In contrast, the RANKL mediated induction of TRAP or cathepsin K, two prominent osteoclast markers, was totally unaffected by CsA.
The expression of a series of chemokines and receptors was investigated. MCP-1 and RANTES were RANKL induced, and this induction was sensitive to CsA. The CC chemokines MCP-1 and RANTES were down regulated by around 10 fold in the presence of CsA. In contrast the RANKL mediated induction of MCP-1 receptor was resistant to CsA. The existence of chemokine and receptor in the same cell provides for a RANKL inducible autocrine loop, suggesting that MCP-1 should act directly on osteoclasts. The fact that the RANKL induction of the MCP-1 receptor, CCR2B, is unaffected by CsA suggests that exogenous MCP-1 should still signal in CsA treated osteoclasts. Addition of either MCP-1 or RANTES to CsA treated cultures resulted in a recovery of 70-80% of the multinuclear TRAP positive phenotype. The MCP-1 and RANTES induced multinuclear cell could not overcome the CsA induced inhibition of bone resorption. Surprisingly, MCP-1 and RANTES induced multinucleation in the absence of RANKL (M-CSF and chemokine treated cells) resulting in 50% of the normal multinucleation present in cells treated with RANKL. The data suggest that chemokines produced by osteoclasts are involved in promoting a multinuclear phenotype. When inhibited by CsA, osteoclasts fail to produce both MCP-1 and RANTES, although their respective receptors are present. This failure to produce MCP-1 and RANTES prevents the formation of an autocrine loop. When provided with MCP-1 or RANTES the CsA inhibited osteoclasts are subsequently able to pass through to the stage of a multinucleated giant cell. Similarly, in the absence of RANKL, chemokines promote the formation of TRAP positive osteoclast-like giant cells visually indistinguishable from osteoclasts. However, the multinuclear cells formed by chemokines in the absence of RANKL were also incapable of bone resorption. In order to determine if chemokines were capable of stimulating bone resorption, after osteoclasts had formed, pre-differentiated mature osteoclasts were plated onto bone and treated with a range of cytokines. The results showed that bone resorption occurred only in cultures that were exposed continuously to RANKL. These data indicate that chemokine induction by RANKL is required for multinucleation but that RANKL is required for bone resorption. The functional testing of genes detected by array analysis proved crucial involvement of both the NFAT pathway and CC chemokines in osteoclast formation knd function. Other genes identified such as GABP and FBP, are likely to be key
factors in the development of a functional osteoclast. Future works investigating
human osteoclast formation should take into strong consideration the genes identified
in this thesis as targets for further functional studies.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Medical Science
Full Text
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Blackbourn, David J. "Molecular studies of cat-86 gene expression". Thesis, University of Newcastle Upon Tyne, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.278764.
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Schwanke, Raquel Cristina. "Clonagem, expressão, purificação e ensaio biológico da proteína GM-CSF: fator estimulador de colônias de granulócitos e macrófagos". Pontifícia Universidade Católica do Rio Grande do Sul, 2008. http://hdl.handle.net/10923/1388.
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Previous issue date: 2008The granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that belongs to a group of glycoprotein that regulates the proliferation and differentiation of hematopoietic cells, more specifically granulocytes and macrophages. The human GM-CSF is a 14. 4 kDa protein consisting of 127 amino acid polypeptide chain and shares 52% of similarities with murine protein. The human protein has 4 cysteine residues that forms two disulphide bonds; only the Cysteine 54 and 96 are required for the biologic activity of it. This biopharmaceutical has been widely used in neutropenic patients who receive high-dose chemotherapy or were transplanted. Therefore, GM-CSF is used to restore hematopoietic dysfunctions, to stimulate the hyper-production of functionally primed effectors cells and to augment host defense against infection and malignant diseases. Its use is associated with significant decreases in chemotherapy-associated infections, antibiotic use, length of hospital day and mortality. The international patent of the biopharmaceutical Molgramostim (generic name) expired in 2006, and thus became an interesting product to pharmaceutical industries, including those settled in Brazil. Presently, Molgramostim is sold in Brazil as an imported biopharmaceutical, which, in turn becomes very costly to the Brazilian government. Therefore, the aim of this work is to develop a methodology for subsequent production of a national Molgramostim. In this work the granulocyte-macrophage colony-stimulating factor gene was assembled by PCR. It was cloned into pET30a(+) expression vector and the best condition for expression of the protein was in the BL21(DE3) strain from Escherichia coli. To the isolation of inclusion bodies an efficient protocol was developed using multi step washing procedure and purification method of the recombinant protein from inclusion bodies using only a cationic and then an anionic exchange column. The immunoassay and N-terminal sequencing confirmed the identity of rhGM-CSF. The result of the biological activity assay, in vitro, showed that the rhGM-CSF produced has an equivalent biological potential to the standard reference. The protein rhGM-CSF was produced through simple, cost effective and economically feasible process and is extremely important to the industrial procedure and healthcare community.
O fator estimulador de colônias de granulócitos e macrófagos (GM-CSF) é uma citocina pertencente a um grupo de glicoproteínas que regula a proliferação e a diferenciação de células hematopoiéticas, mais especificamente macrófagos e granulócitos. O GM-CSF humano é uma proteína de 14,5 kDa constituída de 127 aminoácidos e possui 52% de similaridade com a proteína de rato. A proteína humana possui 4 resíduos de cisteína formando duas pontes dissulfeto, porém apenas as cisteínas 54 e 96 são requeridas para a atividade biológica da mesma. Este biofármaco tem sido usado em pacientes neutropênicos que receberam altas doses de quimioterapia ou transplantados. Além disso, GM-CSF é usado para restabelecer disfunções hematopoiéticas, estimular a hiper-produção de células efetoras pré-induzidas (“primed”) funcionalmente e promover a defesa do hospedeiro contra doenças infecciosas e malignas. O uso dos mesmos está relacionado à redução no número de infecções associadas à quimioterapia, no uso de antibióticos e no tempo total de internação do paciente bem como no número de mortes. A patente internacional do biofármaco Molgramostima (nome genérico) expirou em 2006 e, além disso, tornou-se um interessante produto para indústrias farmacêuticas, inclusive para aquelas estabelecidas no Brasil. Molgramostima é vendida no Brasil como um biofármaco importado, o qual, desta forma, torna-se muito custoso para o governo brasileiro. Contudo, o objetivo deste trabalho é desenvolver uma metodologia para a posterior produção industrial de uma molgramostima a nível nacional. Neste trabalho, o gene para o hGM-CSF foi construído por PCR, clonado no vetor de expressão pET30a(+) e expresso na cepa BL21(DE3) de Escherichia coli na sua forma insolúvel. Para o isolamento e solubilização dos corpos de inclusão um eficiente protocolo foi desenvolvido utilizando múltiplos passos de lavagem e um método de purificação usando somente duas colunas cromatográficas de troca catiônica e aniônica, respectivamente. O teste de atividade biológica in vitro demonstrou que o rhGM-CSF produzido tem potencial equivalente ao padrão internacional utilizado. A proteína foi obtida por meio de um processo simples e econômico, sendo de extrema importância em procedimento industrial e para saúde da população.
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Jiang, Kan. "Regulation of the potential tumorsuppressor gene mad1 by G-CSF". [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=979663180.
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Kovacevic, David. "Physically mapping the CYP2 gene cluster in the domestic cat". Miami University Honors Theses / OhioLINK, 2003. http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1110920381.
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Osborne, Cameron Stuart. "Transcriptional regulation of the GM-CSF gene in T lymphocytes /". Title page, contents and summary only, 1996. http://web4.library.adelaide.edu.au/theses/09PH/09pho81.pdf.
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Heur, J. Martin. "Lysosomal Regulation of Gene Expression". University of Cincinnati / OhioLINK, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1026512116.
