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Academic literature on the topic 'GSP-PCR'
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Journal articles on the topic "GSP-PCR"
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Kim, HJ, MS Kim, YJ Park, et al. "Prevalence of Gs alpha mutations in Korean patients with pituitary adenomas." Journal of Endocrinology 168, no. 2 (2001): 221–26. http://dx.doi.org/10.1677/joe.0.1680221.
Full textAbstract:
The reported frequencies of Gs alpha mutations (gsp mutations) in growth hormone (GH)-secreting pituitary adenomas are variable (ranging from 4.4 to 43%), and the presence of these mutations in the other pituitary adenomas is still a matter of controversy. Previous clinical and biochemical analyses of patients with GH-secreting pituitary adenomas and gsp mutations produced conflicting results and did not demonstrate obvious characteristics. Therefore, we investigated the prevalence of gsp mutations in Korean patients with pituitary adenomas and elucidated the characteristics of these patients. Forty-four GH-secreting adenomas, 7 prolactin (PRL)-secreting adenomas and 32 clinically non-functioning adenomas were examined for the presence of point mutations in codon 201 and 227 of the Gs alpha gene using a nested PCR and direct sequencing of DNA extracted from fresh tissue or paraffin-embedded pituitary adenoma samples. Seven of the 44 GH-secreting pituitary adenomas had point mutations at codon 201 or 227; of these, five mutations were in codon 201 and two were in codon 227. In patients with gsp mutations, mean tumor size was significantly smaller than in patients without gsp mutations (15.9+/-8.7 mm vs. 24.9+/-14.9 mm, P<0.05). Age, sex, basal GH levels, GH response to oral glucose loading, GH response to octreotide and surgical outcome were not different in the two groups. One of the 32 clinically non-functioning pituitary adenomas had a point mutation at codon 201; none of the seven prolactinomas had these mutations. These results show that gsp mutations are not rare in Korean acromegalic patients and mean tumor size is significantly smaller in acromegalic patients with gsp mutations. Our results also confirm the low frequency of gsp mutations in clinically non-functioning pituitary adenomas and the absence of gsp mutations in prolactinoma.
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Fu, Kun, Liqiang Chen, Lifeng Miao, Yan Guo, Wei Zhang, and Yunan Bai. "Grape Seed Proanthocyanidins Protect N2a Cells against Ischemic Injury via Endoplasmic Reticulum Stress and Mitochondrial-associated Pathways." CNS & Neurological Disorders - Drug Targets 18, no. 4 (2019): 334–41. http://dx.doi.org/10.2174/1871527318666190212111650.
Full textAbstract:
Background/Objective: Grape seed proanthocyanidins (GSPs) are a group of polyphenolic bioflavonoids, which possess a variety of biological functions and pharmacological properties. We studied the neuroprotective effects of GSP against oxygen-glucose deprivation/reoxygenation (OGD/R) injury and the potential mechanisms in mouse neuroblastoma N2a cells. Methods: OGD/R was conducted in N2a cells. Cell viability was evaluated by CCK-8 and LDH release assay. Apoptosis was assessed by TUNEL staining and flow cytometry. Protein levels of cleaved caspase-3, Bax and Bcl-2 were detected by Western blotting. CHOP, GRP78 and caspase-12 mRNA levels were assessed by real-time PCR. JC-1 dying was used to detect mitochondrial membrane potential. ROS levels, activities of endogenous antioxidant enzymes and ATP production were examined to evaluate mitochondrial function. Results: GSP increased cell viability after OGD/R injury in a dose-dependent manner. Furthermore, GSP inhibited cell apoptosis, reduced the mRNA levels of CHOP, GRP78 and caspase-12 (ER stressassociated genes), restored mitochondrial membrane potential and ATP generation, improved activities of endogenous anti-oxidant ability (T-AOC, GXH-Px, and SOD), and decreased ROS level. Conclusion: Our findings suggest that GSP can protect N2a cells from OGD/R insult. The mechanism of anti-apoptotic effects of GSP may involve attenuating ER stress and mitochondrial dysfunction.