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Forrest, D. "A study of the myc gene in feline leukaemias". Thesis, University of Glasgow, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.377160.
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Blackwood, Laura. "Transcriptional targeting of gene therapy in the hyperthyroid cat : preliminary investigations". Thesis, University of Glasgow, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.272862.
Pełny tekst źródłaKsiążki na temat "Cst Genes"
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Hubbell, Sue. Shrinking the cat: Genetic engineering before we knew about genes. Boston: Houghton Mifflin, 2001.
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Amram, David. Upbeat: Nine lives of a musical cat. Boulder: Paradigm Publishers, 2008.
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Clauser, Marina, Andrea Grigioni i Mario Landi, red. Peperoncini. Florence: Firenze University Press, 2010. http://dx.doi.org/10.36253/978-88-8453-951-9.
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The chili pepper is a spice and medicinal remedy used since ancient times by the American peoples who were the first to undertake the domestication of 5 species belonging to the genus Capsicum (Solanaceae): Capsicum (Solanaceae): Capsicum annuum, C. baccatum, C. chinense, C. frutescens e C. pubescens. After the sixteenth century the chili pepper became similarly popular in other continents and today the five species number many reference pod-types and over 3,000 varieties. The book describes their uses in the different spheres of cuisine (aromatic, spicy and colourful), medicine (antioxidant and digestive for internal use, rubefacient and anti-rheumatic for external use) and ornamentation (cut branches, floral compositions, border plants, splashes of colour).
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1953-, Carlstedt-Duke J., Eriksson Håkan i Gustafsson Jan-Åke, red. The steroid/thyroid hormone receptor family and gene regulation: Proceedings of the 2nd International CBT Symposium, Stockholm, Sweden, November 4-5, 1988. Basel: Birkhäuser, 1989.
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Hubbell, Sue. Shrinking the Cat: Genetic Engineering Before We Knew About Genes. Mariner Books, 2002.
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Hubbell, Sue. Shrinking the Cat: Genetic Engineering Before We Knew About Genes. Houghton Mifflin, 2001.
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Hubbell, Sue. Shrinking the Cat: Genetic Engineering Before We Knew About Genes. Houghton Mifflin Company, 2002.
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Kühn, Wolfgang, i Gerd Walz. The molecular basis of ciliopathies and cyst formation. Redaktor Neil Turner. Oxford University Press, 2015. http://dx.doi.org/10.1093/med/9780199592548.003.0303.
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Abnormalities of the cilium, termed ‘ciliopathies’, are the prime suspect in the pathogenesis of renal cyst formation because the gene products of cystic disease-causing genes localize to them, or near them. However, we only partially understand how cilia maintain the geometry of kidney tubules, and how abnormal cilia lead to renal cysts, and the diverse range of diseases attributed to them. Some non-cystic diseases share pathology of the same structures. Although still incompletely understood, cilia appear to orient cells in response to extracellular cues to maintain the overall geometry of a tissue, thereby intersecting with the planar cell polarity (PCP) pathway and the actin cytoskeleton. The PCP pathway controls two morphogenetic programmes, oriented cell division (OCD) and convergent extension (CE) through cell intercalation that both seem to play a critical role in cyst formation. The two-hit theory of cystogenesis, by which loss of the second normal allele causes tubular epithelial cells to form kidney cysts, has been largely borne out. Additional hits and influences may better explain the rate of cyst formation and inter-individual differences in disease progression. Ciliary defects appear to converge on overlapping signalling modules, including mammalian target of rapamycin and cAMP pathways, which can be targeted to treat human cystic kidney disease irrespective of the underlying gene mutation.
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Biloshytsky, Vadym, i Roman Cregg. Pioneering use of gene therapy for pain. Redaktorzy Paul Farquhar-Smith, Pierre Beaulieu i Sian Jagger. Oxford University Press, 2018. http://dx.doi.org/10.1093/med/9780198834359.003.0083.
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The landmark paper discussed in this chapter is ‘Gene therapy for pain: Results of a Phase I clinical trial’, published by Fink et al. in 2011. In this study, the first of its kind, researchers studied the efficacy and safety of a modified herpes simplex virus (HSV) vector used to deliver PENK, which encodes proenkephalin, which is cleaved into the enkephalin peptides Met-enkephalin and Leu-enkephalin, which induce analgesia by acting on opioid receptors. The development of the HSV vector was based in part on results studies in which adenovirus, adeno-associated virus, or non-viral vectors were used to overexpress genes. Overexpression of a variety of large molecules leads to a reduction in pain-related behaviour in animals. Gene therapy in the treatment of chronic pain seems to offer a promising alternative to systemic or highly invasive therapies. However, additional research is needed to determine the safety, effectiveness, and cost-efficiency of this approach.
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Fonder, Tom. Business Cat: Hostile Takeovers. Andrews McMeel Publishing, 2019.
Znajdź pełny tekst źródłaCzęści książek na temat "Cst Genes"
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Liu, Guanshu, Jeff W. M. Bulte i Assaf A. Gilad. "CEST MRI Reporter Genes". W Methods in Molecular Biology, 271–80. Totowa, NJ: Humana Press, 2010. http://dx.doi.org/10.1007/978-1-61737-992-5_13.
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Stoffel, Wilhelm. "GalCer Synthase (Ceramide Galactosyltransferase, CGT)". W Handbook of Glycosyltransferases and Related Genes, 51–57. Tokyo: Springer Japan, 2002. http://dx.doi.org/10.1007/978-4-431-67877-9_8.
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Kleopa, Kleopas A., Alexia Kagiava i Irene Sargiannidou. "Gene Therapy for CMT Inherited Neuropathy". W Muscle Gene Therapy, 621–44. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-030-03095-7_35.
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Lev, Zeev, Noa Cohen, Adi Salzberg, Ziva Kimchie, Naomi Halachmi i Orit Segev. "Expression of the Drosophila ras2/cs1 Gene Pair during Development". W The Superfamily of ras-Related Genes, 303–9. Boston, MA: Springer US, 1991. http://dx.doi.org/10.1007/978-1-4684-6018-6_33.
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Subbotin, Sergei A. "Phylogenetic analysis of DNA sequence data." W Techniques for work with plant and soil nematodes, 265–82. Wallingford: CABI, 2021. http://dx.doi.org/10.1079/9781786391759.0265.
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Abstract The goal of phylogenetics is to construct relationships that are true representations of the evolutionary history of a group of organisms or genes. The history inferred from phylogenetic analysis is usually depicted as branching in tree-like diagrams or networks. In nematology, phylogenetic studies have been applied to resolve a wide range of questions dealing with improving classifications and testing evolution processes, such as co-evolution, biogeography and many others. There are several main steps involved in a phylogenetic study: (i) selection of ingroup and outgroup taxa for a study; (ii) selection of one or several gene fragments for a study; (iii) sample collection, obtaining PCR products and sequencing of gene fragments; (iv) visualization, editing raw sequence data and sequence assembling; (v) search for sequence similarity in a public database; (vi) making and editing multiple alignment of sequences; (vii) selecting appropriate DNA model for a dataset; (viii) phylogenetic reconstruction using minimum evolution, maximum parsimony, maximum likelihood and Bayesian inference; (ix) visualization of tree files and preparation of tree for a publication; and (x) sequence submission to a public database. Molecular phylogenetic study requires particularly careful planning because it is usually relatively expensive in terms of the cost in reagents and time.
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Subbotin, Sergei A. "Phylogenetic analysis of DNA sequence data." W Techniques for work with plant and soil nematodes, 265–82. Wallingford: CABI, 2021. http://dx.doi.org/10.1079/9781786391759.0015.
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Abstract The goal of phylogenetics is to construct relationships that are true representations of the evolutionary history of a group of organisms or genes. The history inferred from phylogenetic analysis is usually depicted as branching in tree-like diagrams or networks. In nematology, phylogenetic studies have been applied to resolve a wide range of questions dealing with improving classifications and testing evolution processes, such as co-evolution, biogeography and many others. There are several main steps involved in a phylogenetic study: (i) selection of ingroup and outgroup taxa for a study; (ii) selection of one or several gene fragments for a study; (iii) sample collection, obtaining PCR products and sequencing of gene fragments; (iv) visualization, editing raw sequence data and sequence assembling; (v) search for sequence similarity in a public database; (vi) making and editing multiple alignment of sequences; (vii) selecting appropriate DNA model for a dataset; (viii) phylogenetic reconstruction using minimum evolution, maximum parsimony, maximum likelihood and Bayesian inference; (ix) visualization of tree files and preparation of tree for a publication; and (x) sequence submission to a public database. Molecular phylogenetic study requires particularly careful planning because it is usually relatively expensive in terms of the cost in reagents and time.
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Wagner, Andreas. "On the Energy and Material Cost of Gene Duplication". W Evolution after Gene Duplication, 207–14. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2011. http://dx.doi.org/10.1002/9780470619902.ch11.
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Lu, Bao-Rong. "Assessing environmental impact of pollen-mediated transgene flow." W Gene flow: monitoring, modeling and mitigation, 1–25. Wallingford: CABI, 2021. http://dx.doi.org/10.1079/9781789247480.0001.
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Abstract Potential environmental impact caused by pollen-mediated transgene flow from commercially cultivated genetically engineered (GE) crops to their non-GE crop counterparts and to their wild and weedy relatives has aroused tremendous biosafety concerns worldwide. This chapter provides information on the concept and classification of gene flow, the framework of the environmental biosafety assessment caused by pollen-mediated gene flow, and relevant case studies about transgene flow and its environmental impact. In general, gene flow refers to the movement of genes or genetic materials from a plant population to other populations. Crop-to- crop transgene flow at a considerable frequency may result in transgene 'contamination' of non-GE crops, causing potential food/feed biosafety problems and regional or international trade disputes. Crop-to- wild/weedy transgene flow may bring about environmental impacts, such as creating more invasive weeds, threatening local populations of wild relative species, or affecting genetic diversity of wild relatives, if the incorporated transgene can normally express in the recipient wild/weedy plants and significantly alter the fitness of the wild/weedy plants and populations. It is therefore necessary to establish a proper protocol to assess the potential environmental impacts caused by transgene flow. Three steps are important for assessing potential environment impacts of transgene flow to wild/weedy relatives: (i) to accurately measure the frequencies of transgene flow: (ii) to determine the expression level of a transgene incorporated in wild/weedy populations; and (iii) to estimate the fitness effect (benefit or cost) conferred by expression of a transgene in wild/weedy populations. The recently reported case of non-random allele transmission into GE and non-GE hybrid lineages or experimental populations challenges the traditional method of estimating the fitness effect for the assessment of environmental impacts of transgene flow. Furthermore, case studies of transgenic mitigation (TM) strategies illustrate ways that may reduce the impacts of a transgene on wild/weedy populations if crop-to- wild/weedy transgene flow is not preventable, such as in the case of gene flow from crop rice to its co-occurring weedy rice.
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Wiktor-Jedrzejczak, Wieslaw, i Andrzej Nowicki. "Spontaneous Knockout of CSF-1 Gene in the Mouse as a Model to Study the Organization of the Macrophage System". W Gene Technology, 333–50. Berlin, Heidelberg: Springer Berlin Heidelberg, 1996. http://dx.doi.org/10.1007/978-3-642-61122-3_24.
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Holowiecki, Jerzy. "GM-CSF in Addition to Chemotherapy of ALL for Kinetics Based Protection of Stem Cells and Stimulation of Haemopoiesis.A randomized Study". W Gene Technology, 429–40. Berlin, Heidelberg: Springer Berlin Heidelberg, 1996. http://dx.doi.org/10.1007/978-3-642-61122-3_32.
Pełny tekst źródłaStreszczenia konferencji na temat "Cst Genes"
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Sharma, Jai, i Vidhyacharan Bhaskar. "An Rna Sequencing Analysis of Glaucoma Genesis in Mice". W 12th International Conference on Artificial Intelligence, Soft Computing and Applications. Academy and Industry Research Collaboration Center (AIRCC), 2022. http://dx.doi.org/10.5121/csit.2022.122306.
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Glaucoma is the leading cause of irreversible blindness in people over the age of 60, accounting for 6.6 to 8% of all blindness in 2010, but there is still much to be learned about the genetic origins of the eye disease. With the modern development of Next-Generation Sequencing (NGS) technologies, scientists are starting to learn more about the genetic origins of Glaucoma. This research uses differential expression (DE) and gene ontology (GO) analyses to study the genetic differences between mice with severe Glaucoma and multiple control groups. Optical nerve head (ONH) and retina data samples of genome-wide RNA expression from NCBI (NIH) are used for pairwise comparison experimentation. In addition, principal component analysis (PCA) and dispersion visualization methods are employed to perform quality control tests of the sequenced data. Genes with skewed gene counts are also identified, as they may be marker genes for a particular severity of Glaucoma. The gene ontologies found in this experiment support existing knowledge of Glaucoma genesis, providing confidence that the results were valid. Future researchers can thoroughly study the gene lists generated by the DE and GO analyses to find potential activator or protector genes for Glaucoma in mice to develop drug treatments or gene therapies to slow or stop the progression of the disease. The overall goal is that in the future, such treatments can be made for humans as well to improve the quality of life for human patients with Glaucoma and reduce Glaucoma blindness rates.
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Marco, Anderson, Mario Gazziro i David Martins Jr. "High performance computing architectures analysis for gene networks inference". W XX Simpósio em Sistemas Computacionais de Alto Desempenho. Sociedade Brasileira de Computação, 2019. http://dx.doi.org/10.5753/wscad.2019.8656.
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Modeling and inference of biological systems are an important field in computer science, presenting strong interdisciplinary aspects. In this context, the inference of gene regulatory networks and the analysis of their dynamics generated by their transition functions are important issues that demand substantial computational power. Because the algorithms that return the optimal solution have an exponential time cost, such algorithms only work for gene networks with only dozens of genes. However realistic gene networks present hundreds to thousands of genes, with some genes being hubs, i.e., their number of predictor genes are usually much higher than average. Therefore there is a need to develop ways to speed up the gene networks inference. This paper presents a benchmark involving GPUs and FPGAs to infer gene networks, analysing processing time, hardware cost acquisition, energy consumption and programming complexity. Overall Titan XP GPU achieved the best performance, but with a large cost regarding acquisition price when compared to R9 Nano GPU and DE1-SOC FPGA. In its turn, R9 Nano GPU presented the best cost-benefit regarding performance, acquisition price, energy consumption, and programming complexity, although DE1-SOC FPGA presented much smaller energy consumption.
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Yang, C., E. Zeng, T. Li i G. Narasimhan. "Clustering genes using gene expression and text literature data". W 2005 IEEE Computational Systems Bioinformatics Conference (CSB'05). IEEE, 2005. http://dx.doi.org/10.1109/csb.2005.23.
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Kamble, Pranoti R., Rakhi Wajgi i Manali Kshirsagar. "Identifying Most Significant Genes on Imputed Gene Expression Dataset". W 2015 Fifth International Conference on Communication Systems and Network Technologies (CSNT). IEEE, 2015. http://dx.doi.org/10.1109/csnt.2015.187.
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Stacey, Minviluz. "Utility of CRISPR/Cas in accelerating gene discovery in soybean". W 2022 AOCS Annual Meeting & Expo. American Oil Chemists' Society (AOCS), 2022. http://dx.doi.org/10.21748/rzne1660.