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Luo, Hong, Jian Huang, Wei-Gong Liao, Qing-Yuan Huang, and Yu-Qi Gao. "The antioxidant effects of garlic saponins protect PC12 cells from hypoxia-induced damage." British Journal of Nutrition 105, no. 8 (2010): 1164–72. http://dx.doi.org/10.1017/s0007114510004939.
Full textAbstract:
Hypoxia frequently occurs under several different cellular circumstances. Excess reactive oxygen species that are induced by hypoxia may result in cell injury and dysfunction. Recently, garlic has been found to possess some biological and pharmacological activities. The present study examined the effects of garlic saponins (GSP) on the survival of differentiated PC12 (dPC12) cells and the oxidative–antioxidant system. dPC12 cells were exposed to 2 % O2 in order to establish a neuronal insult model. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction assay and lactate dehydrogenase (LDH) release assay. The expression of selected genes (catalase (CAT), p65 and neuron-specific class III β-tubulin) was evaluated by real-time PCR and immunoblot assays. CAT activity, malondialdehyde (MDA) and 8-hydroxy-deoxyguanosine (8-OH-dG) concentrations were also determined. The data showed that hypoxia dramatically damaged dPC12 cells, while treatment with approximately 5 × 10− 2–10 ng/ml GSP improved cell viability, decreased LDH leakage and caused the cells to maintain neuronal-like characteristics in hypoxia. The production of MDA and 8-OH-dG was attenuated by GSP. CAT activity in dPC12 cells pretreated with GSP was higher than that of the hypoxic control. Moreover, GSP up-regulated CAT expression and decreased the total protein expression as well as the nuclear expression of p65 in hypoxic cells. These data indicate that GSP has antioxidant properties that can protect dPC12 cells from hypoxia-induced damage, which may be related to the up-regulation of CAT expression and activity as well as a decrease in the expression and nucleus distribution of p65 through effects on redox-sensitive signalling pathways.
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Buntjer, J. B., and J. A. Lenstra. "Self-amplification of satellite DNA in vitro." Genome 41, no. 3 (1998): 429–34. http://dx.doi.org/10.1139/g98-036.
Full textAbstract:
We describe a PCR-like reaction in which genomic DNA acts as a template as well as a primer. Interaction between genomic tandem repeat units leads to self-amplification of satellite DNA. This genomic self-priming PCR (GSP-PCR) allowed the rapid amplification of species-specific tandem repeats of horse, cattle, dolphin, and chicken. A novel specific satellite of ostrich with a repeat unit of 60 bp was isolated using this method.Key words: satellite DNA, amplification, isolation, species-specific probes.
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Esapa, Christopher, Sally Foster, Sarah Johnson, J. Larry Jameson, Patricia Kendall-Taylor, and Philip E. Harris. "G Protein and Thyrotropin Receptor Mutations in Thyroid Neoplasia*." Journal of Clinical Endocrinology & Metabolism 82, no. 2 (1997): 493–96. http://dx.doi.org/10.1210/jcem.82.2.3719.
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Abstract The cAMP pathway plays a central role in thyroid follicular cell growth and function. Mutations of the TSH receptor (TSHR) or G proteins (gsp) that activate adenylyl cyclase have been identified in autonomously functioning thyroid nodules. Gsp mutations have been identified also in other forms of thyroid neoplasia, but their reported prevalence has been extremely variable. We have studied the prevalence of gsp mutations and activating mutations of Gi2α (gip) in a series of 66 benign and 34 malignant thyroid tumors. Thirty-six tumors were from Boston and 64 from the UK. In addition, we examined the 64 UK tumors for mutations of the TSHR gene. DNA extracted from fresh-frozen or paraffin-embedded tissue was amplified by PCR and examined for mutations using oligonucleotide-specific hybridization and single-strand conformation polymorphism analysis. No G protein gene mutations were identified in the Boston tumors. One gsp mutation, R201C, in a Hürthle cell adenoma and 1 gip mutation, R179C, in a follicular adenoma were demonstrated in tumors from the UK. Oligonucleotide-specific hybridization and single-strand conformation polymorphism analysis of the UK tumors did not demonstrate any mutations of the TSHR gene. Eleven normal thyroid tissue samples were wild-type for Gsα, Gi2α, and the TSHR gene.