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The use of CRISPR/Cas9 has been successfully applied in various plant species to induce targeted genome editing, including soybean. Soybean is recalcitrant to transformation and thus, plants with stable CRISPR gene edits are costly and take a long time to produce. Moreover, soybean is allotetraploid and editing paralogous genes are often necessary to obtain observable phenotype(s). For each gene target, we designed two gRNAs to increase editing efficiency and allow rapid genotyping by PCR. We also tested the CRISPR reagents in transient hairy root transformation to determine if the Cas9 and gRNAs would perform properly in transgenic soybean plants. Our results showed that we can indeed obtain highly efficient, cost-effective CRISPR/Cas editing in soybean to generate novel genotypes for gene discovery and downstream field propagation and breeding efforts. Examples of CRISPR-edited genes and their associated seed traits will be discussed.
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Heinzen, Rebeca Neves, Maria Eduarda Meyer, Liliane Raupp Gomes Pizatto i Adriana Magalhães de Oliveira Freitas. "PREVALENCE OF VARIANTS OF UNCERTAIN SIGNIFICANCE IN TESTS REQUESTED FOR BREAST CANCER PATIENTS IN A PRIVATE SERVICE". W Scientifc papers of XXIII Brazilian Breast Congress - 2021. Mastology, 2021. http://dx.doi.org/10.29289/259453942021v31s1049.
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Introduction: The genetic mutations test among breast cancer (BC) patients is one of the steps for the diagnosis in the majority of the patients. To identify and manage patients with hereditary predisposition to cancer is also a competence of the breast surgeon. The development of Next Generation Sequence (NGS) has allowed the reduction of the tests’ cost as well as the expansion of the analyzed genes, besides BRCA 1 and 2, and the inclusion of new genes of high and moderate penetrance. There is a concern about the impact of these results because there is not a well-established conduct for all the mutations as well as for the increase of the diagnoses of variant of uncertain significance (VUS) diagnosed in the panels, mostly in patients that did not receive a formal genetic counseling. Studies show that the larger number of analyzed genes is related to a better chance of detecting VUS, reaching 40%, but they are not conduct modifiers. The literature shows that approximately 90% of VUS are reclassified as benign. Objectives: To assess prevalence of VUS in multigenic panels requested by the non-geneticist physician, in private office, performed on patients with BC diagnosis. Methods: A retrospective cross-sectional study was conducted based in data from invasive BC patients or in situ or with high risk for neoplasia that attended a private office and were subjected to multigenic panels requested by the non-geneticist physician from January 2019 to January 2020. Statistical analysis frequency measurements were analyzed in Excel Office®. Results: 147 patients underwent the genetic test of 83 genes with NGS technology. Only one was a male. Among the tests performed, 48 were negative for pathogenic variants and 23 were positive for pathogenic mutations in 22 (15%) patients, the most common being in BRCA2 gene (7 cases), followed by MUTHY (6 cases). 137 VUS occurred in 77 (52.4%) patients, the most common of these being in gene POLE and RECQL4. Conclusions: The data found in our population match the literature, showing more than half of the patients with VUS. This demonstrates the importance of test interpretation as well as inpatient correct orientation.
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Salazar Moscoso, Marcela, Silvia Joly Ruiz Castellanos, Guillem Anglada Escudé i Laia Ribas Cabezas. "Hypergravity induces changes in physiology, gene expression and epigenetics in zebrafish". W Symposium on Space Educational Activities (SSAE). Universitat Politècnica de Catalunya, 2022. http://dx.doi.org/10.5821/conference-9788419184405.044.
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All living organisms that inhabit Earth have evolved under a common value of gravity, which amounts to an acceleration of 9.81 m/s2 at mean sea level. Changes on it could cause important alterations that affect vital biological functions. The crescent interest in spatial exploration has opened the question of how exactly these changes in gravity would affect Earth life forms on space environments. This work is the result of a collaborative co-supervision of a master thesis between experts in the area of space sciences and biology, and it can serve as a case study for training experts in such interdisciplinary environments. In particular, we focus on the effect of gravity as a pressure factor in the development of zebrafish (Danio rerio) in the larval stage as a model organism using up-to-date (genomic and epigenetic) techniques. Given the high cost of any experiment in true low gravity (which would require a space launch), we performed an initial experiment in hypergravity to develop the methodologies and identify good (epi)genetic markers of the effect of gravity in our model organism. Previous studies in zebrafish have shown how alteration in gravity effects the development and the gene expression of important regulatory genes. For this study, we firstly customized a small laboratory scale centrifuge to study changes in fish physiology together with changes at molecular levels. We exposed zebrafish larvae from 0 to 6 days post fertilization to the simulated hypergravity (SHG) (100 rpm 3g). After 6 days of hypergravity exposition the larvae showed changes in their swimming and flotation patterns, and presented corporal alterations. Then, we assessed gene expression of genes implicated in important biological processes, (e.g., epigenetics), and an upregulation were observed when compared to the control. Taken together, these preliminary findings show how gravity alterations could affect some basic biological responses, and illustrate the potential of developing new science cases to be developed by students at postgraduate level (MSc and beyond) in a multidisciplinary environment
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He, Qin, Rubin Wang i Xiaochuan Pan. "This paper presents a two-dimensional histogram shifting technique for reversible data hiding algorithm. In order to avoid the distortion drift caused by hiding data into stereo H.264 video, we choose arbitrary embeddable blocks from 4×4 quantized discrete cosine transform luminance blocks which will not affect their adjacent blocks. Two coefficients in each embeddable block are chosen as a hiding coefficient pair. The selected coefficient pairs are classified into different sets on the basis of their values. Data could be hidden according to the set which the value of the coefficient pair belongs to. When the value of one coefficient may be changed by adding or subtracting 1, two data bits could be hidden by using the proposed method, whereas only one data bit could be embedded by employing the conventional histogram shifting. Experiments show that this two-dimensional histogram shifting method can be used to improve the hiding performance." W 10th International Conference on Software Engineering and Applications (SEAS 2021). AIRCC Publishing Corporation, 2021. http://dx.doi.org/10.5121/csit.2021.110205.
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Arc, one virus-like gene, crucial for learning and memory, was dis-covered by researchers in neurological disorders fields, Arc mRNA’s single directed path and allowing protein binding regional restric-tively is a potential investigation on helping shuttle toxic proteins responsible for some diseases related to memory deficiency. Mean time to switching (MTS) is calculated explicitly quantifying the switching process in statistical methods combining Hamiltonian Markov Chain(HMC). The model derived from predator and prey with typeII functional response studies the mechanism of normals with intrin-sic rate of increase and the persisters with the instantaneous discovery rate and converting coefficients. During solving the results, since the numeric method is applied for the 2D approximation of Hamiltonion with intrinsic noise induced switching combining geometric minimum action method. In the application of Hamiltonian Markov Chain, the behavior of the convertion (between mRNA and proteins through 6 states from off to on ) is described with probabilistic conditional logic formula and the final concentration is computed with both Continuous and Discret Time Markov Chain(CTMC/DTMC) through Embedding and Switching Diffusion. The MTS, trajectories and Hamiltonian dynamics demonstrate the practical and robust advantages of our model on interpreting the switching process of genes (IGFs, Hax Arcs and etc.) with respects to memory deficiency in aging process which can be useful in further drug efficiency test and disease curing. Coincidentally, the Hamiltonian is also well used in describing quantum mechanics and convenient for computation with time and position information using quantum bits while in the second model we construct, switching between excitatory and inhibitory neurons, similarity of qubit and neuron is an interesting object as well. Especially with the interactions operated with phase gates, the excitation from the ground state to excitation state is a well analogue to the neuron excitation. Not only on theoretical aspect, the experimental methods in neuron switching model is also inspiring to quantum computing. Most basic one is as stimulate hippocampus can be identical to spontaneous neural excitation(|g>|e>), pi-pulse is utilized to drive the ground state to the higher state. There thus exists prosperous potential to study the transfer between states with our switch models both classical and quantum computationally.