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Villares Fragoso, Maria Candida B., Ana Claudia Latronico, Filomena Marino Carvalho, et al. "Activating Mutation of the Stimulatory G Protein (gsp) as a Putative Cause of Ovarian and Testicular Human Stromal Leydig Cell Tumors1." Journal of Clinical Endocrinology & Metabolism 83, no. 6 (1998): 2074–78. http://dx.doi.org/10.1210/jcem.83.6.4847.
Full textAbstract:
Activating mutations of the G protein genes have been associated with the development of several endocrine neoplasms. Such activating mutations, gip2, affecting the α-subunit of the Gαi2 protein were previously described by a single group in 30% of ovarian sex cord stromal tumors. Other activating mutations of the α-subunit of the Gs (gsp) have been identified in GH-secreting and nonfunctioning pituitary tumors, autonomous thyroid adenomas, and all affected McCune-Albright tissues, but not in sex cord stromal tumors. In the present study, we investigated the presence of gip2 and gsp mutations in 14 human sex cord stromal tumors. Six Leydig cell tumors (4 ovaries and 2 testes), 2 thecomas, 2 granulosa cell tumors, 3 androblastomas, and 1 gonadoblastoma (sex cord and germ cell) were included in this study. Genomic DNA was obtained from either fresh-frozen tumor tissues or paraffin-embedded sections and in some cases from blood samples. Using PCR, denaturing gradient gel electrophoresis, and direct sequencing, we detected 4 tumors (66.6%) with the gsp mutation (R201C) in our series of ovarian and testicular Leydig cell tumors. In contrast, no gip2 mutations were found in any of the sex cord stromal tumors studied. In conclusion, our findings suggest that the putative oncogene gsp may play a significant role in the molecular mechanism of these tumors.
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Chia, Jean-San, Ya-Yu Lee, Peng-Tu Huang, and Jen-Yang Chen. "Identification of Stress-Responsive Genes in Streptococcus mutans by Differential Display Reverse Transcription-PCR." Infection and Immunity 69, no. 4 (2001): 2493–501. http://dx.doi.org/10.1128/iai.69.4.2493-2501.2001.
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ABSTRACT Streptococcus mutans, which causes dental caries in the human oral cavity and occasionally causes infective endocarditis in the heart, withstands adverse environmental stress through diverse alterations in protein synthesis. Differential gene expression in response to environmental stress was analyzed by RNA fingerprinting using arbitrarily primed PCR with a panel of 11mer primers designed for differential display in Enterobacteriaceae. Dot and Northern blot hybridization confirmed that the transcription of several genes was up- or down-regulated following exposure to acid shock from pH 7.5 to 5.5. RNA of a gene designated AP-185 (acid-stress protein) was induced specifically by acid treatment, while RNA of GSP-781 (general-stress protein) was up-regulated significantly when bacteria were exposed to high osmolarity and temperature, as well as low pH. The deduced amino acid sequence of AP-185 shares homology (78% identity) with branched-chain amino acid aminotransferase. Cloning and sequence analysis of GSP-781 revealed a potential secreted protein of a molecular mass of about 43 kDa and with a pI predicted to be 5.5. Transcriptional levels of another gene, designated AR-186 (acid-repressed protein), which encodes putative aconitase, were repressed by acid treatment but were enhanced by plasma or serum components. Analogous results were identified in icd andcitZ genes, and repression of these genes, along with AR-186, was also observed when they were exposed to high osmolarity and temperature. These results indicate that differential regulation of specific genes at the transcriptional level is triggered by different stress and that genes responsible for glutamate biosynthesis in the citrate pathway are coordinately regulated during the stress response of S. mutans.
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Nuñure O., Jeferson, Nieves Sandoval C., Mauro Estrella O., et al. "Identificación y caracterización de Aeromonas sp patógenas aisladas de truchas arcoíris (Oncorhynchus mykiss) clínicamente enfermas cultivadas en piscigranjas del Perú." Revista de Investigaciones Veterinarias del Perú 32, no. 4 (2021): e20927. http://dx.doi.org/10.15381/rivep.v32i4.20927.
Full textAbstract:
La investigación tuvo como objetivo la caracterización bioquímica e identificación molecular de cepas patógenas de Aeromonas sp aisladas de truchas arcoíris juveniles con signología compatible a aeromoniasis, cultivadas en seis piscigranjas de los departamentos de Junín (2), Pasco (2), Cajamarca (1) y Ancash (1), Perú. Se seleccionaron 60 truchas juveniles de 4-5 meses, con signología sugerente de brotes de aeromoniasis entre 2017 y 2018. Se realizó la eutanasia y necropsia y se registraron las lesiones externas e internas presentes. Se efectuó el aislamiento bacteriano de muestras de bazo y riñón anterior en agar TSA y GSP por 24-48 horas a 25 °C, se caracterizaron las colonias bacterianas mediante pruebas bioquímicas y se identificaron por PCR convencional utilizando cebadores específicos del gen ARNr 16S (Aeromonas spp.), fstA (A. salmonicida) y gyrB (A. hydrophila). Además, se realizó la secuenciación y se confirmó por análisis BLAST (NCBI). Las lesiones externas registradas fueron melanosis, aletas necróticas y hemorrágicas, úlceras en la piel y distensión abdominal. Las lesiones internas fueron hepatomegalia, esplenomegalia y hemorragia petequial en hígado, ciegos pilóricos y grasa peritoneal. Se aislaron 12 cepas en GSP (7 de Junín y 5 de Cajamarca), caracterizadas como Aeromonas spp, siendo negativas en las muestras de Pasco y Ancash. Las pruebas bioquímicas determinaron cuatro cepas como Aeromonas salmonicida y ocho como Aeromonas sp móviles, diferenciándose por pruebas de motilidad e indol. La identificación molecular por PCR determinó cuatro cepas A. salmonicida procedentes de Junín, una cepa A. hydrophila de Cajamarca y siete cepas móviles Aeromonas spp (3 de Junín y 4 de Cajamarca) que fueron negativas a la PCR de A. salmonicida y A. hydrophila. El análisis BLAST (NCBI) confirmó las cepas A. salmonicida y Aeromonas spp (identidad de 98-100%); además, la cepa de Cajamarca (identificada como A. hydrophila por bioquímica) presentó identidad de 95.65% con A. hydrophila subespecie dhakensis. En conclusión, Aeromonas salmonicida, A. hydrophila subespecie dhakensis y otras Aeromonas móviles patógenas causan signología asociada a aeromoniasis en truchas arcoiris juveniles en las piscigranjas de Junín y Cajamarca. Es la primera vez que se detecta A. hydrophila subsp. dhakensis en el país.
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Roy, Avijit, Nandlal Choudhary, John S. Hartung, and R. H. Brlansky. "The Prevalence of the Citrus tristeza virus Trifoliate Resistance Breaking Genotype Among Puerto Rican Isolates." Plant Disease 97, no. 9 (2013): 1227–34. http://dx.doi.org/10.1094/pdis-01-12-0012-re.