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Teixeira, Elaine Calumby, Camila Rodrigues Nepomuceno, Maria Sheila Guimarães Rocha i Eduardo de Paula Estephan. "Charcot-Marie-Tooth type 1F with bi-allelic NEFL mutation". W XIV Congresso Paulista de Neurologia. Zeppelini Editorial e Comunicação, 2023. http://dx.doi.org/10.5327/1516-3180.141s1.628.
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Introduction: Charcot-Marie-Tooth disease (CMT) is the most common hereditary neuropathy, with a diverse phenotypic and genotypic spectrum. The main clinical features are onset during infancy, slowly progressing symptoms and foot deformities, especially if there is a positive family history, although the lack of family awareness can be present. Over 70 distinct genes have been associated, however, their genetic diagnosis can be challenging, especially if we consider the fact that the same gene can transmit disease either dominantly or recessively. The aim to describe a case of CMT1F as a rare case of recessive demyelinating hereditary neuropathy. Clinical case: A 25-year-old woman, born of consanguineous parents, had history of distal weakness and burning sensation in lower limbs, with onset in infancy. Her childhood was marked for abnormal gait and falls, and evolved with foot deformities, requiring surgical corrections. The symptoms progressed slowly and reached upper limbs in few years. On physical evaluation was noted: muscle weakness of upper and lower limbs, predominantly distal, associated with atrophy, foot drop and absent reflexes. Electroneuromyography demonstrated signs of chronic demyelinating polyneuropathy. An initial sequencing analysis of the PMP22 gene was indicated, with normal results. A panel for neuropathies was performed, showing a homozygous frameshift mutation in NEFL (p.Lys362Glufs*2; c.1084_1085delAA), classified as probably pathogenic variant. Conclusion: CMT due to bi-allelic NEFL mutations is a rare condition that should be considered in hereditary demyelinating neuropathy, especially when recessive inheritance is suspected. Our study illustrates this condition and brings attention to the importance of the disponibility of high throughput genetic tests.
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Taha, Kamal, Dirar Mohammad Al Homouz i Hassan Al-Muhairi. "RGRank: Ranking Semantically Related Genes". W 2012 IEEE 15th International Conference on Computational Science and Engineering (CSE). IEEE, 2012. http://dx.doi.org/10.1109/iccse.2012.46.
Pełny tekst źródłaRaporty organizacyjne na temat "Cst Genes"
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Levin, Ilan, Avtar K. Handa, Avraham Lalazar i Autar K. Mattoo. Modulating phytonutrient content in tomatoes combining engineered polyamine metabolism with photomorphogenic mutants. United States Department of Agriculture, grudzień 2006. http://dx.doi.org/10.32747/2006.7587724.bard.
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Fruit constitutes a major component of our diet, providing fiber, vitamins, minerals, and many other phytonutrients that promote good health. Fleshy fruits, such as tomatoes, already contain high levels of several of these ingredients. Nevertheless, efforts have been invested in increasing and diversifying the content of phytonutrients, such as carotenoids and flavonoids, in tomato fruits. Increasing levels of phytonutrients, such as lycopene, is highly justified from the perspective of the lycopene extraction industry due to cost effectiveness reasons. Diversifying phytonutrients, in particular those that contribute to fruit color, could potentially provide an array of attractive colors to our diet. Our major goal was to devise a novel strategy for developing tomato fruits with enhanced levels of phytochemicals known to promote good health with special emphasis on lycopene content. A further important goal was to analyze global gene expression of selected genetic lines produced throughout this study in order is to dissect the molecular mechanisms regulating phytonutrients accumulation in the tomato fruit. To achieve these goals we proposed to: 1. combine, by classical breeding, engineered polyamine metabolism with photomorphogenic high pigment mutants in order generate tomato plant with exceptionally high levels of phytonutrients; 2. use gene transfer technology for genetic introduction of key genes that promote phytonutrient accumulation in the tomato fruit, 3. Analyze accumulation patterns of the phytonutrients in the tomato fruit during ripening; 4. Analyze global gene expression during fruit ripening in selected genotypes identified in objectives 1 and 2, and 5. Identify and analyze regulatory mechanisms of chloroplast disassembly and chromoplast formation. During the 3 years research period we have carried out most of the research activities laid out in the original proposal and our key conclusions are as follows: 1. the engineered polyamine metabolism strategy proposed by the US collaborators can not increase lycopene content either on its own or in combination with an hp mutant (hp-2ᵈᵍ); 2. The hp-2ᵈᵍ affects strongly the transcriptional profile of the tomato fruit showing a strong tendency for up- rather than down-regulation of genes, 3. Ontology assignment of these miss-regulated genes revealed a consistent up-regulation of genes related to chloroplast biogenesis and photosynthesis in hp-2ᵈᵍ mutants throughout fruit development; 4. A tendency for up-regulation was also usually observed in structural genes involved in phytonutrientbiosynthesis; however this up-regulation was not as consistent. 5. Microscopic observations revealed a significantly higher number of chloroplasts in pericarp cells of mature-green hp-2ᵈᵍ/hp-2ᵈᵍ fruits in comparison to their normal fully isogenic counterparts. 6. The relative abundance of chloroplasts could be observed from early stages of fruit development. Cumulatively these results suggest that: 1. the overproduction of secondary metabolites, characterizing hp-2ᵈᵍ/hp-2ᵈᵍ fruits, is more due to chloroplast number rather then to transcriptional activation of structural genes of the relevant metabolic pathways, and 2. The molecular trigger increasing metabolite levels in hp-2ᵈᵍ mutant fruits should be traced at early stage of fruit development.
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Wong, Eric A., i Zehava Uni. Nutrition of the Developing Chick Embryo: Nutrient Uptake Systems of the Yolk Sac Membrane and Embryonic Intestine. United States Department of Agriculture, czerwiec 2012. http://dx.doi.org/10.32747/2012.7697119.bard.
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We have examined the developmental changes in composition, amount, and uptake of yolk nutrients (fat, protein, water and carbohydrates) and the expression ofnutrient transporters in the yolk sac membrane (YSM) from embryonic day 11 (Ell) to 21 (E21) and small intestine from embryonic day 15 (E15) to E21 in embryos from young (22-25 wk) and old (45-50 wk) Cobb and Leghorn breeder flocks. The developmental expression profiles for the peptide transporter 1 (PepTl), the amino acid transporters, EAAT3, CAT-1 and BOAT, the sodium glucose transporter (SGLTl), the fructose transporter (GLUT5), the digestive enzymes aminopeptidase N (APN) and sucraseisomaltase (SI) were assayed by the absolute quantification real time PCR method in the YSM and embryonic intestine. Different temporal patterns of expression were observed for these genes. The effect of in ovo injection of peptides (the dipeptide Gly-Sar, purified peptides, trypsin hydrolysate) on transporter gene expression has been examined in the embryonic intestine. Injection of a partial protein hydrolysate resulted in an increase in expression of the peptide transporter PepT2. We have initiated a transcriptome analysis of genes expressed in the YSM at different developmental ages to better understand the function of the YSM.
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Guy, Charles, Gozal Ben-Hayyim, Gloria Moore, Doron Holland i Yuval Eshdat. Common Mechanisms of Response to the Stresses of High Salinity and Low Temperature and Genetic Mapping of Stress Tolerance Loci in Citrus. United States Department of Agriculture, maj 1995. http://dx.doi.org/10.32747/1995.7613013.bard.
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The objectives that were outlined in our original proposal have largely been achieved or will be so by the end of the project in February 1995 with one exception; that of mapping cold tolerance loci based on the segregation of tolerance in the BC1 progeny population. Briefly, our goals were to 1) construct a densely populated linkage map of the citrus genome: 2) map loci important in cold and/or salt stress tolerance; and 3) characterize the expression of genes responsive to cold land salt stress. As can be seen by the preceding listing of accomplishments, our original objectives A and B have been realized, objective C has been partially tested, objective D has been completed, and work on objectives E and F will be completed by the end of 1995. Although we have yet to map any loci that contribute to an ability of citrus to maintain growth when irrigated with saline water, our very encouraging results from the 1993 experiment provides us with considerable hope that 1994's much more comprehensive and better controlled experiment will yield the desired results once the data has been fully analyzed. Part of our optimism derives from the findings that loci for growth are closely linked with loci associated with foliar Cl- and Na+ accumulation patterns under non-salinization conditions. In the 1994 experiment, if ion exclusion or sequestration traits are segregating in the population, the experimental design will permit their resolution. Our fortunes with respect to cold tolerance is another situation. In three attempts to quantitatively characterize cold tolerance as an LT50, the results have been too variable and the incremental differences between sensitive and tolerant too small to use for mapping. To adequately determine the LT50 requires many plants, many more than we have been able to generate in the time and space available by making cuttings from small greenhouse-grown stock plants. As it has turned out, with citrus, to prepare enough plants needed to be successful in this objective would have required extensive facilities for both growing and testing hardiness which simply were not available at University of Florida. The large populations necessary to overcome the variability we encountered was unanticipated and unforeseeable at the project's outset. In spite of the setbacks, this project, when it is finally complete will be exceedingly successful. Listing of Accomplishments During the funded interval we have accomplished the following objectives: Developed a reasonably high density linkage map for citrus - mapped the loci for two cold responsive genes that were cloned from Poncirus - mapped the loci for csa, the salt responsive gene for glutathione peroxidase, and ccr a circadian rhythm gene from citrus - identified loci that confer parental derived specific DNA methylation patterns in the Citrus X Poncirus cross - mapped 5 loci that determine shoot vigor - mapped 2 loci that influence leaf Na+ accumulation patterns under non-saline conditions in the BC1 population - mapped 3 loci that influence leaf Na+ accumulation paterns during salt sress - mapped 2 loci that control leaf Cl- accumulation patterns under non-saline conditions - mapped a locus that controls leaf Cl- accumulation patterns during salt stress Screened the BC1 population for growth reduction during salinization (controls and salinized), and cold tolerance - determined population variation for shoot/root ratio of Na+ and Cl- - determined levels for 12 inorganic nutrient elements in an effort to examine the influence of salinization on ion content with emphasis on foliar responses - collected data on ion distribution to reveal patterns of exclusion/sequestration/ accumulation - analyzed relationships between ion content and growth Characterization of gene expression in response to salt or cold stress - cloned the gene for the salt responsive protein csa, identified it as glutathione peroxidase, determined the potential target substrate from enzymatic studies - cloned two other genes responsive to salt stress, one for the citrus homologue of a Lea5, and the other for an "oleosin" like gene - cold regulated (cor) genes belonging to five hybridization classes were isolated from Poncirus, two belonged to the group 2 Lea superfamily of stress proteins, the others show no significant homology to other known sequences - the expression of csa during cold acclimation was examined, and the expression of some of the cor genes were examined in response to salt stress - the influence of salinization on cold tolerance has been examined with seedling populations - conducted protein blot studies for expression of cold stress proteins during salt stress and vice versa
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Chejanovsky, Nor, i Bruce A. Webb. Potentiation of pest control by insect immunosuppression. United States Department of Agriculture, lipiec 2004. http://dx.doi.org/10.32747/2004.7587236.bard.
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Our original aims were to elucidate the mechanisms through which the immunosuppressive insect virus, the Campoletis sonorensis polydnavirus (CsV) promotes replication of a well-characterized pathogenic virus, the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in hosts that are mildly or non-permissive to virus replication. According to the BARD panels criticism we modified our short-term goals (see below). Thus, in this feasibility study (one-year funding) we aimed to show that: 1. S. littoralis larvae mount an immune response against a baculovirus infection. 2. Immunosuppression of an insect pest improves the ability of a viral pathogen (a baculovirus) to infect the pest. 3. S. littoralis cells constitute an efficient tool to study some aspects of the anti- viral immune response. We achieved the above objectives by: 1. Finding melanized viral foci upon following the baculoviral infection in S . littoralis larvae infected with a polyhedra - positive AcMNPV recombinant that expressed the GFP gene under the control of the Drosophila heat shock promoter. 2. Studying the effect of AcMNPV-infection in S . littoralis immunosuppressed by parasitation with the Braconidae wasp Chelonus inanitus that bears the CiV polydna virus, that resulted in higher susceptibility of S. littoralis to AcMNPV- infection. 3. Proving that S. littoralis hemocytes resist AcMNPV -infection. 4. Defining SL2 as a granulocyte-like cell line and demonstrating that as littoralis hemocytic cell line undergoes apoptosis upon AcMNPV -infection. 5. Showing that some of the recombinant AcMNPV expressing the immuno-suppressive polydna virus CsV- vankyrin genes inhibit baculoviral-induced lysis of SL2 cells. This information paves the way to elucidate the mechanisms through which the immuno- suppressive polydna insect viruses promote replication of pathogenic baculoviruses in lepidopteran hosts that are mildly or non-permissive to virus- replication by: - Assessing the extent to which and the mechanisms whereby the immunosuppressive viruses, CiV and CsV or their genes enhance AcMNPV replication in polydnavirus- immunosuppressed H. zea and S. littoralis insects and S. littoralis cells. - Identifying CiV and CsV genes involved in the above immunosuppression (e.g. inhibiting cellular encapsulation and disrupting humoral immunity). This study will provide insight to the molecular mechanisms of viral pathogenesis and improve our understanding of insect immunity. This knowledge is of fundamental importance to controlling insect vectored diseases of humans, animals and plants and essential to developing novel means for pest control (including baculoviruses) that strategically weaken insect defenses to improve pathogen (i.e. biocontrol agent) infection and virulence.
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Dubcovsky, Jorge, Tzion Fahima i Ann Blechl. Molecular characterization and deployment of the high-temperature adult plant stripe rust resistance gene Yr36 from wheat. United States Department of Agriculture, listopad 2013. http://dx.doi.org/10.32747/2013.7699860.bard.