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Citrus tristeza virus (CTV) isolates have been grouped into six genotypes: T3, T30, T36, VT, B165, and resistance breaking (RB) based on symptoms, host range, and genomic sequence data. The RB genotype has recently been identified with the novel property of replicating in trifoliate orange trees, a resistant host for the other five genotypes. Puerto Rican CTV isolate B301 caused mild vein clearing symptoms in Mexican lime but did not induce seedling yellows or stem pitting reactions in appropriate indicator Citrus spp., which are typical host reactions of the isolate T30. The isolate B301 was not detected by the genotype specific primer (GSP), which identifies the CTV-T3, -T30, -T36, -VT, and B165 genotypes. A primer pair for reverse transcription polymerase chain reaction (RT-PCR) amplification of the CTV-RB genotype was designed from the heat shock protein (p65) region based on the complete genomic sequences of trifoliate RB isolates from New Zealand available in the GenBank databases. The amplicon sequence from isolate B301 was 98% identical to that of the other trifoliate RB isolates. In addition, B301 was successfully inoculated into ‘Carrizo citrange’ (a trifoliate hybrid) but did not induce any symptoms. Furthermore, the complete genome sequence of B301 followed by the phylogenetic analysis revealed that the isolate is part of the RB clade with other CTV-RB isolates from New Zealand and Hawaii. Additional CTV isolates obtained from Puerto Rico were tested with the RB-GSP and confirmed the presence of trifoliate RB isolates in mixed infection with known CTV genotypes. Although this is the first report of a CTV trifoliate RB genotype from Puerto Rico, this genotype was present there prior to 1992.
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Sozinova, O. I., N. A. Kozub, I. A. Sozinov, and Ya B. Blume. "Genome specificity of primers to puroindoline genes." Faktori eksperimental'noi evolucii organizmiv 22 (September 9, 2018): 191–96. http://dx.doi.org/10.7124/feeo.v22.947.
Full textAbstract:
Aim. Genome specificity of primers for amplification of complete coding sequences of puroindoline genes was studied. Methods. PCR with gene-specific primers was performed for amplification of puroindoline genes of wheat and related species. Results. The primer pair designated PinaDH is not gene-specific as it yields two products of amplification of the genes Pina-1 and Gsp-1. This primers pair is not specific for the Pina-1 gene of the genome V of D. villosum. The primer pairs designated PinaCM and Pinb are gene-specific as they produce one amplification product of respective length. We have demonstrated that the PinaCM primer pair is also specific for the genomes D, E and V, and the Pinb primer pair is not specific for the genome V. Conclusions. Gene and genomic specificity of primers for puroindoline genes was refined. Primers suitable for further direct sequencing of coding parts of puroindoline genes of wheat and related species were chosen.
 Keywords: puroindoline, Triticum aestivum, Aegilops, Dasypyrum, genome specificity.
Dissertations / Theses on the topic "GSP-PCR"
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VĚCHTOVÁ, Pavlína. "Isolation and characterization of highly repetitive fraction of codling moth, \kur{Cydia pomonella}." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-53737.
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Repetitive DNA comprises substantial part of the eukaryotic genome. ?Junk DNA?, as it was originally understood at the beginning of its discovery has attracted a lot of attention lately due to many studies proving its functional perspectives. Analysis of its dynamics, characteristics and distribution has been widely studied in organisms with monocentric chromosomes. Holokinetic system, however, was left behind in these efforts and whole image of repetitive DNA distribution and dynamics in this system remains to be elucidated. In this thesis various approaches were used to isolate and characterise repetitive DNA in the genome of the codling moth, Cydia pomonella. Satellite DNA CPSAT-1 was successfully isolated, characterised with Dot blot and Southern blot and mapped with FISH in the genome of C. pomonella. 17 microsatellite probes were used to localize microsatellite arrays in the genome of C. pomonella. Method of microsatellite FISH revealed distribution of all tested microsatellites in C. pomonella complement.