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Stripe rust, caused by Puccinia striiformis f. sp. tritici is one of the most destructive fungal diseases of wheat. Virulent races that appeared within the last decade caused drastic cuts in yields. The incorporation of genetic resistance against this pathogen is the most cost-effective and environmentally friendly solution to this problem. However, race specific seedling resistance genes provide only a temporary solution because fungal populations rapidly evolve to overcome this type of resistance. In contrast, high temperature adult plant (HTAP) resistance genes provide a broad spectrum resistance that is partial and more durable. The cloning of the first wheat HTAP stripe rust resistance gene Yr36 (Science 2009, 323:1357), funded by our previous (2007-2010) BARD grant, provided us for the first time with an entry point for understanding the mechanism of broad spectrum resistance. Two paralogous copies of this gene are tightly linked at the Yr36 locus (WKS1 and WKS2). The main objectives of the current study were to characterize the Yr36 (WKS) resistance mechanism and to identify and characterize alternative WKSgenes in wheat and wild relatives. We report here that the protein coded by Yr36, designated WKS1, that has a novel architecture with a functional kinase and a lipid binding START domain, is localized to chloroplast. Our results suggest that the presence of the START domain may affect the kinase activity. We have found that the WKS1 was over-expressed on leaf necrosis in wheat transgenic plants. When the isolated WKS1.1 splice variant transcript was transformed into susceptible wheat it conferred resistance to stripe rust, but the truncated variant WKS1.2 did not confer resistance. WKS1.1 and WKS1.2 showed different lipid binding profiling. WKS1.1 enters the chloroplast membrane, while WKS1.2 is only attached outside of the chloroplast membrane. The ascorbate peroxidase (APX) activity of the recombinant protein of TmtAPXwas found to be reduced by WKS1.1 protein in vitro. The WKS1.1 mature protein in the chloroplast is able to phosphorylate TmtAPXprotein in vivo. WKS1.1 induced cell death by suppressing APX activity and reducing the ability of the cell to detoxify reactive oxygen. The decrease of APX activity reduces the ability of the plant to detoxify the reactive H2O2 and is the possible mechanism underlying the accelerated cell death observed in the transgenic plants overexpressing WKS1.1 and in the regions surrounding a stripe rust infection in the wheat plants carrying the natural WKS1.1 gene. WKS2 is a nonfunctional paralog of WKS1 in wild emmer wheat, probably due to a retrotransposon insertion close to the alternative splicing site. In some other wild relatives of wheat, such as Aegilops comosa, there is only one copy of this gene, highly similar to WKS2, which is lucking the retrotransposon insertion. WKS2 gene present in wheat and WKS2-Ae from A. showed a different pattern of alternative splice variants, regardless of the presence of the retrotransposon insertion. Susceptible Bobwhite transformed with WKS2-Ae (without retrotansposon insertion in intron10), which derived from Aegilops comosaconferred resistance to stripe rust in wheat. The expression of WKS2-Ae in transgenic plants is up-regulated by temperature and pathogen infection. Combination of WKS1 and WKS2-Ae shows improved stripe rust resistance in WKS1×WKS2-Ae F1 hybrid plants. The obtained results show that WKS1 protein is accelerating programmed cell death observed in the regions surrounding a stripe rust infection in the wheat plants carrying the natural or transgenic WKS1 gene. Furthermore, characterization of the epistatic interactions of Yr36 and Yr18 demonstrated that these two genes have additive effects and can therefore be combined to increase partial resistance to this devastating pathogen of wheat. These achievements may have a broad impact on wheat breeding efforts attempting to protect wheat yields against one of the most devastating wheat pathogen.
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Dickman, Martin B., i Oded Yarden. Role of Phosphorylation in Fungal Spore Germination. United States Department of Agriculture, sierpień 1993. http://dx.doi.org/10.32747/1993.7568761.bard.
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Spore germination is a common and fundamental event in fungal development and in many instances an essential phase of fungal infection and dissemination. Spore germination is also critical for hyperparasites to function as biocontrol agents as well as in fermentation proceses. Our common objective is to understand the mechanisms which regulated spore germination and identify factors involved in pathogenicity related prepenetration development. Our approach is to exploit the overall similarity among filamentous fungi using both a plant pathogen (Colletotricum trifolii) and a model system that is genetically sophisticated (Neurospora crassa). The simulataneous use of two organisms has the advantage of the available tools in Neurospora to rapidly advance the functional analysis of genes involved in spore germination and development of an economically important fungal phytopathogen. Towards this we have isolated a protein kinase gene from C. trifolii (TB3) that is maximally expressed during the first hour of conidial germination and prior to any visible gene tube formation. Based on sequence similarities with other organisms, this gene is likely to be involved in the proliferative response in the fungus. In addition, TB3 was able to functionally complement a N. crassa mutant (COT-1). Pharmacological studies indicated the importance of calmodulin in both germination and appressorium differentiation. Using an antisense vector from N. crassa, direct inhibition of calmodulin results in prevention of differentiation as well as pathogenicity. Both cAMP dependent protein kinase (PKA) and protein kinase C (PKC) like genes have been cloned from C. trifolii. Biochemical inhibition of PKA prevents germination; biochemical inhibitors of PKC prevents appressorium differentiation. In order to analyze reversible phosphorylation as a regulatory mechanism, some ser.thr dephosphorylative events have also been analyzed. Type 2A and Type 2B (calcineurin) phosphatases have been identified and structurally and functionally analyzed in N. crassa during this project. Both phosphatases are essential for hyphal growth and maintenance of proper hyphal architecture. In addition, a first novel-type (PPT/PP5-like) ser/thr phosphatase has been identified in a filamentous fungus. The highly collaborative project has improved our understanding of a fundamental process in fungi, and has identified targets which can be used to develop new approaches for control of fungal plant pathogens as well as improve the performance of beneficial fungi in the field and in industry. In addition, the feasibility of molecular technology transfer in comparative mycology has been demonstrated.
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Rocheford, Torbert, Yaakov Tadmor, Robert Lambert i Nurit Katzir. Molecular Marker Mapping of Genes Enhancing Tocol and Carotenoid Composition of Maize Grain. United States Department of Agriculture, grudzień 1995. http://dx.doi.org/10.32747/1995.7571352.bard.
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The overall objective of this research was to identify chromosomal regions and candidate genes associated with control of concentration and forms of carotenoids (includes pro-Vitamin A) and tocopherols (Vitamin E), which are both antioxidants and are associated with health advantages. Vitamin A and E are included in animal feeding supplements and the eventual goal is to increase levels of these compounds in maize grain so that the cost of these supplements can be reduced or eliminated. Moreover, both compounds are antioxidants that protect unsaturated fatty acids from oxidation and thus maintaining maize oil quality for longer periods. We identified three SSR markers that are associated with 38% of the variation for total carotenoids and three SSR markers associated with 44% of the variation for total tocopherols in the cross W64a x A632. We identified two candidate genes associated with levels of carotenoids: phytoene synthase and zeta carotene desaturase. Evaluation of (Illinois High Oil x B73) B73 BC 1S1 population for tocopherols detected additional chromosomal regions influencing the level of total tocopherols, and detected a common region on chromosome 5 associated with ratio of the more desirable alpha from to the gamma form of tocopherol. The results suggest molecular marker assisted selection for higher levels of these antioxidants in corn grain should be feasible.
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Or, Etti, Tai-Ping Sun, Amnon Lichter i Avichai Perl. Characterization and Manipulation of the Primary Components in Gibberellin Signaling in the Grape Berry. United States Department of Agriculture, styczeń 2010. http://dx.doi.org/10.32747/2010.7592649.bard.
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Seedless cultivars dominate the table grape industry. In these cultivars it is mandatory to apply gibberellin (GA) to stimulate berry development to a commercially acceptable size. These cultivars differ in their sensitivity to GA application, and it frequently results in adverse effects such as decreased bud fertility and increased fruit drop. Our long term goals are to (1) understand the molecular basis for the differential sensitivity and identify markers for selection of sensitive cultivars (2) to develop new strategies for targeted manipulation of the grape berry response to GA that will eliminate the need in GA application and the undesirable effects of GA on the vine, while maintaining its desirable effects on the berry. Both strategies are expected to reduce production cost and meet growing consumer demand for reduced use of chemicals. This approach relies on a comprehensive characterization of the central components in the GA signaling cascade in the berry. Several key components in the GA signaling pathway were identified in Arabidopsis and rice, including the GA receptors, GID1s, and a family of DELLA proteins that are the major negative regulators of the GA response. GA activates its response pathway by binding to GID1s, which then target DELLAs for degradation via interaction with SLY, a DELLA specific F-box protein. In grape, only one DELLA gene was characterized prior to this study, which plays a major role in inhibiting GA-promoted stem growth and GA-repressed floral induction but it does not regulate fruit growth. Therefore, we speculated that other DELLA family member(s) may control GA responses in berry, and their identification and manipulation may result in GA-independent berry growth. In the current study we isolated two additional VvDELLA family members, two VvGID1 genes and two VvSLY genes. Arabidopsis anti-AtRGA polyclonal antibodies recognized all three purified VvDELLA proteins, but its interaction with VvDELLA3 was weaker. Overexpression of the VvDELLAs, the VvGID1s, and the VvSLYs in the Arabidopsis mutants ga1-3/rga-24, gid1a-2/1c-2 and sly1-10, respectively, rescued the various mutant phenotypes. In vitro GAdependent physical interaction was shown between the VvDELLAs and the VvGID1s, and GAindependent interaction was shown between the VvDELLAs and VvSLYs. Interestingly, VvDELLA3 did not interact with VvGID1b. Together, the results indicate that the identified grape homologs serve as functional DELLA repressors, receptors and DELLA-interacting F-box proteins. Expression analyses revealed that (1) VvDELLA2 was expressed in all the analyzed tissues and was the most abundant (2) VvDELLA1 was low expressed in berries, confirming former study (3) Except in carpels and very young berries, VvDELLA3 levels were the lowest in most tissues. (4) Expression of both VvGID1s was detected in all the grape tissues, but VvGID1b transcript levels were significantly higher than VvGID1a. (5) In general, both VvDELLAs and VvGID1s transcripts levels increased as tissues aged. Unfertilized and recently fertilized carpels did not follow this trend, suggesting different regulatory mechanism of GA signaling in these stages. Characterization of the response to GA of various organs in three seedless cultivars revealed differential response of the berries and rachis. Interestingly, VvDELLA3 transcript levels in the GA-unresponsive berries of cv. Spring blush were significantly higher compared to their levels in the highly responsive berries of cv. Black finger. Assuming that VvDELLA2 and VvDELLA3 are regulating berry size, constructs carrying potential dominant mutations in each gene were created. Furthermore, constitutive silencing of these genes by mIR is underway, to reveal the effect of each gene on the berry phenotype.
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Chamovitz, Daniel A., i Zhenbiao Yang. Chemical Genetics of the COP9 Signalosome: Identification of Novel Regulators of Plant Development. United States Department of Agriculture, styczeń 2011. http://dx.doi.org/10.32747/2011.7699844.bard.
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This was an exploratory one-year study to identify chemical regulators of the COP9 signalosome. Chemical Genetics uses small molecules to modify or disrupt the function of specific genes/proteins. This is in contrast to classical genetics, in which mutations disrupt the function of genes. The underlying concept is that the functions of most proteins can be altered by the binding of a chemical, which can be found by screening large libraries for compounds that specifically affect a biological, molecular or biochemical process. In addition to screens for chemicals which inhibit specific biological processes, chemical genetics can also be employed to find inhibitors of specific protein-protein interactions. Small molecules altering protein-protein interactions are valuable tools in probing protein-protein interactions. In this project, we aimed to identify chemicals that disrupt the COP9 signalosome. The CSN is an evolutionarily conserved eight-subunit protein complex whose most studied role is regulation of E3 ubiquitinligase activity. Mutants in subunits of the CSN undergo photomorphogenesis in darkness and accumulate high levels of pigments in both dark- and light-grown seedlings, and are defective in a wide range of important developmental and environmental-response pathways. Our working hypothesis was that specific molecules will interact with the CSN7 protein such that binding to its various interacting proteins will be inhibited. Such a molecule would inhibit either CSN assembly, or binding of CSN-interacting proteins, and thus specifically inhibit CSN function. We used an advanced chemical genetic screen for small-molecule-inhibitors of CSN7 protein-protein interactions. In our pilot study, following the screening of ~1200 unique compounds, we isolated four chemicals which reproducibly interfere with CSN7 binding to either CSN8 or CSN6.
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Philosoph-Hadas, Sonia, Peter Kaufman, Shimon Meir i Abraham Halevy. Signal Transduction Pathway of Hormonal Action in Control and Regulation of the Gravitropic Response of Cut Flowering Stems during Storage and Transport. United States Department of Agriculture, październik 1999. http://dx.doi.org/10.32747/1999.7695838.bard.
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Original objectives: The basic goal of the present project was to increase our understanding of the cellular mechanisms operating during the gravitropic response of cut flowers, for solving their bending problem without affecting flower quality. Thus, several elements operating at the 3 levels o the gravity-induced signal transduction pathway, were proposed to be examined in snapdragon stems according to the following research goals: 1) Signaling: characterize the signal transduction pathway leading to the gravitropic response, regarding the involvement of [Ca2+]cyt as a mediator of IAA movement and sensitivity to auxin. 2) Transduction by plant hormones: a) Examine the involvement of auxin in the gravitropic response of flower stems with regard to: possible participation of auxin binding protein (ABP), auxin redistribution, auxin mechanism of action (activation of H+-ATPase) mediation by changes in [Ca2+]cyt and possible regulation of auxin-induced Ca2+ action b: calmodulin-activated or Ca2+-activated protein kinases (PK). b) Examine the involvement of ethylene in the gravitropic response of flower stems with regard to auxin-induced ethylene production and sensitivity of the tissue to ethylene. 3) Response: examine the effect of gravistimulation on invertase (associated with growth and elongation) activity and invertase gene expression. 4) Commercial practice: develop practical and simple treatments to prevent bending of cut flowers grown for export. Revisions: 1) Model systems: in addition to snapdragon (Antirrhinum majus L.), 3 other model shoe systems, consisting of oat (Avena sativa) pulvini, Ornithogalun 'Nova' cut flowers and Arabidopsis thaliana inflorescence, were targeted to confirm a more general mechanism for shoot gravitropism. 2 Research topics: the involvement of ABP, auxin action, PK and invertase in the gravitropic response of snapdragon stems could not be demonstrated. Alternatively, the involvement in the gravity signaling cascade of several other physiological mediators apart of [Ca2+]cyt such as: IP3, protein phosphorylation and actin cytoskeleton, was shown. Additional topics introduced: starch statolith reorientation, differential expression of early auxin responsive genes, and differential shoot growth. Background to the topic: The gravitropic bending response of flowering shoots occurring upon their horizontal placement during shipment exhibits a major horticultural problem. In spite of extensive studies in various aboveground organs, the gravitropic response was hardly investigated in flowering shoots. Being a complex multistep process that requires the participation of various cellular components acting in succession or in parallel, analysis of the negative gravitropic response of shoot includes investigation of signal transduction elements and various regulatory physiological mediators. Major achievements: 1) A correlative role for starch statoliths as gravireceptors in flowering shoot was initially established. 2) Differentially phosphorylated proteins and IP3 levels across the oat shoe pulvini, as well as a differential appearance of 2 early auxin-responsive genes in snapdragon stems were all detected within 5-30 minutes following gravistimulation. 3) Unlike in roots, involvement of actin cytoskeleton in early events of the gravitropic response of snapdragon shoots was established. 4) An asymmetric IAA distribution, followed by an asymmetric ethylene production across snapdragon stems was found following gravistimulation. 5) The gravity-induced differential growth in shoots of snapdragon was derived from initial shrinkage of the upper stem side and a subsequent elongation o the lower stem side. 6) Shoot bending could be successfully inhibited by Ca2+ antagonists (that serve as a basis for practical treatments), kinase and phosphatase inhibitors and actin-cytoskeleton modulators. All these agents did not affect vertical growth. The essential characterization of these key events and their sequence led us to the conclusion that blocking gravity perception may be the most powerful means to inhibit bending without hampering shoot and flower growth after harvest. Implications, scientific and agriculture: The innovative results of this project have provided some new insight in the basic understanding of gravitropism in flower stalks, that partially filled the gap in our knowledge, and established useful means for its control. Additionally, our analysis has advanced the understanding of important and fundamental physiological processes involved, thereby leading to new ideas for agriculture. Gravitropism has an important impact on agriculture, particularly for controlling the bending of various important agricultural products with economic value. So far, no safe control of the undesired bending problem of flower stalks has been established. Our results show for the first time that shoot bending of cut flowers can be inhibited without adverse effects by controlling the gravity perception step with Ca2+ antagonists and cytoskeleton modulators. Such a practical benefit resulting from this project is of great economic value for the floriculture industry.